prr.hec.gov.pkprr.hec.gov.pk/jspui/bitstream/123456789/1638/1/886S.pdf · 2018-07-23 · iv ACKNOWLEDGEMENTS All the praises and thanks are for Almighty Allah, the Compassionate,

i

Allelopathic Potential of Sunflower

By

Javed Kamal

M.Sc. (Hons.) AGRNOMY

A thesis submitted in partial fulfillment of the

requirements for the degree of

DOCTORATE OF PHILOSOPHY

IN

PLANT PHYSIOLOGY

DEPARTMENT OF PLANT SCIENCES

FACULTY OF BIOLOGICAL SCIENCES

QUAID-I-AZAM UNIVERSITY,

ISLAMABAD,

(PAKISTAN) 2010.

ii

In the name of Allah,

Most gracious, Most Merciful,

iii

Declaration

I hereby declare that the work presented in the following

thesis is my own effort and that the thesis is my own

composition. No part of this thesis has been previously

presented for any other degree.

Javed Kamal

iv

ACKNOWLEDGEMENTS

All the praises and thanks are for Almighty Allah, the Compassionate, the merciful, the

only creator of the universe and the source of all knowledge and wisdom, Who blessed

me with health, thoughts, talented teachers, helping friends to make some contribution at

the already existing ocean of knowledge. I offer my humblest thanks to the Holy Prophet

(peace be upon him) whose moral and spiritual teachings enlightened my heart and mind

and flourished my thoughts to achieve high ideas of life.

I feel highly privileged to take this opportunity to express my heartiest gratitude and deep

sense of indebt to my worthy Supervisor, Pof. Dr. Asghari Bano, Chairperson,

Department of Plant Sciences, Quaid-I-Azam University, Islamabad for her skillful

guidance, constructive criticism, masterly advice, valuable suggestions and sympathetic

behaviour for the completion of this manuscript.

I am extremely obliged and thankful to Professor Dr. Mir Ajab, Dean of Biological

Sciences, for ever inspiring and encouraging attitude during the course of study. Very

cordial thanks and acknowledgements are due to Dr. Jalal-Ul-Din, Senior Scientific

Officer, institute of crop sciences, National Agriculture Research Center, for his kind co-

operation. I am highly indebted to Dr. Fayyaz Ahmad, Ex- Dean of Biological Sciences.

Very cordial thanks and acknowledgements are due to Dr. Rashid Ahmad Khan,

Associate Professor, Department of Biochemistry.

I am also very thankful to my all friends who encouraged me to complete my work

especially, Mian Sarfraz, Zille Ursh, Tariq Salara, Liaqat Ali, Raja Muhammad Ali,

Rana Aleem Khan, Dr. Mushtaq, Zafar Ahmad, Nazir Ahmad, Fazli, Hamid Ullah,

Ch. Khawar, Faroq Ahmad, Saeed, Ch. Mazhar, Shahid Iqbal, Malik Zaheer,

Zulqarnain, Samee Ullah, Malik Usman, Wahid Jan, Riaz Ahmad, Rana Hafeez-ur-

Rehman, Naveed Ghulam Nabi, Imran Jam, Ch. Amir and Irum Naz bundle of

sweet and sincere people who encouraged me a lot during my life span in the University.

Their nice company is now going to be the sweet part of my life. I am also thankfull to

the Higher Education Commission of Pakistan for the scholarship that made it possible

for me to complete my research work for the doctoral degree. Last but not the least

gratitude to be expressed to my family members, my mother, father, my sisters for their

love, inspiration, good wishes and unceasing prayers for me to achieve higher goals in

life. Their concern in me can never be fully returned but will always be remembered.

(Javed Kamal)

v

DEDICATED TO:

The Holy Prophet (P.B.U.H).

Who is forever a torch of guidance and knowledge of

humanity as whole.

My Mother

By the virtue of her prayers, I have able to reach at this high position

and whose hands are raised for my well-being.

vi

CONTENTS

Title Page No

List of Abbreviations vi- viii

Abstract xxv

Introduction 1-10

Materials and Methods 11-22

Results and discussions 23-86

Conclusions 87

References 88-111

Appendices 112-202

vii

IST OF ABBREVIATIONS

ABA Abscisic acid

°C Degree Centigrade

Ca Calcium

Cm Centimeter

CCM Combined Carbon Medium

CFU Colony forming unit

C2H2 Acetylene

CaC12 Calcium chloride

CaCO3 Calcium carbonate

CuSO4 Copper sulphate

CV Cultivar(s)

DNA Deoxyribonucleic acid

EDTA Ethylene diamine tetra acetic acid

Fe Iron

Fe-EDTA Iron- ethylene diamine tetra acetic acid

g Gram

GA Gibberellic Acid

h Hours

HC1 Hydrochloric acid

H3BO3 Boric acid

H2O2 Hydrogen per oxide

H3PO4 Phosphoric acid

H2SO4 Sulphuric acid

HgCl2 Mercuric chloride

viii

HPLC High pressure liquid chromatograph

IAA Indole acetic acid

K Potassium

Kg Kilogram

KH2PO4 Potasium dihydrogen phospahte

K2HPO4 Di-potassium hydrogen phosphate

K2SO4 Potassium sulphate

L Liter

LB Luria Bertani

m Meter(s)

Mg Magnesium

mg Milligram(s)

mm. Minutes

mL Milliliters

mm Millimeter

MgSO4 Magnesium sulphate

Mn Manganese

MnSO4 Manganese sulphate

N Normal

N Nitrogen

N2 Di-nitrogen

NaC1 Sodium chloride

NaOH Sodium hydroxide

Na2CO3 Sodium carbonate

QTS Quick Test Strip

rpm Revolutions per minute

ix

SDS Sodium Dodecyl Sulphate

TE Tris-EDTA

µ Micro

V Volume

V/V Volume by Volume

Wt. Weight

YMA Yeast Mannitol Agar

Zn Zinc

ZnSO4 Zinc sulphate

x

List of Tables

Table No. Page No.

1st Type of Experiments

1. Physico- chemical characteristics of the soil during the years 2006 and 2007. 24

2. Morphological and biochemical characteristics of bacterial isolates from 25

sunflower roots, rhizosphere soil, and control soil during year 2006.

3. Morphological and biochemical characteristics of bacterial isolates from 26

sunflower roots, rhizosphere soil, and control soil during year 2006.

4. Number of colonies of Rhizobium, Azospirillum and phosphate- solubilizing 30

bacteria from sunflower roots, rhizosphere soil, and control soil.

5. Result of oxidase test from sunflower roots, rhizosphere soil and control soil. 30

6. Result of Gram staining isolates from sunflower roots, rhizosphere soil and 30

control soil.

7. Rf values of sunflower (cv Hysun 38 ) determined by thin layer chromatography. 32

8. Phenols and Flavonoids contents of sunflower (cv-Hysun 38) determined by 32

spectrophotometers.

xi

List of Tables

Table No. Page No.

2nd Type of Experiments

1: Effect of sunflower leaf extract on germination ( % ) of wheat varieties 37

Margalla 99 and Chakwall 97.

2: Effect of sunflower stem extract on germination ( % ) of wheat varieties 37


3: Effect of sunflower root extract on germination (%) of wheat 38

varietiesMargalla 99 and Chakwall 97.

4: Effect of leaf extract of sunflower on root length (cm) of seedlings of wheat 38

varieties Margalla 99 Chakwall 97.

5: Effect leaf extract of sunflower on shoot length (cm) of seedlings of 39

wheat varieties Margalla 99 Chakwall 97.

6: Effect of sunflower leaf extract on fresh weight (g) wheat varieties 39


7: Effect of leaf extract of sunflower on dry weight (g) of seedlings of 39

wheat varieties Margalla 99 and Chakwall 97.

8: Effect of stem extract of sunflower on root length (cm) of seedings of 40


9: Effect of stem extract of sunflower on shoot length (cm) of seedlings 40

of wheat varieties Margalla 99 and Chakwall 97.

10. Effect of stem extract of sunflower on fresh weight (g) of seedings of 40


11. Effect of stem extract of sunflower on dry weight (g) of seedlings of 41


12. Effect of root extract of sunflower on shoot length (cm) of seedlings of 41


13. Effect of root extract of sunflower on fresh weight (g) of seedlings of 41


14. Effect of root extract of sunflower on dry weight (g) of seedlings of 42


15. Effect of root extract of sunflower on root length (cm) of seedlings of 42


xii

List of Tables

Table No. Page No.

2nd Experiments

16. Effect of leaf extract of sunflower on GA (µg g-1) content of leaves in 46


17. Effect of leaf extract of sunflower on GA ( µg g-1

) content of roots 46

in wheat varieties Margalla 99 and Chakwall 97.

18. Effect of leaf extract of sunflower on IAA (µg g-1) content of leaves in 47


19. Effect of leaf extract of sunflower on IAA (µg g-1) contents of roots in 47


20. Effect of leaf extract of sunflower on ABA ( µg g-1

) contents of leaves in 47


21. Effect of leaf extract of sunflower on ABA (µg g-1) content of roots in 48


22. Effect of stem extract of sunflower on GA (µg g-1) content leaves in 48


23. Effect of stem extract of sunflower on GA (µg g-1) content of roots in 48


24. Effect of stem extract of sunflower on IAA (µg g-1) of seedlings of 49


25. Effect of stem extract of sunflower on IAA (µg g-1) content of roots in 49


26. Effect of stem extract of sunflower on ABA (µg g-1) content leaves in 49


27. Effect of stem extract of sunflower on ABA (µg g-1) content roots in 50


28. Effect of root extract of sunflower on GA (µg g-1) content of leaves in 50


xiii

List of Tables

Table No. Page No.

2nd Experiments

29. Effect of root extract of sunflower on GA (µg g-1) content of roots 50


30. Effect of root extract of sunflower on IAA (µg g-1) content of leaves 51


31. Effect of root extract of sunflower on IAA (µg g-1) content of roots in 51


32. Effect of root extract of sunflower on ABA (µg g-1) contents of leaves 51


33. Effect of root extract of sunflower on ABA (µg g-1) content of roots 52


34. Effect of leaf extract of sunflower on DNA (mg/ 100 g F. wt) content 56

of leaves in wheat varieties Margalla 99 Chakwall 97.

35. Effect of stem extract of sunflower of DNA (mg/ 100 g F. wt) content of 56

leaves in wheat varieties Margalla 99 and Chakwall 97.

36. Effect of root extract of sunflower on DNA (mg/ 100 g F. wt) contents 56

of leaves in wheat varieties Margalla 99 and Chakwall 97.

37. Effect of leaf extract of sunflower on chlorophyll (mg/ 100 g F. wt) content 57


38. Effect of stem extract of sunflower on chlorophyll (mg/ 100 g F. wt) 57

content of leaves in wheat varieties Margalla 99 and Chakwall 97.

39. Effect of root extract of sunflower on chlorophyll (mg/ 100 g F. wt) 57


40. Effect of leaf extract of sunflower on proline (mg/ 100 g F. wt) content 58


41. Effect of stem extract of sunflower on proline (mg/ 100 g F. wt) content 58


42. Effect of root extract of sunflower on proline (mg/ 100 g F. wt) contents 58


xiv

List of Tables

Table No. Page No.

2nd Experiments

43. Effect of leaf extract of sunflower on sugar (mg/ 100 g F. wt) 59

contents of leaves in wheat varieties Margalla 99 and Chakwall 97.

44. Effect of stem extract of sunflower on sugar (mg/ 100 g F. wt) 59

contents leaves in wheat varieties Margalla 99 and Chakwall 97.

45. Effect of root extract of sunflower on sugar (mg/ 100 g F. wt) 59


46. Effect of leaf extract of sunflower on protein (mg/ 100 g F. wt) 60


47. Effect of stem extract of sunflower on protein (mg/ 100 g F. wt) 60

activity of leaves in wheat varieties Margalla 99 and Chakwall 97.

48. Effect of root extract of sunflower on protein (mg/ 100 g F. wt) 60


49. Effect of leaf extract of sunflower in superoxidase (mg/ 100 g F. wt) 61


50. Effect of root extract of sunflower on superoxidase (mg/ 100 g F. wt) 61


51. Effect of stem extract of sunflower on superoxidase dismoutase (mg/ 100 61

g F. wt) of leaves in wheat varieties Margalla 99 and Chakwall 97.

52. Effect of root extract of sunflower peroxidase (mg/ 100 g F. wt) activity of 62


53. Effect of stem extract of sunflower on peroxidase (mg/ 100 g F. wt) activity 62


54. Effect of root extract of sunflower on peroxidase (mg/ 100 g F. wt) 62


xv

List of Tables

Table No. Page No.

3rd Experiments

1. Effect of sunflower leaf, stem and root extracts on weed density in wheat 71

30 days after sowing ( wheat varieties Margalla 99 and Chakwall 97).

2. Effect of sunflower leaf, stem and root extracts on fresh weight in wheat 71

40 days after sowing ( Wheat varieties Margalla 99 and Chakwall 97).

3. Effect of sunflower leaf, stem and root extracts on dry weight of 71

weeds in wheat 40 days after sowing ( wheat varieties Margalla 99

and Chakwall 97).

4. Effect of sunflower leaf, stem and root extracts on fresh weight of weeds in 72

wheat 70 days after sowing ( wheat varieties Margalla 99 and Chakwall 97).

5. Effect of sunflower leaf, stem and root extracts on dry weight of weeds in 72


6. Effect of sunflower leaf, stem and root extracts on number of tillers of 72

wheat plants 145 days after sowing ( wheat varieties Margalla 99

and Chakwall 97).

7. Effect of sunflower leaf, stem and root extracts on plant height of 73

wheat plants 145 days after sowing ( wheat varieties Margalla 99 and

Chakwall 97

8. Effect of sunflower leaf, stem and root extracts on 100-grain-wieght 73

of wheat 145 days after sowing (Wheat varieties Margalla 99 and

Chakwall 97).

9. Effect of sunflower leaf, stem and root extracts on fresh wieght of 73

wheat plants 145 days after sowing (Wheat varieties Margalla 99 and

Chakwall 97).

10. Effect of sunflower leaf, stem and root extracts on dry wieght of 74

wheat plants 145 days after sowing (Wheat varieties Margalla 99

and Chakwall 97).

11. Effect of sunflower leaf, stem and root extracts on gibberellic acid 77

contents of wheat seedlings 30 days after sowing (Wheat varieties

Margalla 99 and Chakwall 97).

12. Effect of sunflower leaf, stem and root extracts on indole acetic acid 77



xvi

List of Tables

Table No. Page No.

3rd Experiments

13. Effect of sunflower leaf, stem and root extracts on abscisic acid contents 77

of wheat seedlings 30 days after sowing (Wheat varieties Margalla 99

and Chakwall 97).

14. Effect of sunflower leaf, stem and root extracts on gibberellic acid contents 78

of wheat seedlings root 30 days after sowing (Wheat varieties Margalla 99

and Chakwall 97).

15. Effect of sunflower leaf, stem and root extracts on indole acetic acid 78

contents of wheat seedlings roots 30 days after sowing (Wheat varieties


16. Effect of sunflower leaf, stem and root extracts on abscisic acid contents 78

of wheat seedlings roots 30 days after sowing (Wheat varieties


17. Effect of sunflower leaf, stem and root extracts on chlorophyll contents 81


and Chakwall 97).

18. Effect of sunflower leaf, stem and root extracts on sugar contents of 81

wheat seedlings 30 days after sowing (Wheat varieties Margalla 99

and Chakwall 97).

19. Effect of sunflower leaf, stem and root extracts on protein contents 81


and Chakwall 97).

20. Effect of sunflower leaf, stem and root extracts on proline contents of 82

wheat seedlings 30 days after sowing (Wheat varieties Margalla 99 and

Chakwall 97).

21. Effect of sunflower leaf, stem and root extracts on DNA contents of 84

wheat seedlings 30 days after sowing (Wheat varieties Margalla 99

and Chakwall 97).

22. Effect of sunflower leaf, stem and root extracts on superoxidase dismutase 84

activity of wheat seedlings 30 days after sowing (Wheat varieties Margalla

99 and Chakwall 97).

23. Effect of sunflower leaf, stem and root extracts on peroxidase activity 84


and Margalla 99.

xvii

List of Figures

Table No. Page No.

Fig.1. Results of QTS during year 2006. 27

Fig. 2. Results of QTS during year 2007. 28

APPENDICES

Table No. Page No.

1. Metrological Data of Islamabad 112-113

Appendix ll Yeast extract Mannitol Agar (YMA), medium, (Vincent, 1970) 114

Appendix IIl Glucose Peptone Agar Medium. 115

Appendix IV Combined Carbon Medium (CCM) 116

Appendix V LB (Lubria-Ber (ani) Medium 117

Appendix Vl Gram staining 118

Appendix VIl Soil analysis reagents 119-122

Appendix Vlll Brough ton and D ilworth’s Solutiion 123

Appendix IX Dragendorff Reagent 124-125

xviii

List of FIGURES

Table No. Page No.

APPENDICES

Fig. 1-4. Effect of sunflower leaf extract on germination (%) of wheat 126

varieties Margalla 99 and Chakwall 97.

Fig. 5-8. Effect of sunflower stem extract on germination (%) of wheat 126


Fig. 9-12. Effect of sunflower root extract on germination (%) of wheat varieties 126


Fig 13-16. Effect of leaf extract of sunflower on root length (cm) of seedlings of 126


Fig. 17-20 Effect of leaf extract of sunflower on shoot length (cm) of 127

seedlings of wheat varieties Margalla 99 and Chakwall 97.

Fig. 21- 24. Effect of sunflower leaf extract on fresh weight (g) wheat 127


Fig. 25-28. Effect of leaf extract of sunflower on dry weight (g) of seedlings of 127


Fig. 29-32. Effect of stem extract of sunflower on root length (cm) of seedings 127


Fig. 33-36. Effect of stem extract of sunflower on shoot length (cm) of seedlings of 128


Fig. 37- 40. Effect of stem extract of sunflower on fresh weight (g) of seedings of 128


Fig. 41- 44. Effect of stem extract of sunflower on dry weight (g) of seedings of 128


Fig. 45- 48. Effect of root extract of sunflower on shoot length (cm) of seedlings 128


xix

List of FIGURES

Table No. Page No.

APPENDICES

Fig. 49-52. Effect of root extract of sunflower on fresh weight (g) of seedlings of 129


Fig. 53- 56. Effect of root extract of sunflower on dry weight (g) of seedlings of 129


Fig 57-60. Effect of root extract of sunflower on root length (cm) 129

of seedlings of wheat varieties Margalla 99 Chakwall 97.

Fig. 61- 64. Effect of leaf extract of sunflower on GA (µg g-1) content of leaves 129


Fig. 65-68. Effect of leaf extract of sunflower on GA (µg g-1) content of roots 130


Fig. 69- 72. Effect of leaf extract of sunflower on IAA (µg g-1) content of leaves 130


Fig. 73- 76. Effect of leaf extract of sunflower on IAA (µg g-1) contents of roots in 130


Fig. 77- 80. Effect of leaf extract of sunflower on ABA (µg g-1) contents of leaves in 130


Fig. 81- 84. Effect of leaf extract of sunflower on ABA (µg g-1) content of roots 131


Fig. 85- 88. Effect of stem extract of sunflower on GA (µg g-1) content leaves 131


Fig. 88- 92. Effect of stem extract of sunflower on GA (µg g-1) content of roots in 131


Fig. 92- 96. Effect of leaf extract of sunflower on IAA of seedlings of wheat 131


Fig. 97- 100. Effect of stem extract of sunflower on IAA (µg g-1) content of roots in 132


Fig. 101- 104. Effect of stem extract of sunflower on ABA (µg g-1) content leaves in 132


xx

List of FIGURES

Table No. Page No.

APPENDICES

Fig. 105- 109. Effect of stem extract of sunflower on ABA (µg g-1) content roots in 132


Fig. 110- 113. Effect of root extract of sunflower on GA (µg g-1) content of leaves 132


Fig. 114-117. Effect of root extract of sunflower on GA (µg g-1) content of roots in 133


Fig. 118-121. Effect of root extract of sunflower on IAA (µg g-1) content of leaves 133


Fig. 122- 125. Effect of root extract of sunflower on IAA (µg g-1) content of roots in 133


Fig. 126- 129. Effect of root extract of sunflower on ABA (µg g-1) contents of leaves 133


Fig. 130- 133. Effect of root extract of sunflower on ABA (µg g-1) content of roots in 134


Fig. 134- 138. Effect of leaf extract of sunflower on DNA (mg/ 100 g F. wt) 134

content of leaves in wheat varieties Margalla 99 Chakwall 97.

Fig. 139- 142. Effect of stem extract of sunflower of DNA (mg/ 100 g F. wt) content 134


Fig. 143-147. Effect of root extract of sunflower on DNA (mg/ 100 g F. wt) contents 134


Fig. 148- 151. Effect of leaf extract of sunflower on chlorophyll (mg/ 100 g F. wt) 135


Fig. 152- 155. Effect of leaf extract of sunflower on IAA content of leaves in wheat 135


xxi

List of FIGURES

Table No. Page

No.

APPENDICES

Fig.156-159. Effect of root extract of sunflower on chlorophyll (mg/ 100 g F. wt) 135


Fig. 160-163. Effect of leaf extract of sunflower on praline (mg/ 100 g F. wt) 135


Fig. 164- 167. Effect of stem extract of sunflower on praline (mg/ 100 g F. wt) 136


Fig. 168- 171. Effect of root extract of sunflower on praline (mg/ 100 g F. wt) 136


Fig. 172- 174. Effect of leaf extract of sunflower on sugar (mg/ 100 g F. wt) 136


Fig. 175- 178. Effect of stem extract of sunflower on sugar (mg/ 100 g F. wt) 136

contents leaves in wheat varieties Margalla 99 and Chakwall 97.

Fig.179-182 Effect of root extract of sunflower on sugar (mg/ 100 g F. wt) 137


Fig. 183- 187. Effect of leaf extract of sunflower on protein (mg/ 100 g F. wt) 137


Fig. 188-191. Effect of stem extract of sunflower on protein (mg/ 100 g F. wt) 137


Fig. 192-195. Effect of root extract of sunflower on protein (mg/ 100 g F. wt) 137


Fig. 196- 199. Effect of leaf extract of sunflower in superoxidase (mg/ 100 g F. wt) 138


Fig. 200- 203. Effect of root extract of sunflower on superoxidase (mg/ 100 g F. wt) 138


xxii

List of FIGURES

Table No. Page No.

APPENDICES

Fig. 204-207. Effect of stem extract of sunflower on superoxidase dismoutase 138

(mg/ 100 g F. wt.) of leaves in wheat varieties Margalla 99 and

Chakwall 97.

Fig. 208-211. Effect of root extract of sunflower peroxidase (mg/ 100 g F. wt.) 138


Fig. 212-215. Effect of stem extract of sunflower on peroxidase (mg/ 100 g F. wt.) 139


Fig. 216- 219. Effect of root extract of sunflower on peroxidase (mg/ 100 g F. wt.) 139


Fig. 220- 223. Effect of sunflower leaf, stem and root extracts on weed density in 139

wheat 30 days after sowing ( Wheat varieties Margalla 99 and

Chakwall 97).

Fig. 224-227. Effect of sunflower leaf, stem and root extracts on fresh weight in 139

wheat 40 days after sowing (Wheat varieties Margalla 99 and Chak-

wall 97).

Fig. 228-231. Effect of sunflower leaf, stem and root extracts on dry weight of 140

weeds in wheat 40 days after sowing (wheat varieties Margalla 99

and Chakwall 97).

Fig. 232-235. Effect of sunflower leaf, stem and root extracts on fresh weight of 140

weeds in wheat 70 days after sowing (Wheat varieties Margalla 99

and Chakwall 97).

Fig. 236-239. Effect of sunflower leaf, stem and root extracts on dry weight of 140

weeds in wheat 70 days after sowing (Wheat varieties Margalla 99

and Chakwall 97).

Fig. 240-243. Effect of sunflower leaf, stem and root extracts on number of tillers 140

of wheat plants 145 days after sowing (Wheat varieties Margalla 99

and Chakwall 97).

xxiii

List of FIGURES

Table No. Page No.

APPENDICES

Fig. 244-247. Effect of sunflower leaf, stem and root extracts on plant height of 141

wheat plants 145 days after sowing (wheat varieties Margalla 99

and Chakwall 97).

Fig. 248-251. Effect of sunflower leaf, stem and root extracts on 100-grain-wieght 142

of wheat 145 days after sowing (Wheat varieties Margalla 99 and

Chakwall 97).

Fig. 252-255. Effect of sunflower leaf, stem and root extracts on fresh wieght of 142

wheat plants 145 days after sowing (Wheat varieties Margalla 99

and Chakwall 97).

Fig. 256-259. Effect of sunflower leaf, stem and root extracts on dry wieght of 142

wheat plants 145 days after sowing (Wheat varieties Margalla


Fig. 260-263. Effect of sunflower leaf, stem and root extracts on gibberellic acid 142



Fig. 264-267. Effect of sunflower leaf, stem and root extracts on indole acetic 143

acid contents of wheat seedlings 30 days after sowing

(Wheat varieties Margalla 99 and Chakwall 97).

Fig 268-271. Effect of sunflower leaf, stem and root extracts on abscisic acid 143

contents of wheat seedlings 30 days after sowing


Fig. 272-275. Effect of sunflower leaf, stem and root extracts on gibberellic acid 143

contents of wheat seedlings root 30 days after sowing


Fig. 276-279. Effect of sunflower leaf, stem and root extracts on indole acetic 143

acid contents of wheat seedlings roots 30 days after sowing


xxiv

List of FIGURES

Table No. Page No.

APPENDICES

Fig. 280-283. Effect of sunflower leaf, stem and root extracts on abscisic acid 144

contents of wheat seedlings roots 30 days after sowing


Fig. 284-287. Effect of sunflower leaf, stem and root extracts on chlorophyll 144

Contents of wheat seedlings 30 days after sowing


Fig. 288-291. Effect of sunflower leaf, stem and root extracts on sugar contents 144

of wheat seedlings 30 days after sowing (Wheat varieties


Fig. 292-295. Effect of sunflower leaf, stem and root extracts on protein contents 144



Fig. 296-299. Effect of sunflower leaf, stem and root extracts on proline contents 145



Fig. 300-303. Effect of sunflower leaf, stem and root extracts on DNA contents 145



Fig. 304-307. Effect of sunflower leaf, stem and root extracts on superoxidase 145

dismutase activity of wheat seedlings 30 days after sowing


Fig. 308-311. Effect of sunflower leaf, stem and root extracts on peroxidase 145

activity of wheat seedlings 30 days after sowing (Wheat varieties


xxv

ABSTRACT

Allelopathic effects of a sunflower (Helianthus annus L.) variety, Hysun 38, were studied

on two wheat varieties Margalla 99 and Chakwall 97. For this, three experiments were

conducted. The first experiment was conducted by growing sunflower in pots to evaluate

its effect) on beneficial soil microorganisms (Rhizobium, Azosprillum, and Phosphate-

Solubilizing Bacteria) and 2) on soil physicochemical properties. The quantification of

allelochemicals ( alkaloids, favonoids, and phenols) in leaves, stems and roots of

sunflower were made ; more allelochemicals were found in leaves, followed by roots, and

the least amount of allelochemicals were found in stem. The Rhizobium, Azosprillum,

and phosphate-solubilizing bacterial colonies declined in the soil cultivated with

sunflower. Its effects were shown on the carbon nitrogen utilization pattern of the

microbes as revealed by the Quick testing system (QTS) while no effect was shown on

the gram staining test. The effect of sunflower was also shown on moisture contents of

soil, phosphate, etc., the Ca and Mg contents of soil were increase.

The second experiment was conducted in petri dishes. The aim was to check allelopathic

potential of sunflower on germination rate, fresh weight, dry weight, root length, shoot

length, hormonal contents (Indole acetic acid, Gibberellin and Abscisic acid), and

chlorophyll, protein, proline, sugar, and DNA contents of wheat seedlings. It was noted

that the allelopathic effect was more as the concentrations of extract of sunflower

increased as compared with control. The allelochemical effect decreased the values of

fresh weight, dry weight, root length, shoot length, GA, IAA, Chlorophyll, protein,

proline, sugar, and DNA while increasing the values of ABA. It was also noted that the

allelopathic effect of sunflower leaves was greater than the roots, and the least amount of

effect was noted in stem.

The third experiment was conducted in pots in order to check the effects of sunflower

extracts on the comparative growth of wheat and weed. The fresh and dry weight of

wheat and weed density at 40 days after sowing (DAS) and at 70 DAS were determined

In addition the phytohormone contents, chlorophyll, protein, proline, sugar, DNA

contents and yield of wheat were determined. From the pots experiments, it was

concluded that sunflower leaves extract had decreased weed fresh weight and dry weight

as well as the Gibberrellin and Indole acetic acid contents but increased Abscisic acid

contents of wheat seedling.

1

INTRODUCTION

Since 1960's allelopathy has been increasingly recognized as an important ecological

mechanism which influences plant dominance, succession, formation of plant communities ,

vegetation and crop productivity. It has been related to the problems with weed: crop

interference (Bell and Koeppe, 1972), phytotoxicity in stubble mulch fanning (McCalla and

Haskins, 1964) and in certain types of crop rotations (Conrad, 1927). Rice (1984) incdicated

that allelopathy contributed to weed seed longevity problem through two mechanisms, (a)

chemical inhibitors in the seed prevented their decay by microbes and (b) the inhibitors kept the

seed dormant, although viable for many years.

Allelopathy plays greater role in the tropical and subtropical irrigated regions, where the

climate is inducible for multiple cropping and wide variety of crops and weeds exist together.

Since allelopathy provides basis to sustainable agriculture, therefore, it may be one of the

important strategies to increase the agricultural production under the changing climate.

Phenolics compounds released during red clover decomposition were phytotoxic to wild

mustard seedlings and the bioassay with aqueous red clover extract shoved a linear reduction in

mustard root growth with increasing extract concentration (Ohno et al. 2000).

Allelochemicals

Allelochemicals include mainly the plant secondary metabolites (Levin 1976). They exhibit

allelopathic effect either on the growth and development of the same plant or nearby plants

species. The term allelochemicals include, (a) plant biochemicals that exert their

physiological/toxicological action on plants (allelopathy, autotoxicity or phytotoxicity), (b)

plant biochemicals that exert their physiological/toxicological action on microorganisms

(allelopathy or phytotoxicity) and (c) microbial biochemicals that exert their

physiological/toxicological action on plants (allelopathy and phytotoxicity). About 125 natural

allelopathic compounds were isolated by Macias et al. (2002) from different cultivars of

sunflower, showing phytotoxic effects on growth of many weed species. Macias et al. (2000)

investigated the effect of several compounds isolated from Helianthus annuus on different

dicotyledon and monocotyledon species.

2

Secondary compounds are metabolically active in plants and microorganism, their biosynthesis

and biodegradation play a key role in the ecophysiology of the organism in which they occur

(Waller and Nowacki, 1978; Waller and Dermer, 1981). Some of them are accumulated at

various stages of growth, while, accumulation of some compounds depends upon season.

Classes of Allelochemicals

The allelochemicals are biosynthesized from the metabolism of carbohydrates, fats and amino

acids and arise from acetate or the shikimic acid pathway (Robinson, 1983). So in a review of

the potential use of allelochemicals as herbicides, The allelochemicals isolated from terrestrial

and aquatic plants include : Alkaloids,Benzoxazinones ,Cilmamic acid derivatives, Cyanogenic

compounds,Ethylene and other seed germination stimulants and flavonoids (Putnam ).

Occurrence of allelochemicals

The presence of allelochemicals in higher plant species and microbes has been documented by

several workers. These are originated in upper or lower plant parts or in both and posses

allelopathic impacts on a broad range of plant species. The allelochemicals may be found in all

parts of the plants such as seeds, flowers, pollen, leaves, stems, and roots etc.

Leaves

They are the most important sources of allelochemicals. Specific inhibitors in leaves have been

demonstrated by many workers, root and stem exhibit allelochemicals usually of low potency

and in fewer amounts

Flowers/inflorescence and pollen

There are increasing evidence that the pollen of corn and sunflower have allelopathic

properties.

Fruits and Seeds

Fruits and seeds contain allelochemicals which have been found inhibitory to microbial growth

and seed germination.

Modes of release of allelochemicals

A major pre-requisite of allelopathy is that allelochemical be transferred from a donor plant to

recipient plant; therefore, mode of transfer may play a great role in toxicity and persistence of

3

allelochemicals. Once these chemicals from the donor plants are released into the environment,

they may be either degraded or transformed into other forms, which affect the receiver plants

and may also be toxic to the host plant (autotoxicity).

Volatalization

Allelochemicals may volatilize from the plants to the atmosphere .The volatile vapours may be

absorbed directly from the atmosphere by plants, the adsorbed vapour may condensate in dew

and fall on the ground, these volatile compounds may absorbed on the soil particles and

subsequently taken by plants from the soil solution; the genera which release volatiles are

:Artemisia ,Salvia, Parthenium and Eucalyptus.

Leaching

Many allelopathic compounds both organic and inorganic are leached, such as phenolic acids,

terpenoids and alkaloids. The leaching of mineral nutrients, carbohydrates and phytohormones,

may be beneficial for the growth of associated species; however, mainly toxic effects have been

studied. Although seed leachates may also be important but mainly foliage leachates have been

investigated. Toxin-bearing leachates are important in weed-crop associations and in plant-

plant interactions in grasslands.

Decomposition of plant residues

The decomposition of plant residues adds the largest quantity of allelochemicals to the soil. At

plant death, materials compartmentalized in cells are released into the environment. The nature

of the plant residues, the soil type are important pre requisite for decomposition.

As the roots grow through the soil, at some points they may get in touch with decaying plant

residues and are impacted by allelochemicals. Some of the toxic effects of decomposition

products on plants are: inhibition of seed germination, stunted growth, and inhibition of the

primary root system and increase in secondary roots, inadequate nutrient absorption, chlorosis;

slow maturation and delay or failure of reproduction.

Factors affecting production of allelochemicals

Rice (1984) outlined following factors which affect the amount of allelochemicals produced,

viz., (a) radiation, (b) mineral deficiencies, (c) water stress, (d) temperature, (e) allelopathic

agents, (t) age of plant organs, (g) genetics, (h) pathogens and (i) predators.

4

Mode of action of allelochemicals

Allelopathic agents influence the plant growth (Rice, 1984) through the following physiological

processes viz., (i) cell division and cell elongation, (ii) phytohormone induced growth, (iii)

membrane permeability, (iv) mineral uptake, (v) availability of soil phosphorus and potash, (vi)

gas exchange and process of photosynthesis, (vii) respiration, (viii) protein synthesis and (ix)

changes in lipid and organic acid metabolism, (x) inhibition of porphyrin synthesis, (xi)

stimulation or inhibition of certien specific enzymes, (xii) corking and clogging of xylem

elements, (xiii) conductance of water through stem (xiv) interior water relationships and (xv)

miscellaneous.

