STUDY ON MORPHO-PHYSIOLOGICAL FEATURES AND GROWTH OF

MOKARA SP. ORCHID INFLUENCED BY SPRAY FORMULATION

A Thesis By

Examination Roll No. 09Ag.CBot.JD-22 MSemester: July-December, 2010

Registration No. 35893

Session: 2009-2010

Approved as to style and contents by

(Prof. Dr. M. Obaidul Islam) (Prof. Dr. A.K.M. Azad-ud-doula Prodhan) Supervisor Co-supervisor

(Prof. Dr. Md. Shahidur Rahman)

ChairmanExamination Committee

andHead, Department of Crop Botany

Bangladesh Agricultural University, Mymensingh

November, 2010

STUDY ON MORPHO-PHYSIOLOGICAL FEATURES AND GROWTH OF

MOKARA SP. ORCHID INFLUENCED BY SPRAY FORMULATION

A Thesis By

Examination Roll No. 09Ag.CBot.JD-22 MSemester: July-December, 2010

Registration No. 35893

Session: 2009-2010

MASTER OF SCIENCE (M.S)

IN

CROP BOTANY

DEPARTMENT OF CROP BOTANY

BANGLADESH AGRICULTURAL UNIVERSITY

MYMENSINGH

NOVEMBER, 2010

ACKNOWLEDGEMENT

The author is grateful to the Omnipotent and Almighty Allah, the supreme ruler of the universe, for his immense blessing to keep us alive and successful completion of this research work.

The author likes to express profound gratitude, deepest respect and most sincere appreciation to his supervisor Prof. Dr. M. Obaidul Islam, Department of Crop Botany, Bangladesh Agricultural University (BAU), Mymensingh for his scholastic guidance, supervision, constant encouragement and suggestion and intellectual comments throughout the progress of this research work and preparing the manuscript.

The author expresses deep sense of gratitude and most sincere appreciation to his honorable co-supervisor Prof. Dr. A.K.M. Azad-ud-doula Prodhan Department of Crop Botany, Bangladesh Agricultural University (BAU), Mymensingh for his, suggestion, encouragement, constructive criticism and inspiration during the period of research work

The author expresses special thanks to Md. Iquebal Kabir, Ripon Kumar Gharmi, S. M. Kamrul Hasan, Harun–ar-Rashid, Md.Yousuf Ali, for their kind help during the period of the research work.

Thankfully acknowledgement is due to United State Department of Agriculture (USDA) for giving partly financial support in present research under the project “In vitro regeneration of orchids for commercial production and conservation of endangered species” (Project No. 2008/79/USDA).

Finally, the author expresses his sincere gratitude to beloved parents, brothers and sisters whose sacrifice and encouragement have been paved the way during study period

The Author

ABSTRACT

An experiment was conducted in grill house of the Department of Crop

Botany, Bangladesh Agriculture University (BAU), Mymensingh during the

period of 30 November, 2009 to 30 September, 2010 to investigate the effect of

spray formulation of fertilizer on morpho-physiological feature and growth of

two Mokara sp. orchids. The experiment was laid out in Completely Randomized

Block Design (RCBD) with two cultivars and four spray formulation where each

treatment was replicated three times. Data were collected on plant height, leaf

length, leaf width, leaf area, total leaf area, leaf area index, stem diameter, internal

structure of stem, number of root, internal structure of root, number of leaf,

internal structure of leaf, chlorophyll content, crop growth rate. Results revealed

that the morphological and growth parameters of Mokara Diheard Red and Mokara

Yellow Anne were significantly varied. Mokara Diheard Red was superior in

respect of plant height, leaf wide, leaf area, chlorophyll content and CGR. Mokara

Yellow Anne was superior in respect of leaf length, leaf area index, total lead area,

stem diameter and number of root. It was observed the growth of plant height, leaf

area, leaf length, leaf wide, leaf area index etc. were the highest with spray

formulation-3 (F3) and the lowest were in control (F0). Spray formulation-2 (F2)

showed maximum crop growth rate and relative growth rate over control (F0) and

the ratio of N-P-K as 10:15: 20, 15:20:25, 10:25:25 and 10:25:30 in spray

formulation might be best for growth and development of Mokara sp. orchids.

CONTENTS

CHAPTER TITLE PAGE

ACKNOWLEDGEMENTS I

ABSTRACT II

LIST OF CONTENTS III

LIST OF TABLES VI

LIST OF FIGURES VIII

LIST OF APPENDICES IX

1 INTRODUCTION 1

2 REVIEW OF LITERATURE 5

3 MATERIALS AND METHODS 12

3.1 Experimental Material 12

3.2 Experimental site 12

3.3 Experimental Treatments 12

3.4 Spray Nutrient Formulation 13

3.5 Climate 14

3.6 Water management 14

3.7 Shading 14

3.8 Collection of data 14

3.9 Anatomical investigation 18

3.10 Experimental Design 18

3.11 Statistical analysis 18

CONTENTS (CONTINUED)

CHAPTER TITLE PAGE

4 RESULTS AND DISCUSSION 19

4.1 EXTERNAL MORPHOLOGY 19

4.1.1 Stem 19

4.1.2 Roots 19

4.1.3 Leaves 20

4.1.4 Flowers 21

4.2 INTERNAL MORPHOLOGY 21

4.2.1 Roots 21

4.2.2 Stem 22

4.2.3 Leaves 23

4.3 MORPHOLOGICAL FEATURES 25

4.3.1 Plant Height 25

4.3.2 Individual Leaf Area 26

4.3.3 Leaf Length 28

4.3.4 Leaf Area Index 29

4.3.5 Leaf Width 33

4.3.6 Total Leaf Area 35

4.3.7 Leaf Number 38

4.3.8 Stem Diameter 40

4.3.9 Number of Root 43

CONTENTS (CONTINUED)

CHAPTER TITLE PAGE

4.4 Physiological and Growth Parameters 44

4.4.1 Chlorophyll Content 44

4.4.2 Crop Growth Rate 46

4.4.3 Relative Growth Rate 47

5 SUMMARY AND CONCLUSION 50

REFERENCES 52

APPENDICES 58

LIST OF TABLES

TABLE TITLE PAGE

1 Composition of spray formulation 13

2 Main effect of variety on plant height and leaf area 27

3 Main effect of spray formulation on plant height and

leaf area

27

4 Interaction effect of variety to spray formulation on

plant height and leaf area

28

5 Main effect of variety on leaf area index and leaf

length

30

6 Main effect of spray formulation on leaf area index

and leaf length

30

7 Interaction effect of variety to spray formulation on

leaf area index and leaf length

31

8 Main effect of variety on leaf width 34

9 Main effect of spray formulation on leaf width 34

10 Interaction effect of variety to spray formulation on

leaf width

35

11 Main effect of variety on leaf number and total leaf

area

37

12 Main effect of spray formulation on leaf number and

total leaf area

37

13 Interaction effect of variety to spray formulation on

leaf number and total leaf area

38

14 Interaction effect of variety to spray formulation on

leaf number and total leaf area

41

15 Main effect of spray formulation on number of root

and stem diameter

41

LIST OF TABLES (CONTINUED)

TABLE TITLE PAGE

16 Interaction effect of variety to spray formulation on

number of root and stem diameter

42

17 Main effect of variety on chlorophyll content 45

18 Main effect of spray formulation on chlorophyll

content

45

19 Interaction effect of variety to spray formulation on

Chlorophyll Content

46

20 Main effect of variety on CGR and RGR 48

21 Main effect of spray formulation on CGR and RGR 48

22 Interaction effects of variety to spray formulation on

CGR and RGR

49

LIST OF FIGURES

FIGURE TITLE PAGE

1 External morphology of Mokara Yellow Anne 20

2 External morphology of Mokara Diheard Red 20

3 Flower of Mokara sp. 21

4 Transverse section of leaf of Mokara sp 23

5 Transverse section of stem of Mokara sp. 23

6 Transverse section of root of Mokara sp. 24

7 Showing leaf length of Mokara Yellow Anne 32

8 Showing leaf length of Mokara Diheard Red 32

9 Showing leaf breadth of Mokara Yellow Anne 32

10 Showing leaf breadth of Mokara Diheard Red 32

Chapter I

INTRODUCTION

Orchids are one of the important groups of angiosperms (flowering plants)

belong to the largest and most diverse family Orchidaceae, consists of about

700-800 genera and more than 25,000 species in the world (Singh and Roy,

LIST OF APPENDICES

APPENDIX TITLE PAGE

1 Analysis of variance (Mean square) of plant

height, leaf area and leaf number at different

growth stage of orchid

58

2 Analysis of variance (Mean square) of total leaf

area, leaf area index and leaf length at different

growth stage of orchid

59

3 Analysis of variance (Mean square) of number of

root, stem diameter and leaf width at different

growth stage of orchid

60

4 Analysis of variance (Mean square) of plant

height, leaf area and leaf number at different

growth stage of orchid

61

2004). They are known for their lovely blooms and are found in diverse

habitats. Among the flowering plants, orchids are excellent items for garden

and can be grown in beds, pots, baskets, split hallows of bamboo pieces etc.

Orchids are the most fascinating, varied and beautiful of all flowers due to its

long vase life, attractive structure and excellent color (Singh and Voleti,

1995).

Orchids vary in their growth habit, foliage and flower characteristics.

Knowledge on these characteristics is basic to the understanding of the

different orchid genera and species. According to their growth habit, orchids

are divided in to sympodial and monopodial types (Royer, 2003). The

monopodial orchids have indeterminate terminal growth. This type of orchids

have aerial root and have no pseudo bulb. Whereas sympodial type orchids

have pseudo bulb and have no aerial root (Hawkey, 1978).

Orchids may be propagated either sexually or asexually. Most of the cultivated

orchids are found to be self-sterile. This problem can be overcome by high

frequency plant regeneration from somatic embryo, through tissue culture

technique. When mass propagation of a new hybrid or a variety is needed

within a short time, tissue culture is the only method (Goh et al,. 1992). It has

been estimated that it is possible to obtain more than 4 million plants in a year

from a single explant (Morel, 1964).This high frequency of propagation may

active through tissue culture techniques by using seeds (Stenberg and Kane,

1998), shoot tips (Saiprashed et al., 2002), stem nodes (Pathania et al., 1998),

leaf (Chen et al., 2000), root tips (Chen et al., 2002), lateral buds from young

flower stalks (Ichihashi, 1992) etc.

