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Effect of storage conditions on germinability of Himalayan cedar
(Cedrus deodara Loud.) seeds
by
RAJ KUMAR
Submitted in partial fulfilment of the requirements for the degree of
MASTER OF SCIENCE
in
FORESTRY
(SILVICULTURE)
COLLEGE OF FORESTRY Dr Yashwant Singh Parmar University of Horticulture and
Forestry, Nauni, Solan - 173 230 (H.P.) INDIA
2008
Curriculum vitae
Name : Raj Kumar
Father’s Name : Sh. Man Singh
Date of Birth : 01.12.1983
Sex : Male
Marital status : Single
Nationality : Indian
Educational qualifications:
Certificate/Degree Division Board/University Year
B.Sc. Forestry
First Dr Y.S. Parmar University of Horticulture & Forestry, Nauni- Solan (HP) 173 230
2005
10+2 Second Cenral Board of School Education 2001
Matriculation First HP Board of School Education,
Dharamshala (HP) 1999
Whether sponsored by some state/ : No Central Govt./Univ./SAARC
Scholarship/Stipend/Fellowship, any : Yes, M.Sc. merit scholarship other financial assistance received during the study period
(Raj Kumar)
Dr. R.K. Nayital Department of Silviculture and Agroforestry (Sr. Scientist) College of Forestry Dr. Y.S. Parmar University of Horticulture
and Forestry, Nauni-173 230, Solan (HP)
CERTIFICATE - I This is to certify that the thesis entitled, “Effect of storage conditions on
germinability of Himalayan cedar (Cedrus deodara Loud.) seeds” submitted
in partial fulfilment of the requirements for the award of degree of MASTER OF
SCIENCE in FORESTRY (Silviculture) to Dr. Yashwant Singh Parmar
University of Horticulture and Forestry, Nauni, Solan (HP) is a record of bonafide
research work carried out by Mr. Raj Kumar (F-2005-16-M) under my guidance
and supervision. No part of this thesis has been submitted for any other degree
or diploma.
The assistance and help received during the course of investigations has
been fully acknowledged.
__________________ Place : Nauni-Solan Dr. R.K. Nayital Dated : , 2007 Chairperson
Advisory Committee
CERTIFICATE - II
This is to certify that the thesis entitled, “Effect of storage conditions on
germinability of Himalayan cedar (Cedrus deodara Loud.) seeds” submitted
by Mr. Raj Kumar (F-2005-16-M) to Dr. Yashwant Singh Parmar University of
Horticulture and Forestry, Nauni, Solan (HP) in partial fulfilment of the
requirements for the award of degree of MASTER OF SCIENCE in FORESTRY
(Silviculture) has been approved by the Student’s Advisory Committee after an
oral examination of the same in collaboration with the external examiner.
______________________ _____________________ Dr. G.S. Shamet External Examiner Professor (Co-opted in place of Dr. R.K. Nayital) Chairman Advisory Committee Dept. of Silviculture & Agroforestry
Members, Advisory Committee
______________________ _____________________ Dr. S.D. Bhardwaj Dr. H.P. Sankhyan Professor & Head Scientist Dept. of Silviculture & Agroforestry Dept. of Tree Improvement ______________________ ______________________ Dr. Bhupender Dutt Dean’s Nominee Assistant Professor Dept. of Forest Products
____________________ Professor and Head
Dept. of Silviculture & Agroforestry
____________________ Dean
College of Forestry
CERTIFICATE – III
This is to certify that all the mistakes and errors pointed out by the external
examiner have been incorporated in the thesis entitled, “Effect of storage
conditions on germinability of Himalayan cedar (Cedrus deodara Loud.)
seeds” submitted to Dr. Y.S. Parmar University of Horticulture and Forestry,
Nauni-Solan (HP) by Mr. Raj Kumar (F-2005-16-M) in partial fulfilment of the
requirements for the award of degree of MASTER OF SCIENCE in FORESTRY
(Silviculture).
______________________
Dr. G.S. Shamet Professor
(Co-opted in place of Dr. R.K. Nayital) (Major Advisor)
\
_____________________________________
Professor and Head Dept. of Silviculture & Agroforestry
Dr. Y.S. Parmar, UHF, Nauni-173 230, Solan (HP)
CCOONNTTEENNTTSS
Chapter Title Pages
1 INTRODUCTION 1-4
2 REVIEW OF LITERATURE 5-15
3 MATERIALS AND METHODS 16-21
4 EXPERIMENTAL RESULTS 22-54
5 DISCUSSION 55-65
6 SUMMARY 66-67
7 REFERENCES 68-77
ABSTRACT 78
APPENDIX i-iv
LLIISSTT OOFF TTAABBLLEESS
Table Title Page(s)
1 Germination parameters of freshly collected seed of C. deodara
22
2 Effect of temperature on per cent germination and germinative capacity of C. deodara seed during storage
24
3 Effect of temperature on per cent germinative energy and germination value of C. deodara seed during storage
25
4 Effect of container on per cent germination and germinative capacity of C. deodara seed during storage
28
5 Effect of container on per cent germinative energy and germination value of C. deodara seed during storage
29
6 Interaction effect of temperature and container (T x C) on per cent germination of C. deodara seed during storage
30
7 Interaction effect of temperature and container (T x C) on per cent germinative capacity of C. deodara seed during storage
32
8 Interaction effect of temperature and container (T x C) on per cent germinative energy of C. deodara seed during storage
33
9
Interaction effect of temperature and container (T x C) on germination value of C. deodara seed during storage
34
10 Physio-biochemical parameters of freshly collected seed of C. deodara
35
11 Effect of temperature on moisture content and total sugar of C. deodara seed during storage
37
12 Effect of temperature on reducing and non reducing sugar of C. deodara seed during storage
38
13 Effect of temperature on starch and phenol of C.deodara seed during storage
40
Table Title Page(s)
14 Effect of container on moisture content and total sugar of C. deodara seed during storage
41
15 Effect of container on reducing and non reducing sugar of C. deodara seed during storage
43
16. Effect of container on starch and phenol of C. deodara seed during storage
44
17. Interaction effect of temperature and container (T x C) on Moisture content (%) of C. deodara seed during storage
46
18. Interaction effect of temperature and container (T x C) on total sugar (%) of C. deodara seed during storage
47
19. Interaction effect of temperature and container (T x C) on reducing sugar (%) of C. deodara seed during storage
49
20. Interaction effect of temperature and container (T x C) on non reducing sugar (%) of C. deodara seed during storage
50
21.