Fate of allelochemicals

The biological activity, persistence, movement and fate of natural products in the soil depend

upon their interaction with the soil adsorption complex, soil microbial population and chemical

environment of the soil. Adsorbed allelochemicals may be biologically active or rendered

inactive, depending on nature of the adsorbing surface, but adsorbed molecules are less

available to soil microbes. Some natural products/allelochemicals may be irreversibly bound in

soil humic substances. Thus allelopathic effects in soil depend on the relative rates of

allelochemicals, addition and fixation in the soil.

Crop-to-crop interactions

Promising results were obtained by selecting allelopathic crop types, using allelopathic

companion plants or rotational crops (Weston and Duke 2003). Usually the field crops put in

phytotoxins or allelochemicals to the soils through crop residues and to a certain extent through

root exudates, therefore, their allelopathic effects have been studied most.

Effect of Allelochemicals

Allelochemicals have mostly negative effects on crop plants such as: (a) delayed or complete

inhibition of germination, (b) reduced plant population, (c) stunted and deformed roots and

shoots, (d) deranged nutrient absorption, (e) lack of seedling vigour, (f) reduced tillering, (g)

chlorosis, (h) wilting, (i) increase susceptibility to disease (Walker and Jenkins, 1986; Waller et

5

al., 1987; Oleszek and Jurzysta, 1987; Hicks et al., 1988). However, the main impacts of

phytotoxins on crop plants are: (i) inhibition of nitrification and biological nitrogen fixation, (ii)

predisposing the plants to diseases and (iii) inhibition or stimulation of germination, growth and

yield.

Root exudates

Root exudates of crops influence the germination, development and yield of other crop plants,

therefore, play major role in crop mixtures or intercropping systems. The first report on harmful

effects of root exudates of one plant on the growth of other plants was given by De Cando1le

(1832). Sorghum and maize root exudates inhibited the growth of sesame plants; therefore, its

plants could not be grown closer than 60 cm to live sorghum plants, which released natural

toxins in the growing medium (Fletcher, 1912; Breazeale, 1924; Hawkins, 1925; Comad, 1927;

Mckinley, 1931). Of the buckwheat, alfalfa, red clover, pea, soybean, rye, vetch and blue grass

root exudates, only that of buckwheat reduced the yield of tomato (Alderman and Middleton,

1925).

According to Overland (1966), barley is excellent smother crop due to its extensive root growth

and root exudates, which inhibited the germination and growth of tobacco, chickpea etc.

However, its root exudates had no inhibitory effect on wheat plants. The root exudates from

living plants contained the alkaloid 'gramine' and were more inhibitory than aqueous leachates

of dead roots, this proved active metabolic secretion of allelopathic substances. Root exudates

of rice varieties 'CB-1' and 'Rupsail' inhibited the root and shoot growth of test seedlings of

both these varieties, owing to presence of phenolic compounds also abscisic acid. The

maximum release of inhibitors in root exudates occurred under favourable climatic conditions

for rice growth (Sadhu and Das 1971; Sadhu, 1975). Tobacco root exudates inhibited the

germination of maize, mustard and foxtail seeds and subsequently their seedling growth (Haq

and Hussain, 1979), while that of Chinese cabbage reduced radical growth and dry matter of

Chinese cabbage and mustard (Akram and Hussain, 1987).

The root exudates play significant role in living plants and may inhibit or stimulate the seed

germination or seedling growth of associated weeds. The root exudates of rye (Borner, 1960),

corn (Dzyubenko and Petrenko, 1971; Dzyubenko and Krupa, 1974), oat (Fay and Duke, 1977),

6

wheat and oat (Martin and Rademacher, 1960), sorghum (Forney et al., 1983; A1Saadawi et

al., 1985; Panasuik et al., 1986), alfalfa (Abdul-Rahman and Habib, 1989), lupine (Dzyubenko

and Petrenko, 1971), soybeen (Mas santini et al., 1977; Rose et at., 1984), sunflower (Wilson

and Rice 1968) and buckwheat (Tsuzuki, 1980) inhibited the seed germination and stimulated

the seed germination of red sorrel (Panasuik et at., 1986) and witchweed (Netzy et al., 1988).

Sunflower (Wilson and Rice, 1968; Hall, 1980, Hall et at., 1982, 1983; Leather, 1983a) and

sweet potato (Harrison Jr. and Peterson, 1986) decreased the seed germination and growth of

weeds. Growing crops of barley (Mann and Barnes, 1945, 1947; Prutenskaya, 1974; Putnam

and De Frank, 1983). Rice (1964) reported that aqueous extracts of lambsquarter

(Chenopodium album L.) and crabgrass (Digitaria spp.) inhibited the growth of nitrogen fixing

and nitrifying bacteria. The inhibitors present in prostrate knotweed (Polygonum aviculare L.)

inhibited the growth of Rhizobium and Azotobacter (AI-Saadawi and Rice, 1982). Aqueous

extracts of Avena ludoviciana reduced the seedling growth and nodulation in green gram

(Bhandari et al. 1982).

Phytotoxins produced during the decomposition of crop residues inhibit the nitrification

process in the soil and biological nitrogen fixation in legumes. The maintenance of corn

residues on the soil surface increased the concentration of nitrification inhibitors (ferolic and p-

coumaric acids) in the soil, which decreased the population of nitrosomonas bacteria and thus

increased the concentration of N + over NO3- compared with the soil without corn residues

(Lodhi, 1981). In south Taiwan, soybean following rice, yielded higher when rice residues were

burnt than when decomposed in the field (Asian Vegetables and Research Development Centre,

1978), because phenolics produced from decomposing rice residues inhibited the growth of N

fixing bacteria (Rhizobium japonicum), reduced nodule number and thus decreased biological

nitrogen fixation in soybean (Rice, 1971). Similarly, soil incorporation of vines and storage

root residues of sweet potato reduced the nodulation and nitrogen fixation in cowpea (Walker

and Jel1kins, 1986). In a 8-year study at Los Banos, there was found significant decrease in

plant stand and yield of succeeding green gram crop (cultivated after green gram) was reported

due to allelopathy (Ventura et al. 1984). It was caused by the multiplication of harmful soil

microbe’s viz., fungi, bacteria, nematodes etc. and accumulation of their microbial toxins

which were phytotoxic to seed germination and seedling growth of green gram.

7

Weeds

Weeds cause greater losses in crop yields than either insects or plant diseases. The weeds

reduce the crop yields through (a) allelopathy, i.e., release of inhibitors from seeds, living

plants and plant residues, (b) competition for growth resources (light, nutrients, water and

space) with crops and (c) acting as an alternate host for insects and disease organisms. Recent

reports showed that sunflower posses allelochemicals which could be utilized fort sustainable

weed management (Anjum and Bajwa, 2005). The sunflower extracts inhibited not only the

germination and growth of wheat but also the growth of all species of weeds studied (Shahid, et

al., 2006).

It is necessary to identify the specific concentration of allelochemicals at which response

occurs if allelopathic interaction is to be applied in weed management programme. Moreover,,

different plant parts may differ in their allelopathic potential (Chon and Kim, 2002; Economou

et al., 2002). Jennings and Nelson (2002) have reported that allelochemicals also impact root

morphology in alfalfa autotoxic response

Weed residues

Till now, 129 weed species allelopathic to crops have been indentified (Narwal, 1994b). The

weed residues may exert allelopathic effect on crop plants similar to that of crop residues but

detailed studies are lacking. Allelochemicals released from the weed residues may affect the

crop plants in following manner: (i) inhibition of biological nitrogen fixation, (ii) inhibition of

nutrient uptake and (iii) inhibition of seed germination, growth and yield.

Phytohormones

Gibberellins (GA)

GA's are common and so far and wide in flowering, non-flowering and ferns plants. According

to Davies, (1995); Mauseth, (1991); Raven, (1992); Salisbury and Ross, (1992), many

physiological effects has been shown by active gibberellins, depending on the type of

gibberellins as well as plant species. Some of the physiological processes stimulated by

gibberellins are outlined below. Stimulate stem elongation through cell division stimulation.

8

Stimulates flowering in response to days lenth. Breaks seed dormancy in some plants which

require mechanical stratification or light to persue germination.

Auxins

Besides stem cell elongation auxins generally affect other processes in addition (Mauseth,

1991; Salisbury and Ross, 1992). In combination with cytokinins in tissue culture it stimulates

cell elongation and cell division in the cambium tissues. Auxin synthesis in apical bud induces

apical dominance in plants. It delays fruit ripening and stimulates growth of floral parts.

Alkaloids

Alkaloids are organic compounds possess carbon, hydrogen, nitrogen, and usually oxygen in

their chemical composition. Most alkaloids are derived from a few common amino acids. Most

have physiological activity. Most are basic, but have non- basic forms, such as quaternary

compounds and N-oxides. Many alkaloids give simple color and precipitation reactions, such as

with Dragendorrfs reagent that makes it relatively easy to determine their presence or absence

in plant material. Alkaloids usually contain one or more phenolic or indole rings, with a

nitrogen atom in the ring. The site of the nitrogen atom in the carbon ring varies with different

type of alkaloids and plant species. In a few alkaloids, such as mescaline and ephedrine, the

nitrogen atom is present outside the carbon ring. In fact, it is the position of nitrogen atom that

affects the properties of these alkaloids.

Abscisic Acid (ABA)

ABA is a natural growth inhibitor and well-known growth retardant, also known as "Stress

hormone." It is sesquiterpeniod in nature, synthesized from carotenoids via the mevalonic acid

pathway in leaves and roots (Neales et al., 1989). ABA plays a vital role in the development

and physiology of plants under stress condition (Zeevaart and Creelman, 1988; Salisbury and

Ross, 1992). It is a phytohormone that plays a significant role in the growth of plants and

survival in unfavourable conditions. While exogenous ABA is regarded as a grow1h inhibitor

and affects the morphology of leaves and stem (Sloger and Coldwell 1970. Quarrie, 1982).

ABA application during chickpea seed germination (ABA in germinating media) modified the

mRNA population, introduced new polypeptides and caused novel genes to be expressed in

germinating seeds of chickpea (Colorado et al., 1991, Colorado et al., 1995). ABA alone can

9

induce the synthesis of mRNA for several proteins but their translation requires the presence of

stress (Singh et al., 1987). The naturally occurring enantiomer is S (+) ABA (Oritano and

Yamashita, 1972) as opposed to the synthetic substance.

Wheat crop

Wheat (Tritcum aestivum L.) is the staple food of the people in Pakistan, besides that its straw

is integrated part of daily share for the livestock. Among different constraints including biotic

and abiotic stresses limiting wheat productivity, weed infestation has emerged as serious threat

to its productivity Weeds compete with crop for all growth factors such as nutrients, water,

space, light and carbon dioxide or cause interference in growth of the associated plants through

the production of biomolecule into the rhizosphere. The losses caused by weeds in crop

production vary depending upon the weed density and weed type. Diverse yield losses were

incurred in wheat due to weed infestation, which may very from 15 to 50 percent. In severe

cases weed infestation may cause complete crop failure.

Weeds in wheat are controlled by different methods such as mechanical and chemical. Manual

and mechanical methods are weather dependent whereas former is labour intensive as well.

This situation suggest that efforts should required to develop an substitute technology for weed

control in wheat, which not only effectively controls weeds but is also less labour intensive and

weather dependent as well as environmentally safe. Allelopathy may be suitable possible tools

for this purpose. A vast majority of these compounds are Phenolics, flavonoids and terpenes.

These substances have selective effects depending upon the inhibitory or stimulatory

concentrations to the growth of subsequent crops and weeds (Cheema, 1998). Allelopathy can

be used as technique in weed control by the application of the residues of allelopathic weeds

and crops plants as mulches or growing them better and parting their residues in the field. It has

been reported that crop plants like wheat, sunflower, rye, barley and cucumber have

allelopathic potential on the plants in their close vicinity.

Sunflower (Helianthus annuus. L) is a strongly Allelopathic plant (Hall, 1982). Sunflower is

the most important source of high quality vegetable oil. Chlorogenic acid, isochlorogenic acid,

scopolin and suspected napthol derivatives are phytotoxins identified in sunflower plant

residues. Crop residues of sunflower present mature selective effects on the germination and

growth of weeds. Being a potent allelopathic in nature its effect on subsequent crops and weed

have been reported, but very little information is available regarding its effects on wheat.

10

Alsaadawi (1988) has found that water extracts and decaying residues of root and shoot of

sunflower significantly reduced nitrification. In another experiment (1992), he reported that the

presence of allelopathic potential in sorghum and sunflower against nitrification may help to

augment nitrogen use efficiency of added fertilizer.

11

Role of Plant Stress

Allelopathy also interacts with the stress of the plants, because often a source of water stressed

plants release a greater variety and concentration of allelochemicals, and stressed target plants

may be more susceptible to allelochemicals (Reigosa et al., 2002).

AIMS AND OBJECTIVES

Present attempt was to determine the allelopathic effects of aqueous extracts of different organs

of sunflower on germination and growth of wheat, wheat productivity and weed density and

growth. Both laboratory and field experiments were conducted to achieve the above objectives.

The allelopathic effect of sunflower were evaluated on biochemical parameters, like protein,

proline, sugar, chlorophyll, and antioxidant enzymes viz.Superoxide dismutase, Peroxidase

activities and DNA contents of wheat .

The allelopathic effect of Sunflower was checked on physicochemical properties of soil as well

as on beneficial microorganisms of rhizospheric soil.

12

MATERIALS AND METHODS

Seed Material;

Sunflower (Helianthus annus L.) cv. Hysun 38 and two wheat varieties cv Margalla 99 and cv.

Chakwal 97 used in the experiments were obtained from oil and pulse Department and from

Cereal Crop Research Institute (wheat section) National Agricultural Research Centre (NARC),

Islamabad.

Pot Experiments

Initially, experiments on sunflower cv. Hysun 38 were conducted in pots at the Department of

Plant Sciences, Quaid-e-Azam University, Islamabad. Seeds of Hysun 38 were soaked in

distilled water for 24 h. Prior to sowing each pot was filled with sand, soil and manure (1:3:1)

and were placed under natural climatic conditions, detail of which are given in Table 1 of the

appendix.The diammonium phosphate (2g), urea (1g), and potash (1g) were applied as

recommended. When sunflower crop reached late vegetative stage (40 days after sowing)

plants were harvested and separated into leaves, stems and roots. They were dried, ground and

stored.

Parameters Studied

1. Physico-chemical analysis of soil

2. Study on the survival of beneficial microorganisms

3. Determination of Alkaloids, Flavonoids and Phenolics content of sunflower leaves

Analysis of Soil

According to the method of Nelson and Sommers, (1982) the physical and chemical properties

of soil were determined.

Particle Size Distribution (% Sand, % Silt, %clay)

Fifty grains of soil along with 50 ml of dispersing reagent (2% sodium hexametaphosphate)

were transferred to a stirring cup and left for overnight. The suspension was then stirred for 15

minutes and transferred to a 1000 ml graduated cylinder. Distilled water was added to raise the

volume upto lL. The suspension was stirred vigorously by means of metal plunger. First

13

hydrometer reading (R1) was taken after 40 seconds, followed by a second reading (R2) after 2

hours and temperature reading was noted. Percentage of sand, silt and clay were calculated

after making the temperature corrections as described by Brady (1990)

Calculations

% Separate = wt. of soil taken x 100)

% Separate = Percentage of sand, silt and clay.

CHR = Corrected Hydrometer Reading (after adjusting temperattlre)

Textural class was determined by using textural triangle (US Department of

Agriculture classification system).

Macro and Micronutrients in Soil

Soil samples, 5g each, were collected from the experimental pots at a uniform depth of 5cm,

suspended in 50ml of distilled water, stirred continuousl:y for 20 minutes, and filtered. The

filtrate was used for the analysis.

Electrical Conductivity

Electrical conductivity was detrermined with an EC meter after diluting the sample with an

equal quantity of deionized water.

Soil pH

Soil samples (25g each) were placed in 100 ml beakers; each filled with 25ml of distilled water,

and stirred for 10 minutes before recording the pH with a pH meter (Recommended soil

chemical test procedure, 1988).

Moisture Content

Soil samples (20g each) were taken from a uniform depth of 5cm. Fresh weight of the samples

was recorded. Dry weight of soil was determined after drying the soil in oven for 72h at 70 °C

to constant weight and moisture percentage was calculated.

Fresh Weight and Dry Weight

14

Fresh weight of the seedlings was recorded upon harvest. Dry weight was recorded after drying

the seedlings in an oven at 70 °C for 24 hours.

Determination of Nitrogen, Phosphorus, Potassium, Calcium, Magnesium, Iron and

Manganese

Nitrate-nitrogen was determined following the method described by Soltanpour and Schwab

(1977); K, Mg, Mn, and Ca were extracted from the soil sample as described by Mehlich (1953

and 1984); and concentrations of Fe, Mg, Mn and Zn were determined using an atomic

absorption spectrophotometer (Shimadzu, AA-670).

Solutions for the spectrophotometry were prepared as described by Whitney (1988).

Preparation of Samples

Leaves, stems and roots of sunflower plants were obtained from experimental plot of Quaid-i-

Azam University, Islamabad. They were air dried, ground and stored in a cool place.

Determination of Total Phenols

Total phenols were determined through the method of McDonald et al. 2001). Dilute extract of

each plant part (0.5ml of 1: 1og ml-l) or Gallic acid (Standared phenol compound) was mixed

with Folin Ciocalteu reagent (5ml) and aqueous Na2C03 (4ml, 1M). The mixtures were

incubated for 15 min and total phenols were determined by measuring its O.D. at 765nm. The

standard curve was prepared using different concentration of gallic acid prepared in methanol:

water (50: 50 v/v). Total phenol values were expressed in terms of gallic acid equivalent (mg g-

1 of dry weight).

Total Flavonoids

Total flavonoid was determined following the method of Chm1ge et al., (2002). Each plant

extracts (0.5ml of 1:10g ml-l) in methanol were separately mixed with 1.5ml of methanol, 0.l

ml of 10% aluminum chloride, 0.l M potassium acetate and 2.8ml of distilled water. The

mixture was allowed to stand at room temperature for 30min; the absorbance of reaction

mixture was measured at 415 nm with a double beam Perkin Elmer UV/Visible

spectrophotometer (USA). The quantitative analyses vwas done using quercetin calibration

curve.

15

Alkaloids Detection

Air dried leaves, stem and roots (2g) were extracted with n-hexane (3 x 20ml) followed by

MeOH (3 x 20ml) at room temperature for 24 hours. The metabolic extract was evaporated in

rotary film evaporator (RFE), residue was dissolved in I5ml of distilled waters. The pH of

aqueous phase was acidified to 2.0 by 5% H2SO4 and extracted with dicholoromethane (3x 1/3

of total volume) to obtain non alkaloids mixture. The acidic aqueous solution was basified to

pH 8-10 by using 10% NaOH and extracted with dichloromethane (3x ) to obtain alkaloids

mixture (Ulubelen, 2000). At this stage aqueous phase was discarded and organic phase was

dried in RFE. The residue was then taken in 1 ml dichloromethane and stored till TLC was

made.

Thin Layer Chromatography

Preparation of TLC Plates

The glass for TLC plate measuring 20 x 20 cm was coated (0.25 nm thickness) with silica Gel

HF 254 (Art 7739 Merk) with the help of TLC spreader.

Spotting the Plates

The samples were spotted on TLC plates with the help of micro capillary tubes. The prepared

(spotted) TLC plates were eluted using the solvent systm; Toluene, Ethyle acetate and

Diethylamide (6:2:0.5) (Ulubelen, 2000).

Visualization of the TLC Plates

The air dried plates were studied under UV 254nm and UV 365nm. The spots were marked

with pencil, and Rf value for particular compound were calculated as Distance travelled by the

compound / distance travelled by the solvent system.

Confirmatory Test for Alkaloids

Immediate after visualization under UV light, the plate was sprayed with the Dragendorff spray

reagent.

16

Collection of Soil Samples

The soil samples were collected form rhizosphere of sunflower prior to sowing sunflower and

at harvest rhizosphere soil and roots of sunflower at vegetative phase (35 days after sowing)

were collected for isolation of Rhizobium, Azospirillum and Phosphate solubilising bacteria

(PSB). Three replicates of soil samples were collected from different pots of the sunflower.

Analysis of Soil Samples for Rhizobium

For isolation of Rhizobium from rhizosphere soil of sunflower, 1 g of soil was suspended in

9ml of distilled water. The suspension was diluted ten times with 9ml of distilled water. In this

way decimal dilutions were prepared. Then 20ul from three dilutions was inoculated on yeast

(ty) medium and at 30 C, the colonies of Rhizobium obtained were further transferred to fresh

medium to get pure colonies.

Analysis for Azospirillum

Azospirilum was isolated from rhizosphere soil and roots of sunflower.

Isolation from Rhizosphere Soil

One g rhizosphere soil of sunflower were suspended in 9ml of distilled water, serial dilutions

were made using this stock. Then 10ul from three dilutions (10-1, 10-5 and 10-10) was used to

inoculate vials containing nitrogen free medium (NFM). These vials were incubated at 30 °C

for 48 hours. Vials showing Azospirillum growth were used for inoculation on NFM plates in

order to obtain the pure colonies of Azospirillum species. Single colonies appearing on these

plates were transferred in liquid broth of NFM and on agar slants for further study.

Isolation from Roots

For isolation of Azospirillum 1g roots of these plants were crushed in 9ml of distilled water.

The resulting mixture was centrifuged at 3000 rpm for 15 minutes and the clear suspension was

obtained. Then 100 ul of this solution was inoculated into vials containing nitrogen free

medium. The vials showing growth of Azospirillum were used for study of different

parameters.

17

Isolation of Phosphate Solublizating Bacteria

Phosphate Solubilizing Bacteria were isolated from soil sample by serial dilution and spread

plate method. One gram soil was dispersed in 10 ml of sterile distilled water and thoroughly

shaken. After mixing serial dilutions to 1ml of an aliquot of 0.1 ml from each dilution was

taken with nicropipette and plated on Pikovskaya medium.

Total number of colonies formed was counted after incubation. Total number was determined

as colony forming units (CFU). The P- Solubilizing ability of the isolate on Pikovskaya's agar

medium (Pikovsaya 1948) containing insoluble tricalcuim phosphate was determined.

Identification of Bacterial Isolates

Isolated strains of bacteria were identified on the basis of colony, cell morphology and

biochemical's test. Bacterial isolates from overnight grown cultures in LB broth were spread on

the agar plates of the medium. The morphology of the colonies was noted after 24 h orders to

study the cell motility and shape; single colony from the agar plates was transferred on glass

slide with a drop of sterile water and observed under light microscope (Nikon, Japan). Gram Staining

For observation under light microscope, slides of isolated purified bacterial cultures were

prepared. The Gram staining was done by the Vincent's method (1970). A drop of bacterial

cultures was taken and thin smear was prepared on a glass slides. The smear was air-dried; heat

fixed, stained with crystal violet for one minute and slightly washed with distilled water. The

smear was then flooded with iodine solution for one minute and decolorized with 95% ethanol

for one minute. The smear was again washed with distilled water and counterstained with

safranin. The slide was washed with distilled water, air-dried and observed under light

microscope (Nikon, Japan) at 100x magnification using oil immersion.

Oxidase Test

Oxidase test of bacterial isolate was done using Kovack's reagent (1% N, N, N.N -Tetramethyle

-p - phenylene diamine; Kovacks, 1956) following the method of Steel (1961).

Catalase Test

18

This test was performed according to MacFaddin (1980). One drop of H2O2 (30%) was added

to 24h old bacterial culture. Appearance of gas bubble indicated the presence of catalase

enzyme.

Miniaturized Identification System -QTS 24

Physiological and biochemical tests were performed using QTS 24 miniaturized identification

system (DESTO Laboratories Karachi, Pakistan). The bacterial cultures (24hours old) grown

on LB plates were suspended in saline solution (0.85% NaCl) and were used to inoculate QTS

kits. In case of Rhizobium, colonies were grown on YMA medium.

Plant Material and Growing Conditions

Allelopathic extracts of different parts of sunflower at different concentrations were prepared as

described by Bogatek et al., (2005). The extract was centrifuged at 3000 rpm for l5min and the

supernatant filtered through Whatman No. 42 filter paper. The extracts were stored below 5

°C until use. Seeds of two wheat varieties, namely Margalla 99 and Chakwall 97, were

germinated in Petri dishes (15 seeds in each dish) lined with filter paper moistened with either

distilled water (control) or with different concentrations of the extracts of sunflower Seed were

germinated at 26 °C The experiment consisted of 3 replicates per treatment.

Fresh Weight and Dry Weight

Fresh weight of the seedlings was recorded at harvest. Dry .weight was recorded after drying

the seedlings in oven at 70 °C for 24 h till constant weight.

Endogenous Contents of Phytohormones

The plant leaves or roots (sample of 19 each) were ground in 80% methanol at 4 °C with

antioxidant, (butylated hydroxy toluene (BHT), and kept for 72 h with change of the solvent

each 24 h. The extract was centrifuged and the supernatant was reduced to its aqueous phase

using a rotary thin film evaporator. The pH of the aqueous phase was adjusted to 2.5 -3.0 and

partitioned 4x with 1/3rd

volume of ethyl acetate. The ethyl acetate extract was fully dried using

a rotary thin-film evaporator. The dried sample was re-dissolved in 1ml methanol (100%) and

analysed using HPLC with a UV detector and a C-18 column. Pure IAA, GA and ABA were

19

used as standards for identification and quantification of the plant hormones. These growth

hormones were identified on the basis of retention time and peak area of the standards.

Methanol, acetic acid, and water (30:1:70) were used as the mobile phase. The wavelengths

used for detection were 280nm for IAA (Sarwar et al., 1992) 254nm for ABA and GA (Li et al.,

1994). For ABA the sample was injected onto C 18 column and eluted with linear gradient of

methanol (30-70%) containing 0.0 1% acetic acid, at flow rate of 0.8ml/min. The retention time

of ABA was determined by using authentic standards, monitoring the elution of standard at

254nm (Hansen and Doerffling, 1999).

Pot Experiments

Further experiments were conducted under natural conditions at the Department of Plant

Sciences, Quaid-e-Azam University, and Islamabad. Using the varieties of Margalla 99 and cv-

Chakwal 97. Seeds of both varieties were soaked in distilled water for 24 h. Prior to sowing

each pots (13 x11cm2) were fi11ed with sand soil and manure (1:3:1). During the first week of

November five seeds pot-l were sown and placed in natural climatic conditions. Details of the

climatic conditions are described in (Table1 appendix) for three years; the seedling were

irrigated with tap water.

Application of Sunflower Extracts on Wheat Varieties

In case of pot experiments the sunflowers leaves, roots and stem extract were prepared in the

ratio of Ig of powdered extract mixed with 10 ml distilled water. Then the solution was filtered

through two layer of Whatman paper to remove debris. After 24 days sowing plants were

exposed to the sunflower extract of leaves, stems and roots. After 4 days of application of

sunflower extracts leaves samples of wheat were collected for biochemical analysis. During the

experiments, the following additional parameters were studied during the vegetative and

reproductive growth stages.

Fresh Weight

20

First reading was taken after 30 days application of sunflower extract. For this purpose, the

whole plant was taken and washed thoroughly to remove the soil particle and weighed. The

fresh weight data were taken.

Dry Weight

Dry weight of the plant (roots and shoot) was determined 48h of drying the plant parts in an

oven at 80 °C till content weight.

[Plant Height at Maturity

Five tillers were selected at random from each treatment at maturity at harvest. Their height

were measured from tip to the base

Weed Density

Number of weeds was counted at 40 and 70 days of sowing. Their fresh weight was recorded.

Dry weight of root and shoot were taken after 72 hours at 70 °C.

Macro and Micronutrient Contents of Soil

The soil samples ( 5g) was stirred for 20 min using distilled water ;subsequently filtered

through filter paper and analysed using atomic absorption spectrophotometer.

Determination of Leaf Protein

Measurement of leaf protein was made according to the methods of Lowry et al., (1951) using

BSA as standard. Extraction was made in phosphate Buffer pH 7.5.

Reagent A

2.0g sodium carbonate (Na2CO3)

0.4gNaOH (0.lN) and 1g Na-K tartarate was dissolved in 100ml of distilled water.

Reagent B

The CUSO4. 5H20 (0.5 g) dissolved in 100ml of distilled water.

21

Reagent C

Solution a (50) and solution B (1ml) were mixed together.

Reagent D

Folin phenol reagent was diluted with distilled water in the ratio 1: 1.

Procedure

Fresh leaves (0.1g) were ground with the help of mortar and pestle in 1ml of phosphate buffer

(pH 7.5) with the help of mortar and pestle and was centrifuged for 10 minutes at 3000 rpm.

The supernatant (0.1 ml) was poured in the test tubes. Distilled water was added to make the

total volume of 1ml. 1ml of reagent C was added. After shaking for 10 minute, 100µl of reagent

D was added. The absorbance of each sample was recorded at 650nm after 30min. Incubation.

The concentration of protein content with reference to standard curve made by using standard

BSA (Bovine Serum Albumen) at of different concentrations viz 20, 40, 60, 80, 160, 320 and

640mg. Finally the absorbance of protein extract and BSA was recorded at 650nm.

Determination of Proline content in leaves

Proline was measured from the leaves of wheat following the method of Bates et al.(1973).

Procedure:

Freeze dried ground material (20mg) was suspended in 4 ml sulphosalicylic acid (3%) and

shaken overnight at 5°C. The suspension was centrifuged at room temperature at 3000 rpm for

10 minutes. The supernatant was mixed with 4ml acidic ninhydrin reagent and boiled for 1 h at

100°C in a water bath. After cooling, 4m1 toluene was added to the mixture, which was shaken

for 20s with a whirl mixer to mix the two phases. After the separation of the two phases the

absorbance of the toluene phase was recorded at 520 nm with a spectrophotometer (Shimadzu).

The actual concentration of proline was determined with reference to a proline standard curve.

Chlorophyll content of leaves

The fresh leaves of wheat were collected at 50% flowering for extraction of chlorophyll. The

chlorophyll estimation of leaves was made following the method of Amon (1949) and Kirch

(1968). The crude preparation (1 ml) was mixed with 4 ml of 80% (v/v) acetone and allowed to

22

stand in the dark at room temperature for 10 min. It was centrifuged at 2000 rpm for 5 min to

clear the suspension. The supernatant, which contained soluble pigment, was used to determine

chlorophyll. Absorbance of the solution was read at 645 nm for chlorophyll a and at 663 nm for

chlorophyll b on a spectrophotometer against 80% (v/v) acetone blank. Total chlorophyll was

determined with the equation of Amon (1949).

Total chlorophyll (mg/l) = (20.2 x a 645.A645) + (8.02 x b 663.A663).

Sugar content of leaves

Sugar content of wheat leaves at flowering stage was estimated by the method of Dubo et al.

(1956) as modified by Johnson et al. (1966). Fresh plant material (0.5 g) was homogenized with

10 ml of distilled water in a clean mortar. It was centrifuged at 3000 rpm for 5min. Then to 0.1

ml of supernatant, 1 ml of 5% (v/v) phenol was added. After 1 h incubation at room

temperature, 5 ml of concentrated H2SO4 was added. The absorbance of each sample was

recorded at 420 nm. The concentration of sugar in the unknown sample was calculated with

reference to a standard curve made by using glucose.

DNA analysis

Total DNA was extracted according to Guinn and Brummett 1988 estimated using an UV

spectrophotometer (Spectrophotometer 60l). As adapted by Ogur and Rosen genomic DNA was

extracted from 60 mg of frozen tissue (young leaves ground in liquid nitrogen) using Genomic

prep cells and tissue DNA Isolation Kit (Amersham Pharmacia Biotech Inc.) according to the

instruction manual. The DNA concentration in samples was measured by UV -absorption

spectroscopy at 260 nm at which DNA gives a maximum absorption at a concentration of

approximately 50 µg/g of double- stranded DNA. DNA was treated with 4 restriction enzymes

(Bam HI, EcoRI, Hinfl and SmaI at 5 U per reaction) according to manufacturer's instructions.

Statistical analyses

The data were analysed statistically using MStatC. A completely randomized design was

followed. The data were analyzed statistically by Analysis of Variance technique (Steel

and Torrie, 1980) and comparison among treatment means was made by Duncan’s

Multiple Range Test (DMRT) (Duncan’s, 1955).

23

RESULTS AND DISCUSSION

EXPERIMENT NO.1

Soil Analysis

The results of soil analysis are shown in Table 1. The soil analysis was undertaken three times:

every year: before sowing sunflower, after harvesting sunflower, and after harvesting wheat.

The following parameters were recorded in the second and third analysis: electrical

conductivity, pH, Zn+2

, Pb+4

, K+, Ca

+2, Mg

+2, Fe

+2, Mn

+2, and texture of soil. In years, electric

conductivity (ds/m), Ca+2, P+4, Pb+4, and moisture contents decreased, while the pH, Mn+2,

Fe+2

, Mg+2

, K+, and Zn

+2 were increased.

Isolation, identification, and characterization of microorganisms

The results of QTS of bacterial isolates from sunflower roots, rhizosphere soil, and control soil

are shown in Table 2 and 3. For characterization of the isolates, 24-h-old bacterial cultures

were tested using microbial identification kits QTS 24 and the results recorded following

overnight incubation at 30°C. The QTS results showed that Phosphate solubilising bacteria

(PSB) from each of the three habitats differed in their response to the following tests: VP, Gel,

GIuNO3, MaId, Suc, Inos, Ado, Mel, and Rae. Isolates of PSB from the control and

rhizosphere soils tested positive for VP, whereas those from roots tested negative, while in

2007, isolates of PSB from the control and rhizosphere soil tested positive for Rae, whereas

those from roots tested negative. In the case of Rhizobium, QTS results showed marked

differences between isolates from the control soil (uncultivated soil) and those from the

sunflower rhizosphere soil in two years with respect to ODC, TDA, Ind, VP, Gel, Arab, Rha,

Sor, GluNo3, Mann, the results revealed that isolates from roots and rhizospheric soil tested

negative, whereas those from the control soil tested positive.

24

25

Table 2. Morphological and biochemical characteristics of bacterial solates from

sunflower roots. rhizosphere soil, and control soil during year 2006.

Test Phosphate-

Solubilizing

bacteria (C)

Phosphat-

solublizing

bacteria

(SF)

Phosphate-

Solublizing

bacteria

Rhizobium

(C)

Rhizobium

(SF)

Az. (C) Az.

(SF)

Az.