Orchids grow over a wide range of climatics condition, ranging from the

equator to arctic circle and from low land plains to snow line in mountain areas

(Hatch, 1989). Majority of the cultivated orchids were native of tropical

countries and found to grow to humid tropical forests of south and central

America (Chakrabarti, 1986). They are also distributed through Mexico, India,

Myanmar, china, Thailand, Malaysia, Philippines, New Guinea and Australia

(Rao, 1977)

Orchids having flowers of wonderful beauty and it has very good keeping

qualities. They are the most wonderful items for indoor decoration (Patil,

2001). As cut flower and pot plants, orchid is high demandable item. They are

also used in different purposes as fragrant, glue, medicine, drinks and

flavoring (Goh et al,. 1992). Orchids such as Cymbidium, Dendrobium,

Oncidium are marketed globally and the orchid industry has contributed

substantially to the economy of many Asian countries (Hew, 1994; Laws,

1995). Thailand is now the most important centre for orchid trade and exports

about over 100 million dollars in a year (Singh, 1998).

There is a scope of large scale production of orchid in Bangladesh to meet the

demand of international market and to earn foreign currency through export

(Chowdhury, 1975). Orchids are mainly found in Sylhet, Rangamati, Cox's Bazar,

and Madhupur, Tangail, Bandarban, Sundarban, Chittagong and Hilly areas of

Bangladesh.

Morpho-physiological features are the primary requisite for better understanding

about the potentialities of a variety of orchid. The basic information regarding the

morphological character and desirable physiological traits are the most important

consideration for orchid cultivation. Foliar nutrient application is very common

practice in orchid cultivation as orchid cannot take nutrient significantly from root.

Foliar spray nutrients mainly contain nitrogen (N), phosphorus (P), and potassium

(K) with different concentration. Application of spray nutrient containing N-P-K

with different concentration varied on the basis of growth stage of plants. Urea,

TSP and MP are used as a source of nitrogen (N), phosphorus (P), and potassium

(K) in spray nutrients. Spray nutrients solution is prepared by mixing of Urea, TSP

and MP that plays an important role in growth and development of orchid.

In few years ago, orchids were totally unknown to our country but now it is

becoming familiar day by day and their production is increased in recent years.

To be a successful orchid producer it is pre-requisite to know the morpho-

physiological feature and behavior of orchid. Literature revealed that there

are no research report in Bangladesh on the effect of spray nutrients on

growth and development of orchid.

By considering above facts and background the present study was

undertaken with following objectives.

1. to understand the morpho-physiological features of orchid (Mokara sp.)

under field condition.

2. to determine the effect of spray formulation on morpho-physiological feature

and growth of orchid (Mokara sp.)

3. to select the suitable concentration of N, P and K in spray formulation for

proper growth of orchid (Mokara sp.)

Chapter II

REVIEW OF LITERATURE

Orchids are herbaceous plants posses distinct and special physiological features to

adapt themselves in their habitat. Origin of orchids are mountain, hill and deserts

where lack of soil, water are prevailed and plants adopted in these habitat have

some characters to over come the adverse environmental barrier. Information

regarding morpho-physiological features are available in previous literatures

related to the present experiment are summarised in this chapter as follows.

Razzak (2009) conducted an experiment to study the seasonal effect of

micronutrients on the growth and development of orchid (Dendrobium sp). He

applied 0.2% spray solution of N-P-K on Dendrobium sp. in summer and winter.

Spray nutrient was applied once in a week. After seven months it was found that

the rate of growth of orchids affected with nutrient solution application and it was

better in summer than winter due to temperature variation. It was concluded that

spray nutrient has significant effect concerned to seasonal effect on growth

performance of orchids.

Kabir (2007) investigated the effect of different liquid fertilizers for growth of

orchid. He formulated three liquid fertilizers namely Miracle Grow, Growing

orchid food and Flowering food made of Nitrogen, Phosphorus and Potassium at

different ratio. These three liquid fertilizers applied on orchid’s upto blooming and

it was found that larger flowers are bloomed in those plants which were treated

with flowering food as a source of nutrient.

A pot experiment was conducted by Mohapatra and Saravanan (2006) to determine

the suitable growing medium for orchid (Mokara sp.). Different medium namely

coconut coir, bhusa, rice husk, tile bits, charcoal, cow dung, brick pieces and

groundnut shell single or in combination were used as treatment. They found that

maximum plant height and the highest number of new shoots per plant were on the

medium as mixture of coconut coir + cow dung + bhusa and Gravel + groundnut

shell + cow dung respectively. He also reported the flowering parameters such as

early flower bud emergence, number of spikes per plant, spike length and number

of florets per spike, were also the greatest with gravel + groundnut shell + cow

dung.

Ketsa and Kosonmethakul (2006) studied the effect of aluminum sulfate

[A12(SO4)3] at 50, 100 and 150 mgL-1 and cobalt chloride (CoC12) at 100, 200 and

300 mgL-1 in the holding solution containing 225 mgL-1 8 mgL-1 hydroxyquinoline

sulfate (HQS) and 4% glucose on vase life of Dendrobium 'Sonia Bom Joe' in

comparison with the conventional holding solution containing 225 mgL-1 HQS, 30

mgL-1 AgNO3 and 4% glucose was studied at ambient conditions (30.2°C and 62%

RH) increased vase life and bud opening of orchid flowers effectively. He reported

that the holding solution containing 225 mgL-1 HQS, 50 mgL-1, A12(SO4)3 and 4%

glucose significantly increased vase life and bud opening of Dendrobium 'Sonia

Bom Joe'

Ketsa and Kosonmethakul (2004) observed that application of spray nutrients

named Physan at the concentration of 100, 200, 300, 400, and 500 ppm weekly

interval for 6 months enhanced bud opening and vase life of cut flowers of orchid.

Adding 1-7% sucrose to Physan considerably improved water uptake, bud opening

and vase life of cut flowers of orchid.

Vaz and Kerbauy (2004) studied the effect of nitrogen, phosphorus and potassium

on floral spike induction of Mokara sp. They observed a positive correlation

between the total mineral salt concentration and floral spike induction. He

concluded that increasing phosphorus with reduction of nitrogen in spray solution

have significant effect on floral stimulation.

Ghoshal et al. (1989) investigated the effect of liquid nitrogen on floral

morphology and behavior of some orchids. He used liquid nitrogen with different

concentration and found that those plants are treated with high concentration of

liquid nitrogen produced attractive bloom. He also reported that high concentration

of liquid nitrogen affect on pollen grain sterility.

Wang and Konow (2002) investigated the effect of medium composition on

vegetative growth of orchid. They used bark-peat; saw dust and coconut shell as

growing medium of orchid. He reported that plant height and number of leaves was

maximum on those plants which were grown on saw dust among those medium.

Wang and Konow (2004) determined the interaction effect of water-soluble

fertilizer and growing medium of Dendrobium sp. They used bark peat and

charcoal as growing medium with same concentration of water soluble fertilizer.

After six months they found that more leaves greater fresh weights (FW) and larger

total leaf areas of Dendrobium sp. on bark peat.

Poole and Sheehan (1970) indicated the effect of a low N, high P and K on flower

size, flower longevity of orchids. He applied various concentration of N, P and K

fertilizer on Vanda sp. upto flowering and found that all plants produces more or

less similar size of flowers. It was concluded that Flower size was unaffected by

any treatment in this experiment.

Fleischer (1935) studied on the bud opening and vase life of opened buds. He

found flower buds near the proximal end of the inflorescence were larger and

heavier than those the distal end. He used different chemicals separately or in

combination of them and reported that the optimum holding solution was 200

mgL-1 8-hydroxyquinoline sulphate (HQS) + 50mgL-1 AgNO3 + 8% sucrose

solution as this solution increased the percentage of bud opening and prolonged

vase life .

Furukawa et al. (1989) showed that pretreatments of orchid flower with BA

significantly suppressed ethylene production and prolonged the vase life of cut

florets in Mokara sp. and Dendrobium sp. He found that Combination of BA and

spray of silver thiosulfate (STS) had significant effect on the vase life of Cattleya

sp. It was concluded that ethylene seems to be involved in senescence of the cut

florets but BA suppresses its production which leads to extend vase life.

Wong and Lee (2000) evaluated the effect of potting mixes on the growth of

Dendrobium sp. and the optimum fertilizer requirement for leaf growth. The

different potting media were Cornell Mix (peat moss: vermiculite), Modified Mix

(cocopeat: vermiculite), CVS (cocopeat: vermiculite: sand) and cocopeat which

were treated with 100 ml of foliar fertilizer (N, P and K) weekly at different

concentrations. They reported the average leaf length was higher in Cornell Mix

than other medium.

Webb and Webb (1993) studied on the maturation of pseudobulb of different

orchids as influenced by application of phosphorus (P). They applied different

concentration of phosphatic fertilizer on orchid at weekly interval. After eight

months they found that high concentration of P leads to attain early maturity of

pseudobulb of some orchids.

Matsui et al. (1999) investigated the effect of macronutrients on growth of

Dendrobium sp. to identify the optimum concentration for growth of orchid. The

growth was promoted at 7.5 mM to 15 mM of N per litre with spray solution

weekly whereas growth was suppressed at 0 mM and 30 mM of N per litre spray

solution. There was no effect of phosphorous (P) on growth and flowering between

0.5 mM to 2 mM but growth was superior at 10 mM to 20 mM of K per litre spray

solution. Therefore it was concluded that N-P-K (7.5- 0.5-20 mM L -1) was suitable

for growth of Dendrobium sp.

In an investigation Growth, morphology and anatomy of pseudobulb and

inflorescence of Oncidium 'Gower Ramsey', studied by Sell and Hermann

(1998) in the glasshouse conditions to determine the effect of foliar

nutrients on growth and morphological behavior. They observed that

mature plants could complete two growth cycles per year. Anatomical

observations revealed that a newly elongated vegetative bud contained 13-

14 nodes in which one node was enlarged to form the pseudobulbs. The

pseudobulb contains 5 latent buds with a flower stalk developed from the

first nodes (numbered basipetally from the base). They concluded foliar

nutrients directly affects on growth.Pseudobulb formation occurred

during the unsheathing stage and flower stalks were differentiated when

pseudobulbs were elongating. The length of the flower stalk was 4.3 cm

when the pseudobulb was fully developed. This young inflorescence

contained 23 nodes with floral primardia initiated on the node number of 17 and

above. Differentiation and development of inflorescence branches

occurred rapidly during the period of flower stalk elongation while

differentiation of new floral buds on the main inflorescence apex was slow.

Results suggested that the unsheathing stage was the critical stage for

determining flowering.

Yin-Tung Wang (1995) conducted an experiment to determined the effect of

water-soluble fertilizers on leaf characteristics and applied six water-soluble

fertilizers containing N-P-K at different ratio (10-30-20, 15-10-30, 15-20-25, 20-5-

19, 20-10-20, and 20-20-20 in N2-P2O5-K2O respectively) per litre to young

seedlings of Phalaenopsis sp. After seven months, he found, leaf size, total leaf

area, and shoot and root fresh weight were highly variable among the plants and

concluded that the different N-P-K concentration of nutrient solution had a

significant effect on leaf number and leaf area.

Hlgakl and Imamura (1987) studied on flower production and plant height. He

showed that the flower production and plant height increased linearly with

increasing of P and N concentration respectively in foliar spray nutrients

application.

Komori (2002) conducted an experiment during 15 May 1997 to 31 August

1998 to defermine the effect of low light intensity on the growth of Cymbidium

orchid cultivars Rose Wine 'Fruity Dror' (RF) and Great Katly 'Little

Louransan' (GL). RF and GL plants grown under shaded conditions exhibited

lower fresh weight especially root fresh weight than the control treatment. The

number of leaves and leaf length were also smaller in RF and Gl than the

control. RF and Gl exhibited low development of new leaf than the control.

He found that light intensity affect on leaf characteristics significantly.

Patil (2001) conducted an experiment in a greenhouse for the

characterization of flower and observed that six species (Dendi-obium

densiflorum, Asocentrum ampullaceum, D. nobile, Phaius tankervilleae, D.

pierardii and Ascocentrum ainpullucettin var. Auranticum) and five species

(Spcjlhoglolfis speciosa, Aerides mulfiflorum , D. crepidatum, D. Jenkinsii and

D. primulinuln) flowered during October-December and January-

April respectively. He reported the largest flowers and the longest

spikes (60 cm) were observed in Phaius tankervilleae on April and the

smallest flowers were recorded in D. crepidatum and concluded that

flowering of orchid is affected by growing season.

Lee and Chang (2000) studied the growth of pseudobulb and quality of

flower and inflorescence on Oncidium spot determine the effect of

temperature on pseudobulb formation and flowering longevity. They

observed that high day/night temperature made the pseudobulb thin and long

but lower temperature (13-18°C) made it short and flat. In lower

temperature growth of the pseudobulb was slow and flowering was delayed

but the longevity and floret number of the inflorescence was increased.

Flower stalks emerged in all temperature treatments except at the extreme

temperature. Lower day night temperature favoured inflorescence

development and increased branch and floret numbers.

In an experiment Prasad et al. (1997) studied on the morphological

feature of orchid. They observed that spike length and plant height

had the highest heritability and some leaf characteristics are

most diversied which was also affected by geographical position.

Sobhana and Rajeevan (1993) conducted an experiment on the

performance of' certain epiphytic species in central Kerala as influenced

by spray nutrient.They reported flower characteristics such as

Inflorescence length and the number, size, colour, fragrance and flowering

period are influenced by frequent use of spray nutrients.

Chapter III

MATERIALS AND METHOD

3.1 Experimental material

Two cultivars of Mokara sp orchid were collected from Dipto orchid

nursery Valuka, Mymensingh in the month of November, 2009. The age of

clone was about six months. These clones were separated from mother

plant and cultured for six months with coconut shell, sawdust and

carpenter dust as supporting material.

3.2 Experimental site

The experiment was conducted in grill house of Crop Botany Department,

Bangladesh Agriculture University, Mymensingh during the period of 30

November, 2009 to 30 September, 2010 to determine the effect of spray

nutrients on growth and morph-physiological features of orchid.

3.3 Experimental treatments

There are two factors used in this experiment known as variety and

formulation of spray nutrients. In the present experiment, two cultivars of

Mokara sp were used. As treatment three spray formulations were used

along with a control where only tap water was used.

Factor A: Two orchid species

a. Mokara Diehard Red

b. Mokara Yellow Anne

Factor B: Three levels of spray nutrient with control

a. Spray Formulation 0 (F0)

b. Spray Formulation 1 (F1)

c. Spray Formulation 2 (F2)

d. Spray Formulation 3 (F3)

Thus, the total number of treatments was 8 (4x2) and each treatment was

replicated three times. The whole experimental beds were divided into three

blocks. The size of each block was 2.64 m2.

3.4 Spray nutrient formulation

The spray solutions were prepared by mixing of Urea, Tripple Super

Phosphate (TSP) and Muriate of Potash (MoP). Formulation was prepared by

following the thumb rule of BARI where total amount of fertilizer to be 700 g

per 172 litre water was recommended for use as spray. The formulation used

in present research contained different ratio of N, P and K. The ratio of N, P

and K varied over growth stage of orchid shown in Table 1. Spray

formulation used in early stages contained higher N and gradually reduced in

later stages.

Table 1. Composition of spray formulation.

Name of formulation

Total urea, TSP and MP 70 gm per 17.2 liter water at N : P : K ratio

F0 (Control) 0.0 0.0 0.0 0.0

F1 15:20:25 20:20:25 20:25:30 10:30:25

F2 10:15:20 15:20:25 10:25: 25 10:25:30

F3 5:15:10 10:15:10 10:20:15 15:20:20

Duration of spray

December 15 to January 30

January 31 to

March 15

March 16 to

April 30

May 01 to

June 15

% of N, P and K in Urea, TSP and MP is 46.0, 48.0 and 60.0 respectively. In

each times fresh formulation was prepared and sprayed once weekly with a

hand sprayer at afternoon.

3.5 Climate

The experimental area was located in sub-tropical climatic zone

characterize by moderate rainfall and temperature during research

period.

3.6 Water management

Frequent application of water is essential in orchid cultivation. In November -

March watering was done once per day and in April - July, watering was done

2 times per day. Watering was done by water cane.

3.7 Shading

As an epiphyte, most orchids avoid direct sunlight under natural condition.

Orchids prefer dappled shade. For maintaining shade a hand made shade

was provided by bamboo.

3.8 Collection of data

Data were collected periodically during the growing period of orchid. The data

were recorded on the following morphological parameters.

a. Plant height (cm)

b. Leaf length (cm)

c. Leaf wide (cm)

d. Leaf area (cm2)

e. Total Leaf area (cm2)

f. Leaf area Index (LAI)

g. Stem diameter (cm)

h. Number of root

i. Number of leaf

j. Chlorophyll content in leaf

k. Fresh weight

l. Dry weight

From collected data Crop Growth Rate (CGR) and Relative Growth Rate

(RGR) were calculated.

3.8.1 Plant height

The height of plant was measured in cm from ground level to top of the main

stem by measuring scale at the interval of 45 days during the study period.

3.8.2 Leaf length

The leaf length was measured in cm from leaf base to tip of the leaf by

measuring scale at the interval of 45 days during the study period.

3.8.3 Leaf width

The width of leaf was measured by scale along the middle portion of leaf

transversely and its average was calculated and expressed in cm at the

interval of 45 days during the study period.

3.8.4 Individual leaf area /plant

The Individual leaf area was determined by multiplying leaf length with

leaf width and expressed in cm2.

3.8.5 Total leaf area /plant

The total leaf area was determined by summation of all leaf area and

expressed in cm2.

3.8.6 Leaf Area Index

Leaf Area Index is the ratio of leaf area to ground area. Leaf Area

Index was determined by dividing Individual leaf area to Individual

ground area.

Leaf area Leaf Area Index = Ground area

3.8.7 Stem diameter

Stem diameter was measured by using slide calipers at the middle

portion of stem.

3.8.8 Number of root

Number of Root was estimated by counting all roots in a plant.

3.8.9 Crop Growth Rate

Crop Growth Rate (CGR) defined as the increase of plant material per unit of

time. It is expressed as g of dry matter produced per day. It can be expressed

as following equation.

W 2-W1 Crop Growth Rate = T2-T1

W1 and W2 are dry weights of plants at times T1 and T2 respectively.

3.8.10 Relative Growth Rate

Relative Growth Rate (RGR) defined as the increase of plant material per

unit of time per unit area. This parameter indicates a rate of growth per unit

of dry matter. So, RGR is the unit of plant material per unit of material

present per unit of time. It is expressed as g of dry matter produced by a g of

existing dry matter in a day. RGR is the efficiency of dry matter production.

It can be expressed as following equation.

Log W2-Log W1

Relative Growth Rate =

T2-T1

Where, W1 and W2 are dry weights of plants at times T1 and T2 respectively.

3.8.11 Chlorophyll estimation

Chlorophyll is the most important photosynthetic apparatus in plant.

Chlorophyll content in leaf was estimated at 60 days after transplanting

(DAT) and 90 DAT by following procedure of Arnon (1949). Chlorophyll

content in leaf was expressed as mg/g of a sample by using following equation.

Chl(a)= 0.0127D663-0.00269D645

Chl(b)= 0.0229 D645-0.00468 D663

Where, D645 =Absorbance at 645 nm wave length

D663= Absorbance at 663 nm wave length

Total chlorophyll= Chl. a + Chl. b

3.8.11.1 Steps for chlorophyll estimation

At first 0.5 mg fresh leaf was taken as sample. Each sample was crushed

by mortar and pestle in 10 ml acetone (80%). Then the extract was

centrifuged for 10 minutes at 1000 rpm. Extract of each test tube was

made a final volume of 10 ml by addition of acetone to fulfill

volatilization loss. The reading was recorded at 663 nm and 654 nm in

UV- VIS spectrophotometer.

3.9 Anatomical investigation

To investigate the general anatomical structure of root, stem and leaf thin

(about 10 µM) section were prepared from fresh plant material by sharp

blade. The thin sections were stained in 1.0% safranin and mounted on

slide. A cover sleep was set on thin section and observed under microscope

with high resolution, progressive scan digital camera system (Model

Olympus BX 41 and camera DP 20) and photos were taken.