ONPG

CLT

MALO

LDC

ADH

ODC

H2S

Urea

TDA

IND

VP

Gel

GluNo3

Mald

Suc

Mann

Arab

Rham

Sorv

Inos

Ado

Mel

Rae

Soil

+

-

+

-

-

-

-

-

+

-

+

+

+

+

-

+

-

+

+

+

+

+

+

Soil

+

-

+

-

-

-

-

-

+

-

+

-

-

-

-

+

-

+

+

-

-

-

-

Root

+

-

+

-

-

-

-

-

+

-

-

-

-

-

-

+

-

+

+

-

-

-

-

Soil

+

-

+

-

-

+

-

-

+

-

+

+

+

-

-

+

-

+

+

+

-

-

-

Soil

+

-

+

-

-

-

-

-

+

-

-

-

-

-

-

-

-

+

+

-

-

-

-

Soil

+

-

+

-

-

-

-

-

+

-

+

-

+

-

-

+

-

+

+

-

-

-

-

Soil

+

-

+

-

-

-

-

-

+

-

+

+

-

-

+

-

+

+

+

-

-

-

-

Root

+

-

+

-

-

-

-

-

+

-

-

-

+

-

-

+

-

-

-

-

-

-

Microbial idenlificatioii kits lOTS 24 DES FO Laboratories. Karachi, Pakistani were used for the biochemical

tests. The bacterial cultures were used and results recorded after 18 h of incubation at 30°C. ONPG Ortho nitro

phenyl 1 0- galactopyrarioside, CIT sodium citrate, MALO . sodium malonato LDC — lysine decarboxylase.

ADH argininc dihydrolase. ODC ornithine decarboxytase, H2S — H2S production, URE = urea hydrolysis, IDA =

tryptophan deaminaso, ND — indole, VP — Voges Proskauer ,GEL — getatine hydrolysis, GLU = acid from

glucose, MAL acid froni maltose, SUC — acid from sucrose, MAN — acid from mannitol, ARA = acid from

arabinose, RHA = acid from rhamnose, SOP = acid trom sorbitot, NO — acid from nositot, ADO — acid from

adonitol, MEL acid from melibiose, RAE = acid from raffinose C Control soil, SF — Sunflower rhizosphere soil,

and AZ Azospiri//um.

26

Table 3. Morphological and biochemical characteristics of bacterial solates from

sunflower roots. Rhizosphere soil, and control soil during year 2007.

Test Phosphate-

Solubilizing

bacteria (C)

Phosphate-

solublizing

bacteria (SF)

Phosphate-

Solublizing

bacteria

Rhizobium

(C)

Rhizobium

(SF)

Az. (C) Az. (SF) Az.

ONPG

CLT

MALO

LDC

ADH

ODC

H2S

Urea

TDA

IND

VP

Gel

GluNo3

Mald

Suc

Mann

Arab

Rham

Sorv

Inos

Ado

Mel

Rae

Soil

+

-

+

-

-

-

-

-

+

-

+

+

+

+

-

+

-

+

+

+

+

+

+

Soil

+

-

-

-

-

-

-

-

+

-

+

-

-

-

-

-

-

+

+

-

-

-

-

Root

+

-

-

-

-

-

-

-

+

-

-

-

-

-

-

-

-

+

+

-

-

-

-

Soil

+

-

+

-

-

+

-

-

+

-

+

+

+

-

-

+

-

+

+

+

-

-

-

Soil

+

-

+

-

-

-

-

-

-

-

-

-

-

-

-

-

-

+

-

-

-

-

-

Soil

+

-

+

-

-

-

-

-

+

-

+

-

+

-

-

+

-

+

+

-

-

-

-

Soil

+

-

+

-

-

-

-

-

+

-

+

+

-

-

+

-

+

+

+

-

-

-

-

Root

+

-

+

-

-

-

-

-

-

-

-

-

-

-

-

+

-

-

-

-

-

-

Microbial idenlificatioii kits lOTS 24 DES FO Laboratories. Karachi, Pakistani were used for the biochemical tests. [-or the

tests 24 fiourold bacterial cultures were used and results recorded after 18 h of incubation at 30°C. ONPG Ortho nitro phenyl 1

0- galactopyrarioside, CIT sodium citrate, MALO . sodium malonato LDC — lysine decarboxylase. ADH argininc dihydrolase.

ODC ornithine decarboxytase, H2S — H2S production, URE = urea hydrolysis, IDA = tryptophan doaminaso, ND — indole,

VP — Voges Proskauer = Acetion, GEL — getatine hydrolysis, GLU = acid from glucose, MAL acid froni maltose, SUC —

acid from sucrose, MAN — acid from mannitol, ARA = acid from arabinose, RHA = acid from rhamnose, SOP = acid trom

sorbitot, NO — acid from nositot, ADO — acid from adonitol, MEL acid from melibiose, RAE = acid from raffioseC Control

soil, SF — Sunflower rhizosphere soil, and AZ Azospiri//um.

27

Fig. 1. Results of QTS ( quick test strip ) during year 2006.

28

Fig. 2. Results of QTS (quick test strip) during year 2007.

29

Number of Colonies

The number of colonies of Rhizobium, Azospirillum and Phosphate-Solubilizing Bacteria were

presented in Table 4. Number of Rhizobium colonies was greater in soil sample from the

control soil than in those of the rhizosphere soil. These investigations indicated that soil factors,

such as physicochemicals properties and soil microorganisms, affect the allelopathic activity of

sunflower in soil. The results are in agreement with the general observations that plants that

produce allelochemicals reduce the population of Rhizobium. The number of Rhizobium

colonies was decreased in second year; perhaps this was also due to variation in the moisture

contents. The number of colonies of Azospinllum was highest in the samples from the control

soil, followed by those from the rhizosphere and roots. The number of colonies of PSB

followed similar pattern as that shown by Azospinillum: the highest in the control soil, followed

by that of rhizosphere soil and roots. The number of colonies in the case of Rhizobium,

Azospirillum, and Phosphate-Solubilizing Bacteria were less in the second year.

Oxidase Test

Azospirillum isolates from sunflower roots, rhizosphere and control soil are shown in Table 5.

Azospirillum isolates from the control soil showed a more intense oxidase reaction than that

shown by isolates from the other two habitats namely roots and rhizosphere. Within the two,

the reaction was more intense in isolates from rhizosphere Rhizobium isolates showed

considerable variation in their response to the oxidase while in the second year, Azospirillum

isolate from root showed a more intense oxidase reaction.

30

Table 4: Number of colonies (cfu/g soil or root) of Rhizobium, Azospirillum and

phosphate- solubilizing bacteria from sunflower roots, rhizosphere soil, and control soil.

Rhizobium Azospirillum Phosphate- solubilizing

bacteria Treatments

2006 2007 2006 2007 2006 2007

Control soil 4.95 ×106 4.85 ×10

6 2×10

6 1.5×10

6 1.20×10

6 1.15×10

6

Rhizosphere Soil 4.0×106 3.0×10

6 2×10

6 1.0×10

6 1.25×10

6 1.10×10

6

Roots ----- ----- 1×106 1×10

5 1.0×10

6 1.00×10

6

Table No 5. Result of oxidase test from sunflower roots, rhizosphere soil and control soil.

Treatments Rhizobium Azospirillum Phosphate- solubilizing

bacteria

Control soil + + +

Rhizosphere Soil + + +

Roots + + +

Table No 6: Result of oxidase test from sunflower roots, rhizosphere soil and control soil.

Treatments Rhizobium Azospirillum Phosphate- solubilizing

bacteria

Control soil + + +

Rhizosphere Soil + + +

Roots + + +

Table No. 7: Result of Gram staining test of isolates from sunflower roots, rhizosphere

soil and control soil.

Rhizobium Azospirillum Phosphate- solubilizing

bacteria

Treatments

2006 2007 2006 2007 2006 2007

Control soil ** ** ** ** ** **

Rhizosphere Soil ** * ** * ** *

Roots * * * * ** *

31

Alkaloids

Methanol extracts of sunflower leaves, stems, and roots over two years showed different

banding patterns following TLC coated with Silica gel HF254 and also different Rf values

under UV light at 365 nm (Table 7). All bands from the leaves showed less polarity than those

from the roots and stems. The number of bands was also highest in leaves. Perhaps the

volatilization of the alkaloids was minimal through leaves.

Phenols

Data in Table 8 indicated that the amount of total phenols was found maximum in the leaves,

followed by roots and stems.

Flavonoids

The data on flavonoids (Table 8) showed that the amount of flavonoids was maximal in leaves,

followed by roots and stems, in that order. Similar pattern was followed in the second year. It is

likely that leaves contain the most allelochemicals, because leaching loses those in roots and

those from stems are translocated.

32

Table No. 8: Rf values of sunflower ( cv Hysun 38 ) determined by thin layer

chromatography.

2006 2007 Treatments

Rf values RF values

Leaves 0.98000±0.00577 0.90000±0.0289

Stems 0.7400±0.0153 0.69000±0.00577

Roots 0.8867±0.0418 0.8300±0.0252

Each value is the average of three experiments ± standard deviation.

Table No. 9: Phenols and Flavonoids contents of sunflower (cv-Hysun 38) determined by

spectrophotometers.

Phenols (mg/g) Flavonoids (mg/g) Treatments

2006 2007 2006 2007

Leaves 311.67±4.41 400.00±5.77 83.33±3.76 100±3.61

Stems 200.00±5.77 256.7±12.0 45.00±2.89 65.67±2.96

Roots 270.00±5.77 342.3±24.3 70.00±5.77 85.00±2.89

Each value is the average of three experiments ± standard deviation

33

Discussion

The change in pH was in agreement with those of Periturin (1913). Plant phenolics are capable

of interference with the uptake of nutrients and can also affect rates of nutrient cycling (Izhaki,

2002), which would have a drastic effect upon the growth of surrounding plants.

Microorganisms have been considered to be an important factor affecting allelopathic activity

in soil (Masonsedun and Jessop 1988; May and Ash, 1990). Blum (1998) reported that soil

microorganisms degraded and utilized some allelochemicals in soil; in contrast some

allelochemicals can be produced by microorganisms during the decomposition of plant residue

in soil (Hegde and Miller, 1990; Hoffman et al., 1996 a, 1996b; Ismail and Mah, 191 Jimenez -

Osornio and Gliessman, 1987, Kiton and Yoshida, 1993; Martin and Smith, 1994). Number of

Rhizobium colonies was greater in soil sample from the control soil than in those from the

rhizosphere soil. These investigations indicated that soil factors, such as physicochemical s

properties and soil microorganisms, affect the allelopathic activity of sunflower in soil. The

results are in agreement with the general observations that plants that produce allelochemicals

reduce the population of Rhizobium. The number of Rhizobium colonies was decreased in

second year; this was also due to variation in the moisture contents. The number of colonies of

Azospinllum was the highest in the samples from the control soil, followed by those from the

rhizosphere and from roots. The number of colonies of PSB followed similar pattern as that

shown by Azospinillum: the highest in the control soil, followed by that of rhizosphere soil and

roots. These results agree with those obtained by Rice (1984), who demonstrated that

allelopathy reduces the availability of phosphorous and, therefore, the number of PSB. The

number colonies in case of Rhizobium, Azospirillum, and Phosphate-Solubilizing Bacteria were

more reduced in the second year.

Secondly, alkaloids from roots are lost through leaching and other micro- environmental factors

(Asa and Karlsson, 1998). Differences in chemical composition and diversity may also account

for the differences among the plant parts in terms of their alkaloid contents (Petaraitis et al.,

1989; Roesenzweig and Abransky, 1993; Olanbanji et al.. 1997; Asa and Karlsson, 1998).

34

Experiment no.2

Effect of sunflower leaf, stem and root extract on germination (%) of wheat varieties

Margalla 99 and Chakwall 97

The data regarding germination rate under the effect of sunflower leaf and stem extract were

presented in Table 1 and 2. A perusal of the data revealed that in case of both wheat varieties

Margalla 99 and Chakwall 97, the TI (control) showed maximum germination, followed by T5

(25% extract), whereas minimum germination was shown by T2 (undiluted extract). Similar

pattern was followed in the 2nd

year. The effect of sunflower root extract on germination of

wheat varieties MargaIla 99 and ChakwaIl 97 was shown in Table No. 3. In response to root

extract applied the germination percentage followed the pattern similar to that of leaf and stem

extract, i.e maximum germination was shown by T1 (control) and minimum by T2 (treatment

with undiluted extract of sunflower)

Effect of sunflower leaf extract on root length and shoot length (cm) of seedlings of wheat

varieties Margalla 99 and ChakwaIl 97

The data given in Table No. 4 and 5 revealed that in case of both wheat varieties Margalla 99

and Chakwall 97, Tl (control) showed the maximum root and shoot length, followed by T5

(25% extract) < T4 (50% extract) < T3 (75% extract), and T2 (undiluted extract), in the second

year, the results were the same.

Effect of sunflower leaf extract on fresh and dry weight (g) of seedlings of wheat varieties


Fresh weight is a function of accumulated effect of growth parameters, like final plant height.

The data on the fresh weight are given in Table 6. A perusal of the table 6 revealed that fresh

weight of wheat was affected significantly by various extracts of sunflower. In both wheat

varieties Margalla 99 and Chakwall 97, the ranking of the fresh weight among the treatments

were TI (control) > T5 (25% extract > T4 (50% extract)> T3 (75% extract)>and T2 (undiluted

extract). From the Table No. 7, it was evident that in two years the wheat variety Margalla 99

gave better response to the sunflower extracts in terms of dry weight, and in the first year the

results were better.

35

Effect of sunflower stem extract on root length and shoot length (cm) of seedlings of

Wheat varieties Margalla 99 and Chakwall 97

Root length indicates the productive efficiency of a crop. The higher the root length, the greater

the efficiency and vice versa. The data in Table 8 revealed that in the case of both first and

second year, wheat variety Margalla 99 showed better response. In two wheat varieties Tl

(control) showed maximum response, followed by T5 (25% extract), whereas T2 (undiluted

extract) showed minimum response. In the case of Margalla 99, T4 was at par with T3 (75%

extract). While in the case of Chakwall 97, T4 (50% extract) was at par with T2 (undiluted

extract). The data on shoot length indicated (Table No.9) that in first year, both wheat varieties

Margalla 99 and Chakwall 97, Tl (control) showed the maximum, followed by T5 (25%

extract), whereas T2 (undiluted extract) showed minimum increase . Results were found similar

in the second year.

Effect of sunflower stem extract on fresh and dry weight (g) of seedlings of wheat varieties


The data on fresh and dry weight of seedlings of wheat varieties Margalla 99 and Chakwall 97

are presented in Tables 10 and 11. A perusal of the table revealed that in both wheat varieties

Margalla 99 and Chakwall 97, TI (control) showed the maximum, followed by T5 (25%

extract), whereas T2 (undiluted extract) showed minimum increase in fresh and dry weight.

Results were found similar in the second year.

Effect of sunflower root extract on shoot length and root length (cm) of seedlings of wheat

varieties Margalla 99 and Chakwall 97

Shoot length is an important growth component in cereals and was to be influenced by the

sunflower extract. The data given in Table 12 and 15 revealed that the shoot and root length in

the case of both wheat varieties Margalla 99 and Chakwall 97, was variously affected. The Tl

(control) showed the maximum, followed by the T5 (25% extract), minimum result was found

in T2 (undiluted extract). Similar result was found in second year.

36

Effect of sunflower root extract on fresh and dry weight (g) of seedlings of wheat varieties


Fresh weight is a function of the combination of its individual growth components, which are

likely to be influenced by the genetic as well as environmental factors. A perusal of the Table

No. 13 and 14 revealed that, Tl (control) showed the maximum fresh and dry weight in both

wheat varieties followed by T5 (25% extract), whereas T2 (undiluted extract) showed

minimum fresh and dry weight. Similar pattern was followed in the second year.

37

Table 1: Effect of sunflower leaf extract on germination (% ) of wheat varieties


2006 2007 Treatement

V1 V2 V1 V2

T1 13.67 A 12.00 A 14.00 A 12.33 A

T2 10.00 C 7.00 C 9.00 C 6.00 C

T3 11.00 BC 8.00 C 10.00 BC 7.00 C

T4 10.67 BC 9.667 B 9.667 BC 8.667 B

T5 12.00 AB 10.33 B 11.00 B 9.33 B

Values followed by the same letter within a column are not significantly different.

Table 2: Effect of sunflower stem extract on germination ( % ) of wheat varieties



V1 V2 V1 V2

T1 13.66 A 12.00 A 14.00 A 12.33 A

T2 11.00 C 9.333 C 11.33 B 8.00 C

T3 12.00 BC 10.00 C 11.67 B 8.667 C

T4 12.33 B 10.33 BC 12.00 B 9.66 BC

T5 12.67 AB 11.67 AB 12.33 B 10.33 B


T1 = Control (distilled water; DW); T2 = Undiluted extract (9g extract + 10ml DW); T3 = 75%

extract + 25% DW (7.5g + 10ml); T4 = 50% extract + 50% DW (5g +10ml); T5 = 25% DW

(2.5g + 10ml).

38

Table 3: Effect of sunflower root extract on germination (%) of wheat varieties



V1 V2 V1 V2

T1 13.67 A 12.00 A 14.00 A 12.33 A

T2 10.33 C 9.00 C 9.333 C 6.667 C

T3 11.67 BC 9.667 C 10.67 BC 7.667 C

T4 12.00 ABC 10.67 B 11.00 B 9.333 B

T5 12.67 AB 11.33 A 11.67 B 9.667 AB


Table 4: Effect of leaf extract of sunflower on root length ( cm ) of seedlings of



V1 V2 V1 V2

T1 12.33 B 11.67 AB 14.00 A 13.00 A

T2 10.67 C 8.667 D 9.50 C 8.700 C

T3 11.67 BC 9.667 CD 10.20 C 9.500 C

T4 12.67 AB 10.67 BC 12.00 B 11.08 B

T5 13.67 A 12.50 A 13.00 AB 12.60 A


T1 = Control (distilled water; DW); T2 = Undiluted extract (9g extract + 10ml DW); T3 =

75% extract + 25% DW (7.5g + 10ml); T4 = 50% extract + 50% DW (5g +10ml); T5 =

25%

Table 5: Effect leaf extract of sunflower on shoot length (cm) of seedlings of wheat



V1 V2 V1 V2

T1 18.30 A 17.50 A 18.43 A 17.00 A

T2 8.700 E 12.30 E 8.267 E 11.91 E

T3 9.800 D 13.20 D 9.140 D 12.95 D

T4 11.20 C 13.90 C 11.01 C 13.53 C

T5 13.27 B 15.30 B 12.59 B 14.90 B


39

Table 6: Effect of sunflower leaf extract on fresh weight (g) wheat varieties



V1 V2 V1 V2

T1 0.9100 A 0.8900 A 0.9267 A 0.8300 A

T2 02000 E 0.6200 E 0.1833 E 0.59.00 E

T3 0.3000 D 0.6600 D 0.2733 D 0.6200 D

T4 0.6000 C 0.7200 C 0.5700 C 0.6800 C

T5 0.8100 B 0.8033 B 0.7567 B 0.7700 B




25% DW (2.5g + 10ml).

Table 7: Effect of leaf extract of sunflower on dry weight (g) of seedlings of wheat



V1 V2 V1 V2

T1 0.8700 A 0.8600 A 0.9833 A 0.8100 A

T2 0.1700 E 0.4200 E 0.1533 E 0.5600 E

T3 0.2700 D 0.4767 D 0.2433 D 0.5900 D

T4 0.5700 C 0.5133 C 0.5300 C 0.6540 C

T5 0.7700 B 0.5867 B 0.7500 B 0.7400 B


Table 8: Effect of stem extract of sunflower on root length (cm) of seedings of



V1 V2 V1 V2

T1 14.00 A 14.00 A 14.67 A 14.69 A

T2 10.00 C 10.33 CD 10.83 D 10.03 CD

T3 11.33 B 9.250 D 11.17 CD 8.917 D

T4 12.20 B 11.25 C 12.07 BC 10.87 C

T5 13.50 A 12.58 B 12.98 B 12.20 B




(2.5g + 10ml).

40

Table 9: Effect of stem extract of sunflower on shoot length (cm ) of seedlings of wheat



V1 V2 V1 V2

T1 18.30 A 18.30 A 18.43 A 18.43 A

T2 11.50 C 12.32 D 11.33 11.64 D

T3 12.49 CD 13.23 CD 12.35 C 13.03 CD

T4 13.15 C 13.85 C 12.58 C 13.40 C

T5 14.58 B 15.67 B 13.98 B 15.35 B


Table 10: Effect of stem extract of sunflower on fresh weight (g) of seedings of wheat



V1 V2 V1 V2

T1 0.9100 A 0.9100 A 0.9267 A 0.9100 A

T2 0.5100 C 0.7200 D 0.4900 C 0.6900 D

T3 0.5767 C 0.7800 C 0.5433 C 0.7433 C

T4 0.6933 B 0.8300 B 0.6733 B 0.8000 B

T5 0.8600 A 0.8900 A 0.8367 A 0.8600 A




(2.5g + 10ml).

Table 11: Effect of stem extract of sunflower on dry weight (g) of seedings of wheat



V1 V2 V1 V2

T1 0.8700 A 0.8600 A 0.8933 A 0.8933

T2 0.4667 C 0.4700 E 0.4500 C 0.5833 D

T3 0.5267 C 0.5167 D 0.5100 C 0.6567 C

T4 0.5433 C 0.5500 C 0.5200 C 0.7000 B

T5 0.7533 B 0.6400 B 0.7367 B 0.7567


41

Table 12: Effect of root extract of sunflower on shoot length (cm ) of seedlings of



V1 V2 V1 V2

T1 18.30 A 17.50 A 18.43 A 17.00 A

T2 11.86 D 12.50 C 11.64 D 12.92 B

T3 12.66 CD 14.20 B 13.03 CD 13.17 B

T4 13.19 C 12.13 C 13.40 C 13.70 B

T5 14.63 B 14.70 B 15.35 B 16.32 A




(2.5g + 10ml).

Table 13: Effect of root extract of sunflower on fresh weight ( g ) of seedlings of


2006 2007 Trtement

V1 V2 V1 V2

T1 0.8900 A 0.8900 A 0.9100 A 0.8300 A

T2 0.2333 E 0.7500 B 0.1900 E 0.7100 B

T3 0.3200 D 0.8300 A 0.2900 D 0.7900 A

T4 0.6333 C 0.8667 A 0.6000 C 0.8233 A

T5 0.8300 B 0.8733 A 0.7500 B 0.8400 A

Values followed by the same letter within a column are not significantly ifferent

Table 14. Effect of root extract of sunflower on dry weight ( g ) of seedlings of wheat



V1 V2 V1 V2

T1 0.8600 A 0.8600 A 0.8933 A 0.8333 A

T2 0.1833 E 0.5433 C 0.1633 E 0.5167 D

T3 0.2833 D 0.6300 B 0.2633 D 0.6133 C

T4 0.5833 C 0.6700 B 0.5567 C 0.6467 BC

T5 0.7833 B 0.6833 B 0.7533 B 0.6700 B




(2.5g + 10ml).

42

Table 15: Effect of root extract of sunflower on root length (cm) of seedlings of

wheat varieties Margalla 99 Chakwall 97


V1 V2 V1 V2

T1 13.00 A 17.50 A 12.57 A 17.00 A

T2 9.633 C 13.20 B 8.283 C 11.88 C

T3 10.50 BC 13.67 B 9.950 B 13.68 B

T4 10.90 B 14.20 B 11.65 A 12.01C

T5 12.40 A 16.96 A 12.30 A 13.98 B


43

Effect of leaf extract of sunflower on Gibberellic Acid contents of wheat seedlings


A perusal of the data in Table 16 and 17 revealed that both in roots and leaves of wheat

seedlings TI (control) showed the maximum GA, followed by T5 (25% extract)., GA content

was found minimum in T2 (undiluted extract). Results were found similar in the following

year.

Effect of leaf extract of sunflower on Indole Acetic Acid (µg g-l) contents of seedlings of

wheat varieties Margalla 99 and Chakwall 97

The data regarding IAA content in leaves of seedlings were presented in Table 18. A perusal of

table revealed that both the wheat varieties exhibited maximum IAA contents in leaves of TI

(control) followed by T5 (25% extract) which showed significant decrease in leaves IAA. The

treatment with the T2 (undiluted extract) showed the least IAA. The data regarding IAA

content in root presented in Table No. 19 revealed that in case of wheat variety Margalla 99 and

Chakwall 97, Tl (control) showed the maximum which was at par with T5(25%) , while T2

(undiluted extract) showed the minimum amount of IAA. Similar results were found in second

year.

Effect of leaf extract of sunflower on Abscisic Acid (µg g-l) contents of leaves and roots of


Data regarding ABA contents in leaves of wheat presented in Table 20 and 21 revealed that in

case of both wheat varieties Margalla 99 and Chakwall 97, maximum ABA contents in both

leaves and roots samples were found in T2 (undiluted extract), followed by T3 (75% extract)

and T4 (50% extract), whereas minimum ABA content was found in Tl (control). Results were

found similar in the second year.

Effect of stem extract of sunflower on Gibberellic Acid (µg g-l) contents of leaves and roots

in wheat varieties Margalla 99 and Chakwall 97

The data on the GA content in leaves were given in Table 22. The maximum GA contents in

case of wheat variety Margalla 99 were given by T5 (25% extract) which was at par with T1

44

(control), the minimum amount of GA was found in T2 (undiluted extract), while in the second

year Tl (control) showed the maximum amount of GA and T2 (undiluted extract) showed the

minimum amount of GA. In th wheat variety Chakwall 97, in both years, Tl (control) showed

the maximum and T2 (undiluted extract) showed the minimum quantity of GA in wheat leaves.

The data regarding GA content in root samples presented in Table No. 23 revealed that in both

years and in both the varieties, the maximum GA contents were recorded in Tl (control) and

the minimum in T2 (undiluted extract).

Effect of stem extract of sunflower on Indole-3-acetic Acid content of leaves and roots in


The data regarding IAA contents in leaves are presented in Table No. 24. A perusal of the table

revealed that in the case of both wheat varieties Margalla 99 and Chakwall 97, the maximum

IAA contents in leaves samples were calculated in TI (control) and the least amount were found

in T2 in the first year as well as second year. From the Table No. 25, it is revealed that in the

case of the root sample of wheat variety Margalla 99, TI (control) showed the maximum IAA

content, followed by T5 (25% extract), T4 (50% extract), T3 (75% extract), and T2 (undiluted

extract). In the case of wheat variety Chakwal197, Tl (control) showed the maximum values

which non significantly differed with T5 (25% extract) and T2 showed the minimum values;

similar results were found in the second year.

Effect of stem extract of sunflower on Abscisic Acid (µg g-l) content of leaves and root in


The data regarding ABA contents in leaves presented in Table No. 26 revealed that in both the

wheat varieties Margall 99 and Chakwall 97, maximum amount of ABA was recorded in T2

(undiluted extract) and the minimum in Tl (control). Similar result was found in the second

year. From the Table No. 27, it was revealed that in the case of wheat varieties, T2 (undiluted

extract) showed the maximum ABA contents in the root sample of wheat varieties and T5 (25%

extract) showed the minimum ABA contents.In the second year result was found similar.

45

Effect of root extract of sunflower on GA (µg g-l) content of leaves and root in wheat


The data regarding GA contents in leaves and roots samples were presented in Table 28 and 29.

A perusal of the tables regarding GA contents in leaves and root samples of wheat revealed that

in the case of both wheat varieties Margalla 99 and Chakwall 97, TI (control) showed the

maximum, while T2 (undiluted extract) showed the minimum values, whereas in the second

year result were found similar.

Effect of root extract of sunflower on IAA (µg g-l) content of leaves and roots in wheat

varieties Margalla 99 and Chakwall97

From the Table No. 30, it is revealed that in the case of leaves samples of both wheat varieties

Margalla 99 and Chakwall 97, T1 showed the maximum values, followed by T5 (25% extract),

and T4 (50% extract) whereas T2 (undiluted extract) showed minimum values. Results were

found similar in the second year. The data regarding IAA contents in root samples were

presented in Table No. 31. The data on IAA contents in both wheat varieties Margalla 99 and

Chakwall 97 revealed that T2 (undiluted extract) showed the maximum IAA content and Tl

(control) showed the minimum IAA contents in root sample in the first and second year of

experiment.

Effect of root extract of sunflower on Abscisic Acid (µg g-l) content of leaves and root in


The data regarding ABA contents in leaves and roots samples presented in Table No. 32 and 33

revealed that in the case of both wheat varieties Margalla 99 and Chakwall 97, T2 (undiluted

extract) showed the maximum and T1 (control) showed the minimum quantity of ABA

contents. Similar result was found in the second year.

46

Table 16: Effect of leaf extract of sunflower on GA ( µµµµg g-1

) content of leaves in wheat



V1 V2 V1 V2

T1 500.0 A 490.0 A 503.3 A 492.0 A

T2 382.7 D 340.3 B 380.0 D 335.3 B

T3 392.0 D 380.0 B 298.0 CD 375.0 B

T4 408.3 C 387.0 B 405.3 C 382.0 B

T5 446.7 B 430.0 AB 442.3 B 421.7 AB




25% DW (2.5g + 10ml).

Table 17: Effect of leaf extract of sunflower on GA ( µµµµg g-1

) content of roots in



V1 V2 V1 V2

T1 414.7 A 398.3 A 415.0 A 500.0 A

T2 303.7 C 344.3 B 301.3 C 382.7 D

T3 345.7 B 377.3 AB 340.7 B 392.0 CD

T4 357.7 C 393.7 A 353.0 B 408.3 C

T5 405.0 A 417.7 A 402.7 A 446.7 B


Table 18: Effect of leaf extract of sunflower on IAA ( µµµµg g-1

) content of leaves in



V1 V2 V1 V2

T1 163.0 A 132.0 A 164.7 A 133.0 A

T2 92.00 E 100.7 D 90.33 E 99.33 C

T3 120.0 D 105.7 CD 118.0 D 103.7 C

T4 139.0 C 110.7 BC 137.0 C 108.3 BC

T5 152.00 B 118.0 B 149.7 B 115.0 B




(2.5g + 10ml).

47

Table 19: Effect of leaf extract of sunflower on IAA (µµµµg g-1

) contents of roots in



V1 V2 V1 V2

T1 190.0 A 130.0 A 191.3 A 131.3 A

T2 113.3 E 90.33 D 111.3 E 87.67 D

T3 125.0 D 105.00 C 123.0 D 103.00 C

T4 135.0 C 103.0 C 131.7 C 107.0 C

T5 145.0 B 116.0 B 141.7 B 114.0 B


Table 20: Effect of leaf extract of sunflower on ABA (µµµµg g-1

) contents of leaves in



V1 V2 V1 V2

T1 68.00 E 90.30 E 66.33 E 91.61 E

T2 182.00 A 185.7 A 182.7 A 186.7 A

T3 162.00 B 167.3 B 163.3 B 168.3 B

T4 142.00 C 146.3 C 143.00 C 147.3 C

T5 108.00 D 110.3 D 109.00 D 111.7 D




(2.5g + 10ml).

Table 21: Effect of leaf extract of sunflower on ABA ( µµµµg g-1




V1 V2 V1 V2

T1 100.0 E 81.33 E 98.67 E 82.33 E

T2 168.7 A 122.00 A 169.7 A 123.00 A

T3 138.7 B 112.7 B 140.3 B 113.7 B

T4 117.7 C 100.0 C 119.3 C 101.1 C

T5 108.7 D 90.33 D 110.7 D 91.33 D


48

Table 22: Effect of stem extract of sunflower on GA ( µµµµg g-1

) content leaves in wheat



V1 V2 V1 V2

T1 500.0 A 490.0 A 503.3 A 492.0 A

T2 435.7 D 392.7 C 432.7 D 390.7 C

T3 459.0 C 404.0 C 456.7 C 399.3 C

T4 486.0 B 408.3 C 476.0 B 402.7 C

T5 504.0 A 458.3 B 497.0 A 452.0 B




(2.5g + 10ml).

Table 23: Effect of stem extract of sunflower on GA ( µµµµg g-1




V1 V2 V1 V2

T1 515.0 A 414.7 A 415.0 A 414.7 A

T2 394.3 E 340.0 D 340.0 D 312.0 C

T3 405.0 D 358.3 C 358.3 C 352.7 B

T4 415.3 C 385.7 B 385.7 B 363.7 B

T5 475.0 B 414.3 A 414.3 A 417.7 A


Table 24: Effect of leaf extract of sunflower on IAA (µµµµg g-1

) of seedlings of wheat



V1 V2 V1 V2

T1 163.0 A 132.00 A 144.0 A 191.3 A

T2 98.33 A 105.3 C 109.3 E 115.0 D

T3 127.00 C 110.3 BC 115.3 D 130.0 CD

T4 145.7 B 117.3 B 121.0 C 136.0 C

T5 158.00 A 127.3 A 133.7 B 167.0 B




(2.5g + 10ml).

49

Table 25: Effect of stem extract of sunflower on IAA (µµµµg g-1




V1 V2 V1 V2

T1 190.0 A 130.00 A 164.7 A 133.00 A

T2 118.0 D 100.0 C 96.33 E 100.3 C

T3 135.0 CD 110.0 C 124.00 D 106.3 CD

T4 141.7 C 118.0 B 140.7 C 112.3 C

T5 171.7 B 129.00 A 155.00 B 124.0 B


Table 26: Effect of stem extract of sunflower on ABA (µµµµg g-1

) content leaves in



V1 V2 V1 V2

T1 68.00 D 90.30 D 66.33 D 91.61 D

T2 141.0 A 145.0 A 143.0 A 147.0 A

T3 108.0 B 109.0 B 110.7 B 111.0 B

T4 103.00 B 101.3 BC 104.00 C 103.7 BC

T5 81.00 C 96.33 CD 84.00 D 98.33 CD




(2.5g + 10ml).

Table 27: Effect of stem extract of sunflower on ABA (µµµµg g-1

) content roots in wheat



V1 V2 V1 V2

T1 100.00 C 81.33 D 98.67 C 82.33 D

T2 117.00 A 99.67 A 119.00 A 101.7 A

T3 108.0 B 91.33 C 110.3 B 93.67 B

T4 102.3 C 85.33 C 105.0 C 88.00 C

T5 94.00 D 80.00 D 97.33 D 81.67 D

Values followed by the same letter within a column are not significantly different

50

Table 28: Effect of root extract of sunflower on GA ( µµµµg g-1




V1 V2 V1 V2

T1 500.0 A 490.00 A 503.0 A 492.00 A

T2 404.00 D 351.00 C 402.00 D 348.7 C

T3 418.7 C 388.3 BC 416.7 C 386.0 BC

T4 527.0 C 394.3 BC 424.7 C 391.7 BC

T5 480.0 B 437.0 AB 477.0 B 434.0 AB




(2.5g + 10ml).