3.10 Experimental design

The experiment was laid out in Completely Randomized Block Design

(RCBD) with 8 treatments and each treatment was replicated three times.

3.11 Statistical analysis

The collected data analyzed by following the analysis of variance (ANOVA)

technique and mean differences were adjudged by Duncan’s Multiple Range

Test (DMRT) (Gomez and Gomez, 1984) using a computer operated

programme named MSTAT-C (Russel, 1986).

Chapter IV

RESULTS AND DISCUSSION

The results obtained from the experiment are described and discussed in this

chapter. The morphological and physiological parameters of orchid have been

presented and discussed under separate heads and sub-heads with tables and

figures.

4.1 External morphological features of Orchid

Orchids easily identified by its leaves, stems and roots, flowers, fruits and

seeds. They are herbaceous plants having distinct characteristics. The present

cultivars namely Mokara Diheard red and Mokara Yellow Anne grew as

epiphyte on sawdust, coconut coir and carpenter dust. The general

morphological features of the species under study were as follows.

4.1.1 Stem

The stem of Mokara sp. was erected, cylindrical having nodes and internodes.

Basal parts of the stem were enclosed by leaf sheath. The lower most part of

the stem contained 3 types of roots. The color of stem was light green to dark

green.

4.1.2 Roots

Roots of Mokara sp. under study were thin and cylindrical. There were three

types root found in Mokara sp. namely clinging root, absorbing root and

aerial root. The color of root was brown to purple. Tip of the root is blunt and

smooth in aerial root. Orchid roots arouse from the base of pseudobulb, node.

Fig.1. External morphology of Mokara Yellow Anne

Fig. 2. External morphology of Mokara Diheard Red

4.1.3 Leaves

Leaves were simple, alternate and very thick. Leaves of Mokara sp. orchid

have parallel venation. The shapes of orchid leaves were palmate, ovate,

obovate.

Fig. 3. Flower of Mokara sp.

4.1.4 Flower

Orchid flowers were zygomorphic. The inflorescence of Mokara sp. is

racemos. Their segments were arranged in several whorls. The outermost

whorl consists of three sepals which together make up the calyx. The second

whorl of Mokara sp consists of three petals and it is known as corolla. Calyx

and corolla together produced perianth. The anthers of orchids are finger

shaped which partially fused with the filaments of stamens and individual

pollen grains are monads.

4.2 Anatomical features

4.2.1 Root

The orchid under study had stout roots that were circular in transverse

section. The epidermis functions as protective layer and consists of

rectangular cells with lignified wall. The cortex remains inside of epidermis

which functions as storage and transportation (Fig. 6). An endodermis bounds

the cortex on the inside and encloses the stele. The endodermis cell had

distinct casperian strips and was rectangular to oblong in shape. The vascular

bundles were aerial in which protoxylem distributed throughout the

periphery and metaxylem throughout the centre. The velamen tissue consists

of 4-6 layers of polygonal radially elongated cells which transport water and

nutrient from soil to leaf.

4.2.2 Stem

The stem under microscope was showed different components of tissue.

Epidermis of Mokara sp orchid was observed as single layered with cuticle. A

few stomata were present in the epidermis. Inside the epidermis 7 layers of

round, oval, elliptical or irregular parenchyma cells were found known as

cortex (Fig. 5). Interior to the epidermis one layer of irregularly shaped

parenchymatous cell was observed and called hypodermis. Ground tissue was

mostly isodiametric cells. Vascular bundles are distributed throughout the

ground tissue. There was no cambium in vascular bundle. The metaxylem

present in peripheral region and protoxylem present in centre of stem under

microscope.

4.2.3 Leaf

The leaf of orchid under study was thick and fleshy. A transverse section of

orchid leaf was observed under microscope and different component of tissue

was found. In Mokara sp. a thick cuticle covers the epidermis which consists

of small heavy walled cells. A multilayered palisade parenchyma was seen

bellow the epidermis. The epidermis was two types known as upper epidermis

and lower epidermis. The palisade layer is located bellow the hypodermis and

was two or three layers of thick elongated cell. The spongy mesophyll bellow

the palisade parenchyma was round. Both the palisade layer and spongy

mesophyll are photosynthetic tissue. Vascular bundles were collateral and

closed (Fig. 4).

Epidermis

Fig. 4. Transverse section of leaf of Mokara sp.

Cuticle

Spongy parenchyma

Vascular bundle

Palisade parenchyma

Corte

Vascular Bundle

Epidermis

Cuticle

Fig. 5. Transverse section of stem of Mokara sp.

Fig. 6. Transverse section of root of Mokara sp.

Cuticle

Xylem vessels

Epidermis (Radially

Cortrex

Endodermis

Velamen

Epidermis

4.3 Morphological features

4.3.1 Plant height

Plant height varied significantly between the two cultivars of orchids. At 45

DAP, plant height of Mokara Diheard Red and Mokara Yellow Anne were

24.88 cm and 19.75 cm respectively. At 90 DAP, it was 27.23 cm and 22.28

cm, respectively. Plant height of Mokara Diheard Red and Mokara Yellow

Anne were 29.68 cm and 24.95 cm , respectively at 135 DAP. Maximum plant

height of Mokara Diheard red and Mokara Yellow Anne were 32.19 cm and

27.44 cm, respectively at 180 DAP. It was observed that plant height increased

progressively with the advancement of age and growth stages. Plant height

increased rapidly in Mokara Diheard Red than Mokara Yellow Anne

comparatively (Table 2).

Different nutrients as spray have significant effects on plant height. At 45

DAP, the highest plant height was 23.15 cm recorded in F1 and lowest was

21.63 cm in F3. At 90 DAP and 135 DAP, maximum plant height 25.91 cm and

28.8 cm respectively were recorded in F1 and minimum plant height was 24.2

cm and 25.51 cm in control (F0). At 180 DAP, maximum plant height was 31.9

cm in F1 and minimum was 26.88 cm in F0 (Table 3). The plant height

increased slowly in control (F0) and comparatively rapid in F1 indicated that

different formulation was effective in plant height.

The interaction effect of variety to formulation revealed that plant height

affected significantly. Maximum plant height was 26.8 cm in Mokara Diheard

Red at formulation-1 (V1F1) and minimum was 19.03 cm in Mokara Yellow

Anne with formulation-2 (V2F2) at 45 DAP. Similar trend in plant height was

observed with increasing cultivation period. At 180 DAP, maximum plant

height was 32 cm that was found in V1F1 and minimum was 22.93 cm recorded

in V2F0 (Table 4). The present study support the report of Wang and Konow

(2004)

4.3.2 Individual leaf area

Varietal effect of leaf area was significant. The leaf area ranged from 50.5-

77.1 cm2 and 44.2 - 73.2 cm2 in Mokara Diheard red and Mokara Yellow

Anne, respectively over growing period. The Mokara Diheard red (V1) showed

maximum leaf area of 60.03 cm2 over leaf area 53.18 cm2 of Mokara Yellow

Anne (V2) at 45 DAPS. At 90 DAP, the leaf area of V1 was 60.03 cm2 and of V2

Was 53.18. At 135 DAP, the leaf area of V1 and V2 was 68.82 cm2 and 63.58

cm2, respectively. At 180 DAP, the leaf area of V1 was 77.14 cm2 and of V2 Was

73.20 cm2 (Table 2). From table it was observed that leaf area increment was

higher in V2 than V1.

Spray formulation was effective in enhancing the leaf area. At 45 DAP,

maximum leaf area was 51.81 cm2 with F2 and minimum was 46.89 cm2 was

with F1. At 90 DAP, maximum leaf area was 59.52 cm2 in F2 and minimum leaf

area of 52.67 cm2 in control (F0). Leaf area gradually increased with

increasing age of the plant. At 135 DAP, maximum leaf area was 70.16 cm2 in

F3 and minimum leaf area was 58.93 cm2 in control (F0). At 180 DAP,

maximum leaf area 80.60 cm2 was found in F3 and minimum 66.85 cm2 in F0

(Table 3). The trend of leaf area increment over vegetative growth was rapid

in F3.

Leaf area varied significantly due to the interaction effect between variety and

treatments. At 45 DAP, the highest leaf area was 51.29 cm2 recorded in V1F1

and the lowest was 41.94 cm2 in V2F3. At 90 DAP, the highest leaf area was

62.77 cm2 and the lowest was 48.54 cm2 recorded in V1F3 and V2F0,

respectively. At 135 DAP, the highest leaf area was 74.03 cm2 and the lowest

was 54.98 cm2 that recorded in V1F3 and V2F0, respectively. V1F3 and V2F0 also

shows the highest 82.84 cm2 and the lowest 63.94 cm2, respectively at 180 DAP

(Table 4). The variation in leaf area might occur due to athe vriation in

number of leaves and their expansion. The result obtained from the present

study is consistent with the results of Sharma and Haloi (2001) who stated at

variation in leaf area might be attributed to the difference in number of

leaves.

Table 2. Main effect of variety on plant height and leaf area.

Variety Plant Height (cm) at DAP Leaf area (cm2) at DAP

45 90 135 180 45 90 135 180

V1 24.88 27.23 29.68 32.19 50.54 60.03 68.828 77.1

V2 19.75 22.28 24.95 27.44 44.25 53.18 63.58 73.20

CV 16.91 14.63 12.88 12.09 10.89 11.06 10.59 10.12

Table 3. Main effect of spray formulation on plant height and leaf area.

Spray

formulation

Plant height (cm) at DAP Leaf area (cm2) DAP

45 90 135 180 45 90 135 180

F0 22.76 b 24.20 25.51 b 26.88 b 46.69 52.67 58.93 b 66.85 b

F1 24.15 a 25.91 28.8 a 31.90 a 46.89 56.94 66.81 a 77.17 a

F2 21.71 b 24.45 27.71 a 30.58 a 51.81 59.52 68.90 a 76.08 a

F3 21.63 b 24.46 27.25 a 29.9 a 47.79 57.27 70.16 a 80.60 a

CV 16.91 14.63 12.88 12.09 10.89 11.06 10.59 10.12

Legend

V1 = Mokara Diheard Red F0 = Control

V2 = Mokara Yellow Anne F1 = Spray Formulation-1

DAP = Days After Planting F2 = Spray Formulation-2

F3 = Spray Formulation-3

Variety x

spray

formulatio

n

Plant height (cm) at DAP Leaf area (cm2) at DAP

45 90 135 180 45 90 135 180

V1F0 25.46 a 26.96 a 28.10 a 29.56 b 49.87ab 56.81ab62.87

ab

69.76

ab

V1F1 26.80 a 29.53 a 32.00 a 35.1 a 51.29 a 62.41 a 71.18 a 81.97 a

V1F2 24.40 a 26.93 a 30.o a 32.86 a 47.76 ab 58.12ab67.22

ab74.01 ab

V1F3 22.86 ab 25.50 a 28.63ab 31.23 a 53.26 ab 62.77 a 74.03 a 82.84 a

V2F0 20.06 b 21.43 b 22.93 b 24.20 b 44.07 ab 48.54 b 54.98 b 63.94 b

V2F1 19.50 ab 22.3 b 25.6 ab 28.70 b 42.89 b 51.48ab62.45

ab72.37 ab

V2F2 19.03 b 21.96 b 25.43ab 28.30 b 48.11 ab 60.93 a 70.58 a 78.15 ab

V2F3 20.40 b 23.43 ab 25.86ab 28.56 b 41.94 b 51.78ab66.29

ab78.36 ab

CV 16.91 14.63 12.88 12.09 10.89 11.06 10.59 10.12

Table 4. Interaction effect of variety to spray formulation on plant height and

leaf area.

4.3.3 Leaf length

Average leaf length recorded at 45, 90 , 135 and 180 DAP was varied

significantly between Mokara Diheard Red and Mokara Yellow Anne .The

average leaf length of V1 was 22.84 cm and V2 was 22.65 cm at 45 DAP. At 90

DAP, the average leaf length of V1 was 23.63 cm and V2 was 23.15 cm. The

average leaf length was 24.07 cm and 23.6 cm at 135 DAP in V1 and V2

respectively. At 180 DAP, the average leaf length of V1 was 24.75 cm and V2

was 24.34 cm (Table 5). From table it was observed that in all day’s leaf length

was more in V1 than V2. So, Mokara Diheard Red (V1) was superior to

Mokara Yellow Anne (V2) in respect of leaf length.

At 45 DAP, the highest leaf length was 23.23 cm recorded in F2 and the lowest

of 22.44 cm in F3. Similar trend in leaf length was observed with increasing

cultivation period. At 180 DAP, maximum leaf length was 25.13 cm that

recorded in F2 and minimum 24.03 cm was in F3 (Table 6). Leaf length

increased at slow rate with advancement of time and growth stage. The N, P

and K concentration gradually changed with culture period and found less

little effective on leaf length. The lowest leaf length was observed in F3 which

contained the lowest concentration of N, P and K. Results revealed that

increasing the concentration of N, P and K in spray formulation leads to

increase leaf length.

The interaction effect of variety and treatments on leaf length was significant.

At 45 DAP, the highest leaf length was 23.9 cm recorded in V1F0 and the

lowest 21.78 cm was in V1F3. At 90 DAP, the highest leaf length was 24.78 cm

in V1F0 and the lowest 22.2 cm was in V1F3. At 135 DAP, the highest leaf

length 25.23 cm was recorded in V1F0 and the lowest 22.66 cm was in V1F3. At

180 DAP, the highest leaf length 26.03 cm was recorded in V1F0 and the lowest

23.2 cm was in V1F3 (Table 7).

4.3.4 Leaf area index

Leaf Area Index (LAI) recorded at 45 DAP, 90 DAP, 135 DAP and 180 DAP

was significantly varied between two cultivars of orchids. In V1, the average

leaf area index was 0.47, 0.66, 0.94 and 1.21 at 45, 90,135 and 180 DAP

respectively. The average leaf area index of V2 was 0.50, 0.74, 1.05 and 1.39 at

45, 90,135 and 180 DAP respectively (Table 5). In comparison between two

varieties degree of increasing of leaf area index was more in V2 than V1. Thus

varietal effect on leaf area index was significant and V2 may be considered as

superior over V1 in this respect.

At 45 DAP, the highest leaf area index was 0.50 recorded in F2 and the lowest

was 0.45 found in F3. At 90 DAP, the highest leaf area index was 0.76 recorded

in F2 and the lowest was 0.63 in F0. At 135 DAP, the highest and lowest leaf

area index were 1.05 and 0.89 observed in F2 and F0, respectively. At 180 DAP,

the highest leaf area index was 1.37 and the lowest was 1.17 found in F3 and

F0, respectively (Table 6). It was observed that degree of increasing of leaf

area index was more in F3. The variation in leaf area index might occur due to

the variation in number of leaves and their expansion. The results obtained

from the present study was consistent with the result of Sharma and Haloi

(2001) who stated that variation in leaf area index could be attributed to the

change in leaf number. The results obtained from the present study also

supported by Chandra and Das (2000).

Table 5. Main effect of variety on leaf area index and leaf length.

Variety Leaf area index at DAP Leaf length (cm) at DAP

45 90 135 180 45 90 135 180

V1 0.47 0.66 0.94 1.21 22.84 23.63 24.07 24.75

V2 0.5 0.74 1.05 1.39 22.65 23.15 23.60 24.34

CV 11.62 14.25 12.61 12.77 7.65 7.52 7.49 7.13

Table 6. Main effect of spray formulation on leaf area index and leaf length

Spray

formulation

Leaf area index at DAP Leaf length (cm) at DAP

45 90 135 180 45 90 135 180

F0 0.46 a 0.63 b 0.89 b 1.17 b 23.00 a 23.60 a 24.0 a 24.65 a

F1 0.49 a 0.71 a 1.02 a 1.34 a 22.33 a 23.24 a 23.66 a 24.36 a

F2 0.5 a 0.76 a 1.05 a 1.33 a 23.23 a 23.83 a 24.40 a 25.13 a

F3 0.45 a 0.68 a 1.02 a 1.37 a 22.44 a 22.88 a 23.3 a 24.03 a

CV 11.62 14.25 12.61 12.77 7.65 7.52 7.49 7.13

Legend

V1 = Mokara Diheard Red F0 = Control

V2 = Mokara Yellow Anne F1 = Spray Formulation-1

DAP = Days After Planting F2 = Spray Formulation-2

F3 = Spray Formulation-3

Variety x

Spray

formulation

Leaf area index at DAP Leaf length (cm) at DAP

45 90 135 180 45 90 135 180

V1F0 0.55 ab 0.68 b 0.93 bc 1.14 b 23.9a 24.78 a 25.23 a 26.03 a

V1F1 0.43 c 0.64 b 0.92 bc 1.24 ab 22.76a 24.18 a 24.56 a 25.36 b

V1F2 0.44 c 0.62 b 0.88 bc 1.15 b 22.93a 23.36 a 23.83 a 24.4 bc

V1F3 0.46 bc 0.69 b 1.02 bc 1.33 ab 21.78b 22.2 b 22.66 b 23.2 c

V2F0 0.45 bc 0.59 b 0.85 c 1.21 ab 22.1 a 22.43 b 22.76 b 23.26 c

V2F1 0.56 a 0.78 ab1.13

ab1.44 ab 21.9 b 22.3 b 22.76 b 23.36 c

V2F2 0.56 a 0.90 a 1.21 a 1.51 a 23.53 24.3 a 24.96 a 25.86 a

V2F3 0.45 bc 0.67 b 1.03 bc 1.41 ab 23.1 a 23.56 a 23.93 a 24.86 bc

CV 11.62 14.25 12.61 12.77 7.65 7.52 7.49 7.13

Table 7. Interaction effect of variety to spray formulation on leaf area index

and leaf length.

Fig. 7.

Showing leaf length of Mokara Yellow Anne

Fig. 8 . Showing leaf length of Mokara Diheard Red

Fig. 9. Showing leaf breadth of Mokara Yellow Anne

Fig. 10. Showing leaf breadth of Mokara Diheard Red

4.3.5 Leaf width

Leaf width ranged from 2.22 - 3.12 cm and 1.95 - 3.02 cm in Mokara Diheard

red and Mokara Yellow Anne respectively during the growing period. At 45

DAP, leaf width of V1 was 2.22 cm and 1.95 cm in V2. At 90 DAP, leaf width of

V1 was 2.5 cm and 2.29 cm in V2. At 135 DAP, leaf width of V1 was 2.84 cm and

2.68 cm in V2. At 180 DAP, leaf width of V1 was 3.12 cm and 3.02 cm in V2

(Table 8). From table it was observed that rate of increasing of leaf width is

more in Mokara Yellow Anne than Mokara Diheard red. Results revealed

that Mokara Yellow Anne is superior to Mokara Diheard red in respect of leaf

width.

Spray formulation significantly affected the leaf width. At 45 DAP, the highest

leaf wide was 2.13 cm recorded in F3 and the lowest was 2.05 cm found in

control (F0). At 90 DAP, the highest leaf width (2.51 cm) was found in F3 and

the lowest (2.23 cm) in control (F0). Similar trend was observed at 135 DAP.

At 180 DAP, the highest leaf width was 3.36 cm was found in F3 and the lowest

was 2.71 cm recorded in control F0 (Table 9). Data revealed that leaf width

increased at rapidly in F3 than other treatments. Maximum leaf width always

recorded in F3 and minimum in control (F0) which revealed that Spray

nutrients containing N, P and K have little effect on leaf wide.

Leaf width varied significantly by the interaction between variety and

treatments. At 45 DAP, the highest leaf width was 2.45 cm recorded in

Mokara Diheard red with formulation 1 (V1F3) and the lowest was 1.81 cm

found in Mokara Yellow Anne with formulation 3 (V2F3). Similar trend in

leaf width was observed with increasing cultivation period (Table 10). At 180

DAP, the highest leaf width (3.56 cm) was recorded in V1F3 and the lowest

(2.68 cm) was in V1F0.

Table 8. Main effect of variety on leaf width.

Variety Leaf width (cm) at DAP

45 90 135 180

V1 2.22 2.5 2.84 3.12

V2 1.95 2.29 2.68 3.02

CV 7.95 7.63 7.14 6.73

Table 9. Main effect of spray formulation on leaf width.