Table 29: Effect of root extract of sunflower on GA ( µµµµg g-1




V1 V2 V1 V2

T1 505.3 A 422.3 A 407.0 A 419.7 A

T2 392.3 B 318.0 C 301.0 E 313.0 E

T3 399.0 B 360.0 B 349.7 D 349.0 D

T4 411.7 B 364.7 B 364.3 C 362.0 C

T5 482.3 A 417.0 A 417.0 A 412.3 B


Table 30: Effect of root extract of sunflower on IAA ( µµµµg g-1




V1 V2 V1 V2

T1 172.7 A 131.0 A 190.0 A 132.0 A

T2 100.0 E 102.0 E 113.0 E 96.00 D

T3 127.0 D 107.0 D 127.0 D 106.0 C

T4 139.3 C 114.0 E 139.0 C 107.0 C

T5 160.0 B 122.0 B 149.0 B 121.0 B




(2.5g + 10ml).

51

Table 31: Effect of root extract of sunflower on IAA ( µµµµg g-1




V1 V2 V1 V2

T1 193.0 A 164.0 A 164.0 A 134.0 A

T2 117.0 D 94.00 E 96.00 E 93.30 D

T3 129.0 C 120.0 D 125.7 D 109.0 C

T4 135.0 C 139.0 C 140.3 C 113.0 C

T5 155.7 B 153.0 B 155.0 B 123.0 B


Table 32: Effect of root extract of sunflower on ABA ( µµµµg g-1




V1 V2 V1 V2

T1 68.00 E 90.30 E 66.33 E 91.61 E

T2 177.3 A 177.3 A 180.3 A 180.0 A

T3 156.0 B 159.0 B 159.0 B 162.3 B

T4 133.7 C 132.7 C 136.3 C 135.0 C

T5 98.00 D 102.0 D 100.7 D 104.0 D




(2.5g + 10ml).

Table 33: Effect of root extract of sunflower on ABA ( µµµµg g-1




V1 V2 V1 V2

T1 100.0 C 81.33 D 98.67 C 82.33 D

T2 151.0 A 113.7 A 153.0 A 116.3 A

T3 130.0 B 106.0 B 132.0 B 108.0 B

T4 105.7 C 96.67 C 107.7 C 98.67 C

T5 100.3 C 84.00 D 102.7 C 86.00 D


52

Effect of leaves, stem and root extracts of sunflower on Deoxyribonucleic Acid content of

leaves in wheat varieties Margalla 99 and Chakwall 97

The data regarding DNA were presented in Table 34, 35 and 36. The data pertaining to the

DNA contents in leaves of wheat varieties were also affected significantly by leaf, stem and

root extract of sunflower. In both wheat varieties Margalla 99 and Chakwall 97, the maximum

DNA was observed in TI (control) and the minimum in T2 (undiluted extract) in the first as

well as second year of experiment under the effect of leaves, stem and root extract of

sunflower.

Effect of leaves, stem and root extracts of sunflower on chlorophyll content (mg/g) of

leaves in wheat varieties Margalla 99 and Chakawall 97

The data regarding effect of leaf stem and root extract on chlorophyll contents were presented

in Table 37, 38 and 39. From the tables, it is revealed that the maximum chlorophyll contents

under the effect of leaf, stem and root extract of sunflower were recorded in Tl (control) and the

minimum chlorophyll content was found in T2 (undiluted extract) in both wheat varieties

Margalla 99 and Chakwall 97. In the second year, results were found similar.

Effect of leaves, stem and root extract of sunflower on proline content (mg/g) of leaves in


Data presented in Table 40, 41 and 42 revealed that proline contents of leaves under the effect

of leaf, stem and root extract were affected significantly by sunflower extract. A perusal of the

tables revealed that in both the wheat varieties, T2 (undiluted extract) showed the maximum

proline contents under the effect of leaf, stem and root extract and TI (control) showed the

minimum proline contents. Results were found similar in the second year of experiment.

Effect of leaves, stem and root extract of sunflower on sugar content (mg/g) of leaves of


Effect of leaf and stem extract of sunflower on sugar content of leaves in wheat varieties was

shown in Table 43 and 44. The data regarding sugar contents revealed that in both the wheat

varieties Margalla 99 and Chakwall 97 in two years, T2 (undiluted extract) showed the

maximum sugar content while Tl (control) showed the minimum amount of sugar. Effect of

53

root extract of sunflower on sugar content of leaves in wheat varieties was given in Table No.

45. A perusal of the table showed that in the case of wheat variety Margalla 99, T2 (undiluted

extract) showed the maximum while T5 (25% extract) showed the minimum amount of sugar

contents; this is on par with TI (control), and TI (control) showed the minimum values in the

Chakwall 97. Similar result was found in the second year of experiment.

Effect of leaves, stem and root extract of sunflower on protein content (mg/g) in leaves of


Effect of leaf and root extract of sunflower on protein content in leaves of wheat varieties was

shown in Table No. 46 and 48. A perusal of the table revealed that in the case of both wheat

varieties Margalla 99 and Chakwall 97, T2 (undiluted extract) showed the maximum contents

of protein under the effect of leaf and root extract, which was followed by T3 (75% extract),

and TI (control) showed minimum amount of protein. Similar results were shown in second

years. Table No. 47 revealed the effect of stem extract on the protein content under the effect of

stem extract in the case of wheat variety Margalla 99, T2 (undiluted extract) showed the

maximum and T5 (25% extract) showed the minimum, which was at par with TI (control),

while in the case of wheat variety Chakwall 97, Tl (control) showed the maximum values,

followed by T2 (undiluted extract), minimum amount was found in T5 (25% extract), and in

the second year in the case of wheat variety Margalla 99, T2 showed the maximum and TI

(control) showed the minimum amount of protein contents. In the case of wheat variety

Chakwall 97, T2 (undiluted extract) showed the maximum and TI (control) showed the

minimum.

Effect of leaf, stem and root extract of sunflower on Superoxide dismutase (mg/100g fresh

weight) of leaves in wheat varieties Margalla 99 and Chakwall 97

From the Table 49 and 50 it is revealed that in the case of both wheat varieties Margalla 99 and

Chakwal1 97, T2 (undiluted extract) showed the maximum Superoxide dismutase (SOD)

content under the effect of sunflower leaf and stem extract, followed by T3 (75% extract), and

T4 (50% extract) whereas TI (control) showed minimum amount of SOD. Similar results were

found in the second. From the Table No. 51, it is revealed that in the case of wheat variety

Margalla 99, T2 (undiluted extract) showed the maximum SOD content under the effect of root

54

extract, (which was at par with T3 ;75% extract) and T4 (50% extract), followed by T5 (25%

extract) and TI (control) showed minimum amount of SOD, while in the second year T2

(undiluted extract) showed the maximum, which was on par with T3 (75% extract), followed

by T4 (50% extract), T5 (25% extract), and T1 (control). In the case of wheat variety Chakwall

97, T2 (undiluted extract) showed the maximum, followed by T3 (75% extract) and T4 (50%

extract) while T1 (control) showed minimum amount of SOD. Similar result was shown in

second year.

Effect of leaf, stem and root extract of sunflower on Peroxidase (POD) content (mg/100g

fresh weight) of leaves in wheat varieties Margalla 99 and Chakwall 97

From the Table No. 52 and 53, it is revealed that in the case of both wheat varieties Margalla

99 and Chakewall 97, T2 (undiluted extract) showed the maximum POD content under the

effect of leaf and stem extract of sunflower, which was followed by T3 (75% extract) and T4

(50% extract), whereas Tl (control) showed minimum amount of POD. Similar result was

shown in the second year. From the Table No. 54, it is revealed that in the case of wheat variety

Margalla 99, T2 (undiluted extract) showed the maximum POD activity under the effect of root

extract, followed by T3 (75% extract), T4 (50% extract), T5 (25% extract), and Tl (control). In

the case of wheat variety Chakwal197, T2 showed the maximum values, which was on par with

T3 (75% extract), T4 (50% extract), T5 (25% extract), and T1 (control). In the case of wheat

variety Chakwall 97, T2 (undiluted extract) showed the maximum values, which was on par

with T3 (75% extract), T4 (50% extract), and T5 (25% extract), followed by Tl (control), while

in the second year result was found similar.

55

Table 34: Effect of leaf extract of sunflower on DNA ( mg/ 100 g F. wt ) content of leaves

in wheat varieties Margalla 99 Chakwall 97.


V1 V2 V1 V2

T1 513.00 A 485.3 A 515.00 A 487.0 A

T2 135.6 D 93.67 E 133.4 D 92.00 E

T3 221.00 C 193.3 D 218.7 C 190.7 D

T4 256.7 C 249.3 C 254.7 C 247.0 C

T5 376.7 B 367.3 B 374.7 B 365.3 B




25% DW (2.5g + 10ml).

Table 35: Effect of stem extract of sunflower of DNA ( mg/ 100 g F. wt ) content of



V1 V2 V1 V2

T1 513.0 A 485.3 A 514.6 A 488.0 A

T2 248.7 C 242.0 D 247.0 C 239.3 D

T3 386.0 B 378.3 C 383.7 B 374.0 C

T4 420.7 B 415.3 B 417.3 B 412.7 B

T5 490.3 A 466.0 A 488.0 A 464.3 A


Table 36: Effect of root extract of sunflower on DNA ( mg/ 100 g F. wt ) contents of



V1 V2 V1 V2

T1 512.9 A 485.3 A 515.0 A 487.0 A

T2 150.4 D 101.3 E 148.7 D 99.33 E

T3 239.0 C 206.7 D 237.7 D 204.7 D

T4 272.3 C 269.3 C 269.0 C 266.7 C

T5 398.3 B 396.7 B 395.3 B 393.7 B




(2.5g + 10ml).

56

Table 37: Effect of leaf extract of sunflower on chlorophyll ( mg/ 100 g F. wt)



V1 V2 V1 V2

T1 109.7 A 107.0 A 111.9 A 108.0 A

T2 83.43 D 81.67 D 82.47 D 80.33 D

T3 92.10 C 90.33 BC 91.09 C 89.71 C

T4 98.00 BC 96.33 BC 96.97 BC 95.45 BC

T5 103.0 AB 103.0 AB 102.0 B 102.6 AB


Table 38: Effect of leaf extract of sunflower on IAA content of leaves in wheat



V1 V2 V1 V2

T1 107.0 A 109.7 A 111.9 A 108.0 A

T2 90.43 C 98.00 C 96.00 D 89.77 C

T3 96.33 BC 103.0 B 101.3 BC 95.00 BC

T4 103.7 AB 104.7 B 102.3 BC 102.7 AB

T5 106.00 A 106.3 AB 104.7 AB 105.00 A




(2.5g + 10ml).

Table 39: Effect of root extract of sunflower on chlorophyll ( mg/ 100 g F. wt)



V1 V2 V1 V2

T1 109.7 A 107.0 A 111.9 A 108.0 A

T2 87.34 D 84.00 D 86.66 D 83.00 D

T3 94.34 CD 93.00 C 93.00 AB 92.00 C

T4 100.00 BC 98.67 BC 99.00 AB 97.67 BC

T5 105.3 AB 103.3 AB 104.3 A 102.3 AB


57

Table 40: Effect of leaf extract of sunflower on proline ( mg/ 100 g F. wt ) content of



V1 V2 V1 V2

T1 82.70 E 65.06 D 81.00 E 64.63 D

T2 143.3 A 132.7 A 145.0 A 133.7 A

T3 131.8 B 119.3 B 133.7 B 122.1 B

T4 122.5 C 114.1 C 124.6 C 115.5 B

T5 106.1 D 97.67 C 109.1 D 102.4 C




(2.5g + 10ml).

Table 41: Effect of stem extract of sunflower on proline ( mg/ 100 g F. wt ) content



V1 V2 V1 V2

T1 82.70 C 65.06 C 81.00 C 64.63 C

T2 118.0 A 111.3 A 119.8 A 112.3 A

T3 112.0 A 102.7 A 114.2 A 106.7 A

T4 94.67 B 88.00 B 96.00 B 89.00 B

T5 90.34 B 73.34 C 91.33 B 74.34 C


Table 40: Effect of root extract of sunflower on proline ( mg/ 100 g F. wt ) contents



V1 V2 V1 V2

T1 82.70 E 65.06 D 81.00 E 64.63 D

T2 130.0 A 119.7 A 131.3 A 121.0 A

T3 121.0 B 112.4 A 123.0 B 113.4 A

T4 114.7 C 101.3 B 116.7 C 102.3 B

T5 96.00 D 91.67 C 97.00 D 92.67 C




(2.5g + 10ml).

58

Table 43: Effect of leaf extract of sunflower on sugar ( mg/ 100 g F. wt ) contents of



V1 V2 V1 V2

T1 175.0 C 170.0 C 174.0 C 169.0 C

T2 225.0 A 217.7 A 226.0 A 218.7 A

T3 205.0 B 200.0 :B 206.0 B 201.0 B

T4 181.7 C 176.7 C 182.7 C 177.7 C

T5 178.3 C 173.7 C 179.3 C 174.7 C


Table 44: Effect of stem extract of sunflower on sugar ( mg/ 100 g F. wt ) contents



V1 V2 V1 V2

T1 175.0 C 170.0 C 174.0 C 169.0 C

T2 213.3 A 207.7 A 214.3 A 208.7 A

T3 199.0 B 192.3 B 200.0 B 193.7 B

T4 176.7 C 173.3 C 177.7 C 175.0 C

T5 173.0 C 169.0 C 174.0 C 170.0 C




(2.5g + 10ml).

Table 45: Effect of root extract of sunflower on sugar ( mg/ 100 g F. wt ) content of



V1 V2 V1 V2

T1 175.0 C 180.0 CD 174.0 C 169.0 C

T2 235.0 A 226.0 A 222.0 A 212.3 A

T3 215.0 B 210.0 B 202.7 B 197.0 B

T4 190.0 C 184.3 C 178.0 C 175.3 C

T5 182.0 :E 177.7 D 176.0 C 172.0 C


59

Table 46: Effect of leaf extract of sunflower on protein (mg/ 100 g F. wt ) contents of



V1 V2 V1 V2

T1 1685 E 1723 AB 1690 E 1732 AB

T2 1725 A 1737 A 1731 A 1742 A

T3 1720 B 1725 AB 1726 B 1731 ABC

T4 1715 C 1720 B 1721 C 1725 BC

T5 1705 D 1711 B 1711 D 1717 C




(2.5g + 10ml).

Table 47: Effect of stem extract of sunflower on protein (mg/ 100 g F. wt ) activity



V1 V2 V1 V2

T1 1685 AB 1723 A 1690 C 1755 D

T2 1700 A 1695 B 1725 A 1792 A

T3 1683 AB 1677 C 1718 A 1784 B

T4 1670 B 1665 C 1710 B 1772 C

T5 1670 B 1665 C 1696 C 1765 C


Table 48: Effect of root extract of sunflower on protein (mg/ 100 g F. wt ) activity of



V1 V2 V1 V2

T1 1685 D 1766 B 1753 D 1675 C

T2 1723 A 1785 A 1783 A 1725 A

T3 1720 A 1775 AB 1775 B 1760 B

T4 1712 B 1772 B 1768 BC 1755 A

T5 1695 C 1765 B 1761 A 1692 C




(2.5g + 10ml).

60

Table 49: Effect of leaf extract of sunflower in superoxidase (unit/g f.w) activity of leaves



V1 V2 V1 V2

T1 2.670 B 2.330 D 3.670 B 3.000 B

T2 8.330 A 7.670 A 7.330 A 6.670 A

T3 8.000 A 7.000 AB 7.000 A 5.670 A

T4 7.000 A 6.330 B 6.000 A 5330 A

T5 4.000 B 3.670 C 3.000 B 2.670 B


Table 50: Effect of root extract of sunflower on superoxidase (unit/g f.w) of leaves in



V1 V2 V1 V2

T1 2.670 C 3.000 B 3.000 B 3.000 BC

T2 6.000 A 5.000 A 5.330 A 5.000 A

T3 5.000 AB 4.000 AB 5.000 A 4.330 AB

T4 3.000 C 2.670 B 4.670 A 3.670 ABC

T5 3.330 BC 2.330 B 3.000 B 2.330 C




(2.5g + 10ml).

Table 51: Effect of stem extract of sunflower on superoxidase dismoutase (unit/g f.w) of



V1 V2 V1 V2

T1 2.670 B 2.330 C 3.670 BC 2.000 A

T2 7.330 A 6.670 A 6.330 A 5.670 A

T3 7.000 A 6.000 AB 6.000 A 4.670 A

T4 6.000 A 5.330 B 5.000 AB 4.330 A

T5 3.000 B 2.670 C 2.000 C 1.670 B


61

Table 52: Effect of root extract of sunflower peroxidase (OD/min/g f.w) activity



V1 V2 V1 V2

T1 8.000 C 7.000 D 8.000 B 6.000 D

T2 15.33 A 13.67 A 14.33 A 12.67 A

T3 14.33 AB 12.33 AB 13.33 A 11.33 AB

T4 13.33 AB 10.67 BC 12.33 A 9.670 BC

T5 11.67 B 9.000 CD 12.00 A 8.000 CD




(2.5g + 10ml).

Table 53: Effect of stem extract of sunflower on peroxidase (OD/min/g f.w) activity of



V1 V2 V1 V2

T1 8.000 C 7.000 C 7.000 C 6.000 C

T2 13.67 A 12.00 A 12.67 A 11.67 A

T3 12.67 AB 10.00 AB 11.67 AB 10.67 AB

T4 11.00 ABC 9.330 ABC 10.33 AB 9.330 AB

T5 9.670 BC 8.670 BC 8.670 BC 7.670 BC


Table 54: Effect of root extract of sunflower on peroxidase (OD/min/g f.w)



V1 V2 V1 V2

T1 8.000 C 7.000 B 7.670 C 6.00 B

T2 14.67 A 13.00 A 15.33 A 14.00

T3 13.67 AB 11.00 A 14.67 A 11.67 A

T4 12.33 AB 10.67 A 13.33 AB 11.33 A

T5 10.67 BC 10.33 A 11.00 B 8.00 C




(2.5g + 10ml).

62

DISCUSSION

The allelopathic phenomenon has received much attention, as shown by numerous reports on

the subject (Narwal e1 1998; Anaya, 1999; Callaway and Aschhoug, 2000;Singh et al., 2001;

Weston and Duke, 2003; Harper et al., 2005: Khan et al., 2005;Reigosa et al., 2006).

Recently, allelopathy is exploiting as a weed control strategy, alternative to the commercial

herbicide dominated programs (Bhowmik and Inderjit 2003). Sunflower (Helianthus annuus

L.) can actively influence the growth of surrounding plants due to its high allelopathic potential

(Azania et al. 2003). Significant inhibition in growth of mustard plants due to the presence in

the soil both sunflower roots exudates and/or tiny not possible to collect roots and root hairs

was detected by Ciarka et al. 2004. It was suggested that the reduced growth and yield of crops

can be attributed to the release of phytotoxic phenolics from decomposing sunflower residues,

since the soil collected from sunflower fields was rich in phenolics (Batish et al. 2002).

The production of allelochemicals in crop plants and their release into the soil could influence

the germination and growth of plant species (Rice, 1984). These effects are selective,

depending upon the concentrations and residue type, either inhibitory or stimulatory to the

growth of companion or subsequent crops or weeds (Cheema et al., 2004; Jalili et al., 2007

Naseem et al., 2003 ;). The present study has shown that sunflower (Helianthus annus) extracts

from leaves, stems, and roots) affected seed germination significantly. It indicates that

reduction in these parameters (fresh weight, dry weight, root length, shoot length) might have

been the result of water-soluble allelochemicals in the extract and their inhibitory effect or

phytotoxicity on the measured parameters. Results showed that sunflower plant parts varied in

their allelopathic activity against wheat seed extracts of leaves, followed by roots, which

showed a higher allelopathic potential on test plants as compared to stems. These results are in

accordance with previous studies reporting that allelopathy may vary among plant parts (Turk

and Tawaha 2002, Peng al., 2004). For all extracts, allelopathic activity increased with

increases in extract concentrations. This is in agreement with the finding that under certain

conditions, rate of an elementary reaction is positively related to the reactant concentration.

Previous studies have shown that the phytotoxicity of extracts was significantly increased as

the concentration increased (Rice 1984, Putnam 1994, Sinkkonen 2001, Peng et al., 2004)

63

Wheat seed germination was reduced by all extracts of sunflower as compared with untreated

control. The observed inhibition of the two wheat varieties, namely Margalla 99 and Chakwall

97, for germination of seeds could be attributable to a contribution (allelochemicals released

from the sunflower extracts of leaves, stems, and roots, as the allelochemicals are water-soluble

and can accumulate upon release in seeds in direct contact with bioactive concentrations. There

are several reports from the literature showing that addition or incorporation of plant residues

into the growth environment of another plant can result in growth inhibition (Patterson 1981,

Qasem 1994, Chung and Miller 1995, AI-Khatib et al., 1997). Seedling chlorosis was observed

at high concentration of sunflower extracts; this may be due to a sunflower phenolic

allelochemical effect. The effect of higher concentration of extracts of sunflower suggests that

reduction in measured parameters may have the result of either or both the osmotic potential of

the extracts or the presence of allelochemicals in the extracts. Beside possible allelochemicals,

higher chemical concentrations in the extracts might possibly cause an osmotic stress during

seed germination and seedling growth. The inhibition of seed germination and seedling growth

was concentration- dependent; present results are similar to those obtained by other workers,

who observed inhibition of seed germination and seedling growth on other crops and weeds by

sunflower extracts (Batish et al., 2002: Bogatek et al., 2005; Anjum et al., 2005). The

allelochemicals inhibit germination and seedling growth, probably by affecting the cell division

and elongation process that are very important at this stage or by interfering with enzymes

involved in the mobilization of nutrients necessary for germination. Levizou et al. (2002) found

that mitosis in root apex of lettuce retarted with leaf extracts. Growth hormones, like GA and

IAA, are very important in agriculture. The growth-inhibiting substances of agricultural

importance have received considerable research attention these days (Siquira et al., 1991;

Inderjit, 1996). These compounds, which affect phytohormones, are phenols with allelopathic

characteristics (Schenk et al., 1999; Callaway and Aschhoug, 2000). In plants exposed to

sunflower extracts changes in ABA were also recorded. In the leaves, the ABA content

increased. In the case of roots, levels of ABA were very low in control plants, and in treated

plants they were high. This was probably due to the fact that in roots, as being initially affected

by allelochemicals, cell death would occur. In roots with gradually dying cells, most of the

already synthesized ABA is probably transported to the shoot, as a stress signal messenger. The

increase of ABA in leaves correlates well with changes described above in the plant water

64

status; i.e., loss of turgor, lowered osmotic potentials and relative water content, and the

increase in leaf diffusive resistance. The involvement of ABA in plants coping with abiotic and

biotic stresses, especially those dealing with cell water deficit, is well documented in the

literature (Davies and Jones, 1991). This study clearly showed that ABA also plays an

important role in the defense, processes against allelopathic stress. An increase of ABA levels

in seedlings of wheat exposed to allelopathy stress was observed in another study (Bernat et al.

2003). However, in this study, conversely to this work, the increase was recorded not only in

leaves but also in roots. Different patterns of ABA accumulation in these two growth stages by

wheat plants due to the same stress suggest that there are differences in mechanisms of coping

with allelopathy stress or in sensitivity to this stress. Results presented in this work

demonstrated that sunflower cv. Hysun 38 are the donors of allelopathic compounds, which

affect acceptor plants via changes in many physiological processes and that these changes,

being mostly statistically significant, showed a dose- and time- dependent relation. Recently,

some progress has been made in the study of molecular processes involved in morphological

and physiological adaptation of plants exposed to allelopathic chemicals of other plants. The

present research was carried out to study the allelopathic effects of leaf stem and root on

morphological, biochemical, and molecular criteria of wheat leaves. These extracts can be used

in biological control as natural herbicides to reduce the risk of manufactured herbicides. With

respect to the total soluble protein and the nucleic acids contents, there was a highly significant

increase in the level of DNA in all treated samples of wheat seedlings. Wheat seedlings treated

with sunflower extracts showed a highly significant increase in the level of DNA. These results

were confirmed partially by Duhan et al., (1995) who revealed a drastic increase in the level of

nucleic acids and decrease in the level of soluble proteins in legume crops in response to plant

extracts. In accordance with these results, (Baziramakenga et al., 1997), reported that many

phenolic acids reduced the incorporation of phosphorus into DNA in soybean. The present

results are in controversy with those obtained by (Padhy et al, 2000), who reported that

chlorophyll synthesis in leaves, as well as protein, carbohydrate, and nucleic acid (DNA and

RNA) contents, in both shoots and roots of seedling was also decreased with increases in

extracts concentrations. It was found that the activities of SOD and POD of many plants were

affected by allelochemicals. Numerous studies have shown that the degree of injury caused by

allelochemicals was negatively correlated to the increase of activities of SOD and POD

65

(Dhindsa and Matow, 1981; Chowdhury and Choudhuri, 1985). In this study, it was found that

SOD and POD activities in roots and leaves increased with allelochemicals. These differences

may result in different treatment methods. In this study, the allelochemicals were applied

during the whole growth period. Allelochemicals probably acted directly on roots, whereas

leaves could reduce water transpiration by curling and stomatal regulation. Furthermore, the

degree of leaf injury caused by allelochemicals could be lessened through morphological

adaptation and regulation of stem and sheath water content, stem diameter, or plant height. This

showed that the ability of crops to resist allelochemicals was connected to the activities of

protective enzymes and their defensive function. These mechanisms may be the main

physiological action and defense from injury of crops under allelochemicals. There were

significant differences in endogenous concentrations of GA and .IAA in both leaves and roots

of two wheat varieties during their growth when different concentrations of extracts of

sunflower were applied. The GA and .IAA concentration in leaves as well as roots decreased

when different concentrations of extracts of sunflower were applied, while a higher reduction in

both IAA and GA concentration was found when higher concentrations of extracts of sunflower

were applied in 2 wheat varieties. This decrease in leaves' GA and IAA may be attributed to

slow transport of growth-promoting hormones from vegetative organs. Phytohormones may

also enhance root development under growing conditions. Kuroha et al. (2002) reported that

plant hormones stimulate adventitious root formation. Similar findings have been reported

previously (Oda et al., 2003). The ability of the plants to absorb nutrients efficiently can be

attributed to a higher number of adventitious root formations, and GA promotes this (Wahyuni

et al., 2003; Kono, 1995; Watanabe, 1997). Root initiation and early development of root are

also stimulated by auxin (Bellamine et al., 1998; Pan and Tian, 1999). There were significant

decreases in endogenous concentrations of GA and IAA in the T1 and T5 treatments in both

varieties (Margalla 99 and Chakwall 97). In controlling stomatal resistance, the most important

signal is considered to be the concentration of plant hormones in xylem sap (Borel et al., 1997).

Extracts of sunflower (leaves, stems, and roots) decreased IAA and GA and increased ABA

concentration in both leaves and roots compared to control. Plant responses to allelopathy

(stress) can also be determined by the variations in IAA, GA, and ABA concentration (Naqvi,

1999). A rapid decrease in leaf ABA concentration observed with low concentrations of

allelochemicals. The GA concentration was decreased in plants under water stress conditions

66

(Aharaoni, 1979; Guinn-Brummet, 1988; Yang et al., 2001; Xie et al., 2003). The increase in

ABA may possibly be attributed to an induction in ABA synthesis. Abscisic acid contents in

seedlings of leaves treated with extracts of sunflower were greater than those of control (Yang

et at., 2001, 2004). During the present investigation, it was found that extracts of sunflower

significantly increased the accumulation of soluble sugars in both wheat varieties. In these

experiments, extracts of leaves, followed by root extracts and stem extracts, significantly

increased the soluble sugar contents of wheat varieties Margalla 99 and Chakwall 97.

Allelochemical treatment (TI) markedly increased sugar content in leaves. This increase may

be due to the positive effect of ABA on assimilate translocation. Assimilate translocation to the

developing seeds is reported to be under the control of ABA ('Brenner and Cheikh 1995; Yang

et al., 1999, 2004). Allelochemicals decreased the accumulation of sugars markedly in leaves.

The allelochemical treatments also showed an increase in sugar contents both in leaves.

Ahmadi and Bakicer (1999) reported that ABA is involved in osmolyte regulation under

moisture stress conditions. Mahajan and Tuteja (2005) reported that under severe drought,

growth was inhibited by high concentrations of ABA and sugar, whereas low concentrations

promote growth. Increased rates of photosynthesis and higher chlorophyll content might cause

accumulation of sugars due to ABA or allelochemical treatments (Dong et al., 1995; Ndung et

al., 1997). Drought tolerance in plants is enhanced by ABA caused by allelochemical treatment,

possibly due to the accumulation of osmolytes, such as sugars. The present investigation

indicated that application extracts of sunflower parts, like leaves, stems, and roots, caused

accumulation of protein in both wheat varieties, Margalla 99 and Chakwall 97. Nevertheless,

the magnitude of increase was higher in Margalla 99 and Chakwall 97. The application of

extracts of sunflower significantly increased the protein accumulation in both wheat varieties

Margalla 99 and Chakwal197. In these experiments, the leaf extracts was more effective,

followed by root extracts, and the lowest effect was observed in the case of stem extract. It was

observed that allelochemicals increased protein content in leaves. This may be due to a positive

role of ABA present in the extract on protein accumulation. Guerrero and Mullet (1986) and

Schmitz et al. (2000) reported that protein synthesis in developing seeds is induced by ABA.

Zhang et al. (2001) reported that protein phosphorylation is enhanced under water stress due to

increased concentration of ABA. Bartels and Sunkar (2005) and Ingram and Bartels (1996)

investigated late embryogenesis- abundant (LEA) proteins induced in vegetative tissues of

67

plants in response to osmotic stress that may interact with carbohydrates to prevent cellular

damage during dehydration. Which might bear similar to allelochemicals effect. In the present

study, it is shown that allelopathy of sunflower caused accumulation of proline in both wheat

varieties Margalla 99 and Chakwal1 97. The intensity of increase was higher in Margalla 99.

The application of extracts of sunflower significantly increased the proline accumulation in

both wheat varieties. However, a significantly higher increase was recorded in Margalla 99.

The efficiency of sunflower leaves, followed by root extracts, to increase proline contents was

higher than root and stem extracts. Applications of allelochemical treatments markedly

increased proline content in leaves of two wheat varieties, Margalla 99 and Chakwall 97. This

magnitude of the increase was similar to that under drought stress. Under drought stress, in

addition to its role as an osmoregulator, proline, like other soluble organic compounds, may

also act as osmo-protectants (Kamelie and Lose, 1995). ABA was considered to be involved in

the accumulation of proline (Hare et al., 1999; Hose et al., 2000; Trotel Aziz et al., 2000;

Nayyar and Walia, 2003), carbohydrates (Ahmadi and Baker, 2001), and other osmolytes in

plants (Popova et al., 2000). Allelochemical-caused stress increased the accumulation of

proline significantly in leaves. ABA enhanced the accumulation of proline contents in leaves.

This increase may be due to the role of ABA, which may stimulate proline accumulation under

water deficit conditions. Root length, shoot length, fresh weight, and dry weight of wheat

seedling were significantly different in both the varieties with respect to different

concentrations of extracts of sunflower; indicating difference in response.

68

THIRD EXPERIMENT

Effect of sunflower leaf, stem and root extracts on weed density in wheat field 30 days

after sowing (wheat varieties Margalla 99 and Chakwall 97)

Data in Table 1 revealed that in the case of both wheat varieties Margalla 99 and Chakwal1 97,

minimum weeds were counted in T2 (leaf extract), followed by T4 (root extract), The effect of

T3 (stem extract) was at par with the untreated control (Tl) in both the year of experimentation.

Effect of sunflower leaf, stem, and root extracts on fresh weight and dry weight (g) in

wheat 40 days after sowing (wheat varieties Margalla 99 and Chakwall 97)

The data regarding fresh and dry weight of weeds were presented in Table 3 and 4. A perusal of

the table revealed that fresh and dry weight of weeds was affected significantly by various

extracts of sunflower. Data indicated that in case of both wheat varieties Margalla 99 and

Chakwall 97, T2 (leaf extract) showed the minimum weeds fresh and dry weight, followed by

T4 and T3 (stem extract), and maximum weeds fresh and dry weight was counted in Tl

(control). Result was found similar in the second year.

Effect of sunflower leaf, stem, and root extracts on fresh weight and dry weight (g) of

weeds in wheat 70 days after sowing (wheat varieties Margalla 99 and Chakwall 97)

From the Table No.5 and 6, it is revealed that in the case of wheat variety Margalla 99,

maximum fresh and dry weight was counted in TI (control) under the effect of leaf, stem and

root extract, followed by T4 (root extract), T3 (stem extract), and T2 (leaf extract). Results

were found similar in the second year. In the case of wheat variety Chakwall 97, minimum

weeds fresh weight was counted in TI (control), followed by T3 (stem extract), T4 (root

extract), and T2 (leaf extract), while in the second year in the case of wheat variety Chakwall

97, maximum dry weight was counted in Tl (control) followed by T3 (stem extract), T4 (root

extract), and T2 (leaf extract).

Effect of sunflower leaf, stem and root extracts on number of tillers of wheat 145 days


A perusal of the Table No.7 showed that the number of tillers was affected significantly by

69

various extracts of sunflower. Data indicated that in the case of both wheat varieties Margalla

99 and Chakwall 97, T2 (leaf extract) give the highest number of tillers, followed by T4 (root

extract) and T3 (stem extract), while T1 (control) showed minimum number of tillers. In the

second year results was found similar.

Effect of sunflower leaf, stem and root extracts on plant height (cm) of wheat 145 days


From the Table No.7, it is revealed that plant height of wheat was affected significantly by

various extracts of sunflower. In the case of both wheat varieties Margalla 99 and Chakwall 97

maximum plant height was recorded in T2 (leaf extract) in the first as well as second year of

experiment, followed T4 (root extract) and T1 (control) showed minimum hight.

Effect of sunflower leaf, stem, and root extracts on l00-seed weight (g) of wheat 145 days

after sowing (Wheat varieties Margalla 99 and Chakwall 97)

Data presented in Table No.8 showed that 100-seed weight was affected by extract of

sunflower leaves. The maximum 100-seed weight in both wheat varieties Margalla 99 and

Chakwall 97 was recorded in the case of T2 (leaf extract), followed by T4 (root extract),

whereas T1 (control) showed minimum seed weight. Results were found similar in the first and

second year.