Spray

formulation

Leaf width (cm) at DAP

45 90 135 180

F0 2.05 a 2.23 b 2.45 c 2.71 c

F1 2.1 a 2.44 a 2.81 ab 3.15 ab

F2 2.05 a 2.4 a 2.75 b 3.07 b

F3 2.13 a 2.51 a 3.01 a 3.36 a

CV 7.95 7.63 7.14 6.73

Legend

V1 = Mokara Diheard red F0 = Control

V2 = Mokara Yellow Anne F1 = Spray Formulation-1

DAP = Days After Planting F2 = Spray Formulation-2

F3 = Spray Formulation-3

Table 10. Interaction effect of variety to spray formulation on leaf width.

Variety x spray

formulation

Leaf width (cm) at DAP

45 90 135 180

V1F0 2.10 bc 2.30 bc 2.50 cd 2.68 d

V1F1 2.25 ab 2.58 ab 2.90 b 3.23 ab

V1F2 2.08 bc 2.31 bc 2.70 bcd 3.03 bcd

V1F3 2.45 a 2.83 a 3.26 a 3.56 a

V2F0 2.0 bc 2.16 c 2.41 d 2.75 cd

V2F1 1.95 bc 2.3 bc 2.73 bcd 3.06 bcd

V2F2 2.03 bc 2.5 ab 2.81 bc 3.11 bc

V2F3 1.81 c 2.2 c 2.76 bcd 3.16 b

CV 7.95 7.63 7.14 6.73

4.3.6 Total leaf area

The development of total leaf area over time in orchid was significantly

affected by varieties. The total leaf area ranged from 523.55-1346.96 cm2 and

565.16 - 1540.38 cm2 in Mokara Diheard red and Mokara Yellow Anne,

respectively over growing period. At 45 DAP, the total leaf area of V1 was

523.55 cm2 and of of V2 was 565.16 cm2. At 90 DAP, the average total leaf area

of V1 was 735.01 cm2 and of V2 was 809.60 cm2. At 135 DAP, average total leaf

area of V1 was 1029.82 cm2 and of V2 was 1169.59 cm2. At 180 DAP average

total leaf area of V1 was 1346.96 cm2 and of V2 Was 1540.38 cm2. (Table 11).

From table it was observed degree of leaf area increment was higher in

Mokara Yellow Anne than Mokara Diheard red and spray formulation

affected leaf width.

Spray formulation significantly affected the leaf width. At 45 DAP, maximum

average total leaf area was 562.3 cm2 was found in control (F0) and minimum

average total leaf area was 508.09 cm2 in F3. At 90 DAP, maximum total leaf

area 844.59 cm2 was found in F2 and minimum 691 cm2 in control (F0). At 135

DAP, maximum average total leaf area 1160.46 cm2 was found in F2 and

minimum was 965.90 cm2 found in control (F0). At 180 DAP, maximum

average total leaf area was 1524.03 cm2 found in F3 and minimum was 1298.03

cm2 observed in F0 (Table 12). From table it was observed that trend of leaf

area increased over vegetative growth was relatively higher in F3. In all

treatments, average total leaf area increased in later growth stage during

which phosphorus (P) and potassium (K) concentration was increased in

spray solution which indicated that P, and K in spray nutrients increases leaf

area.

Total leaf area varied significantly by the interaction between variety and

formulation. At 45 DAP, the highest total leaf area was 627.50 cm2 recorded in

V2F2 and the lowest was 478.0 cm2 observed in V1F1. At 90 DAP, the highest

and lowest leaf area were 1000.54 cm2 and 625.8 cm2 recorded in V2F2 and

V2F0 respectively. At 135 DAP, the highest and lowest total leaf area were

1341.67 cm2 and 947.66 cm2 in V2F2 and V2F0, respectively. V2F3 and V1F0 also

showed the highest (1665.61 cm2) and lowest (1256.78 cm2) total leaf area

respectively at 180 DAP (Table 13). The variation in leaf area might occur due

to the variation in number of leaves and their expansion. The result obtained

from the present study is consistent with the results of Sharma and Haloi

(2001) who stated that variation in leaf area might be attributed to the

difference in number of leaves.

Table 11. Main effect of variety on leaf number and total leaf area.

Variety Leaf number at DAP Total leaf area (cm2) at DAP

45 90 135 180 45 90 135 180

V1 10.25 12.41 15.00 17.58 523.55 735.01 1029.82 1346.96

V2 12.66 15.33 18.41 21.25 565.16 809.60 1169.59 1540.38

CV 14.69 11.77 10.37 8.48 11.62 14.41 12.91 12.08

Table12. Main effect spray formulation leaf number and total leaf area.

Spray

formulation

Leaf number at DAP Total leaf area (cm2) at DAP

45 90 135 180 45 90 135 180

F0 11.83 a 13.50 16.50

a

19.50 562.3 a 691 b 965.9 b 1298.03

c

F1 12.83 a 14.16 17.86

a

19.33 549.01

a

789.06

ab

1135.1

ab

1482.42

a

F2 11.66 a 14.50 17.16

a

19.83 558.02

a

844.59 a 1160.46

a

1470.2 a

F3 10.50 b 13.33 15.oo b 19.00 508.09

b

764.57

ab

1137.37

ab

1524.03

a

CV 14.69 11.77 10.37 8.48 11.62 14.41 12.91 12.08

Legend

V1 = Mokara Diheard Red F0 = Control

V2 = Mokara Yellow Anne F1 = Spray Formulation-1

DAP = Days After Planting F2 = Spray Formulation-2

F3 = Spray Formulation-3

Variety x

spray

formulation

Leaf number at DAP Total leaf area (cm2) at DAP

45 90 135 180 45 90 135 180

V1F0 12.0

abc

13.33

bc

15.66

b

18.0

cd

613.7

ab

756.19

bc

984.15

bc

1256.78

b

V1F1 9.0 c 11.33

c

14.33

b

16.66

d

478.0 c 708.82

bc

1022.99

bc

1369.14

ab

V1F2 10.33

bc

12.66

c

15.33

b

17.66

cd

488.54

c

688.64

bc

979.26

bc

1274.78

b

V1F3 9.66 c 12.33

c

14.66

b

18.0

bcd

513.96

abc

786.39

bc

1132.9

abc

1487.13

ab

V2F0 11.66

abc

13.66

bc

17.33

ab

21.0

ab

510.9

abc

625.80

c

947.66 c 1339.29

ab

V2F1 14.66

a

17.0

a

20.0 a 22.0

a

620.02

ab

869.31

ab

1247.20

ab

1595.7

ab

V2F2 13.0

ab

16.33

ab

19.0

a

22.0

a

627.50

a

1000.54

a

1341.67

a

1665.61

a

V2F3 11.33

bc

14.33

abc

17.33

ab

20.0

abc

502.22

bc

742.75

bc

1141.85

abc

1560.93

ab

CV 14.69 11.77 10.37 8.48 11.62 14.41 12.91 12.08

Table 13. Interaction effect of variety to spray formulation on leaf number

and total leaf area.

4.3.7 Leaf number

The number of leaves per plant was statically significant at 45, 90,135,180

DAP. The leaf number ranged from 10.25-17.58 and 12.66-21.25 in Mokara

Diheard red and Mokara Yellow Anne respectively over growing period. At

45 DAP, the average leaf number of Mokara Diheard red was 10.25 and of

Mokara Yellow Anne was 12.66. In Mokara Diheard red, the average number

of leaves per plant was 12.41, 15 and 17.58 at 90,135 and 180 DAP

respectively. In Mokara Yellow Anne, the average number of leaves per plant

was 15.53, 18.41 and 21.25 at 90,135 and 180 DAP respectively (Table 11).

Though both varieties were treated with same spray nutrients but production

of leaf was higher in Mokara Yellow Anne than Mokara Diheard Red. Results

revealed that Mokara Yellow Anne might be superior over Mokara Diheard

Red in respect of leaf production.

At 45 DAP, the averave highest leaf number was 12.83 recorded in F1 and the

lowest was 10.5 found in F3. At 90 DAP, the highest leaf number was 14.5 in F2

and the lowest was 13.33 in F3. At 135 DAP, the average highest leaf number

was 17.86 was in F1 and the lowest was 15 found in F3. At 180 DAP, the highest

(19.83) and the lowest (19) leaf number was also in F2 and F3, respectively

(Table 12). Since level of nutrient concentrations in spray solution was altered

over growth stage and number of leaves also increased with alteration of

nutrient concentrations in spray formulation. It might be concluded that

spray nutrients was t effective on leaf production. The present result supports

the report of Singh and Kohli (1999).

Leaf number varied significantly due to the interaction effect between variety

and formulation. At 45 DAP, the highest leaf number was 14.66 and the lowest

leaf number was 9 in V2F1 and V1F1, respectively. The highest leaf number at

90,135, 180 DAP were 17, 20 and 22, respectively and recorded in of V2F1. The

lowest leaf number at 90,135, 180 DAP were 11.33, 14.33, 16.66, respectively

and in V1F1 (Table 13). The variation in total number of leaves might be

concerned with genetically variation, physiological function and growth

characters of plant.

4.3.8 Stem diameter

Varietal effect of stem diameter was significantly varied. In Mokara Diheard

Red, the average stem diameter per plant was 0.81, 0.97, 1.1, 1.25 at 45, 90,

135 and 180 DAP, respectively. In Mokara Yellow Anne, the average stem

diameter per plant was 0.96, 1.11, 1.27 and 1.43 at 45, 90,135 and 180 DAP,

respectively (Table 14). In comparison between two varieties degree of

increasing of stem diameter was more in Mokara Yellow Anne than Mokara

Diheard Red.

At 45 DAP, the highest stem diameter was 0.96 cm recorded at spray

formulation 1 (F1) and the lowest was 0.81 cm in control (F0). Spray

formulation-3 (F3) showed maximum stem diameter (1.11 cm) and F2 showed

minimum stem diameter (0.97 cm) at 90 DAP. At 180 DAP, maximum (1.41`

cm) and minimum (1.27 cm) stem diameter was observed in F3 and F0,

respectively (Table 15). At 90 and 135 DAP, stem diameter in control plants

was more than F1 and F2 which indicates that spray nutrients containing N, P,

and K have little effect on stem diameter.

The interaction between variety and formulation on stem diameter was varied

significantly. At 45 DAP, the highest stem diameter was 1.05 cm and the

lowest was 0.75 cm recorded in V2F1 and V1F0, respectively. At 90 DAP, the

highest and the lowest stem diameter was 1.2 cm and 0.91 cm also recorded in

V2F3 and V1F2, respectively. At 135 DAP, the highest stem diameter was 1.35

cm and the lowest was 1.08 cm recorded in V2F3 and V1F1, respectively. At 180

DAP, the highest stem diameter was 1.5 cm and the lowest was 1.2 cm

observed in V2F3 and V1F0, respectively (Table 16).

Table 14. Main effect of variety on number of root and Stem diameter.

Variety Number of root at DAP Stem diameter (cm) at DAP

45 90 135 180 45 90 135 180

V1 1.08 3.08 5.25 7.58 0.81 0.97 1.10 1.25

V2 2.08 3.91 5.75 8.66 0.96 1.11 1.27 1.43

CV 14.66 12.47 20.99 19.62 13.56 11.23 8.37 8.12

Table 15.Main effect of spray formulation on number of root and stem

diameter.

Spray

formulati

on

Number of root at DAP Stem diameter (cm) at DAP

45 90 135 180 45 90 135 180

F0 1.33 b 3.0 5.33 8.0 0.81 1.0 1.16 1.27

F1 1.5 ab 3.50 5.50 8.33 0.96 1.08 1.21 1.37

F2 2.33 a 4.16 5.66 8.33 0.83 0.97 1.13 1.3

F3 1.16 b 3.33 5.50 7.83 0.93 1.11 1.25 1.41

CV 14.66 12.47 20.99 19.62 13.56 11.23 8.37 8.12

Legend

V1 = Mokara Diheard Red F0 = Control

V2 = Mokara Yellow Anne F1 = Spray Formulation-1

DAP = Days After Planting F2 = Spray Formulation-2

F3 = Spray Formulation-3

Table16. Interaction effect of variety to spray formulation on number of root

and Stem diameter.

Variety x

spray

formulatio

n

Number of root at DAP Stem diameter (cm) at DAP

45 90 135 180 45 90 135 180

V1F0 1.33

b

3.0 ab 5.66 8.66 0.75 b 0.93 b 1.11 b 1.2 b

V1F1 1.0 b 3.33 5.0 7.0 0.88 1.0 ab 1.08 b 1.26 b

ab ab

V1F2 1.33

b

3.33

ab

5.33 7.33 0.76 b 0.91 b 1.08 b 1.2 b

V1F3 0.66

b

2.66 b 5.0 7.33 0.85

ab

1.03

ab

1.15 b 1.33 ab

V2F0 1.33

b

3.0 ab 5. 0 7.33 0.88

ab

1.06

ab

1.21 ab 1.35 ab

V2F1 2.0 b 3.66

ab

6.0 9.66 1.05 a 1.16 a 1.35 a 1.48 a

V2F2 3.33

a

5.0 a 6.0 9.33 0.90

ab

1.03 ab 1.17 ab 1.4 ab

V2F3 1.66

b

4.0 ab 6. 0 8.33 1.01 a 1.2 a 1.35 a 1.5 a

CV 14.66 12.47 20.99 19.62 13.56 11.23 8.37 8.12

4.3.9 Number of root

The number of root varied from 1.08 - 7.58 and 2.08 - 8.66 during growth

stage in Mokara Diheard red and Mokara Yellow Anne, respectively. The

average number of root of Mokara Diheard red was 1.08, 3.08, 5.25 and 7.58

at 45, 90,135 and 180 DAP, respectively. In Mokara Yellow Anne average

number of root were 2.08, 3.91, 5.75 and 8.66 at 45, 90, 135 and 180 DAP,

respectively (Table 14). Though both varieties were treated with same

treatments but from table it was observed that degree of root production was

higher in Mokara Yellow Anne than Mokara Diheard red. So, varietal effect

on number of root was significant.

Different spray formulations have significant effects on number of roots. At

45 DAP, maximum average number of roots was 2.33 recorded in F2 and

minimum was 1.16 in F3. At 90 DAP, the highest number of roots was 4.16

observed in F2 and the lowest was 3.0 found in control (F0). At 135 DAP, the

highest (5.66) number of roots showed in F2 and the lowest (5.33) was found in

F0. At 180 DAP, the highest number of roots was 8.33 showed in F1 and F2 and

the lowest was 7.83 found in F3 (Table 15). From table it was observed that

root development increased rapidly at later growth stage than early growth

stage.

At 45 DAP, the highest and the lowest number of roots was 3.33 and 0.66 in

V2F2 and V1F3, respectively. Similar trend in number of root was observed

with increasing cultivation period. At 180 DAP, the highest number of roots

was 9.66 recorded in V2F1 and the lowest number of roots was 7 investigated

in V1F1 (Table 16). The interaction effect between variety and formulation on

root development was varied significantly. The similar results also reported

by Nandi (1999).

4.4 Physiological and growth parameters

4.4.1 Chlorophyll content

Chlorophyll content in leaf of Mokara Diheard Red and Mokara Yellow Anne

was varied significantly. At 45 DAP, average Chlorophyll a, Chlorophyll b

and total Chlorophyll content of Mokara Diheard Red were 11.47, 5.25 and

16.72 (mg/g), respectively. At 45 DAP, average Chlorophyll a, Chlorophyll b

and total Chlorophyll content of Mokara Yellow Anne were 8.12, 5.20 and

13.32 (mg/g), respectively. The average Chlorophyll a, Chlorophyll b and total

Chlorophyll content of Mokara Diheard Red were 13.09, 6.02 and 19.12

(mg/g), respectively at 90 DAP. At 90 DAP, average Chlorophyll a,

Chlorophyll b and total Chlorophyll content of Mokara Yellow Anne were

10.94 , 6.65 and 17.9 (mg/g ), respectively (Table 18). The total chlorophyll

content of Mokara Diheard Red was relatively higher than Mokara Yellow

Anne which indicated varietal effect on chlorophyll content was significant.

At 45 DAP, the average highest chlorophyll a and chlorophyll b were 13.21

(mg/g) and 6.01(mg/g) were observed in F3. The average lowest chlorophyll a

and chlorophyll b were 5.56 (mg/g) and 3.46 (mg/g) obtained in control (F0) at

45 DAP. At 45 DAP, the highest (19.22 mg/g) and the lowest (9.023 mg/g) total

chlorophyll content were also found in F3 and control (F0), respectively. At 90

DAP, the average highest chlorophyll a and chlorophyll b were 15.66 (mg/g)

and 8.0 (mg/g) observed in F3 and F1, respectively. At 90 DAP, the average

lowest chlorophyll a and chlorophyll b were 7.69 (mg/g) and 4.28 (mg/g),

observed in control (F0). At 90 DAP, the total chlorophyll content (22.73 mg/g)

was highest in F3 and the lowest was 11.96 mg/g in control (F0), respectively

(Table 17). In all treatments chlorophyll content was lowest in control plants

indicated that N, P and K in spray formulation was effective on chlorophyll

content.

The interaction between variety and spray formulation on total chlorophyll

content was varied significantly. At 45 DAP, the highest total chlorophyll

content was 25.3 mg/g found in V1F3 and the lowest was 8.68 mg/g recorded in

V1F0. At 90 DAP, the highest total chlorophyll content was 26.31 mg/g and the

lowest chlorophyll content was 9.88 mg/g found in V1F3 and V1F0, respectively

(Table 19).

Table 17. Main effect of variety on Chlorophyll content.

Spray

formulation

Chlorophyll content (mg/g)

45 DAP 90 DAP

Chl a Chl b Total

Chlorophyll

Chl a Chl b Total

Chlorophyll

F0 5.56 d 3.46 d 9.02 d 7.69 d 4.28 d 11.96 d

F1 11.30 b 5.80 b 17.11 b 13.04 b 8.0 a 21.05 b

F2 9.11 c 5.64 c 14.75 c 11.68 c 6.0 c 17.80 c

F3 13.21 a 6.01 a 19.22 a 15.66 a 7.06 b 22.73 a

CV 0.36 0.81 0.44 2.45 0.67 2.03

Table 18. Main effect of spray formulation on Chlorophyll content.

Variety Chlorophyll content (mg/g)

45 DAP 90 DAP

Chl a Chl b Total Chl. Chl a Chl b Total Chl.

V1 11.47 5.25 16.72 13.09 6.02 19.11

V2 8.12 5.20 13.32 10.94 6.65 17.9

CV 0.36 0.81 0.44 2.45 0.67 2.03

Legend

V1 = Mokara Diheard red F0 = Control

V2 = Mokara Yellow Anne F1 = Spray Formulation-1

DAP = Days After Planting F2 = Spray Formulation-2

Chl. = Chlorophyll F3 = Spray Formulation-3

Table 19. Interaction effect of variety to spray formulation on Chlorophyll

content of leaf.

Variety x

Spray

formulation

Chlorophyll content (mg/g)

45 DAP 90 DAP

Chl. a Chl. b Total Chl. Chl. a Chl. b Total Chl.

V1F0 5.92 f 2.76 h 8.68 h 6.77 e 3.11 h 9.88 g

V1F1 12.07 b 5.63 d 17.7 b 13.12 c 7.39 c 20.52 c

V1F2 10.58 c 4.63 e 15.21 d 14.58 b 5.19 g 19.79 d

V1F3 17.31 a 7.99a 25.3 a 17.89 a 8.42 b 26.31 a

V2F0 5.19 g 4.17 f 9.36 g 8.61 d 5.45 f 14.05 f

V2F1 10.54 c 5.97 c 16.51 c 12.96 c 8.62 a 21.58 b

V2F2 7.65 e 6.65 b 14.30 e 8.78 d 6.82 d 15.82 e

V2F3 9.10 d 4.03 g 13.13 f 13.44 c 5.71 e 19.15 d

CV 0.36 0.81 0.44 2.45 0.67 2.03

4.4.2 Crop growth rate (CGR)

The crop growth rate (CGR) derived in Mokara Diheard red and Mokara

Yellow Anne assessed from 45 to 180 DAP and it was observed that growth

rate of orchid was very low. The crop growth rate of Mokara Diheard red was

0.29 (g plant-1/day) and of Mokara Yellow Anne was 0.17 (g plant-1/day)

during research period. Mokara Diheard red might be considered superior

over Mokara Yellow Anne in respect of growth rate. Varietal effect of crop

growth rate was varied significantly (Table 20).