Effect of sunflower leaf, stem and root extracts on fresh weight and dry weight (g) of

wheat plant 145 days after sowing (wheat varieties Margalla 99 and hakwa1l 97)

Data presented in Table No. 9 and 10 showed that fresh and dry weight of wheat varieties were

significantly affected by leaf, stem and root extracts of sunflower. In the case of both wheat

varieties Margalla 99 and Chakwall 97, T2 (leaf extract) showed the maximum fresh and dry

weight, followed by T4 (root extract), whereas Tl (control) showed minimum fresh and dry

weight. Similar results were found in the following year.

70

Table 1: Effect of sunflower leaf, stem and root extracts on weed density in wheat 30

days after sowing ( wheat varieties Margalla 99 and Chakwall 97).


V1 V2 V1 V2

T1 5.000 A 6.670 A 6.00 A 7.330 A

T2 2.330 B 3.330 C 3.010 C 4.020 C

T3 4.330 A 5.000 B 5.00 AB 5.150 B

T4 3.670 AB 4.000 C 4.00 C 4.480 C


Table 2. Effect of sunflower leaf, stem and root extracts on fresh weight (g) in

wheat 40 days after sowing ( Wheat varieties Margalla 99 and Chakwall 97).


V1 V2 V1 V2

T1 8.490 A 6.270 A 6.000 A 7.330 A

T2 5.000 B 3.6000 D 2.970 C 4.020 C

T3 6.490 B 4.980 B 4.330 B 5.150 B

T4 5.900 AB 4.9000 C 3.670 B 4.480 C


Distilled water: T0V1, control ( DW ); T1V1, leaf extract ( 1g + 10 ml DW ); T2V1, stem

extract ( 1g + 10 ml DW ); T3V1, root extract ( 1g + 10 ml DW ); V1, Margalla 99; Distilled

water: T0V2, control ( DW ); T1V2, leaf extract ( 1g + 10 ml DW ); T2V2, stem extract ( 1g +

10 ml DW ); T3V2, root extract ( 1g + 10 ml DW ); V2, Chakwall 97;

Table 3: Effect of sunflower leaf, stem and root extracts on dry weight (g) of Weeds in

wheat 40 days after sowing (wheat varieties Margalla 99 and Chakwall 97).


V1 V2 V1 V2

T1 3.210 A 5.310 A 2.280 A 3.060 A

T2 2.100 B 2.640 C 1.730 D 2.330 B

T3 2.960 A 3.090 B 1.990 B 3.010 A

T4 2.910 A 3.070 B 1.820 C 2.920 A


71

Table 4: Effect of sunflower leaf, stem and root extracts on fresh weight (g) of

Weeds in wheat 70 days after sowing (wheat varieties Margalla 99 and Chakwall 97).


V1 V2 V1 V2

T1 8.200 A 10.00 A 7.150 A 8.100 A

T2 5.620 C 8.000 D 4.400 C 6.050 D

T3 6.340 B 9.050 B 5.370 B 7.370 B

T4 5.970 BC 8.640 C 4.940 B 6.590 C






Table 5: Effect of sunflower leaf, stem and root extracts on dry weight (g) of weeds in



V1 V2 V1 V2

T1 4.010 A 6.040 A 3.210 A 4.030 A

T2 2.490 C 3.890 B 3.000 C 3.230 B

T3 2.890 B 4.080 B 3.190 A 3.940 A

T4 2.700 BC 3.970 B 3.090 B 3.310 B


Table 6. Effect of sunflower leaf, stem and root extracts on number of tillers of

wheat plants 145 days after sowing ( wheat varieties Margalla 99 and Chakwall 97).


V1 V2 V1 V2

T1 14.00 C 12.00 C 15.00 C 13.33 B

T2 18.00 A 17.00 A 19.00 A 18.00 A

T3 16.00 B 14.00 B 17.00 B 15.00 B

T4 17.00 AB 15.00 B 17.67 AB 17.67 A






72

Table 7: Effect of sunflower leaf, stem and root extracts on plant height (cm) of

wheat plants 145 days after sowing ( wheat varieties Margalla 99 and Chakwall 97


V1 V2 V1 V2

T1 72.67 C 85.44 C 74.97 C 90.98 C

T2 117.0 A 120.0 A 123.3 A 130.0 A

T3 109.0 B 112.3 B 111.0 B 117.7 A

T4 116.0 A 119.3 A 121.0 A 127.7 A


Table 8: Effect of sunflower leaf, stem and root extracts on 100-grain-wieght (g)

of wheat 145 days after sowing (Wheat varieties Margalla 99 and Chakwall 97).


V1 V2 V1 V2

T1 4.000 C 3.980 B 5.000 C 4.390 B

T2 4.210 A 4.140 A 5.920 A 5.050 A

T3 4.170 B 4.130 A 5.170 B 4.480 B

T4 4.180 AB 4.137 A 5.880 A 5.010 A






Table 9: Effect of sunflower leaf, stem and root extracts on fresh weight (g) of

wheat plants 145 days after sowing (Wheat varieties Margalla 99 and Chakwall 97).


V1 V2 V1 V2

T1 4.000 D 3.960 D 4.490 B 4.270 C

T2 4.900 A 4.800 A 5.080 A 4.990 A

T3 4.450 C 4.600 C 4.650 B 4.570 B

T4 4.710 B 4.700 B 5.010 A 4.950 A


73

Table 10.Effect of sunflower leaf, stem and root extracts on dry weight (g) of

wheat plants 145 days after sowing (Wheat varieties Margalla 99 and Chakwall 97).


V1 V2 V1 V2

T1 0.7800 B 0.7000 C 0.9900 D 0.9200 D

T2 0.8900 A 0.8500 A 1.890 A 1.843 A

T3 0.8000 B 0.7700 B 1.213 C 1.153 C

T4 0.8700 A 0.8200 A 1.440 B 1.393 B






74

Effect of sunflower leaf, stem, and root extracts on Gibberellic Acid (µg g-l) contents of

wheat seedlings and roots 30 days after sowing (wheat varieties Margalla 99 and

Chakwall 97)

From the Table number 11 and 14, it is revealed that gibberellic acid contents were affected by

leaf, stem, and root extracts of sunflower. The maximum gibberellic acid contents in wheat

seedling in both wheat varieties Margalla 99 and Chakwall 97 were found in Tl (control)

followed by T3 (stem extract) and T2 (leaf extract) showed minimum in the first year of

experiment. While in the second year in the case of Margalla 9; maximum gibberellic acid

contents were counted in T2 (leaf extract), followed by T4 (root extract), whereas T3 (stem

extract) showed minimum. The data on GA contents of roots in the second year in case of

wheat variety Chakwall 97 revealed that T2 (leaf extract) showed the maximum GA contents,

which is on par with T4 (root extract) and T3, followed by Tl (control).

Effect of sunflower leaf, stem, and root extracts on Indole Acetic Acid (µg g-l) contents of

wheat seedlings and roots 30 days after sowing (wheat varieties Margalla 99 and

ChakwaI197)

The data of indole acetic acid contents in weat seedlings and root are presented in Table No.12

and 15. From the table, it is revealed that maximum indole acetic acid contents in wheat

seedling and root of wheat variety Margalla 99 were recorded in TI (control), followed by T3

(stem extract), whereas T4 (root extract) showed minimum in the first year. Result was found

similar in IAA content in case of Margalla 99 seedling in second year, while in root T2 (leaf

extract) showed maximum IAA content and TI (control) showed minimum values. In the case

of seedling of wheat variety Chakwall 97, T2 (leaf extract) showed the maximum values,

followed by T4 (root extract), T3 (stem extract), and TI (control). Whereas in root samples TI

(control) showed the maximum IAA content in wheat variety Chakwall 97, followed by T4

(root extract), T3 (stem extract) and T2 (leaf extract) in the first year, while in the second year

in the case of wheat variety Chakwal1 97, T2 (leaf extract) showed the maximum values,

followed by T4 (root extract), T3 (stem extract), and Tl (control).

75

Effect of sunflower leaf, stem, and root extracts on Abscisic Acid (µg g-l) contents of wheat

seedlings and roots 30 days after sowing (wheat varieties Margalla 99 and Chakwall 97)

From the Table number 13 and 16, it is revealed that abscisic acid contents in seedlings were

affected by extract of sunflower. The maximum abscisic acid contents in both wheat varieties

Margalla 99 and Chakwall 97 were found in T2 (leaf extract), followed by T4 (root extract),

and TI (control) showed minimum in the first year in both root and seedling samples. Result

was similar in the following year in case of seedling samples. While in the case of wheat

variety Margalla 99, TI (control) showed maximum values in root sample in the second year,

followed by T3 (stem extract), T2 (leaf extract), and T4 (root extract). In case of wheat variety

Chakwall 97, TI (control) showed the maximum values followed by T3 (stem extract), T4 (root

extract), and T2 (leaf extract).

76

Table 11. Effect of sunflower leaf, stem and root extracts on gibberellic acid (µµµµg g-1

)

contents of wheat seedlings 30 days after sowing (Wheat varieties Margalla 99 and

Chakwall 97).


V1 V2 V1 V2

T1 510.0 A 580.0 A 529.0 B 585.0 B

T2 500.0 C 570.0 B 547.3 A 593.3 A

T3 508.0 A 575.0 AB 527.7 B 582.7 B

T4 504.0 B 572.0 B 537.7 AB 590.0 AB


Table 12. Effect of sunflower leaf, stem and root extracts on indole acetic acid (µµµµg g-

1) contents of wheat seedlings 30 days after sowing (Wheat varieties Margalla 99

and Chakwall 97).


V1 V2 V1 V2

T1 265.0 A 200.0 D 195.0 A 190.0 A

T2 230.0 C 290.0 A 186.7 B 118.0 B

T3 242.3 B 230.0 C 194.0 A 141.7 A

T4 221.1 D 266.7 C 192.0 AB 171.7 AB






Table 13. Effect of sunflower leaf, stem and root extracts on abscisic acid (µµµµg g- 1

) contents

of wheat seedlings 30 days after sowing (Wheat varieties Margalla 99 and hakwall

97).


V1 V2 V1 V2

T1 82.00 B 100.00 B 101.7 B 119.0 D

T2 86.00 A 104.0 A 106.3 A 124.3 A

T3 82.00 B 102.3 AB 100.7 B 121.0 C

T4 84.00 AB 103.0 A 104.3 AB 123.0 B


77

Table 14. Effect of sunflower leaf, stem and root extracts on gibberellic acid (µµµµg g-1

)

contents of wheat seedlings root 30 days after sowing (Wheat varieties Margalla 99

and Chakwall 97).


V1 V2 V1 V2

T1 505.7 A 565.0 A 515.0 C 599.3 B

T2 494.0 B 543.0 B 520.0 A 615.0 A

T3 500.7 A 548.0 B 517.0 B 610.0 A

T4 501.7 A 562.7 A 518.7 A 614.0 A






Table 15. Effect of sunflower leaf, stem and root extracts on indole acetic acid (µµµµg g-

1) contents of wheat seedlings roots 30 days after sowing (Wheat varieties Margalla



V1 V2 V1 V2

T1 255.0 A 200.0 A 285.0 C 225.0 B

T2 217.7 C 290.0 C 302.0 A 263.3 A

T3 227.7 B 230.0 B 292.0 AC 221.7 B

T4 214.3 C 266.7 C 297.7 AB 255.0 A


Table 16. Effect of sunflower leaf, stem and root extracts on abscisic acid (µµµµg g-1

)

contents of wheat seedlings roots 30 days after sowing (Wheat varieties Margalla 99

and Chakwall 97).


V1 V2 V1 V2

T1 62.00 B 54.00 B 58.67 A 76.00 A

T2 66.00 A 60.00 A 56.67 AB 69.67 C

T3 62.00 B 51.67 B 58.00 A 74.00 B

T4 63.33 AB 58.67 A 55.00 B 70.33 C






78

Effect of sunflower leaf, stem, and root extracts on Chlorophyll contents (mg/g) of wheat

seedlings 30 days after sowing (wheat varieties Margalla 99 and Chakwall 97)

The data of chlorophyll contents are presented in Table No. 17. A perusal of the table revealed

that chlorophyll contents of wheat were affected significantly by various extracts of sunflower,

like leaves, roots, and stem. From the table, it is revealed that in case of both wheat varieties

Margall 99 and Chakwall 97 maximum chlorophyll contents were recorded in T2 (leaf extract).

T1 (control) showed minimum in Margall 99, and T3 (stem extract) in Chakwall. In the second

year in the case of wheat variety Margall 99, T4 (root extract) showed the maximum

chlorophyll contents; T2 (leaf extract) and T3 (stem extract) are on par with each other and

minimum chlorophyll contents were counted in T1 (control). In the case of wheat variety

Chakwal1 97, T2 (leaf extract) showed maximum chlorophyll contents, followed by T4 (root

extract), T3 (stem extract), and T1 (control).

Effect of sunflower leaf, stem, and root extracts on sugar contents (mg/g) of leaves in

wheat seedlings 30 days after sowing (wheat varieties Margalla 99 and Chakwall 97)

Sugar contents of wheat were influenced by the genetic and environmental factors and various

extracts of sunflower (leaves, stems, and roots). The data regarding sugar contents of wheat are

presented in Table No. 18. A perusal of the table revealed that the maximum sugar contents in

the case of both wheat varieties Margalla 99 and Chakwall 97 were recorded in T2 (leaf

extract) in the first as well as second year of experiment. T3 (stem extract) showed minimum

sugar content in the first year and T1 (control) in the second year in both wheat varities


Effect of sunflower leaf, stem, and root extracts on protein contents (mg/g) of leaves in


The data regarding protein contents are presented in Table 19. A perusal of the table revealed

that protein contents of wheat were affected by various extracts of sunflower. In case of both

wheat varieties Margalla 99 and Chakwall 97, T2 (leaf extract) showed the maximum protein

contents; T1 (control) showed minimum in the first year. Chakwall 97 showed similar result in

the second year. In case of wheat variety Margalla 99 in the second year, TI showed the

maximum values, followed by T3 (stem extract), and T2 (leaf extract) showed minimum.

79

Effect of sunflower leaf, stem, and root extracts on proline contents (mg/g) of leaves in


The data regarding proline contents are presented in Table No. 20. A perusal of the table

revealed that proline contents of wheat were affected by various extracts of sunflower. In the

case of both wheat varieties Margalla 99 and Chakwall 97, T2 (leaf extract) showed the

maximum values, and proline content was found minimum in Tl (control). Results were found

similar in the following year.

80

Table 17. Effect of sunflower leaf, stem and root extracts on chlorophyll (mg/ 100 g

F. wt) contents of wheat seedlings 30 days after sowing (Wheat varieties Margalla



V1 V2 V1 V2

T1 92.00 B 81.67 A 100.00 C 86.00 C

T2 95.33 A 83.00 A 103.3 B 100.0 A

T3 94.33 A 80.00 A 103.3 B 90.67 B

T4 94.67 A 82.33 A 105.7 A 97.67 B


Table 18. Effect of sunflower leaf, stem and root extracts on sugar (mg/ 100 g F. wt

) contents of wheat seedlings 30 days after sowing (Wheat varieties Margalla 99 and

Chakwall 97).


V1 V2 V1 V2

T1 182.0 B 177.3 B 194.0 B 180.0 C

T2 192.3 A 185.0 A 205.3 A 196.3 A

T3 174.0 C 174.0 B 200.0 AB 185.0 BC

T4 185.7 B 176.7 B 202.7 A 192.0 AB






Table 19. Effect of sunflower leaf, stem and root extracts on protein (mg/ 100 g F.

wt) contents of wheat seedlings 30 days after sowing (Wheat varieties Margalla 99

and Chakwall 97).


V1 V2 V1 V2

T1 1732 B 1613 C 1700 A 1615 B

T2 1781 A 1663 A 1635 C 1635 A

T3 1732 B 1630 BC 1692 AB 1615 B

T4 1764 A 1651 AB 1645 BC 1628 A


81

Table 20. Effect of sunflower leaf, stem and root extracts on proline (mg/ 100 g F.

wt) contents of wheat seedlings 30 days after sowing (Wheat varieties Margalla 99

and Chakwall 97).


V1 V2 V1 V2

T1 112.0 C 102.0 C 99.67 C 105.0 C

T2 126.0 A 112.0 A 109.0 A 122.0 A

T3 118.0 B 106.0 B 103.7 B 115.7 B

T4 124.3 A 109.3 A 106.3 AB 121.0 AB






82

Effect of sunflower leaf, stem, and root extracts on DNA contents of leaves in wheat

seedlings 30 days after sowing (wheat varieties Margalla 99 and Chakwall 97)

[The data regarding praline contents of wheat are presented in Table No. 21. A perusal of the

table revealed that DNA contents of wheat were affected by various extracts of sunflower

(leaves, stems, and roots). In the case of both wheat varieties Margalla 99 and Chakwall 97, T2

(leaf extract) showed high values of DNA in the first as well as second year of experimen,

while Tl (control) showed minimum in the first year. While in the second year in the case of

wheat variety Margalla 99, T3 (stem extract) and in the case of wheat variety Chakwall 97 Tl

(control) showed minimum.

Effect of sunflower leaf, stem, and root extracts on Superoxide Dismutase contents of


The data regarding SOD contents are presented in Table No. 22. A perusal of the table revealed

that SOD contents of wheat were affected by various extracts of sunflower (leaves, stems, and

roots). In case of both wheat varieties Margalla 99 and Chakwall 97, T2 (leaf extract) showed

the maximum SOD activities, and Tl (control) showed minimum SOD activities in the first

year. Results were found similar in the second year.

Effect of sunflower leaf, stem, and root extracts on Peroxidase contents of wheat seedlings

30 days after sowing (wheat varieties Margalla 99 and Chakwall 97)

The data regarding POD activity are presented in Table No. 23. A perusal of the table revealed

that POD contents of wheat were affected by various extracts of sunflower. In the case of both

wheat varieties Margalla 99 and Chakwall 97, T2 (leaf extract) showed the maximum values

and Tl (control) showed minimum values in the first as well as second year of experiment.

Effects of growing wheat treated with sunflower extracts on the soil physicochemical

properties

Results presented in Table 24 revealed that in both the varieties soil EC and pH decreased as

compared to control soil ( where untreated wheat plant was grown)but Mn, Fe, K ,Zn and Pb

were increased in both the varieties while Mg and Ca were decreased.

83

Table 21.Effect of sunflower leaf, stem and root extracts on DNA (mg/ 100 g F. wt )

contents of wheat seedlings 30 days after sowing (Wheat varieties Margalla 99 and

Chakwall 97).


V1 V2 V1 V2

T1 497.3 C 490.0 B 485.0 B 502.7 B

T2 516.0 A 498.3 A 501.7 A 522.3 A

T3 504.0 BC 492.0 B 463.7 C 506.3 B

T4 512.0 AB 497.7 A 490.7 AB 517.3 A


Table 22. Effect of sunflower leaf, stem and root extracts on superoxidase dismutase

(unit/g f.w) activity of wheat seedlings 30 days after sowing (Wheat varieties



V1 V2 V1 V2

T1 4.00 B 3.000 C 5.000 B 4.000 B

T2 11.00 A 8.670 A 11.67 A 9.000 A

T3 9.000 A 5.000 B 10.00 A 6.000 B

T4 10.00 A 7.670 A 11.00 A 8.670 A


Table 23. Effect of sunflower leaf, stem and root extracts on peroxidase (OD/min/g f.w)

activity of wheat seedlings 30 days after sowing (Wheat varieties Margalla 99 and

Chakwall 97).


V1 V2 V1 V2

T1 11.67 B 10.00 C 10.67 C 9.00 C

T2 18.00 A 15.00 A 17.00 A 14.00 A

T3 14.00 D 12.67 B 13.00 B 12.00 B

T4 16.67 A 14.00 AB 16.00 A 13.33 AB


84

DISCUSSION

Investigations of the intensity of carbon dioxide assimilation (an essential feature of

photosynthesis) are very significant. Allelochemicals results in water deficiency and suppress

photosynthetic absorption of carbon dioxide; the degree of inhibition, however, vary with

species and variety, being dependent on the amplitude of water balance and plant age. The 2

wheat varieties used are stress-tolerant varieties, and so they produced different results; the

decrease in the rate of photosynthesis caused by allelochemicals was not pronounced in these

varieties, but weeds were affected by allelochemicals. Increased chlorophyll contents in

treatment treated with extract from leaves, followed by that from roots and then stems, were

due to control of weeds in these treatments compared with controls, because weeds compete

with the crop for light, water, shelter, and nutrients. The chlorophyll contents were increased in

the leaf-extract treatment, followed by roots and stems. This was due to leaves of sunflower

extracts having more allelochemicals, followed by roots and stem; thus, they have the ability to

control more weeds. This is possibly why chlorophyll contents increased in leaf extract

treatment, followed by root extracts and stem extracts. Rice (1974) has reported earlier that

application of sunflower water extract suppressed the growth of weeds, and the content of

allelochemicals was higher in leaves, followed by roots and stems (Rice, 1984). Aqueous

extracts of sunflower leaves, stems, and roots inhibited germination of weeds (Leather 1983).

However, it appears that many varieties of cultivated sunflower have retained the genetic

information necessary for allelochemical production; these observations are good evidence of

sunflower allelopathic potential and suggest that appropriate management strategies can be

used to successfully exploit sunflower allelopathy, especially in the area of weed control. So,

increased chlorophyll contents in leaf extracts were possibly due to control of weeds. The

phenolics cause reduction in chlorophyll contents; the effect of allelochemicals (e.g., vanillic,

ferulic, alkaloids, and benzoic acid) on DNA has been found by various workers. The

stimulation was obvious in wheat seedlings treated with aqueous extract at (I g extract of

leaves, stems, and roots), respectively. These findings are agreement with those of

Bazirramakenga et al, (1997), who reported that the low concentrations of the allelochemicals

vanillic and ferulic acids stimulated the biosynthesis of nucleic acids by increasing the

85

incorporation of 32P into DNA in seedlings. Inversely, allelochemical treatments at higher

concentrations gradually reduced the contents of DNA. Additionally, Seigler (1996)

demonstrated that allelopathic compounds interact with nucleic acid metabolism, causing

modification of DNA. The obvious reduction in the DNA levels of applied allelochemicals

coincides with its depressive effect on the activity of amylase enzyme. This agrees with the

general knowledge of enzyme activity and contents of nucleic acids (Finer et al., 1969). The

affect of allelochemicals on weed control also caused increased protein contents. Activity

levels of SOD and POD showed progressive, significant increases with increased

allelochemicals, compared to controls. This is likely related to stress, because allelopathy

causes stress. Receiver-plants, as well as donor- plants, are active during the stress conditions.

These results are in agreement with those of Fridovich (1986), who found that stress enhanced

the activities of leaf mitochondrial Mn-SOD and chloroplastic Cu/Zn-SOD. In the

detoxification of reactive oxygen species, SOD converts superoxide to H202 and 02 and thus

protects the macromolecules from superoxide induced oxidative stress. During present

investigation, SOD and POD activity increased with increase in concentration of

allelochemicals. The sugar content was higher in leaf extract treatments, followed by root

extract and the minimum impact was shown by extracts prepared from stem. Moreover, cv.

Margalla 99 performed better than cv. Chakwall 97. The increase in sugar content in treatments

T2, T4, and T3 might be attributed to the control of weeds. Varieties may differ in their sugar

contents and be influenced by management and environmental conditions. The sugar is the

major photoassimilate in leaves (Koch, 1996). The synthesis of enzyme stimulated for

mobilization of seed reserves in germinating grains is caused by gibberellic acid, which also

stimulate the growth of nearby plants (Salisbury and Ross, 1992; Arteca, 1995; John and Mitra,

2001). During present study, allelochemicals enhanced the accumulation of proline in leaves,

and similar effect of ABA is also reported to increase proline accumulation in leaves.

Therefore, the greater proline accumulation might be due to higher abscisic acid content, the

production of which may be attributed to the effect of allelochemicals. The aqueous extracts

prepared from various parts of the sunflower (leaves, stems, and roots) showed stimulatory

effects on shoot length, root length, fresh weight, and dry weight of wheat seedlings. The lower

concentration of leaves, shoot, and root extract exhibited non-significant effects. Nevertheless,

86

the inhibitory effect of the rest of the concentrations was found to be high. The inhibitory

impact of extracts usually improved with increases in concentration.

87

CONCLUSION

From first experiment, it was concluded that in sunflower, the contents of allelochemicals were

maximum in leaves, followed by roots and stem, in that order. Stress stimulated the production

of allelochemicals. From the experiment, it was also concluded that sunflower roots lowered

the number of colonies of the group of microorganisms studied, namely Azospirillum,

Phosphate-Solubilizing Bacteria, and Rhizobium. The effects of the three habitats were also

apparent in the Gram Staining and Oxidase tests.

From the 2nd type of experiments these were concluded: allelochemicals secreted by sunflower

inhibited germination and lowered the level of hormones, GA, IAA, Shoot length, Root length,

fresh weight, dry weight, Chlorophyll contents, and DNA, while they increased the values of

ABA, Protein, Proline, Sugar, Sod, and POD contents in wheat seedlings.

From the 3rd experiments it was inferred that allelochemicals secreted by sunflower lowered

the level of ABA, weeds density, and fresh and dry weight of weeds, while they increased the

values of GA, IAA, Protein, Proline, Sugar, SOD, POD, Fresh weight, dry weight, shoot length,

root length, and 100-seed weight of wheat plants, In sunflower, the content of allelochemicals

was the maximum in leaves, followed by roots and stems, in that order. Stress stimulated the

production of allelochemicals.

88

REFERENCES

Aharoni, N., Blummenfeld,, Richmond, A.E., 1979. Hormonal activity in detachec lettuce

leaves as affected by leaf water content. Plant Physiol. 59: 1169-1173.

Ahmadi, A., Baker, D.A., 1999. Effects of abscisic acid (ABA) on grain filling processes in

wheat. Plant Growth Regul. 28: 187-197.

Asa, M. H. and Karsson, P. S., 1998. Altitudinal variation in size effects on plant eproductive

effect and somatic costs of reproduction. Journal of Ecology, 5: 112- 118.

Ahmadi, A., Baker, D. A., 2001. The effect of water stress on the activities of key regulatory

enzymes of the sucrose to starch pathway in wheat. Plant Growth Regul. 35: 81-91.

Amon, D.J., (1949). Copper enzymes in isolated chloroplast phenol oxidase in Beta vulgaris.

Plant Physiol. 24: 1-15.

AI-Khatib, K, Libbey C, Boydston R.,1997. Weed suppression with Brassica green manure

crops in green pea. Weed Sci. 45: 439-445.

Anjum, T., Bajwa, R., and Javaid, A., 2005. Biological Control of Parthenium I: Effect of

Imperata cylindrica on distribution, germination and seedling growth of Parthenium

hysterophorus L. Int. J. Agric. Biol., 7(3): 448-450.

Anaya, A.L., 1999. Allelopathy as a tool in the management of biotic resources in agrosystems.

Crit. Rev. Plant Sci., 18: 697- 739.

Abdul-Rahman, A.A., Habib, S.A. 1989. Allelopathic effect of alfalfa {Medicago sativa) on

bladygrass (Imperata cylindrica). J. Chem. Ecol 15: 22 89- 2300.

Akram, M., Hussain, F. 1987. The possible role of allelopathy exhibited by root extracts and

exudates of Chinese cabbage in hydroponics. Pakistan Journal of Science and industrial

Research 30: 918- 920.

89

Alderman, W.H. , Middletonl, J.A. 1925. Toxic relations of other crops to tomatoes.

Proceedings, American Society of Horticultural Science 22: 307- 308.

AI-Saadawi, I.S. and Ricel, E.L., 1982. Allelopathic effects ofPolygonum aviculare L. 1.

Vegetational pattering. J. Chem. Ecol 8: 993-1010.

AI-Saadawi, I.S., AI-Ugaili, J.K., AI-Hadithy, S.A.M., AI-Rubeaa, 1976. J.Asian Vegetables

and Research Development Centre (1978). A vrdc, Shanhua, Taiwan, China.134

Arteca, RN., 1995. Plant growth substances; Principles and applications. Chapman Hall, New

York, USA, p. 332.

Arteca, R., 1996. Plant Growth Substances: Principles and Applications. Chapman & Hall New

York.

Addocott, F.,Lyon, J. L., 1961. Physiology of abscisic acid and related substances. Ann. Rev.

Plant Physiol. 20: 139-164.

Anjum, T., Stevenson. P., Hall. D., Bajwa. R., 2005. Allelopthic potential of sunflower

(Helianthus annus L.) as a natural herbicide. In proceedings of the 4th World Congress on

Allelopathy. Harper, J. D. I., H. Wu and J.H. Kent (eds), 26 August 2005, International

Allelopathy Society.

Anonymous., 1996. International Allelopathy Society 1996. First World Congress on

Allelopathy: A science for future, Cadiz, Spain.

Azania, A.A.P.M., Azania, C.A.M., Alives, P.L.C.A., Palaniraj, R., Kadian, H.S., Sati, S.C.,

Rawat, L.S., Dahiya, D.S., Narwal, S.S.: Allelopathic plants. 7. Sunflower (Helianthus annuus

L.). - Allelopathy J. 11: 1-20, 2003.

Batish, D.R., Tung, P., Singh, H.P., Kohli, R.K.: Phytotoxicity of sunflower residues against

some summer season crops. - J. Agron. Crop Sci. 188: 19-24, 2002.

Bhowmik, P.C., Inderjit.: Challenges and opportunities in implementing allelopathy for natural

90

weed management. -

Crop Protect. 22: 661-671, 2003.

Ba. M., Lutts, S., Kinet, J., 2001. Water deficit effects on solute contribution to osmotic

adjustment as a function leaf aging in three durum wheat (Triticum durum Desf.) cultivars

performing differently in arid conditions. Plant Sci. 160: 699-681.

Bartels, D., Sunkar, R., 2005. Drought and salt tolerance in plants. Crit. Rev. in Plant Sci. 24:1-

36.

Bellanmine, J., Penel, C., Greppin, H., Gaspar, T., 1998. Confirmation of the role of auxin and

calcium in the late phases of adventitious root formation. Plant Growth Regul. 26: 191-194.

Borel, C., Simonneau, T., This, D., Tardieu, F., 1997. Stomatal conductance and ABA

concentration in the xylem sap of barley lines and constrasting genetic origins. Aust. J. Plant

Physiol. 24: 607-615.

Brenner, M.L., Chiekh, N., 1995. The role of hormones in photosynthate partitioning and seed

filling. In P.J. Davies (ed.) Plant hormones, physiology, biochemistry and molecular biology.

Kluwer Academic Publ., Dordrecht, the Netherlands. P: 649-670.

Bell, D.T. and Koeppe, D.E., 1972. Non competitive effects of giant foxtail on the growth of

com. Agronomy 64: 32 1-325.

Bhandari, S.C., Khurana, A.S. Bhatia, R.K., 1982. Geobios 9: 261-262. Bioprodukt., 1984.

Agrostemin-Gfl of Nature Novi Dani Beograd, Yugoslavia. Bode, H.R., 1940. Planta 30: 567.

Bonner, J., 1960. Liberation of organic substances from higher plants and their role in

soil sickness problem. Botanical Review, 26: 393-424.

Bradow, J .M., Connick Jr., W.J., 1987. Allelochemicals from palmer amaranth,

(Amarnathus palmeri S. Wats). Journal of Chemical Ecology 13: 185-202.

Bradow, J.M. Connick Jr., W.J., 1988a. Seed-germination inhibition by volatile alcohols and

other compounds associated with Amaranthus palmeri residues. Journal of Chemical Ecology,

91

14: 1633-1648.

Bradow, J.M. and Connick Jr., W.J., 1988b. Volatile methyl ketone seed germination inhibitors

from Amaranthuspalmeri S. Wats. Journal of Chemical Ecology 14: 16 17- 1630.

Breazeale, J.F., 1924. The injurious after-effects of sorghum. Journal of American Society of

Agronomy, 16: 689-700.

Bates, L.S. Waldren RP, Teare 1973. Rapid determination of free proline for water stress

studies. Plant soil, 39: 205-207.

Beauchamp C., Fridovich I., 1971. Superoxide dismutase: improved assays and an assay

applicable to acrylamide gels. Anal. Biochem. 44: 276-287.

Beyer W, Fridovich I., 1987. Assaying for superoxide dismutase activity: some large

consequences of minor changes in conditions, Anal. Biochem. 161: 559- 566.

Bogatek R, Gniazdowska A, Zakrzewska W, Oracz K, Gawroski, SW (2005). Allelopathic

effects of sunflower extracts on mustard seed germination and seedling growth. BioI. Plant

Oubo SM, Giles KA, Hmilton JK, Robers P A, Smith FA., 1956. Calorimetric method for

determination of sugar and related substances. Anal. Chern. 28: 350-353.

Blum, U. Shafer, S.R., Microbial populations and phenolics acids in soil. Soil BioI. Biochem.

20: 793, 1988.

Brady, N.C., 1990. The nature and properties of soils. 151 edition 621 pp. Macmillan

Publishing Co., New York, N.Y.

Batish, O.R., Singh, H.P., Saxena, D.B., Kohil, R.K., 2002. Weed suppressing ability of

parthenin -A sesquiterpene lactone from Parthenium hysterophorus. New Zealand Plant

Protection. 55: 218-221.

Bernat W., Gawaronska H., Janowiak F., Gawronski S.W 2003. The effect of sunflower

allelopathics on germination and seedling vigor of winter wheat and mustard. Abstract of Fifth

92

International Conference, Ecophysiological Aspects of plants responses to stress factor,

Cracow, Poland. Acta Physiol Plant. 25 : 24- 25.

Bangerth, F., 1982. Changes in the ratio of cis-trans- abscisic acid during ripening of apple

fruits, Plants. 155: 199-203.

Barbham, D.E., Biggs, R.H., 1981. Cis-trans photo -isomerisation of abscisic acid. Phytochem

and photobiol. 34: 33- 37.

Baziramakenga, R., heroux, G.D., Simard R. R., Nadeau., P., 1997. Allelopathic effects of

phenolic acids on nucleic acids on nucleic acids and protein levels in soybeans seedlings. Can.

J. Bot., 75: 445- 450.

Brian, P.W., Elson, G.W., Hemming, H.G., Radley, M., 1954. "The plant growth promoting

properties of gibberellic acid, a metabolic product of

the fungus Gibberella fujikuroi".J. Sci. Food. Agr. 5: 602-612.

Bennet R.C., Wallsgrove RM. 1994. Secondary metabolites in plant defence mechanisms,

Tansley Review No. 72. New Phytol, 127: 617-633.

Bogatek, R., Gniazdowska, A., Zakrzewska, W., Oracz K, Gawro_ski SW., 2005. Allelopathic

effects of sunflower extracts on mustard seed germination and seedling growth. BioI. Plant. (In

Press).

Cheema, Z. A., A. Khaliq and S. Saeed. 2004. Weed control in maize Zea mays L.) through

sorghum allelopathy. J. Sustainable Agric. 23(4) 73-86.