Different spray formulations have significant effect on growth rate. The crop

growth rate among different treatments ranged from 0.14-0.29 g plant-1/day.

Maximum crop growth rate was 0.29 g plant-1/day found in F2 and minimum

crop growth rate was 0.14 g plant-1/day was in control (F0) (Table 21) which

indicated that spray nutrients formulation containing N, P and K directly

affected on crop growth rate.

The interaction between variety and treatments on crop growth rate was

varied significantly. Maximum crop growth rate was 0.37 g plant-1/day

recorded in V1F2 and minimum crop growth rate was 0.12 g plant -1/day

recorded in V2F0 (Table 22).

4.4.3 Relative growth rate (RGR)

The relative growth rate (RGR) of Mokara Diheard red and Mokara Yellow

Anne assessed from 45 to 180 DAP have been presented in Table 20. The

relative growth rate of Mokara Diheard red was 6.88 (mg g-1/day) and of

Mokara Yellow Anne was 5.89 (mg g-1/day).

Different spray formulations have significant effect on relative growth rate.

The relative growth rate among different treatments ranged from 6.04-6.76

mg g-1/day. Maximum relative growth rate was 6.76 mg g-1/day was found in

F2 and minimum crop growth rate was 6.04 mg g-1/day recorded in control

(Table 21) which indicated that spray nutrients formulation containing N, P

and K directly affect on relative growth rate.

The interaction between varieties to spray formulation on relative growth rate

was varied significantly. Maximum relative growth rate was 7.14 mg g -1/day

found in V2F3 and minimum growth rate was 4.96 mg g-1/day recorded in

control V1F0 (Table 22).

Table 20. Main effect of variety on CGR and RGR.

Table 21. Main effect of spray formulation on CGR and RGR.

Spray formulation CGR (g plant-1/day) RGR (mg g-1/day)

F0 0.14 c 6.04 d

F1 0.21 b 6.12 c

F2 0.29 a 6.76 a

F3 0.28 a 6.62 b

CV 13.2 6.50

Legend

V1 = Mokara Diheard red F0 = Control

V2 = Mokara Yellow Anne F1 = Spray Formulation-1

DAP = Days After Planting F2 = Spray Formulation-2

F3 = Spray Formulation-3

Variety CGR (g plant-1/day) RGR (mg g-1/day)

V1 0.29 6.88

V2 0.17 5.89

CV 17.82 8.50

Table 22. Interaction effect of variety to spray formulation on CGR and RGR.

Variety x spray

formulation

CGR (g plant-1/day) RGR (mg g-1/day)

V1F0 0.17 de 4.96 g

V1F1 0.28 bc 6.03 f

V1F2 0.37 a 6.48 c

V1F3 0.35 ab 6.1 e

V2F0 0.12 e 7.13 a

V2F1 0.15 de 6.21 d

V2F2 0.22 cd 7.05 b

V2F3 0.21 cd 7.14 a

CV 17.82 5.50

Chapter V

SUMMARY AND CONCLUSION

An experiment was conducted in grill house of the Department of Crop

Botany, BAU, during the period of 30 November, 2009 to 30 September,

2010 to determine the effect of spray nutrients on morpho-physiological

features and growth of orchid. The experiment comprised of two orchid

cultivars and four levels of spray nutrients with control. The experiment

was laid out in Completely Randomized Block Design (RCBD) with 8

treatments and each treatment was replicated three times. Data were taken

on some selected parameters and statistical analyses were performed by

using MSTAT-C program. The results of the experiment were summarized

below.

All the studied parameters were significantly influenced by spray formulation

consists of N, P and K. Morpho-physiological features like plant height, leaf

length, leaf wide, leaf area, leaf number, leaf area index, stem diameter,

number of root and growth parameters viz. crop growth rate, relative growth

rate, chlorophyll content in leaf were affected by spray formulation. The effect

of cultivars on those morpho-physiological and growth features also

significant. Increasing plant height, leaf wide, leaf area, leaf number, leaf area

index, stem diameter, number of root and RGR was higher in Mokara Yellow

Anne. CGR, leaf length, total chlorophyll content were higher in Mokara

Diheard red.

Different spray formulation with different ratio of N, P and K also

significantly affected morpho-physiological features and growth parameters.

Leaf length was slightly affected by spray formulation. Degrees of increasing

leaf length, number of root were higher in spray formulation-1 (F1). Rate of

increasing of plant height, crop growth rate, relative growth rate were more

in spray formulation-2 (F2). Rate of increasing of leaf width, total leaf area,

leaf number, leaf area index, stem diameter and total chlorophyll content

were more in spray formulation-3 (F3).

Plant height, stem diameter, total leaf area and CGR were best in V2F2 among

all treatments. Leaf area, leaf area index and RGR were best in V2F3 among

all treatments. Leaf length and leaf width were best in V2F1 and V2F3,

respectively. Chlorophyll content was superior in V1F3 among all treatments.

Based on experimental results it may be concluded that

i) the morphological and physiological attributes was significantly affected

by spray formulation.

ii) spray formulation-2 (F2) was the best for growth of orchid.

iii) the ratio of N, P and K at 10:15: 20, 15:20:25, 10:25:25, and 10:25:30

for spray formulation might be the best for growth of orchid.

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132.

Analysis of variance

Degree of freedom

Plant height Leaf area Leaf number

45 DAP 90 DAP 135 DAP 180 DAP 45 DAP 90 DAP 135 DAP 180 DAP 45 DAP 90 DAP 135 DAP 180 DAP

Factor A (Variety)

1158.1

**147.01* 133.95 ** 135.37** 237.57* 281.12** 165.27 * 93.18 * 35.04**

51.04**

70.04**

80.66**

Factor B (Spray

formulation) 3 3.44 * 3.66 ns 11.2 * 27.07 ** 1.22 ns 49.12 ns 152.45 * 207.08 * 2.486* 1.81 ns

1.93*

0.72 ns

Interaction(AxB)

3 5.96 ** 6.93 * 3.43 * 3.73 * 37.02 ** 64.46 * 49.76 ** 50.56 ** 9.37* 7.81 * 4.37 ** 3.2 *

Error

16 14.24 13.12 12.3812.98

26.67 39.21 49.19 57.9 2.83 2.66 3.0 2.7

Appendix 1. Analysis of variance (Mean square) of plant height, leaf area and leaf number at different growth stage of orchid

Legend * Indicates significant at 5 % level

* * Indicates significant at 1 % level

Ns Indicates non -significant

58

Appendix 2. Analysis of variance (Mean square) of total leaf area, leaf area index and leaf length at different growth stage of orchid

Analysis of variance

Degree of

freedom

Total leaf area Leaf area index Leaf length

45 DAP 90 DAP 135 DAP 180 DAP 45 DAP 90 DAP 135 DAP 180 DAP 45 DAP 90 DAP 135 DAP 180 DAP

Factor A (Variety)

110388.35

**33383.50

*117214.88 *

224479.37 **

0.007 ** 0.038 * 0.082* 0.18 ** 0.21 ns 1.4 ns 1.3 ns 1.0 ns

Factor B (Spray

formulation)3

3690.55 *

24353.24 **

48532.35 *59745.47

*0.003 ns 0.016 * 0.03 * 0.04 ** 1.13 ns 1.04 ns 1.32 ns 1.3 ns

Interaction(AxB)

321629.82

**59844.41

**52437.71 *

33338.43 *

0.017** 0.043* 0.051 * 0.02 ** 2.97 * 5.43 * 5.67 * 7.95 *

Error

16 4001.67 12392.89 20144.04 30407.56 0.003 0.01 0.016 0.026 3.03 3.09 3.18 3.06

Legend* Indicates significant at 5 % level

* * Indicates significant at 1 % level Ns Indicates non -significant

59

Appendix 3. Analysis of variance (Mean square) of number of root, stem diameter and leaf width at different growth stage of orchid

Analysis of variance

Degree of freedom

Number of root Stem diameter Leaf width

45 DAP 90 DAP 135 DAP 180 DAP 45 DAP 90 DAP 135 DAP 180 DAP 45 DAP 90 DAP 135 DAP 180 DAPFactor A (Variety) 1 6.0 ** .16 ns 1.5 ns 7.04 * 0.13 ** 0.12 ** 0.16 ** 0.20 ** 0.44 ** 0.28 ** 0.15 ns 0.06 ns

Factor B (Spray

formulation)3 1.61 * 1.44 ns 0.11 ns 0.375 ns 0.033 ns 0.027 ns 0.017 ns 0.026 ns 0.009 ns 0.08 * 0.32 ** 0.43 **

Interaction(AxB) 3 1.0 ns 0.94 ns 0.94 ns 4.59 ns 0.001 * 0.001 ** 0.01 ** 0.001 ** 0.1 * 0.17 ** 0.09 ** 0.07 **

Error

16 0.50 1.29 1.33 2.54 0.014 0.014 0.01 0.012 0.02 0.03 0.03 0.04

Legend

* Indicates significant at 5 % level

* * Indicates significant at 1 % level Ns Indicates non -significant

60

Appendix 4. Analysis of variance (Mean square) of plant height, leaf area and leaf number at different growth stage of orchid

Analysis of variance

Degree of

freedom

Chlorophyll content (45DAP) Chlorophyll content(90DAP)CGR RGR

Chl(a) Chl(b) Total chlorophyll Chl(a) Chl(b) Total

chlorophyllFactor A (Variety)

1 67.19 ** 0.014** 69.21 ** 27.57** 2.319** 13.061** 0.084** 5.9 **

Factor B (Spray

formulation )3 64.73 ** 8.43 ** 116.06 ** 66.37 ** 15.29 ** 134.98 ** 0.028** 0.76 **

Interaction(AxB) 3

17.005 ** 10.92 ** 52.32 ** 19.2 ** 7.71 ** 38.34 ** 0.003 ns 1.1 **

Error16 0.001 0.002 0.004 0.087 0.002 0.139 0.002 0.001

Legend

* Indicates significant at 5 % level

* * Indicates significant at 1 % level Ns Indicates non -significant

61