Chowdhury, R.S., Chowdhuri M. A. 1985. Hydrogen peroxide metabolism as index of water

stress tolerance injute. Phylogica Plantrum, 65: 503- 507.

Chon, S.-U., Kim, J.-D. (2002). Biological activity and quantification of suspected

allelochemicals from alfalfa plant parts. J. Agron. Crop Sci. 188.

Cochrane, V.W., Elliott, L.F., Papendick, R.I., 1977. The production of phytotoxins from

93

surface crop residues. Soil Science Society of American Journal 41: 903-908.

Conrad, J.P., 1927. Soome causes of the injurious after-effects of sorghum and suggested

remedies. Journal of American Society of Agronomy, 19: 1091-1110.

Colorado, P., Nicholas, G., Rodirguez, D., 1995. Convergent effects of stress and ABA on

expression during germination of chickpea seeds. Physiol. Plant. 146: 535-540.

Callaway RM, Aschehoug ET .M, 2000. Invasive plant versus their new and old neighbours: a

mechanism for exotic invasion. Science. 290: 521-523.

Ciarka, D., Gawrońska, H., Małecka, M., Gawroński, S.W.: Allelopathic potential of sunflower

roots and root exudates. - Zesz. probl. Post. Nauk roln. 496: 301-313, 2004.

Dong, B., Rengel, Z., Graham, R. D., 1995. Root morphology of wheat genotypes differing in

Zn efficiency. J. Plant Nutr. 18: 2761-2773.

Dixon RA, Paiva NL. 1995. Stress-induced phenylpropanoid metabolism. Plant Cell 7: 1085-

1097.

Dathe, W., Schneider, G. and Sembdner, D., 1978. Endogenous gibberilines and inhibitors in

caryopases of rye. Phytochemis. 17: pp. 963- 966.

Davies W. J., Jones H.G. 1991. Abscisic acid, physiology and biochemistry. Bios Scientific

Publishers Limited. St Thomas House, Backet Street, Oxford OXI ISJ, UK: 63: 77, 137-149,

189-197, 201-225.

De Candolle, M.A.P., 1832. Physiologie Vegetale. Tome III, Bechet Jeune, Paris. Pp.

1474-1475.

Dzyubenko, N.N. and Krupa, L.l., 1974. On interactions of cultivated plants vegetation and

weeds in agrophytocenoses. In: Physiological and Biochemical Interactions Between the Plants

in Phytocenosis (Ed., A.M. Grodzinsky) Vol. 5: 55- 56. Naukova Dumka, Kiev, USSR.

94

Dzyubenko, N.N. and Petrenko, N .1., 1971. On biochemical interactions of cultivated plants

and weeds. In: Physiological and Biochemical Interactions Between the Plants in Phytocenosis

(Ed., A.M. Grodzinsky) Vol. 2: 60-66. Naukova Dumka, Kiev, USSR.

Dhindsa R.S, Matow E. W. 1981. Drought tolerance in two mosses: correlated with enzyme

defence against lipid perioxidation. Journal of Exprimental Botany, 32: 79- 91.

Duhan, J. S. and K. hakshinarayand, 1995. Allelopathic effect of Acacia nilotia on cereal and

legume crop grown in field. All. J., 21: 93- 98.

Dixon RA, Paiva NL. 1995.Stress-induced phenylpropanoid metabolism. Plant Cell 7: 1085-

1097.

Dathe, W., Schneider, G. and Sembdner, D., 1978. Endogenous gibberilines and inhibitors in

caryopases of rye. Phytochemis. 17: pp. 963- 966.

Davies W. J., Jones H.G. 1991. Abscisic acid, physiology and biochemistry. Bios Scientific

Publishers Limited. St Thomas House, Backet Street, Oxford OXI ISJ, UK: 63: 77, 137-149,

189-197,201- 225.

Davies, P. J., 1995. Plant Hormones: Physiology, Biochemistry and Molecular Biology.

Dordrecht: Kluwer.

Economou, G., Tzakou, O., Gani, A., Yannitsaro, A. and Bilalis, D.(2002). Allelopathic effect

of Conyza albida. Ecol. 17: 2021-2034.

Fukui, H., koshimizu, K., Usuda, S. and Yamazaki, Y., 1977. Isolation of plant growth

regulators from seeds of cucurbite. Pepol. Agric. Bioi. Chern. 41: pp. 175-180.

Fridovich, I., 1986. Biological effects of the superoxide radical. Arch. Biochem. Biophys.247:

1-11.

Finer P, Wray JL, Varner JE., 1969. Enzyme induction in higher plants. Science, 165: 358.

95

Fay, P.K. and Duke, W.B., 1977. An Assessment of allelopathic potential of Avena germplasm.

Weed Science 25: 224-228.

Fukui, H., koshimizu, K., Usuda, S. and Yamazaki, Y., 1977. Isolation of plant growth

regulators from seeds of cucurbite. Pepol. Agric. BioI. Chern. 41: pp. 175-180.

Grechkanev, O.M. and Rodionov, V.I., 1971. Interactions of the components of the nector

fodder mixture with wild heliotrope. In: Physiological and Biochemical Interactions Between

the Plants in Phytocenosis (Ed., A.M. Grodzinsky). Vol. 2: 88- 94. Naukova Dumka, Kiev,

USSR.

Guerrero, F., Mullet, J.E., 1986. Increased abscisic acid biosynthesis during plant dehydration

requires transcription. Plant Physiol. 80: 588-591.

Guinn, G., Brummett, D.L. 1988. Abscisic acid in young cotton furits and their abscission

zones in relation to fruit retention during and after moisture stress. Plant Physiol. 86: 28-31.

Guenzi, W.O. and Mccalla, T .M., 1962. Inhibition of germination and seedling development

by crop residues. Soil Science Society of America Proceedings 26: 456-458.

Guenzi, W.O. and Mccalla, T .M. and Norstadt, F., 1967. Presence and persistence of

phytotoxic substances in wheat, oat, corn and sorghum residues. Agronomy Journal 59: 163-

165.

Goldschmidt, E. E.; Goren, R.; Evenchen, Z. and Bittner, S., 1973. Increase in free and bound

ABA during natural and ethylene induced senescence of citrus fruit peel. Plant Physiol. 51: pp.

879-82.

Hedge R.S., Miller D.A. 1990. Allelopathy and autotoxicity in alfalfa: Characterization and

effects of preceding crops and residue incorporation. Crop Sci. 30: 1255-1259.

Hose, E., Studle, E., Hartung, W., 2000. Abscisic acid and hydraulic conductivity of maize

roots: a study using cell-n and root pressure probes. Planta. 211: 874- 882.

Ingram, J., Bartels. D., 1996. The molecular basis of dehydration tolerance in plants. Annu.

96

Rev. Plant Physiol. Plant Mol. BioI. 47: 377-403.

Hall, A.B., Blum, U. and Fites, K.C., 1982. Stress modification of allelopathy of He/ianthus

annuus L. debris on seed germination. American Journal of Botany 69: 776-89.

Hall, A.B., Blum, U. and Fites, R.C., 1983. Stress modification of allelopathy of He/ianthus

annuus L. debris on seedling bio mass production of Amaranthus retroflexus L. Journal of

Chemical Ecology 9: 1213- 1222.

Harrison, H.F. and PETERSON, J.K., 1986. Evidence that sweet potato (Ipomea batatas L.) is -

allelopathic to yellow nutsedge (Cyperus esculentus). Weed Science. 39: 308-312.

Hicks, S.K., Wendt, C. W. and Gannaway, J .R., 1988. Allelopathic Effects of Wheat Straw on

Cotton Germination and Seedling Development. The Texas Agricultural Experiment Station.

The Texas A&M University, College Station. Texas. Bulletin pp.1-5.

Holm, L., Pancho, J.V., Herberger, J. P. and Plucknett, D.L., 1979. A Geographical Atlas of

World Weeds, John Wiley, New York.

Horricks, J.S., 1969. Influence of rape residue on cereal production. Canadian Journal of Plant

Science. 49: 632-634.

Haddock EA, Gupta RK, AI-Shafi SMK, Layden K, 1-taslam E, Magnolato D. 1982. The

metabolism of gallic acid and hexahydroxydiphenic acid in plants: biogenetic and molecular

taxonomic considemtions. Phytochem 21: 1049-1062.

Harborne JB. The flavonoids: recent advances. In: Goodwin TW, ed. Plant Pigments. London,

England: academic Press, 1988, p. 299-343.

Hasam E. Vegetable tannins. In: StumpfPK, Conn EE, eds. The Biochemistry of Plants. New

York, Academic Press, 1981, P. 527-556.

Haslam E. 1982. The metabolism of gallic acid and hexahydroxydiphenic acid in higher plants.

Fortschr Chern Naturs 41: 1-46.

97

Hoffman, M. L., L. A. Weston, J. C. Synder and E. E. Regnier 1996b. Allelopathic influence of

germinating seeds and seedlings of cover crops on weed species. Weeds Sci. 44(3): 579-584.

Hoffman, M., L., L. A. Weston, J. C. Snyder and E. E. Regnier 1996a. Separating the effects of

sorghum ( Sorghum bico/ar L. ) and rye ( Seca/e cerea/e L. ) root and shoot residues on weed

development. Weed Sci. 44(2): 402- 407.

Izhaki, I., Tsahar, E., Palury, I. and Friedman, J., 2002 withen-population variation and

interrelationships between morphology, nutritional contents, and secondry compounds of

Rhamnus alaternus fruits. New Phytologist 156: 217- 223.

Inderjit., 1996. Plant phenolics in allelopathy. Botanical Review 62, 182-202.

Iturbe- Ormatexe I, Escuredo R. P, Arrese Igor C, Becana B. 1998. Oxidative damage in pea

plant exposed to water deficit or paraquat. Plant Physiology, 119: 173-181.

Ingram, J., Bartels., D., 1996. The molecular basis of dehydration tolerance in plants. Annu.

Rev. Plant Physiology. Plant Mol. BioI. 47: 377- 403.

Jimenez- Osornio, J. J. and S. R. Gliessman 1987. Allelopathic interference in a wild

mustard ( Brassica campestris L. ) and broccoli ( Brassica oleracea L. var. italica L. ) intercrop

agroecosystems. In Allelochemicals: Role in agriculture and forestry, ed. By G. R. Waller,

American Chemical Socity, Washington, D. C., 262- 274.

Jalili A., F. Abbassi and M. Bazoobandi. 2007. Allelopathic influence of canola on germination

of five weeds of canola fields. Absts. International Workshop on Allelopathy- current trends

and future applications, Univ. of Agri., Faisalabad, Pakistan.

Rice, E. L. 1984. Allelopathy 2nd Ed. Acad. Press, Inc. Orlando, Florida.

Jennings, J.A. and Nelson, C.J. (2002). Zone of autotoxic influence around established alfalfa

plants. Agron. J. 94: 1104-1111.

Jones, W. W.; Coggins, C. W. and Embleton, T. W., 1976. Endogenous abscisic acid in relation

to bud wowth in alternate bearing 'Valencia' orange, Plant Physiol. 58: pp. 681-682.

98

Johnson RP, Balwani TL, Johnson LJ, Meclure KE, Denority BA., 1966. Carbon plant naturity

II Effect on in vitro cellular digestibility and soluble carbohydrate content, Anim. Sci. 25: 617-

619.

Johri MM, Mitra D., 2001. Action of plant hormones. Curr. Sci. 80: 2-25.

Jimenez- Osornio, J. J. and S. R. Gliessman 1987. Allelopathic interference in wild mustard

(Brassica campestris L.) and broccoli (Brassica oleracea L. var. italica ) intercrop

agroecosystem. In Allelochemicals: Role in agriculture and forestry ed. BY G. R. Waller,

American Chemical Society, Washington, D. C., pp. 262- 274.

Kovac N., 1956. Identification of Pseudomonas pyocyaned by the oxidase reaction. Nature pp.

178-703.

Kitou, M. and S. Yoshida 1993. Difference of phytotoxicity between undecomposed and

microbially decomposed plant residues. Weed Res., 38 ( 1 ): 43- 46.

Kamelie, A., Losel, D. M., 1995. Contribution of carbohydrates and solutes to osmotic

adjutment in wheat leaves under water stress. Plant Physiol., 145: 363-366.

Kumazawa, R. Ishii, K. Ishihara, and H. Hirata (Eds.). Science of the Rice. Vol. II. Physiology.

Food and Agricultur Policy Research Center, Tokyo, Japan.

Kimber, R. W. L., 1973a. Phytotoxicity from plant residue II. The effects of time of rotting

straw from grasses and legumes on the growth of wheat seedling. Plant and Soil. 38: 347-361. '

Kimber, R. W. L., 1973b. Phytotoxicity from plant residue LI. The relative effects of toxins and

nitrogen immobilization on the germination and growth of wheat. Plant and Soil. 38: 347-361.

Kuo, C.G., CHO, M.H. and PARK, H.G., 1981. Effect of Chinese cabbage residue on

mungbean. Plant and Soil. 61: 473-477.

Khanh, T. D., M.I. Chung, T. D. Xuan and S. Tawanta, 2005. The exploitation of crop

alleloopathy in sustainable agriculture production. J. Agron. Crop Sci., 188: 19- 24.

99

Kuroha, T., Kato, H., Asami, T., Yoshida, S., Kamada, H.; Satoh, S., 2002. A transzeatin

riboside in root xylem sap negatively regulates adventitious root formation on cucumber

hypocotyls. J. Exp. Bot. 53: 2193- 2200.

Kirch, JTO., 1968. Studies on the dependence of chlorophyll synthes.is on protein synthesis in

Euglena agraci/lis together with a monogram for determination of chlorophyll concentration.

Planta, 78: 200-207.

Koch KE., 1996. Carbohydrate modulated gene expression in plants. Annu. Rev. Plant physiol.

Plant mol. BioI. 47: 509-540.

Laughlin, R.G., Munyon, R. L., Ries, S. K and Wert, V. F., 1983. Science 219: 12: 19-1220.

Leather, G. R., 1983a .Sunflower (Helianthus annus L. ) are allelopathic to weeds. Weeds

Sciences 31: 37- 42.

Levin, D. A., 1976. Annual Review of Ecological System 7: 121-157.

Lodhi, M.A.K., 1981. Accelerated soil mineralization, nitrification and revegetation of

abandoned fields due to the removal of crop-soil phytotoxicity. Journal of Chemical Ecology 7:

685-693.

Lovett, J. V., 1982 .Allelopathy and self-defence in plants. Australian Weeds 2: 33- 36.

Lovett, J. V ., 1986. Allelopathy: The Australian experience. In: The Science of Allelopathy

(Eds., A. R. Putnam and C.S. Tang). pp. 75-99. John Wiley & , Sons Inc., New York.

Lynch, J.M. and Penn, D.J., 1980. Damage to cereals caused by decaying weed residues.

Journal of the Science of Food and Agriculture 31: 32 1-324.

Li JC, Shi J, Zhao XL, Wang GYO, Ren YJ., 1994. Separation and determination of three kinds

of plant hormone by high performance liquid chromatography. Fenxi-Huaxue 22: 801-804.

Leshem, Y.; Philosoph, S. Wurzburger, J., 1974. Glycosylation of free trans-trans abscisic acid,

a contributing factor in bud dormancy breaks Biochem. Biophys. Res. And Commun. 59: pp.

100

526-531.

Levizou, E., P. Karageorgou, G. K. Psaras and Y. Manetas" 2002. Inhibitory effects of water

soluble leafleachates from Dittrichia viscose on lettuce root growth, statocyte development and

graviperception. Flora, 197: 152- 157.

Lowery OH, Rosenbrough NJ, Farr AL, Randall RJ., 1951. Protein measurements with the

Folin Phenol Reagent. J. BioI. Chem. 27:265-275.

Macias, F.A., Varela, R.M., Torres, A., Galindo, J.L.G., Molinillo, J.M.G.: Allelochemicals

from sunflowers: chemistry, bioactivity and applications. - In: Inderjit, Mallik, A.U. (ed.):

Chemical Ecology of Plants: Allelopathy in Aquatic and Terrestial Ecosystems. Pp. 73-87.

Birkhäuser Verlag, Basel - Boston - Berlin 2002.

Macias, F.A., Varela, R.M., Torres, A., Molinillo, J.M.G.: Potential allelopathic activity of

natural plant heliannanes: a proposal of absolute configuration and nomenclature. - J. chem.

Ecol. 26: 2173-2186, 2000.

Mauseth, J. D., 1991. Botany: An Introduction to Plant Biology. Philadelphia: Sa!lnders. pp.

348-415.

May, F.E. and J.E. Ash. 1990. An assessment of the allelopathic potential of Eucalyptus.

Australian J. Bot. 38(3): 245-254.

Mauseth, J. D., 1991. Botany: An Introduction to Plant Biology. Philadelphia: Saunders. pp.

348-415.

Mann, H.H. and Barnes, T.W., 1945. The competition between barley and certain weeds under

controlled conditions. Annals of Applied Biology 32: 15-22.

Mann, H.H. and Barnes, T. W., 1947. The competition between barley and certain weeds under

controlled conditions. II. Competition with Holcus mollis. Annals of Applied Biology 34: 252-

267.

101

Mann, H.H. and Barnes, T. W., 1950. The competition between barley and certain weeds under

controlled conditions. V. Competition with Stellaria media. Annals of Applied Biology 37: 139-

148.

Martin, P. and Rademacher, B., 1960. Studies on the mutual influences of weeds and crops.

Symposium of British Ecological Society 1: 143-152.

Massantini, F., Caporali, F. and Zellini, G., 1977. EWRS Symposium on Different Methods of

Weed Control and their Integration 1: 23-30.

Martin, L. D. and A. E. Smith., 1994. Allelopathic potential of some warm seasiol grasses.

Crop Protection, 13 (5): 388- 392.

Mason- Sedun, Wand R. S. Jessop 1988. Differential phytotoxicity among specie~ and

cultivars of the genus Brassica to wheat. Plant and Soil, 107: 69 - 80.

May, F.E. and J.E. Ash. 1990. An assessment of the allelopathic potential of Eucalyptus.

Australian J. Bot. 38(3): 245-254.

Mehlich A, 1953 determination of P, Ca, Mg, K, Na and NH4. North Carolina Soil test

Division (Mimeo).Mehlich A (1984). Mehlich 3 soil test extractant. A modification of the

Mehlich 2 extractant. 15: 1409-1416.

Nelson DW, Sommers LE., 1982. Total carbon, organic carbon and organic matter. In: Page

AL, Miller RH, Keeny DR ( eds,), Methods of Soil Analysis. Part 2: Chemical and

Microbiological properties, 2nd ed. American Society of Agronomy, Madison, WI, pp. 538-580

Mehlich A., 1984. Mehlich 3 soil test extractant: a modification of the Mehlich 2 extractant.

Soil Sci. Plant Anal. 15: 1409-1416.

Mehlich,-A., 1953. Determination ofP, Ca, Mg, K, Na and NH4. North Carolina Soil test

Division (Mimeo). Nelson DW, Sommers LE (1982). Total carbon, organic carbon and organic

matter. In: Page AL, Miller RH, Keeny DR (eds), Methods of Soil Analysis. Part 2: chemical

and microbiological properties, 2nd ed. Am. Soc. Agron. Madison WI. pp. 538-580.

102

Mccalla, T.M. and Duley, F.L., 1949. Stubble mulch studies. III. Influence of soil

microorganisms and crop residues on the germination, growth and direction of root growth of

corn seedlings. Soil Science Society of America Proceedings

14: 196-199.

Macheix J-J, Fleuriet A, Billot J. Fruit Phenolics. Boca Raton, USA: CRC Press,

1990.

Milborrow, B.V., 1970. The metabolism ofabscisic acid. J. Exp. Bot. 21: pp. 17-29. Mahajan,

S., Tueja, N., 2005. Cold, salinity and drought stresses: An overview.

Archives of Biochemistry and Biophysics. 444, 139-158.

Naqvi, S.S. M., 1999, Plant hormones and stress phenomena. -In: Pressarakli, M. (ed.):

Handbook of Plant and Crop Stress. pp. 709-730. Marcel Dekker, New York-Basel.

Naseem, M., Z. A. Cheema and S. A. Bazmi. 2003. Allelopathic effects of sunflower aqueous

extracts on germination of wheat and some important wheat weeds. Pak. J. Scientific Res. 55

(3-4): 71-75.

Naseem, M., Z. A. Cheema and S. A. Bazmi. 2003. Allelopathic effects of sunflower aqueous

extracts on germination of wheat and some important wheat weeds. Pak. J. Scientific Res. 55

(3-4): 71-75.

Nayyar, H., Walia, D.P., 2003. Water stress induced praline accumulation in contrasting wheat

genotypes as affected by calcium and abscisic acid. BioI. Plant. 46, 275-279.

Ndung'u, C. K., Shimizu, M., Okamoto, G., Hirano, K., 1997. Abscisic acid, carbohydrates, and

nitrogen contents of Kyoho grapevines in relation to bud break induction by water stress.

Amer.J. Enol. Vitic. 48, 115-120.

Norstadt, F.A. and Mccalla, T.M., 1963. Phytotoxic substances from a species of Penicillium.

Science 140: 410-411.

Neales, T.F.; Maria, H.; Zhang, H. and Davies, W.J., 1989. The effects of partially drying parts

of root system of Helianthus annus on the ABA contents in the roots, xylem Sap and Leaves, J.

103

Exp. Botany. 26: pp. 1113-1120.

Narwal, S. S., R.E. Hoagland, R. H. Dilchy and M. J. Reigosa, 1998. Allelopathy in Ecological

Agriculture and Forestry. In proceedings of the 3rd International Congress on Allelopathy in

Ecological Agriculture and Forestry, 18-21 August, 1998. Dharawad, India.

Ohno, T., Doolan, K.I., Zibilske, L.M., Liebman, M., Gollandt, E.R., Berube, C.: Phytotoxic

effect of red clover amended soils on wild mustard seedling growth. - Agricult. Ecosyst.

Environ. 78: 187-192, 2000. Rice, E.L.: Allelopathy. 2nd Ed. - Academic Press, New York

1984.

Olanbanji , SO, O. V. Makanji, D. Ceccato, M. C. Buoso, A. M. Haqu, R. Cherubini and G.

Moschini.1997. PIGE-PIXE analysis of medicinal plants and vegetables of pharmacological

importance, BioI. Trace Elem., Res., 58 (3):

223-236.

Oritano, T. and Yamashita, K., 1972. Synthesis of optically active abscisic acid and its

analogues. Tetrahedron Lett, pp. 2521.

Oleszek, W., 1987. Allelopathic effects of volatiles from some cruciferae species on lettuce,

barnyard grass and wheat growth. Plant and Soil 102: 271-273.

Osvald, H., 1950. On antagonism between plants. In: Proceedings, 7th International Congress

on Botany, Stockholm

Overland, L., 1966. The role of allelopathic substances in the barley crop. American Journal of

Botany 53: 423-432.

ada, A., Sakuta, C., Masuda, S., Mizoguchi, T., Kamoda, H., Satoh, S., 2003. Possible

involvement of leaf gibberellins in the clock-controlled expression of XSP 30, a gene encoding

a xylem sap lectin, in cucumber root. Plant Physiol. 133, 1779- 1790.

Pan, R., Tian, X., 1999. Comparative effect ofIBA, BSAA, and 5,6-ClAA-Me on the rooting

104

ofhYPocotyls in mungbean. Plant Growth Regul. 27, 91-98.

Petaraitis. P. S. R. E. Latham and R. A. Niesenbaun 1989. The maintenance of species diversity

by disturbance. Quarterly Re iew of Biology 64: 393-418.

Popova, L. P., Outlaw, W.H., Aghoram, K., Hite, D. R. C., 2000. Abscisic acid and intra leaf

water-stress signal. Physiol. Plant. 108,376-381.

Petaraitis, P. S. R. E. Latham and R. A. Niesenbaum 1989. The maintenance of species

diversity by disturbance. Quarterly Review of Biology 64: 393- 418.

Putter, J., 1974. Peroxidase, In: Ed. BergemeyerHU, Methods of Enzymatic Analysis Vol. II.

Acadmic Press, London, pp. 685-690.

Panasuik,. Bills, D. D. and Leather, G. R., 1986. Allelopathic influence of sorghum bicolor on

weeds during germination and early development of seedlings.Journal of Chemical Ecology

12:1533-44.

Patrick, l.A., 1971. Phytotoxic substances associated with the decomposition of plant residues

in soil. Soil Science 111: 13-18.

Prutenskaya"N.J., 1974. Effect of barley leachates and germinating seeds of crops on wild

mustard. In: Physiological and Biochemical Basis of Plant Interactions in Phytocenosis (Ed.,

A.M. Grodzinsky) 3: 73-75. Naukova Dumka, Kiev, USSR.

Putnam; A. R. and Defrank, J., 1983. Use of phytotoxic plant residues for selective weed

control. Crop Protection 2: 173-181.

Putnam, A.R. and Tang, C.S., 1986. The Science of Allelopathy. Wiley Interscience, New

York.

Peng, S. L., J. Wen and Q. F. Guo., 2004. Mechanism and active variety of allelochemicals.

Act. Bot. Sinica, 46: 757-766.

Putnam, A.R., J. De Frank and J.P. Barnes. 1994. Exploitation of allelopathy for weed control

105

in annual and perennial cropping system. J. Chern. Ecol. 9; 1001-1010.

Patterson, D. T., 1981. Effects of allelopathic chemicals on growth and physiological responses

of soybean (Glycine max). Weed Sci. 29: 53-59.

Padhy, B. B., P. K. Patanaik and A. K. Tripathy" 2000. Allelopathic potential of Eucalyptus

leaf litter leachates on germination and seedling growth of fingermillet. Allel. J., 7: 69- 78.

Pikovskaya, R.L., 1948, Mobilization of phosphorus in soil in connection with the vital activity

of some microbial species, Mikrobiologiya. 17: 362-370.

Periturin, F.T., 1913. Izv.Mosk. S-kh.Inst.kn.4. Recommended soil chemical test procedure for

North region (1988) North Regional Publication No. 221 (NDSU Bull. No. 499. WP5. Estlt ests

04/11/91. '

Putnam AR, Weston LA., 1986. Adverse impacts of allelopathy in agricultural systems. In The

Science of Allelopathy, eds. Putnam AR, Tang CS. John Wiley and Sons, New York. pp. 43-

56.

Quarrie, S. A., 1982. The role of abscisic acid in control of spring wheat growth and develop.

In plant growth substance. (P.F-Wareing ed.), pp. 609-619.

Qasem, J.R. 1994. Allelopathic effect of white top (Lepidium draba) on wheat and barley.

Allelopathy J. 1:29-40.

Rice, E.L., 1964. Inhibition of nitrogen-fixing and nitrifying bacteria by seed plants. Ecology

45: 824- 837.

Rice, E.L., 1971. Inhibition of nodulation of inoculated legumes by leaf leachates of pioneer

plant species from abandoned fields. American Journal of Botany 58: 368-371.

Reigosa, M. J., M. Pedrol, L. Gonzales and S.S. Narwal., 2006. Allelopathy in Ecologial

Sustainable Agriculture. In: Allelopathy: A physiological Process with Ecological Implications.

106

Reigosa, M. J., M. Pedro I and L. Gonzales ( Eds. ), Springer, Netherlands, pp: 537-564.

Rice, E.H., 1974. Allelopathy Academic Press, New York.

Roesenzweig, M. L. Z. Abransky., 1993. How is diversity and productivity related? pp. 52---65

in R. E. Ricklefs & D. Schluter (eds). Species diversity in ecological communities. University

of Chicago Press.

Reigosa, M. J., N. Pedrol, A. M.Sanchez-Moreiras, and L. Gonzales. 2002. Stress and

allelopathy. In: Allelopathy, from Molecules to Ecosystems, M.J. Reigosa and N. Pedrol, Eds.

Science Publishers, Enfield, New Hampshire.

Rice, E.L., 1984. Allelopathy, 2nd ed. New York: Academic Press.

Robinson, T., 1983. The Organic Constituents of Higher Plants. 5tI Edition. Cordus Press,

North Amherst, Massachusetts.Rose, S.J., Burnside, D.C. Specht, J.E. and Swisher, B.A.

(1984). Agronomy Journal 76: 523-528.

Raven, P. H., Evert, R. F., and Eichhorn, S. E., 1992. Biology of Plants. New York: Worth. pp.

545-572.

Sadhu, M. K. and DAS, T. M., 1971. Root exudates of rice seedlings. The Influence of one

variety on another. Plant and Soil 34: 54 1-546.

Schmitz, N., Abrams, S. R., Kermode, A. R., 2000. Changes in the abscisic acid content and

embryo sensitivity to (+)-abscisic acid during the termination of dormancy of yellow cedar

seed. J. Exp. Bot. 51, 1159-1162.

Sadhu, M.K., 1975. Nature of inhibitory substances in the root exudates of rice seedlings.

indian Journal of Experimental Biology 13: 5 77-579.

Smith, G.L. and Secoy, D., 1975. Journal of Agricultural and Food Chemistry 23:1050- 1055.

Shahid. M., B.Ahmad, R.A.Khattak, G. Hassan and H. Khan. 2006. Response of wheat and its

weeds to different allelopathic plant water extract. Pak. J. Weed Sci. Res. 12(1-2):61-68.

107

Shaybany, B.; Weinbunm, S. A. and Martien, G. C., 1977. Identification of ABA stereomers In

French prune seeds and association of ABA with ethylene- enhanced prune abscission. J. Am.

Soc. Hort, Sci. 102: pp.501-503.

Singh, N. K.; LaRosa, P.C.; Handa, A.K.; Hasegawva, P.M. and Bressan, R. A. ,1987.

.Hormonal regulation of protein synthesis associated with salt tolerance in plant cell. Proc.

Natt. Acad. Sci. USA. 84: pp. 739-743.

Singh, T .B., 1993. Effects of growth regulators on nodulation and N2 fixation in Urd bean

(Vigna mungo L.). Compo Physiol. Ecol. 18: (3) pp. 79-82.

Sloger, C. and Coldwell, B. F., 1970. Response of cultivars of soybean to synthetic abscisic

acid. Plant Physiol. 46: pp. 634-635.

Salisbury, F. B. and Ross, C. W., 1992. Hormones and growth regulators: auxin and GA. In:

Plant Physiology. (Eds.). FB Salisbury, F. B. and Ross, C.W. pp.

372-407.

Schuster B, Herrmann K. Hydroxybenzoic and hydroxycinnamic acid derivatives in soft fruits.

Phytochem 1985; 24: 2761-2764.

Shahidi, F, Naczk M. Food Phenolics. Sources, Chemistry, Effects, Applications. Lancaster,

USA: hnomic Publishing Company, Inc., 1995. Starke H, Herrmann K. The phenolics of fruits.

VIII. Changes in flavonol concentrations during fruit development. Z Lebensm-Unters Forsch

1976; 161: 131-135..

Strack, D. Phenolic metabolism. In: Dey PM, Harborne JB, eds. Plant Biochemistry. London,

UK:Academic Press, 1997, p. 387-416.

Singh, H. P., R. K. Kohli and D. R. Batish, 2001. Allelopathy in agrosystems: An overview. J.

Crop Prod., 4: 1- 41.

Strube, M, Dragsted La, Larsen JC. Naturally occurring antitumourigens. I. Plan phenols.

Nordiske Seminar- og Arbejdsrapporter 605. Copenhagen, Denmark Nordic Council of

Ministers, 1993.

108

Sinkkonen, A., 2001. Density-dependent chemical interference~an extension of th( biological

response model. J Chern EcoI27:1513-1523.

Sarwar M, Arshad M, Martens DA, Frankenberger WT Jr., 1992. Tryptophan- dependent

biosynthesis of auxins in soil. Plant Soil 147: 207-215.

Soltanpour PN, Schwab AP., 1977. A new soil test for simultaneous extraction of macro and

micro nutrients in alkaline soils. Commun.Soil Sci. Plant Anal. 8: 195-207.

Salisbury FB, Ross CW., 1992. Plant physiology. Fourth Edition. Wadworth Publ. Co.,

Belmont. p. 682.

Steel Mac Faddin, JF., 1980. Biochemical tests for identification of medical Bacteria. William

and Baltimore Wilkins, USA. pp. 51-54.

Steel KJ., 1961. The oxidase reaction as a toxic tool. J. Gen. Microbial.25: 297-306.

Siqueira, J.O., Safir G.R. & Nair, M.G., 1991. Stimulation of vesicular-arbuscular mycorrhiza

formation and growth of white clover by flavonoid compounds. New Phytologist 118, 87-93.

Schenk HJ, Callaway RM, Mahal BE., 1999. Spatial root segregation: are plants t erritorial.

Adv. Ecol. Res. 28: 146-180.

Seigler, D.S., 1996. Chemistry and mechanisms ofallelopathic interactions. Agron. J. 88: 876-

885.

Salisbury, F. B., and Ross, C. W., 1992. Plant Physiology. Belmont,CA: Wadsworth. pp. 357-

407, 531-548.

Soltanpour PN, Schwab AP., 1977. A new soil test for simultaneous extraction of macro and

micro nutrients in alkaline soils. Commun. Soil Soc. Plant Anal. 8: 195-207.

Toussoun, TA., Weinhold, A.R., Linderman, R.G. and Patrick, Z.A., 1968. Phytopathology 58:

41-45.

109

Tsuzuki, E., 1980. Studies on the allelopathy in buckwheat plants. Buckwheat Symposium,

Ljublajana, Yugoslavia. Supplementary Volume. Pp. 13-23.

Trotel-Aziz, P., Niogret, M. F., Larher, F., 2000. Proline level is partly under the control of

abscisic acid in canola leaf discs discs during recovery from hyper-osmotic stress in the adult

maize leaf. J .Exp. Bot. 54, 2177-2186.

Ulubelen, A., 2000. Terpenoids in genus Salvia. Sage; the genus Salvia 55-68. Kintzioss. E.

Amsterdam. Harwood Acadmic Publishers.

Vincent, J.M., 1970. In: A manual for the practical study of the r06t nodule bacteria. Burgess

and Son Ltd. Great Britain, p. 45.

Ventura, W., Watanable, I., Komada, H., Nishio, M., Cruz, A.D. and Castillo, M. , 1984. IRRI

Research Paper Series No. 99. International Rice Research Institute, Los Banos, Philippines.

Wall, M.K. and Iverson, L.R., 1978. Abstract of the 144th National American Association of

Advance Science Meeting, Washington, D.C. pp. 12 1-122.

Walker, D.W. and Jenkins, D.D., 1986. Influence of sweet potato plant residue on growth of

sweet potato vine cuttings and cowpea plants. Hort Science 21: 426-428.

Waller, G.R. and Dermer, O.C., 1981. Enzymology of alkaloid metabolism in plants and

microorganisms. The Biochemistry of Plants 7: 317-402.

Waller, G.R., Kenzer, F.G. and Mcpherson, J.K., 1987. Allelopathic compounds in soil from no

tillage versus conventional tillage in wheat production. Plant and Soil 98: 5-15.

Waller, G.R. and Nowacki, E.K., 1978. The role of alkaloids in plants. In: Alkaloid Biology

and Metabolism in Plants (Eds., G.R. Wailer and E.K. Nowacki). Pp. 143- 181. Plenum Press,

New York.

110

Weston, L.A., Duke, S.O.: Weed and crop allelopathy. - Crit. Rev. Plant Sci. 22: 367-389, 2003

White, R.H., Worsham, A.D. and Blum, O. (1989). Allelopathic potential of legume debris and

aqueous extracts. Weed Science 37: 674-679.

Wilson, R.E. and Rice, E.L., 1968. Allelopathy as expressed by Hetianthus annuus and its role

in old- field succession. Buttetin of the Torrey Botanicat Ctub 95: 432-448.

Weston, L. A and S.O. Duke, 2003. Weed and crop allelopathy. Crit. Rev. Plant Sci., 22: 367-

389.

Wang J, Ll U (J, UULS. 2002. The response to water stress of the antioxidant system in maize

seed;ing root with different drought resistance. Acta Botanica Boreali- Occidentalia Sinica, 22,

285- 290. ( In Chinese ).

Winkel-Shirley, B. Evidence for enzyme complexes in the phenylpropanoid and flavonoid

pathways. Physiol Plant 1999; 107: 142-149.

Wardle DA, Ahmad M, Nicholson KS., 1991. Allelopathic influence of nodding thistle (Cardus

nutans L.). seed on germination and radicle growth of pasture plants. N. Z. J. Agric. Res. 34:

185-191.

Whitney, DA., 1988. Micronutrients soil tests for zinc, iron, manganese and copper. pp. 20- 22.

In recommended chemical soil test procedure for the North Chemical region. Ed. WC Dahnke.

pp. 117- 130. North Dakota agric. Exp. Stn Bull. No. 499.

Wardle DA, Ahmad M, Nicholson KS., 1991. Allelopathic influence of nodding thistle (Cardus

nutans L.) seed on germination and radicle growth of pasture plants. New Zealand J. Agric.

Res. 34: 185-191.

Wahyuni, S., Sinniah, U. R., Amarthalingam, R., Yusop. M. K., 2003. Enhancement of

seedling establishment in rice by selected growth regulators as seed treatment. Jum Penelitian

Pertanian Pangan. 22, 51-55.

111

Watanabe, T., 1997. Lodging resistance. P. 567-577. in T. Y. Futsuhara, F. Kikuchi, and H.

Yamaguchi (Eds.). Science of the rice Plant. Vol. III. Genetics. Food and Agriculture Policy

Research Center, Tokyo, Japan.

Xie, Z., Jiang, D., Cao, W., Dai, T., Jing, Q., 2003. Relationships of endogenous plant

hormones to accumulation of grain protein starch in winter wheat under different post- anthesis

soil water statuses. Plant Growth Regul. 41, 117-127.

Yang, J., Wang, Z., Zhu., Q., Lang Y., 1999. Regulation of ABA and GA to rice grain filling.

Acta Agron Sin. 25, 341-348.

Yang J., Zhang, J., Wang, Z., Zhu, Q., Wang. W., 2001. Hormonal changes in the graings of

rice subjected to water stress during grain filling. Plant Physiol. 127,315-323.

Zhang, X., Miao, Y.C., An, G.Y., Zhou, Y., Shangguan, Z.P., Gao, J.F., Song, C.P., 2001. K+

channels inhibited by hydrogen peroxide mediate abscisic acid signalling in guard cells. Cell

Res. 11, 195-202.

Zeevaart, J.A.D. and Creelman, R.A. (1988). Metabolism and physiology of abscisic acid. Ann.

Rev. Plant Physiol. Plant Mol. BioI. 39: pp. 439-473.

112

APPENDICES

Appendix I

Metrological Data of Islamabad

Years : 2006-2007

Air Temperature (Co) Month / Year

Minimum Maximum

Average R.H. % Average Rainfall

(mm

Jan – 2006 3 17 78 53.69

Feb 8 24 70 22.94

March 9.41 24.16 71 51.81

April 12.75 31.46 45 20.56

May 21.2 38.5 40 41.26

June 21.6 37 46 61.98

July 24 35 76 19.11

Aug 23.17 31.77 82 312.15

Sep 19.6 32.8 67 13.42

Oct 15.38 30.48 66 35.07

Nov 9.3 23.53 73 14.89

Dec 7.16 22.10 70 10.90

Jan-2007 1.6 18.9 59 73.21

Feb 6.6 18.9 81 93.56

March 9.06 22.88 73 178.99

April 14.6 33.3 54 3.02

May 17.22 34.17 46 57.76

June 22.6 37.6 54 104.82

113

July 22.70 34.27 77 33.5

Aug 23.3 33.32 81 22.81

Sep 19.9 32.16 76 133.13

Oct 11.16 30.80 57 103.03

Nov 7.20 25.23 67 13.35

Dec 2.58 18.93 70 7.98

Jan-2008 4 16 76 8.20

Feb 6 20 75 17.10

March 10 25 73 18.20

April 14 29 70 30.25

May 19 40 38 38.20

June 19.10 36 44 60.10

114

Appendix ll

Yeast extract Mannitol Agar (YMA), medium, (Vincent, 1970).

K2HPO4

Mannitol

MgSO4.7H20

NaCI

Yeast extract

Agar”

Distilled water

0.5g

10.Og

0.2g

l.Og

0.1 g

20.0 g

l000rnL

Yeast inannitol agar medium consists of’ the following components:

Congo-red Medium:

Congo-red (2.5 mL/1 of 1% solution) is incorporated into yeast extract mannitol medium.

Congo-red solution is sterilized separately and added to the sterilized medium.

YMA Medium with Bromothymol Blue (BTB).

In YMA medium 0.5 mL (0.5% in absolute alcohol) Brornothyinol blue was added.

115

Appendix IIl

Glucose Peptone Agar Medium:

Glucose peptone agar medium was consisted of the following components;

Glucose

Peptone

Agar

I3romo cresol purple (I % alcoholic solution)

Distilled water

5.Og

10.Og

15.Og

I 5.Og

I 000rnL.

116

Appendix IV

Combined Carbon Medium (CCM)

CCM was prepared according to Rennie (1981) as follows:

Sucrose

Mannitol

Sodium lactate

K2HPO

KH2PO4

MgSO4.7H20

CaCI2.2H20

NaCI

Yeast extract

Na2MoO4.2H20

Na2Fe-EDTA

Biotin

Paraminobenzoic

acid

5g/L

5gIL

60% v/v

O.8gm/L

0.2g/L

0.2g/L

0.OGg/L

lg/L

lg/L

0.O25gJL

0.028g/L

5ig/L

10ig/L

IL

117

Appendix V

LB (Lubria-Ber (ani) Medium

Trypton

Yeast extract

NaCI

Agar

H20

pH (Final)

lOg

5g

lOg

18g

l000mL

7.0

118

Appendix Vl

Gram staining

Preparation of solutions

Crystal violet (Hucker’s)

• SoIution A

Crystal violet (90% dye content)

Ethyl alcohol (95%)

• Solution B

Ammonium oxalate

Distilled water

Mix solutions A and B

• Grams’ iodine

Iodine

Potassium iodine

Distilled water

• EEhyl Alcohol (95%)

Ethyl alcohol (100%)

Distilled water

• Safranin

Safranin 0(2.5% solution in 95% ethyl alcohol)

Distilled water

2 gm

20 rnL

0.8 gm

80 rnL

1 gm

2 gm

300

mL

95 rnL

5 mL

10 mL

100

119

Appendix VIl Soil analysis reagents

The soil nitrogen content and total extractable P was determined by the method described by

Soltanpour (1985).

1. N03-N determination

N03-N was determined according to the method of Soltanpour and Schwab (1977).

• Extraction procedure

Oven dried soil (10.0 g) was taken in 50 rnL conical flask. Extracting solution (ABD1’PA; 20

mL) was added. Samples were shaken on a shaker for 15 minutes and filtered through

Whatman No. 40 filter paper. The extracting aliquots were kept in storing bottles for analyses.

• Reagents required for N03-N

i. Hydrazinc sulphate stock solution

Hydrazine (NH2NH2H2SO4 27.0 g) was dissolved in I L warm distilled water to prepare stock

solution. For preparation of working solution, 22.5 mL hydrazine sulphate from stock solution

was taken and diluted to 1 L with distilled water.

ii. CuSO4 stock solution

Stock solution was prepared by dissolving 3.9 g CuSO4.5H20 in 1 L distilled water. From

stock solution, 6.25 mL solution was taken and diluted up to I L for the preparation of working

solution.

iii. NaOH stock (1.5 N) working solution

To prepare I .5 N NaOH stock solution 60.0 g NaOH was dissolved in I L distilled water. For

the preparation olworking solution 200 rnL of the stock solution were diluted to I L.

120

iv. Stock N03-N solution for standard

For the standard preparation 3.6090 g KNO3 was dissolved in I L distilled water. From this

stock solution, working solution was prepared by mixing 25 mL stock solution in 1 L distilled

H2O.

• Standard for N03-N

By using working N03-N, the standards containing 0, 0.5, I, 1.5, 2, 2.5 and 3 mg N03-N per

liter were prepared.

Preparation of sample for analyses

Samples along with standards were processed as follows: one ml. of sample (or standard

solutions), two ml. of working CuSO4 solution and one niL of hydrazine sulphate solution were

mixed and placed in a water bath for 20 minutes at 38°C. After adding 3 mL of colour reagent,

samples were analyzed on Spectronic 21 at 540 nrn.

• Preparation of color reagent

Sulfanilamide (5.0 g) and Naphthyl-ethylendiamine dihydrochloride (0.25 g) were dissolved in

300 mL distilled H2O. After the addition of 50 mL H3 P04, the volume was made upto 500 ml.

with distilled water.

2. Determination of P contents

• Preparation of mixed reagent

Mixed reagent was prepared by mixing I L of 5 N H2S04 with I L distilled water containing

potassium tartarate (0.2908 g).

• Working color reagent

For the preparation of working colour reagent, 0.74 g of ascorbic acid were dissolved in 140

ml. of mixed reagent.

121

• Preparation of standard solutions

H2PO4 solution (100 ppm) was prepared by diluting stock (1000 ppm) solution. From this (1 00

ppm) solution, 0, 0.5, 1, 1 .5, 2, 2.5 and 3.0 ppm solutions were prepared.

Samples preparation

Samples along with standard were prepth-ed as follows: One mL of sample (or standard

solutions), 9.0 mL of distilled water and 2.5 mL of working colour reagent (colour reagent +

ascorbic acid) were mixed and analyzed after 1 5 to 20 minutes on Spectronic 21 at 880 nm.

3. Determination of K, Ca++

and Mg++

ions

Reagents

i. Lanthanum diluting solution

Lanthanum oxide (La2O3: 5.9 g) was dissolved in 20 mL distilled H2O in a 500mL flask and

placed in a cold water bath. Concentrated HCI (10.5 mL) and HNO3, (14 mL) were added to

100 mL flask containing lanthanum oxide. The final volume was diluted with 200 rnL distilled

H20.

ii. High stock solutions

i. K’ = (2000 ppm): 3.815 g KCI diluted to volume (1L) with distilled H20.

ii. Ca’’ = (1 0,000 ppm): 24.97 g CaCO3 dissolved in 1 L distilled H2O.

iii. Mg’‘= (1000 ppm): 1.0 g Mg ribbon dissolved in IL distilled H2O

III. Low stock solutions

Employing following salts low stock solutions were made;

i. K’ (100 ppm): 0.1907 g of KCI dissolved in IL distilled H2O.

ii. Ca’’ (500 ppm): 1 .25 g CaCO3 dissolved in IL distilled H20

iii. Mg (500 ppm): 0.829 g MgO in 10 mL ofHNO3 and final volume was made I L.

Low stock solutions were added in 1 00 mL flask and final volume was made with appropriate

extracting solutions.

Extraction ofK’, Mg’ Mn++

, Ca’, from the soil samples were done accordng to

Mehlich 1953 and 1984.

Procedure

Aliquot (1.5 mL) of’ each working standard and all soil extracts were diluted with the

122

CaCO3 working solution to final volume of’ 15 mL. K, Ca and Mg’ were measured

by Atomic Absorption Spectrophotometer (Shimidzu, AA-670) at the wavelength of

766.5, 422.7, and 285.2 nm, receptively.

4. Analysis of Fe++

, Mn++

and Zn++

Reagents

i. Redistilled 6 M HCI

ii. Stock standard solution

iii. 0.1 M HCI extracting solution

iv. Working standard

Procedure

i. Sieved 5.0 g dried soil was taken in 50 mL flask.

ii. 20 mL of extracting solution was added to each flask and shaken for 30 minutes at 180 rpm.

iii. The suspension was filtered through a medium pore size filter in 30 niL beaker

123

iv. Concentrations of Fe++

, Mn++

and Zn++

were determined with Atomic Absorption

Spectrophotometer (Shimidzu, AA-670).

All the solutions were prepared according to Whitney (1988).

Appendix Vlll

B rough ton and D ilworth’s Solti tion

The composition of the modified Broughton and Dilworth (1970) nitrogen free mineral solution

was:

Stock Compound Amount gIL Final solution concentration

I CaCI2.2H20 204.1 1.00mM

2 KH2PQ1 136.1 0,50mM

3 MttSQ4.7H2Q4 123.3 0.25 mM

4

KSO4

MnSO4.H2O

H.1BO.

ZnSO4.7H20

(‘uSO4.5H20

‘NaMoO2.2H20

CoSO1.7H20

87.0

0.338

0.247

0.288

0.1

0.048

0.056

0.25mM

I.00tM

0.30p.M

0.50tM

0.20iM

0.Ol1iM

0.01tM

5 Fe citrate+ 5.4 1 0.00M

* For each liter of lull-strength solution added 0.5 rnL from each of the live stock solutions.

124

Appendix IX

Dragendorff Reagent

Take 1.7 g basic bismuth nitrate is dissolved in a mixture of and 20 ml acetic acid and

80 ml water (= solution a); 16 g potassium iodide in 40 ml water (solution b); a and b

are mixed 1: 1 (v/v) (stock solution - may be kept several months in refrigerator); for

spraying: 1 ml stock solution is mixed with 2 ml acetic acid and 10 ml water. Color

reaction: orange-brown spots on a yellow background.

Appendix X

KNO3 stock solution:

For preparation of lOOppm solution of Potassium, 0.0258 gm of Potassium nitrate (KNO3) was

dissolved in five ml of 1% nitric acid and volume was made to 100 ml by adding distilled

water.

MgSO4 stock solution:

100 ppm stock solution of Magnesium sulphate was prepared by dissolving 0.0495gm

MgSO4 in five ml of 1% nitric acid and volume was made to lOOmI by adding distilled

water.

CaC12 stock solution:

0.027 gm of Calciuni chloride was dissolved in five ml of 1% HNO3 and volume was raised up

to 1 OOml by adding distilled water.

FeSO4 stock solution:

100 ppm stock solution of FeSO4 was made by dissolving 0.0497 gm of FeSO4 in five ml of

1% nitric acid and volume were raised up to lOOmi by adding distill water.

ZnSO4 stock solution: ZnSO4 lOOppm stock solution was prepared by adding 0.0439 gm in

five ml of 1% nitric acid and volume was raised up to lOOmI by adding distilled water.

125

PbNO3 stock solution:

PbNO3 100 ppm stock solution was prepared by adding 0.0159 gm in five ml of 1%

nitric acid and volume were raised up to lOOm! by adding distill water

126

V1 2006

13.67

1011 10.67

12

0

5

10

15

Treatments

Germ

ination %

T1 T2 T3 T4 T5

`

V2 2006

12

78

9.667 10.33

0

5

10

15

Treatments

Germ

ination %

T1 T2 T3 T4 T5

`

V1 2007

14

910 10.67

12

0

5

10

15

Treatments

Germ

ination %

T1 T2 T3 T4 T5

`

V2 2007

12.33

67

8.77 9.33

0

5

10

15

Treatments

Germ

ination %

T1 T2 T3 T4 T5

`

Fig. 1-4. Effect of sunflower leaf extract on germination (%) of wheat varieties


127

V1 2006

13.66

1112 12.33 12.67

0

5

10

15

Treatments

Germ

ination %

T1 T2 T3 T4 T5

`

V2 2006

12

9.33310 10.33

11.67

0

5

10

15

Treatments

Germ

ination %

T1 T2 T3 T4 T5

`

V1 2007

14

11.33 11.67 12 12.33

0

5

10

15

Treatments

Germ

ination %

T1 T2 T3 T4 T5

`

V2 2007

12.33

8 8.6679.66 10.33

0

5

10

15

Treatments

Germ

ination %

T1 T2 T3 T4 T5

`

Fig. 5-8. Effect of sunflower stem extract on germination (%) of wheat varieties


128

V1 2006

13.67

10.3311.67 12 12.67

0

5

10

15

TreatmentsGerm

ination %

T1 T2 T3 T4 T5

`

V2 2006

12

99.667

10.6711.33

0

5

10

15

Treatments

Germ

ination %

T1 T2 T3 T4 T5

`

V1 2007

14

9.3310.67 11

11.67

0

5

10

15

Treatments

Germ

ination %

T1 T2 T3 T4 T5

`

V2 2007

12.33

6.6677.667

9.333 9.667

0

5

10

15

Treatments

Germ

ination %

T1 T2 T3 T4 T5

`

Fig. 9-12. Effect of sunflower root extract on germination ( % ) of wheat varieties


129

V1 2006

12.3310.67

11.6712.67

13.67

0

5

10

15

Treatments

Root Length (cm)

T1 T2 T3 T4 T5

`

V2 2006

11.67

8.679.667

10.67

12.5

0

5

10

15

Treatments

Root Length (cm)

T1 T2 T3 T4 T5

`

V1 2007

14

9.5 10.212

13

0

5

10

15

Treatments

Root Length (cm)

T1 T2 T3 T4 T5

`

V2 2007

13

8.7 9.511.08

12.6

0

5

10

15

Treatments

Root Length (cm)

T1 T2 T3 T4 T5

`

Fig. 13-16. Effect of leaf extract of sunflower on root length ( cm ) of seedlings of wheat


130

V1 2006

18.3

8.7 9.811.2

13.27

0

5

10

15

20

Treatments

Shoot Length (cm)

T1 T2 T3 T4 T5

`

V2 2006

17.5

12.3 13.2 13.915.3

0

5

10

15

20

Treatments

Shoot Length (cm)

T1 T2 T3 T4 T5

`

V1 2007

18.43

8.267 9.1411.01

12.59

0

5

10

15

20

Treatments

Shoot Length (cm)

T1 T2 T3 T4 T5

`

V2 2007

17

11.91 12.95 13.5314.9

0

5

10

15

20

Treatments

Shoot Length (cm)

T1 T2 T3 T4 T5

`

Fig. 17-20. Effect of leaf extract of sunflower on shoot length (cm) of seedlings of wheat


131

V1 2006

0.91

0.20.3

0.6

0.81

0

0.5

1

Treatments

Fresh weight (g)

T1 T2 T3 T4 T5

`

V2 2006

0.89

0.62 0.660.72

0.8033

0

0.5

1

Treatments

Fresh W

eight (g)

T1 T2 T3 T4 T5

`

V1 2007

0.9267

0.18330.2733

0.57

0.7567

0

0.5

1

Treatments

Fresh W

eight (g)

T1 T2 T3 T4 T5

`

V2 2007

0.83

0.59 0.62 0.680.77

0

0.5

1

Treatments

Fresh W

eight (g)

T1 T2 T3 T4 T5

`

Fig. 21-24. Effect of sunflower leaf extract on fresh weight ( g ) wheat varieties


132

V1 2006

0.87

0.170.27

0.57

0.77

0

0.5

1

Treatments

Dry W

eight (g)

T1 T2 T3 T4 T5

`

V2 2006

0.86

0.42 0.4767 0.51330.5867

0

0.5

1

Treatments

Dry W

eight (g)

T1 T2 T3 T4 T5

`

V1 2007

0.9833

0.15330.2433

0.530.75

0

0.5

1

1.5

Treatments

Dry W

eight (g)

T1 T2 T3 T4 T5

`

V2 2007

0.810.56 0.59 0.654 0.74

0

0.5

1

Treatments

Dry W

eight (g)

T1 T2 T3 T4 T5

`

Fig. 25-28. Effect of leaf extract of sunflower on dry weight ( g ) of seedlings of wheat


133

V1 2006

14

1011.33

12.213.5

0

5

10

15

TreatmentsRoot Length (cm)

T1 T2 T3 T4 T5

`

V2 2006

14

10.339.25

11.2512.58

0

5

10

15

Treatments

Root Length (cm)

T1 T2 T3 T4 T5

`

V1 2007

14.67

10.83 11.17 12.07 12.98

0

10

20

Treatments

Root Length (cm)

T1 T2 T3 T4 T5

`

V2 2007

14.69

10.03 8.91710.87 12.2

0

10

20

Treatments

Root Length (cm)

T1 T2 T3 T4 T5

`

Fig. 29-32. Effect of stem extract of sunflower on root length (cm) of seedings of wheat


134

V1 200618.3

11.5 12.49 13.1514.58

0

5

10

15

20

Treatments

Shoot Length (cm)

T1 T2 T3 T4 T5

`

V2 2006

18.3

12.32 13.23 13.8515.67

0

5

10

15

20

Treatments

Shoot Length (cm)

T1 T2 T3 T4 T5

`

V1 2007

18.43

11.33 12.35 12.5813.98

0

5

10

15

20

Treatments

Shoot Length (cm)

T1 T2 T3 T4 T5

`

V2 2007

18.43

11.64 13.03 13.415.35

0

10

20

Treatments

Shoot Length (cm)

T1 T2 T3 T4 T5

`

Fig. 33-36. Effect of stem extract of sunflower on shoot length (cm) of seedlings of


135

V1 2006

0.91

0.510.5767

0.69330.86

0

0.5

1

TreatmentsFresh weight (g)

T1 T2 T3 T4 T5

`

V2 2006

0.91

0.72 0.78 0.83 0.89

0

0.5

1

Treatments

Fresh W

eight (g)

T1 T2 T3 T4 T5

`

V1 2007

0.9267

0.49 0.54330.6733

0.8367

0

0.5

1

Treatments

Fresh W

eight (g)

T1 T2 T3 T4 T5

`

V2 2007

0.910.69 0.7433 0.8 0.86

0

0.5

1

Treatments

Fresh W

eight (g)

T1 T2 T3 T4 T5

`

Fig. 37-40. Effect of stem extract of sunflower on fresh weight (g) of seedings of wheat


136

V1 2006

0.87

0.4667 0.5267 0.5433 0.5733

0

0.5

1

Treatments

Dry W

eight (g)

T1 T2 T3 T4 T5

`

V2 2006

0.86

0.47 0.5167 0.550.64

0

0.5

1

Treatments

Dry W

eight (g)

T1 T2 T3 T4 T5

`

V1 2007

0.8933

0.45 0.51 0.52

0.7367

0

0.5

1

Treatments

Dry W

eight (g)

T1 T2 T3 T4 T5

`

V2 2007

0.89330.5833 0.6567 0.7 0.7567

0

1

Treatments

Dry W

eight (g)

T1 T2 T3 T4 T5

`

Fig. 41-44. Effect of stem extract of sunflower on dry weight (g) of seedings of wheat


137

V1 2006

18.3

11.86 12.66 13.1914.63

0

10

20

TreatmentsShoot Length (cm)

T1 T2 T3 T4 T5

`

V2 2006

17.5

12.514.2

12.1314.7

0

10

20

Treatments

Shoot Length (cm)

T1 T2 T3 T4 T5

`

V1 2007

18.43

11.6413.03 13.4

15.35

0

10

20

Treatments

Shoot Length (cm)

T1 T2 T3 T4 T5

`

V2 2007

1712.92 13.17 13.7

16.32

0

10

20

Treatments

Shoot Length (cm)

T1 T2 T3 T4 T5

`

Fig. 45-48. Effect of root extract of sunflower on shoot length (cm) of seedlings of


138

V1 2006

0.89

0.2333 0.32

0.6333

0.83

0

0.5

1

Treatments

Fresh weight (g)

T1 T2 T3 T4 T5

`

V2 2006

0.89

0.750.83

0.8667 0.8733

0.6

0.8

1

Treatments

Fresh W

eight (g)

T1 T2 T3 T4 T5

`

V1 2007

0.91

0.190.29

0.60.75

0

0.5

1

Treatments

Fresh W

eight (g)

T1 T2 T3 T4 T5

`

V2 2007

0.830.71

0.79 0.8233 0.84

0.5

1

Treatments

Fresh W

eight (g)

T1 T2 T3 T4 T5

`

Fig. 49-52. Effect of root extract of sunflower on fresh weight (g) of seedlings of wheat varieties


139

V1 2006

0.86

0.1833 0.2833

0.58330.7833

0

0.5

1

Treatments

Dry W

eight (g)

T1 T2 T3 T4 T5

`

V2 2006

0.86

0.5433 0.63 0.67 0.6833

0

0.5

1

Treatments

Dry W

eight (g)

T1 T2 T3 T4 T5

`

V1 2007

0.8933

0.16330.2633

0.55670.7533

0

0.5

1

Treatments

Dry W

eight (g)

T1 T2 T3 T4 T5

`

V2 2007

0.8333

0.51670.61330.6467 0.67

0

0.5

1

Treatments

Dry W

eight (g)

T1 T2 T3 T4 T5

`

Fig. 53-56. Effect of root extract of sunflower on dry weight (g) of seedlings of wheat varieties Margalla 99 Chakwall 97.

140

V1 2006

13

9.63310.5 10.9

12.4

0

5

10

15

Treatments

Root Length (cm)

T1 T2 T3 T4 T5

`

V2 2006

17.5

13.2 13.67 14.2

16.96

0

5

10

15

20

Treatments

Root Length (cm)

T1 T2 T3 T4 T5

`

V1 2007

12.57

8.823 9.511.65 12.3

0

5

10

15

Treatments

Root Length (cm)

T1 T2 T3 T4 T5

`

V2 2007

14.69

10.03 8.91710.87

12.2

0

10

20

Treatments

Root Length (cm)

T1 T2 T3 T4 T5

`

Fig 57-60. Effect of root extract of sunflower on root length (cm) of seedlings of wheat


141

V1 2006

500

382.7 392 408.3446.7

0

200

400

600

Treatments

GA (µµ µµg g-1)

T1 T2 T3 T4 T5

`

V2 2006

490

340.3380 387

430

0

200

400

600

Treatments

GA (µµ µµg g-1)

T1 T2 T3 T4 T5

`

V1 2007

380298

405.3442.3503.3

0

200

400

600

Treatments

GA ( µµ µµg g-1)

T1 T2 T3 T4 T5

`

V2 2007

335.3 375 382421.7492

0

200

400

600

Treatments

GA (µµ µµg g-1)

T1 T2 T3 T4 T5

`

Fig. 61-64. Effect of leaf extract of sunflower on GA ( µµµµg g-1



142

V1 2006

414.7

303.7345.7 357.7

405

0

200

400

600

Treatments

GA (µµ µµg g-1)

T1 T2 T3 T4 T5

`

V2 2006

398.3344.3 377.3 393.7 417.7

0

200

400

600

Treatments

GA (µµ µµg g-1)

T1 T2 T3 T4 T5

`

V1 2007

301.3 340.7353

402.7415

0

200

400

600

Treatments

GA ( µµ µµg g-1)

T1 T2 T3 T4 T5

`

V2 2007

382.7 392 408.3 446.7500

0

200

400

600

Treatments

GA (µµ µµg g-1)

T1 T2 T3 T4 T5

`

Fig. 65-68. Effect of leaf extract of sunflower on GA (µµµµg g-1

) content of roots in wheat


143

V1 2006163

92120

139 152

0

50

100

150

200

Treatments

IAA ( µµ µµg g-1)

T1 T2 T3 T4 T5

`

V2 2006

132

100.7 105.7 110.7 118

0

50

100

150

Treatments

IAA (µµ µµg g-1)

T1 T2 T3 T4 T5`

V1 2007

90.33118

137149.7164.7

0

50

100

150

200

Treatments


T1 T2 T3 T4 T5

`

V2 2007

99.33 103.7108.3 115133

0

50

100

150

Treatments


T1 T2 T3 T4 T5

`

Fig. 69-72. Effect of leaf extract of sunflower on IAA (µµµµg g-1



144

V1 2006190

113.3125

135145

0

50

100

150

200

Treatments


T1 T2 T3 T4 T5

`

V2 2006

130

90.33105 103

116

0

50

100

150

Treatments


T1 T2 T3 T4 T5

`

V1 2007

111.3 123 131.7 141.7191.3

0

100

200

300

Treatments


T1 T2 T3 T4 T5

`

V2 2007

87.67 103 107 114131.3

0

100

200

Treatments


T1 T2 T3 T4 T5

`


) contents of roots in wheat


145

V1 2006

68

182162

142

108

0

50

100

150

200

Treatments

ABA ( µµ µµg g-1)

T1 T2 T3 T4 T5

`

V2 2006

90.3

185.7167.3

146.3

110.3

0

50

100

150

200

Treatments

ABA (µµ µµg g-1)

T1 T2 T3 T4 T5

`

V1 2007

182.7163.3

143

109

66.33

0

50

100

150

200

Treatments


T1 T2 T3 T4 T5

`

V2 2007

186.7168.3

147.3

111.7

91.61

0

50

100

150

200

Treatments


T1 T2 T3 T4 T5

`

Fig. 77-80. Effect of leaf extract of sunflower on ABA (µµµµg g-1



146

V1 2006

100

168.7138.7

117.7 108.7

0

50

100

150

200

Treatments


T1 T2 T3 T4 T5

V2 2006

81.33

122 112.7100 90.33

0

50

100

150

Treatments


T1 T2 T3 T4 T5

`

V1 2007

169.7140.3

119.3 110.798.67

0

50

100

150

200

Treatments


T1 T2 T3 T4 T5

`

V2 2007

123 113.7101.1 91.33

82.33

0

50

100

150

Treatments


T1 T2 T3 T4 T5

`

Fig. 81-84. Effect of leaf extract of sunflower on ABA (µµµµg g-1



147

V1 2006

500

435.7459

486504

400

450

500

550

Treatments

GA (µµ µµg g-1)

T1 T2 T3 T4 T5

`

V2 2006

490392.7 404 408.3 458.3

0

200

400

600

Treatments

GA (µµ µµg g-1)

T1 T2 T3 T4 T5

V1 2007

432.7456.7

476497503.3

350

400

450

500

550

Treatments

GA (µµ µµg g-1)

T1 T2 T3 T4 T5

`

V2 2007

390.7 399.3 402.7452492

0

200

400

600

Treatments

GA (µµ µµg g-1)

T1 T2 T3 T4 T5

`

Fig. 85-88. Effect of stem extract of sunflower on GA (µµµµg g-1

) content leaves in wheat


148

V1 2006

515

394.3 405 415.3475

0

200

400

600

Treatments

GA (µµ µµg g-1)

T1 T2 T3 T4 T5

`

V2 2006

414.7340 358.3 385.7

414.3

0

200

400

600

Treatments

GA (µµ µµg g-1)

T1 T2 T3 T4 T5

V1 2007

340 358.3385.7414.3415

0

500

Treatments

GA (µµ µµg g-1)

T1 T2 T3 T4 T5

`

V2 2007

312 352.7 363.7417.7

414.7

0

200

400

600

Treatments

GA (µµ µµg g-1)

T1 T2 T3 T4 T5

`

Fig. 88-92. Effect of stem extract of sunflower on GA (µµµµg g-1



149

V1 2006

163

98.33127

145.7 158

0

50

100

150

200

Treatments

IAA (cm)

T1 T2 T3 T4 T5

V2 2006

132

105.3 110.3 117.3127.3

0

50

100

150

Treatments

IAA (cm)

T1 T2 T3 T4 T5

`

V1 2007

109.3 115.3 121133.7144

0

50

100

150

200

Treatments

IAA (cm)

T1 T2 T3 T4 T5

V2 2007

115 130 136167191.3

0

100

200

300

Treatments

IAA (cm)

T1 T2 T3 T4 T5

`


) of seedlings of wheat


150

V1 2006

190

118135 141.7

171.7

0

50

100

150

200

Treatments


T1 T2 T3 T4 T5

`

V2 2006

130

100 110 118129

0

50

100

150

Treatments


T1 T2 T3 T4 T5

V1 2007

96.33124

140.7155164.7

0

100

200

Treatments


T1 T2 T3 T4 T5

V2 2007

100.3 106.3 112.3124133

0

50

100

150

Treatments


T1 T2 T3 T4 T5

Fig. 97-100. Effect of stem extract of sunflower on IAA (µµµµg g-1



151

V1 2006

68

141

108 103

81

0

50

100

150

Treatments


T1 T2 T3 T4 T5

V2 2006

90.3

145

109 101.3 96.33

0

50

100

150

200

Treatments


T1 T2 T3 T4 T5

V1 2007

143

110.7 10484

66.33

0

50

100

150

200

Treatments

ABA ( µµ µµ

g g-1)

T1 T2 T3 T4 T5

V2 2007

147

111 103.7 98.3391.61

0

50

100

150

200

Treatments


T1 T2 T3 T4 T5

Fig. 101-104. Effect of stem extract of sunflower on ABA (µµµµg g-1

) content leaves in


152

V1 2006

100117

108 102.3 94

0

50

100

150

Treatments


T1 T2 T3 T4 T5

V2 2006

81.3399.67 91.33 85.33 80

0

50

100

150

Treatments


T1 T2 T3 T4 T5

V1 2007

119 110.3 105 97.3398.67

0

50

100

150

Treatments


T1 T2 T3 T4 T5

V2 2007

101.7 93.7 88 81.6782.33

0

50

100

150

Treatments


T1 T2 T3 T4 T5

Fig. 105-109. Effect of stem extract of sunflower on ABA (µµµµg g-1

) content roots in


153

V1 2006

500

404 418.7

527480

0

200

400

600

Treatments

GA (µµ µµg g-1)

T1 T2 T3 T4 T5

V2 2006

490

351388.3 394.3

437

0

200

400

600

Treatments

GA (µµ µµg g-1)

T1 T2 T3 T4 T5

V1 2007

402 416.7424.7477503

0

200

400

600

Treatments

GA (µµ µµg g-1)

T1 T2 T3 T4 T5

V2 2007

348.7 386 391.7434492

0

200

400

600

Treatments

GA (µµ µµg g-1)

T1 T2 T3 T4 T5

Fig. 110-113. Effect of root extract of sunflower on GA (µµµµg g-1



154

V1 2006

505.3

392.3 399 411.7482.3

0

200

400

600

Treatments

GA (µµ µµg g-1)

T1 T2 T3 T4 T5

V2 2006

422.3318 360 364.7

417

0

200

400

600

Treatments

GA (µµ µµg g-1)

T1 T2 T3 T4 T5

V1 2007

301349.7364.3

417407

0

200

400

600

Treatments

GA (µµ µµg g-1)

T1 T2 T3 T4 T5

V2 2007

313 349 362412.3

419.7

0

200

400

600

Treatments

GA (µµ µµg g-1)

T1 T2 T3 T4 T5

Fig. 114-117. Effect of root extract of sunflower on GA (µµµµg g-1



155

V1 2006172.7

100127 139.3

160

0

50

100

150

200

Treatments


T1 T2 T3 T4 T5

V2 2006131

102 107 114 122

0

50

100

150

Treatments


T1 T2 T3 T4 T5

V1 2007

113139 149

190

127

0

50

100

150

200

Treatments


T1 T2 T3 T4 T5

V2 2007

96 106 107121132

0

50

100

150

Treatments


T1 T2 T3 T4 T5

Fig. 118-121. Effect of root extract of sunflower on IAA (µµµµg g-1



156

V1 2006

193

117 129 135 155.7

0

100

200

300

Treatments


T1 T2 T3 T4 T5

V2 2006

164

94120

139 153

0

100

200

Treatments


T1 T2 T3 T4 T5

V1 2007

96

140.3155164

125.7

0

50

100

150

200

Treatments


T1 T2 T3 T4 T5

V2 2007

93.3109 113

123134

0

50

100

150

Treatments


T1 T2 T3 T4 T5

Fig. 122-125. Effect of root extract of sunflower on IAA (µµµµg g-1



157

V1 2006

68

177.3156

133.7

98

0

50

100

150

200

Treatments


T1 T2 T3 T4 T5

V2 2006

90.3

177.3159

132.7102

0

100

200

Treatments


T1 T2 T3 T4 T5

V1 2007

180.3

136.3

100.7

66.33

159

0

50

100

150

200

Treatments


T1 T2 T3 T4 T5

V2 2007

180162.3

135

104

91.61

0

50

100

150

200

Treatments


T1 T2 T3 T4 T5

Fig. 126-129. Effect of root extract of sunflower on ABA (µµµµg g-1



158

V1 2006

100

151130

105.7100.3

0

50

100

150

200

Treatments


T1 T2 T3 T4 T5

V2 2006

81.33

113.7 106 96.67 84

0

50

100

150

Treatments


T1 T2 T3 T4 T5

V1 2007

153

107.7 102.798.67

132

0

50

100

150

200

Treatments


T1 T2 T3 T4 T5

V2 2007

116.3 108 98.6786

82.33

0

50

100

150

Treatments


T1 T2 T3 T4 T5

Fig. 130-133. Effect of root extract of sunflower on ABA (µµµµg g-1



159

V2 2007

135.6221 256.7

376.7513

0

200

400

600

Treatments

DNA (mg/ 100 g

F. wt)

T1 T2 T3 T4 T5

V2 2007

93.67193.3

249.3

367.3485.3

0

200

400

600

Treatments

DNA (mg/ 100 g

F. wt)

T1 T2 T3 T4 T5

V2 2007

133.4218.7 254.7

374.3515

0

200

400

600

Treatments

DNA (mg/ 100 g

F. wt)

T1 T2 T3 T4 T5

V2 2007

92

190.7247

365.3487

0

200

400

600

Treatments

DNA (mg/ 100 g F.

wt)

T1 T2 T3 T4 T5

Fig. 134-138. Effect of leaf extract of sunflower on DNA (mg/ 100 g F. wt) content of

leaves in wheat varieties Margalla 99 Chakwall 97.

160

V2 2007

248.7

386 420.7490.3513

0

200

400

600

Treatments

DNA (mg/ 100 g

F. wt)

T1 T2 T3 T4 T5

V2 2007

242

378.3415.3466485.3

0

200

400

600

Treatments

DNA (mg/ 100 g

F. wt)

T1 T2 T3 T4 T5

V2 2007

247

383.7 417.3488514.6

0

200

400

600

Treatments

DNA (mg/ 100 g

F. wt)

T1 T2 T3 T4 T5

V2 2007

239

374412.7

464.3488

0

200

400

600

Treatments

DNA (mg/ 100 g

F. wt)

T1 T2 T3 T4 T5

Fig. 139-142. Effect of stem extract of sunflower of DNA (mg/ 100 g F. wt) content of


161

V2 2007

150.4239 272.3

398.3512.9

0

200

400

600

Treatments

DNA (mg/ 100 g

F. wt)

T1 T2 T3 T4 T5

V2 2007

101.3206.7

269.3

396.7485.3

0

200

400

600

Treatments

DNA (mg/ 100 g

F. wt)

T1 T2 T3 T4 T5

V2 2007

148.7237.7 269

395.3515

0

200

400

600

Treatments

DNA (mg/ 100 g

F. wt)

T1 T2 T3 T4 T5

V2 2007

99.33

204.7266.7

393.7487

0

200

400

600

Treatments

DNA (mg/ 100 g

F. wt)

T1 T2 T3 T4 T5

Fig. 143-147. Effect of root extract of sunflower on DNA (mg/ 100 g F. wt) contents of


162

V2 2007

81.6790.3396.33 103107

0

50

100

150

Treatments

Chlorphyll (mg/

100 F. wt)

T1 T2 T3 T4 T5

V2 2007

82.47 91.09 96.97 102111.9

0

50

100

150

Treatments

Chlorphyll (mg/

100 F. wt)

T1 T2 T3 T4 T5

V2 2007

80.3389.71 95.45 102.6

108

0

50

100

150

Treatments

DNA (mg/ 100

g F. wt)

T1 T2 T3 T4 T5

Fig. 148-151. Effect of leaf extract of sunflower on chlorophyll (mg/ 100 g F. wt) content


V2 2007

83.43 92.198 103109.7

0

50

100

150

Treatments

Chlorphyll (mg/

100 F. wt)

T1 T2 T3 T4 T5

163

V2 2007

98103 104.7 106.3

109.7

90

100

110

120

Treatments

IAA

T1 T2 T3 T4 T5

V2 2007

96101.3 102.3

104.7111.9

80

90

100

110

120

Treatments

IAA

T1 T2 T3 T4 T5

V2 2007

89.77 95 102.7 105108

0

50

100

150

Treatments

IAA

T1 T2 T3 T4 T5




V2 2007

90.43

96.33

103.7106

107

80

90

100

110

Treatments

IAA

T1 T2 T3 T4 T5

164

V1 2006

87.34 94.34 100 98.67109.7

0

50

100

150

Treatments

Chlorophyll

(mg/100 g F. wt)

T1 T2 T3 T4 T5

V2 2006

84 93 98.67103.3

107

0

50

100

150

Treatments

Chlorophill (m

g/

100 g F. wt)

T1 T2 T3 T4 T5

V1 2007

86.66 93 99 104.3111.9

0

50

100

150

Treatments

Chlorophill (m

g/

100 g F. wt)

T1 T2 T3 T4 T5

V2 2007

83 92 97.67102.3108

0

50

100

150

Treatments

Chlorophill (m

g/

100 g F. wt)

T1 T2 T3 T4 T5

Fig. 156-159. Effect of root extract of sunflower on chlorophyll (mg/ 100 g F. wt) content


165

V1 2006

143.3131.8

122.5106.1

82.7

0

50

100

150

200

Treatments

Praline (mg/100 g

F. wt)

T1 T2 T3 T4 T5

V2 2006

132.7119.3 114.1

97.67

65.06

0

50

100

150

Treatments

Pralinel (m

g/ 100 g

F. wt)

T1 T2 T3 T4 T5

V1 2007

145133.7

124.6109.1

81

0

50

100

150

200

Treatments

Praline (mg/ 100 g F.

wt)

T1 T2 T3 T4 T5

V2 2007

133.7122.1 115.5

102.4

64.63

0

50

100

150

Treatments

Pralinel (m

g/ 100 g

F. wt)

T1 T2 T3 T4 T5

Fig. 160-163. Effect of leaf extract of sunflower on proline (mg/ 100 g F. wt) content of


166

V1 2006

118 11294.67 90.34

82.7

0

50

100

150

Treatments

Praline (mg/100 g

F. wt)

T1 T2 T3 T4 T5

V2 2006

111.3 102.788

73.3465.06

0

50

100

150

Treatments

Pralinel (m

g/ 100

g F. wt)

T1 T2 T3 T4 T5

V1 2007

119.8 114.2

96 91.33

81

0

50

100

150

Treatments


wt)

T1 T2 T3 T4 T5

V2 2007

112.3 106.789

74.3464.63

0

50

100

150

Treatments

Pralinel (m

g/ 100

g F. wt)

T1 T2 T3 T4 T5

Fig. 164-167. Effect of stem extract of sunflower on proline (mg/ 100 g F. wt) content of


167

V2 2006

119.7 112.4101.3 91.67

65.06

0

50

100

150

Treatments

Pralinel (m

g/ 100

g F. wt)

T1 T2 T3 T4 T5

V1 2007

131.3123 116.7

97

81

0

50

100

150

Treatments


wt)

T1 T2 T3 T4 T5

V2 2007

121 113.4102.3 92.67

64.63

0

50

100

150

Treatments

Pralinel (m

g/ 100

g F. wt)

T1 T2 T3 T4 T5

Fig. 168-171. Effect of root extract of sunflower on proline (mg/ 100 g F. wt) contents of


V1 2006

130121 114.7

96

82.7

0

50

100

150

Treatments

Praline (mg/100 g

F. wt)

T1 T2 T3 T4 T5

168

V2 2006

217.7200

176.7 173.7170

0

100

200

300

Treatments

Sugarl (mg/

100 g F. wt)

T1 T2 T3 T4 T5

V1 2007

226 206182.7 179.3

174

0

100

200

300

Treatments

Sugar (m

g/ 100 g F.

wt)

T1 T2 T3 T4 T5

V2 2007

218.7 201177.7174.7

169

0

100

200

300

Treatments

Sugar (m

g/ 100

g F. wt)

T1 T2 T3 T4 T5

Fig. 172-174. Effect of leaf extract of sunflower on sugar (mg/ 100 g F. wt) contents of


V1 2006

225 205181.7178.3

175

0

100

200

300

Treatments

Sugar (m

g/100 g

F. wt)

T1 T2 T3 T4 T5

169

V1 2006

213.3 199 176.7 173175

0

100

200

300

Treatments

Sugar (m

g/100 g

F. wt)

T1 T2 T3 T4 T5

V2 2006

207.7 192.3 173.3 169170

0

200

400

Treatments

Sugarl (mg/

100 g F. wt)

T1 T2 T3 T4 T5

V1 2007

214.3 200177.7 174

175

0

100

200

300

Treatments

Sugar (m

g/ 100 g F.

wt)

T1 T2 T3 T4 T5

V2 2007

208.7193.7 175 170169

0

100

200

300

Treatments

Sugar (m

g/ 100

g F. wt)

T1 T2 T3 T4 T5

Fig. 175-178. Effect of stem extract of sunflower on sugar (mg/ 100 g F. wt) contents


170

V1 2006

235 215190 182

175

0

100

200

300

Treatments

Sugar (m

g/100 g F.

wt)

T1 T2 T3 T4 T5

V2 2006

226 210184.3177.7

180

0

100

200

300

Treatments

Sugarl (mg/ 100 g

F. wt)

T1 T2 T3 T4 T5

V1 2007

222 202.7178 176

174

0

100

200

300

Treatments

Sugar (m

g/ 100 g

F. wt)

T1 T2 T3 T4 T5

V2 2007

212.3197175.3172169

0

100

200

300

Treatments

Sugar (m

g/ 100

g F. wt)

T1 T2 T3 T4 T5

Fig. 179-182. Effect of root extract of sunflower on sugar (mg/ 100 g F. wt) content of


171

V2 2006

1737

17251720

17111723

1680

1700

1720

1740

Treatments

Protein (mg/ 100

g F. wt)

T1 T2 T3 T4 T5

V1 2007

17311726

1721

1711

1690

1660

1680

1700

1720

1740

Treatments

Protein (mg/ 100

g F. wt)

T1 T2 T3 T4 T5

V2 2007

1742

17311725

1717

1732

170017101720173017401750

Treatments

Protein (mg/ 100

g F. wt)

T1 T2 T3 T4 T5

Fig. 183-187. Effect of leaf extract of sunflower on protein (mg/ 100 g F. wt) contents of


V1 2006

17251720

1715 1711

1685

1660

1680

1700

1720

1740

Treatments

Proteinr (m

g/100

g F. wt)

T1 T2 T3 T4 T5

172

V2 2006

16951677

16651665

1723

1600

1650

1700

1750

Treatments

Protein (mg/ 100

g F. wt)

T1 T2 T3 T4 T5

V1 2007

17251718

1710

1696

1690

1660

1680

1700

1720

1740

Treatments

Protein (mg/ 100

g F. wt)

T1 T2 T3 T4 T5

V2 2007

17921784

17721765

1755

1720

1740

1760

1780

1800

Treatments

Protein (mg/

100 g F. wt)

T1 T2 T3 T4 T5

Fig. 188-191. Effect of stem extract of sunflower on protein (mg/ 100 g F. wt) activity of


V1 2006

1700

16831670 1670

1685

1640

1660

1680

1700

1720

TreatmentsProteinr

(mg/100 g F. wt)

T1 T2 T3 T4 T5

173

V2 2006

178517751772

17651766

1740

1760

1780

1800

Treatments

Protein (mg/ 100

g F. wt)

T1 T2 T3 T4 T5

V1 2007

17831775

17681761

1753

1720

1740

1760

1780

1800

Treatments

Protein (mg/

100 g F. wt)

T1 T2 T3 T4 T5

V2 2007

1725

1760 1755

1692

1675

1600

1650

1700

1750

1800

Treatments

Protein (mg/

100 g F. wt)

T1 T2 T3 T4 T5

Fig. 192-195. Effect of root extract of sunflower on protein (mg/ 100 g F. wt) activity of


V1 2006

172317201712

1695

1685

1660

1680

1700

1720

1740

Treatments

Proteinr

(mg/100 g F. wt)

T1 T2 T3 T4 T5

174

V1 2006

8.33 87

4

2.67

0

2

4

6

8

10

Treatments

Superoxidase

(mg/100 g F. wt)

T1 T2 T3 T4 T5

V2 2006

7.677

6.33

3.67

2.33

0

2

4

6

8

10

Treatments

Superoxidase (mg/

100 g F. wt)

T1 T2 T3 T4 T5

V1 2007

7.33 76

33.67

0

2

4

6

8

Treatments

Superoxidase (mg/

100 g F. wt)

T1 T2 T3 T4 T5

V2 2007

6.675.67 5.33

2.673

0

2

4

6

8

Treatments

Superoxidase (mg/

100 g F. wt)

T1 T2 T3 T4 T5

Fig. 196-199. Effect of leaf extract of sunflower in superoxidase (mg/ 100 g F. wt)


175

V1 2006

65

3 3.332.67

02468

Treatments

Superoxidase

(mg/100 g F. wt)

T1 T2 T3 T4 T5

V2 2006

54

2.67 2.333

0

2

4

6

Treatments

Superoxidase

(mg/ 100 g F. wt)

T1 T2 T3 T4 T5

V1 2007

5.33 5 4.67

33

0

2

4

6

Treatments

Superoxidase (mg/

100 g F. wt)

T1 T2 T3 T4 T5

V2 2007

54.33

3.67

2.333

0

2

4

6

Treatments

Superoxidase (mg/

100 g F. wt)

T1 T2 T3 T4 T5

Fig. 200-203. Effect of root extract of sunflower on superoxidase (mg/ 100 g F. wt) of


176

V1 2006

7.33 76

32.67

0

2

4

6

8

Treatments

Superoxidase

(mg/100 g F. wt)

T1 T2 T3 T4 T5

V2 2006

6.676

5.33

2.672.33

0

2

4

6

8

Treatments

Superoxidase

(mg/ 100 g F. wt)

T1 T2 T3 T4 T5

V1 2007

6.33 65

23.67

0

2

4

6

8

Treatments

Superoxidase

(mg/ 100 g F. wt)

T1 T2 T3 T4 T5

V2 2007

5.674.67 4.33

1.672

0246

Treatments

Superoxidase

(mg/ 100 g F. wt)

T1 T2 T3 T4 T5

Fig. 204-207. Effect of stem extract of sunflower on superoxidase dismoutase (mg/ 100 g

F. wt.) of leaves in wheat varieties Margalla 99 and Chakwall 97.

177

V1 2006

15.3314.3313.3311.67

8

05101520

Treatments

Peroxidase

(mg/100 g F. wt)

T1 T2 T3 T4 T5

V2 2006

13.6712.33

10.679

7

0

5

10

15

Treatments

Peroxidase (mg/

100 g F. wt)

T1 T2 T3 T4 T5

V1 2007

14.33 13.3312.33 12

8

0

5

10

15

20

Treatments

Peroxidase (mg/

100 g F. wt)

T1 T2 T3 T4 T5

V2 2007

12.6711.33

9.678

6

0

5

10

15

Treatments

Peroxidase (mg/

100 g F. wt)

T1 T2 T3 T4 T5

Fig. 208-211. Effect of root extract of sunflower peroxidase (mg/ 100 g F. wt.) activity of


178

V1 2006

13.6712.6711 9.67

8

051015

Treatments

Peroxidase

(mg/100 g F. wt)

T1 T2 T3 T4 T5

V2 2006

1210 9.33 8.67

7

0

5

10

15

Treatments

Peroxidase (mg/

100 g F. wt)

T1 T2 T3 T4 T5

V1 2007

12.67 11.6710.33

8.67

7

0

5

10

15

Treatments

Peroxidase (mg/

100 g F. wt)

T1 T2 T3 T4 T5

V2 2007

11.6710.67

9.337.67

6

0

5

10

15

Treatments

Peroxidase (mg/

100 g F. wt)

T1 T2 T3 T4 T5

Fig. 212-215. Effect of stem extract of sunflower on peroxidase (mg/ 100 g F. wt.)


179

V2 2006

1311 10.6710.33

7

0

5

10

15

Treatments

Peroxidase (mg/

100 g F. wt)

T1 T2 T3 T4 T5

V1 2007

15.3314.6713.33

11

7.67

0

10

20

Treatments

Peroxidase (mg/

100 g F. wt)

T1 T2 T3 T4 T5

V2 2007

1411.6711.33

8

6

0

5

10

15

Treatments

Peroxidase (mg/

100 g F. wt)

T1 T2 T3 T4 T5

Fig. 216-219. Effect of root extract of sunflower on peroxidase (mg/ 100 g F. wt.) activity


V1 2006

14.6713.6712.3310.678

05101520

Treatments

Peroxidase

(mg/100 g F. wt)

T1 T2 T3 T4 T5

180

V1 2006

5

2.33

4.333.67

0

1

2

3

4

5

6

Treatments

Weed Density

T1 T2 T3 T4

V2 2006

6.67

3.33

54

0

2

4

6

8

Treatments

Weed Density

T1 T2 T3 T4

V1 2007

6

3.01

54

0

5

10

Treatments

Weed Density

T1 T2 T3 T4

V2 2007

7

4.025.15

4.48

0

2

4

6

8

Treatments

Weed Density

T1 T2 T3 T4

Fig. 220-223. Effect of sunflower leaf, stem and root extracts on weed density in wheat 30

days after sowing ( Wheat varieties Margalla 99 and Chakwall 97).

181

V1 2006

8.49

5

6.495.9

0

2

4

6

8

10

Treatments

Fresh weight

T1 T2 T3 T4

V2 2006

6.27

3.6

4.98 4.9

0

2

4

6

8

Treatments

Fresh W

eight

T1 T2 T3 T4

V1 2007

6

2.97

4.333.67

0

2

4

6

8

Treatments

Fresh W

eight

T1 T2 T3 T4

V2 2007

7.33

4.02

5.154.48

0

2

4

6

8

Treatments

Fresh W

eight

T1 T2 T3 T4

Fig. 224-227. Effect of sunflower leaf, stem and root extracts on fresh weight (g) in wheat

40 days after sowing (Wheat varieties Margalla 99 and Chakwall 97).

182

V2 2006

5.31

2.643.09 3.07

0

2

4

6

Treatments

Dry W

eight

T1 T2 T3 T4

V1 2007

2.28

1.731.99

1.82

0

1

2

3

Treatments

Dry W

eight

T1 T2 T3 T4

V2 2007

3.06

2.33

3.01 2.92

0

1

2

3

4

Treatments

Dry W

eight

T1 T2 T3 T4

Fig. 228-231. Effect of sunflower leaf, stem and root extracts on dry weight (g) of weeds

in wheat 40 days after sowing (wheat varieties Margalla 99 and Chakwall

97).

V1 2006

3.21

2.1

2.96 2.91

0

1

2

3

4

Treatments

Dry weight

T1 T2 T3 T4

183

V2 2006

10

89.05 8.64

0

5

10

15

Treatments

Fresh weight

T1 T2 T3 T4

V1 2007

7.15

4.4

5.374.94

0

2

4

6

8

Treatments

Fresh weight

T1 T2 T3 T4

V2 2007

8.1

6.05

7.376.59

0

2

4

6

8

10

Treatments

Fresh weight

T1 T2 T3 T4

Fig. 232-235. Effect of sunflower leaf, stem and root extracts on fresh weight (g) of

weeds in wheat 70 days after sowing (Wheat varieties Margalla 99 and

Chakwall 97).

V1 2006

8.2

5.626.34 5.97

0

2

4

6

8

10

Treatments

Fresh weight

T1 T2 T3 T4

184

V2 2006

6.04

3.89 4.08 3.97

0

2

4

6

8

Treatments

Dry W

eight

T1 T2 T3 T4

V1 2007

3.21

3

3.19

3.09

2.8

2.9

3

3.1

3.2

3.3

Treatments

Dry W

eight

T1 T2 T3 T4

V2 2007

4.03

3.23

3.94

3.31

0

1

2

3

4

5

Treatments

Dry W

eight

T1 T2 T3 T4

Fig. 236-239. Effect of sunflower leaf, stem and root extracts on dry weight (g) of weeds

in wheat 70 days after sowing (Wheat varieties Margalla 99 and Chakwall

97).

V1 2006

4.01

2.492.89 2.7

0

1

2

3

4

5

Treatments

Dry W

eight

T1 T2 T3 T4

185

V2 2006

12

17

1415

0

5

10

15

20

Treatments

Number of Tillers

T1 T2 T3 T4

V1 2007

15

1917 17.67

0

5

10

15

20

Treatments

Number of

Tillers

T1 T2 T3 T4

V2 2007

13.33

18

1517.67

0

5

10

15

20

Treatments

Number of Tillers

T1 T2 T3 T4

Fig. 240-243. Effect of sunflower leaf, stem and root extracts on number of tillers of


Chakwall 97).

V1 2006

14

1816

17

0

5

10

15

20

Treatments

Number of Tillers

T1 T2 T3 T4

186

V2 2006

85.44

120 112.3 119.3

0

50

100

150

Treatments

Height

T1 T2 T3 T4

V1 2007

74.97

123.3111 121

0

50

100

150

Treatments

Height

T1 T2 T3 T4

V2 2007

90.98

130117.7

127.7

0

50

100

150

Treatments

Height

T1 T2 T3 T4

Fig. 244-247. Effect of sunflower leaf, stem and root extracts on plant height (cm) of

wheat plants 145 days after sowing (wheat varieties Margalla 99 and

Chakwall 97).

V1 2006

72.67

117109 116

0

50

100

150

Treatments

Height

T1 T2 T3 T4

187

V2 2006

3.93

4.14 4.13 4.137

3.8

3.9

4

4.1

4.2

Treatments

100 Grain W

eight

T1 T2 T3 T4

V1 2007

5

5.92

5.17

5.88

4.5

5

5.5

6

Treatments

100 Grain W

eight

T1 T2 T3 T4

V2 2007

4.39

5.05

4.48

5.01

4

4.5

5

5.5

Treatments

100 Grain W

eight

T1 T2 T3 T4

Fig. 248-251. Effect of sunflower leaf, stem and root extracts on 100-grain-wieght (g) of

wheat 145 days after sowing (Wheat varieties Margalla 99 and Chakwall

97).

V1 2006

4

4.214.17 4.18

3.8

3.9

4

4.1

4.2

4.3

Treatments

100 Grain W

eight

T1 T2 T3 T4

188

V1 2006

4

4.94.45 4.71

0

2

4

6

Treatments

Fresh W

eight

T1 T2 T3 T4

V2 2006

3.96

4.8 4.6 4.7

0

2

4

6

Treatments

Fresh W

eight

T1 T2 T3 T4

V1 2007

4.49

5.08

4.65

5.01

4

4.5

5

5.5

Treatments

Fresh W

eight

T1 T2 T3 T4

V2 2007

4.27

4.99

4.57

4.95

3.5

4

4.5

5

5.5

Treatments

Fresh W

eight

T1 T2 T3 T4

Fig. 252-255. Effect of sunflower leaf, stem and root extracts on fresh weight (g) of


Chakwall 97).

189

V1 2006

0.78

0.89

0.8

0.87

0.7

0.75

0.8

0.85

0.9

Treatments

Dry W

eight

T1 T2 T3 T4

V2 2006

0.7

0.850.77

0.82

0

0.2

0.4

0.6

0.8

1

Treatments

Dry W

eight

T1 T2 T3 T4

V1 2007

0.99

1.89

1.2131.44

0

0.5

1

1.5

2

Treatments

Dry W

eight

T1 T2 T3 T4

V2 2007

0.92

1.843

1.1531.393

0

0.5

1

1.5

2

Treatments

Dry W

eight

T1 T2 T3 T4

Fig. 256-259. Effect of sunflower leaf, stem and root extracts on dry weight (g) of wheat

plants 145 days after sowing (Wheat varieties Margalla 99 and Chakwall

97).

190

V1 2006

510

500

508

504

495

500

505

510

515

Treatments

Gibberellic Acid

T1 T2 T3 T4

V2 2006

580

570

575

572

565

570

575

580

585

Treatments

Gibberellic Acid

T1 T2 T3 T4

V1 2007

529

547.3

527.7

537.7

500

520

540

560

Treatments

Gibberellic Acid

T1 T2 T3 T4

V2 2007

585

593.3

582.7

590

575

580

585

590

595

Treatments

Gibberellic Acid

T1 T2 T3 T4

Fig. 260-263. Effect of sunflower leaf, stem and root extracts on gibberellic acid (µµµµg g-1

)



191

V1 2006

265

230

242.3

221.1

180

200

220

240

260

280

Treatments

Indole Acetic Acid

T1 T2 T3 T4

V2 2006

200

290

230266.7

0

100

200

300

400

Treatments

Indole Acetic Acid

T1 T2 T3 T4

V1 2007

195

186.7

194192

180

185

190

195

200

Treatments

Indole Acetic Acid

T1 T2 T3 T4

V2 2007

190

118141.7

171.7

0

50

100

150

200

Treatments

Indole Acetic Acid

T1 T2 T3 T4

Fig. 264-267. Effect of sunflower leaf, stem and root extracts on indole acetic acid

(µµµµg g-1

) contents of wheat seedlings 30 days after sowing (Wheat

varieties Margalla 99 and Chakwall 97).

192

V1 2006

82

86

82

84

80

82

84

86

88

Treatments

Absisic Acid

T1 T2 T3 T4

V2 2006

100

104

102.3103

98

100

102

104

106

Treatments

Absisic Acid

T1 T2 T3 T4

V1 2007

101.7

106.3

100.7

104.3

96

98

100

102

104

106

108

Treatments

Absisic Acid

T1 T2 T3 T4

V2 2007

119

124.3

121

123

116

118

120

122

124

126

Treatments

Absisic Acid

T1 T2 T3 T4

Fig 268-271. Effect of sunflower leaf, stem and root extracts on abscisic acid (µµµµg g-1

)

contents of wheat seedlings 30 days after sowing (Wheat varieties Margalla


193

V2 2006

565

543548

562.7

520

540

560

580

Treatments

Gibberellic Acid

T1 T2 T3 T4

V1 2007

515

543 548562.7

450

500

550

600

Treatments

Gibberellic Acid

T1 T2 T3 T4

V2 2007

515

520

517

518.7

512

514

516

518

520

522

Treatments

Gibberellic Acid

T1 T2 T3 T4

Fig. 272-275. Effect of sunflower leaf, stem and root extracts on gibberellic acid (µµµµg g-1

)

contents of wheat seedlings root 30 days after sowing (Wheat varieties


V1 2006

505.7

494

500.7 501.7

485

490

495

500

505

510

Treatments

Gibberellic Acid

T1 T2 T3 T4

194

V1 2006

255

217.7227.7

214.3

180

200

220

240

260

TreatmentsIndole Acetic Acid

T1 T2 T3 T4

`

Fig. 276-279. Effect of sunflower leaf, stem and root extracts on indole acetic acid (µµµµg g-1

)



V2 2006

200

290

230266.7

0

100

200

300

400

Treatments

Indole Acetic

Acid

T1 T2 T3 T4

V1 2007

285

302292

297.7

260

280

300

320

Treatments

Indole Acetic Acid

T1 T2 T3 T4

V2 2007

225

263.3

221.7

255

200

220

240

260

280

Treatments

Indole Acetic Acid

T1 T2 T3 T4

195

V1 2006

62

66

62

63.33

60

62

64

66

68

Treatments

Abscisic Acid

T1 T2 T3 T4

V2 2006

54

60

51.67

58.67

45

50

55

60

65

Treatments

Abscisic Acid

T1 T2 T3 T4

V1 2007

58.67

56.67

58

55

52

54

56

58

60

Treatments

Abscisic Acid

T1 T2 T3 T4

V2 2007

76

69.67

74

70.33

66

68

70

72

74

76

78

Treatments

Abscisic Acid

T1 T2 T3 T4

Fig. 280-283. Effect of sunflower leaf, stem and root extracts on abscisic acid (µµµµg g-1

)



196

V1 2006

92

95.3394.33 94.67

90

92

94

96

TreatmentsChlorophyll

T1 T2 T3 T4

V2 2006

81.67

63

80 82.33

0

50

100

Treatments

Chlorophyll

T1 T2 T3 T4

V1 2007

100

103.3 103.3

105.7

95

100

105

110

Treatments

Chlorophyll

T1 T2 T3 T4

V2 2007

86

100

90.6797.67

70

80

90

100

110

Treatments

Chlorophyll

T1 T2 T3 T4

Fig. 284-287. Effect of sunflower leaf, stem and root extracts on chlorophyll (mg/ 100 g F.

wt) contents of wheat seedlings 30 days after sowing (Wheat varieties


197

V1 2006

182

192.3

174

185.7

160

170

180

190

200

TreatmentsSugar

T1 T2 T3 T4

V2 2006

177.3

185

174176.7

160

170

180

190

Treatments

Sugar

T1 T2 T3 T4

V1 2007

194

205.3200

202.7

180

190

200

210

Treatments

Sugar

T1 T2 T3 T4

V2 2007

180

196.3

185

192

170

180

190

200

Treatments

Sugar

T1 T2 T3 T4

Fig. 288-291. Effect of sunflower leaf, stem and root extracts on sugar (mg/ 100 g F. wt)



198

V1 2006

1732

1781

1732

1764

1700

1720

1740

1760

1780

1800

Treatments

Protein

T1 T2 T3 T4

V2 2006

1613

1663

16301651

1550

1600

1650

1700

Treatments

Protein

T1 T2 T3 T4

V1 2007

1700

1635

1692

1645

1600

1650

1700

1750

Treatments

Protein

T1 T2 T3 T4

V2 2007

1615

1635

1615

1628

1600

1610

1620

1630

1640

Treatments

Protein

T1 T2 T3 T4

Fig. 292-295. Effect of sunflower leaf, stem and root extracts on protein (mg/ 100 g F. wt)



199

V1 2006

112

126

118

124.3

100

110

120

130

Treatments

Praline

T1 T2 T3 T4

V2 2006

102

112

106109.3

90

100

110

120

Treatments

Praline

T1 T2 T3 T4

V1 2007

99.67

109

103.7

106.3

95

100

105

110

Treatments

Praline

T1 T2 T3 T4

V2 2007

105

122115.7

121

90

100

110

120

130

Treatments

Praline

T1 T2 T3 T4

Fig. 296-299. Effect of sunflower leaf, stem and root extracts on proline (mg/ 100 g F. wt)



200

V1 2006

497.3

516

504512

480

490

500

510

520

Treatments

DNA

T1 T2 T3 T4

V2 2006

490

498.3

492

497.7

485

490

495

500

Treatments

DNA

T1 T2 T3 T4

V1 2007

485501.7

463.7

490.7

400

450

500

550

Treatments

DNA

T1 T2 T3 T4

V2 2007

502.7

522.3

506.3517.3

480

500

520

540

Treatments

DNA

T1 T2 T3 T4

Fig. 300-303. Effect of sunflower leaf, stem and root extracts on DNA (mg/ 100 g F. wt)



201

V1 2006

4

119 10

0

5

10

15

Treatments

Superoxidase

T1 T2 T3 T4

V2 2006

3

8.67

5

7.67

0

5

10

Treatments

Superoxidase

T1 T2 T3 T4

V1 2007

5

11.6710 11

0

5

10

15

Treatments

Superoxidase

T1 T2 T3 T4

V2 2007

4

9

6

8.67

0

5

10

Treatments

Superoxidase

T1 T2 T3 T4

Fig. 304-307. Effect of sunflower leaf, stem and root extracts on superoxidase dismutase

(mg/ 100 g F. wt) activity of wheat seedlings 30 days after sowing (Wheat

varieties Margalla 99 and Chakwall 97).

202

V1 2006

11.67

18

1416.67

0

10

20

Treatments

Peroxidase

T1 T2 T3 T4

V2 2006

10

1512.67 14

0

10

20

Treatments

Peroxidase

T1 T2 T3 T4

V1 2007

10.67

1713

16

0

10

20

Treatments

Peroxidase

T1 T2 T3 T4

V2 2007

914 1213.33

0

10

20

Treatments

Peroxidase

T1 T2 T3 T4

Fig. 308-311. Effect of sunflower leaf, stem and root extracts on peroxidase activity (mg/

100 g F. wt) of wheat seedlings 30 days after sowing (Wheat varieties


Documents

prr.hec.gov.pkprr.hec.gov.pk/jspui/bitstream/123456789/1638/1/886S.pdf · 2018-07-23 · iv ACKNOWLEDGEMENTS All the praises and thanks are for Almighty Allah, the Compassionate,