Interaction effect of temperature and container (T x C) on starch (%)of C. deodara seed during storage
52
22. Interaction effect of temperature and container (T x C) on phenol (mg/g) of C. deodara seed during storage
53
23 Correlation of per cent germination with germinable and physio-biochemical parameters of deodara seed.
54
LLIISSTT OOFF PPLLAATTEESS
Plates Title Between Page(s)
1 Determination of biochemicals
2 Seed storage at different temperatures
Acknowledgements
Research is an evolving concept. Any endeavour, in this regard is challenging as well as exhilarating. It implies the testing of our nerves. It brings to light our patience, vigour and dedication.
Every result arrived at is a modest beginning for a higher goal my work in the same spirit, is just a step in the ladder. It is a drop of ocean. No work can be turned as a one-man show. It needs the close cooperation of friends and colleagues and the guidance of experts in the field to achieve something worthwile and substantial.
With the blessings of Good Hearts, I could bring this piece of work into light. I shall like to pen down my immense gratitudes for all those who directly or indirectly helped me in this endeavour.
With great reverence, I express my warmest feelings with deep sense of gratitude to the Chairperson of my advisory committee, Dr. R .K. Nayital. I have no words to express my heartful thanks to him for illuminating guidance, unfailing encouragement, scholarly suggestions, unique supervision, constructive criticism, sympathetic attitude and keen interest during the course of this investigation and in the preparation of this manuscript.
I seize this opportunity to express my sincere regard to the Members of my Advisory Committee, Dr.S.D.Bhardwaj (Professor and Head, Department of Silviculture and Agroforestry), Dr.Bhupinder Dutt and Dr. H.P. Sakhyan for their valuable suggestions during the entire degree programme.
I am also grateful to Dr. N.K Gupta, M.Prabhakar, Dr.(Mrs) Vidya Thakur, Dr. N.B.Singh, Dr. D.R.Bhardwaj, Dr. K.S.Panth, Dr. K .S.Verma, Dr. G.S.Shamet ,Dr. P.S.Thakur Dr.I.K.Thakur and Dr.(Mrs) Menu Sood for their Herculean encouragement.
Yes, I believe in god, that is my parents. This flesh, these bones and each drop of my blood belongs to them. Any and every good quality which seems to be of myself is actually theirs. I thank the Supernatural for giving me such caring and sacrificial parents to whom I owe all that is mine. I thank them from the depths of my existence for all that which helped me achieve this goal.
Emotions of heart find new boundaries to express my deep feelings for ‘my grand mother, my father, mother, uncle, aunty, brothers and cousin sisters for their understanding attitude, resplendent nature and untiring help to complete my work. Thanks for always being besides me and encouraging me for doing noble works.
I would like to avail this opportunity to extend my heartiest thanks to Dr.Tara chand, Dr. Rashid Dr. Ashok Vijay Minj and Dr.Vikas Thakur for his kind guidance and suggestions.
I express my personal regards to my friends; Asif, Manjeet, Marrry, Dinesh, Sivani, Sanjeev, Ratan, Vishwesh, Bilal, Vivek, Nitin, Anish, Subhash, Ngura, Vijay , Dhram , Rana, Munde, Surinder, Balbir, Aman, Sudershan, A. Mittal, Chandru, Suhail, Irfan, Millo, Vinod, Mishra Sir, Subhash Sir, Yogi Sir, Pervej Sir, Vinay sir, Ajay sir, Pradeep sir, Dinesh sir, Denu sir, Sanjeev sir, Navneet sir, Rakesh sir , and for their motivation and support when they were needed most.
The cooperation and help rendered by Sh.Padam Singh and other laboratory staffs, the office bearers of the Department of Silviculture and Agroforestry is also duly acknowledged.
My sincere thanks are due to Sh. Basnsal ji and Sh. Vinod ji (Swastika computers) for giving a worthful shape to this manuscript.
Needless to say errors and omissions are mine.
Place: Nauni, Solan Date: January, 2008 (Rajkumar)
Chapter-I
INTRODUCTION
Cedrus deodara Loud. belongs to family Pinaceae and is one of the
most important timber species of western Himalaya. It occurs between 68o to
800E longitude and 30o to 36oN latitude. The species is distributed from
Afghanistan to Garhwal with an altitude range of 1200 to 3000 m (Gamble,
1881; Brandis, 1906; Troup, 1921 and Champion and Seth, 1968). Cedrus
deodara is a major plant species found in the forest type 12/C1c (Moist deodar
forest) and is an associated species in the forest sub types of 12/C1b and
12/C1d of Himalayan moist temperate forest (Champion and Seth, 1968).
Deodar occurs on all important geological formations. Occurrence of
good deodar has been reported on metamorphic rocks on Tehri Garhwal
Himalaya (Biswas, 1985). Besides that, it has also been occurring on granite,
gneiss, mica, shale, limestone, quartzite and conglomerate. The best growth
is attained on deep, fairly porous and fertile soil with an annual rainfall of 1000
to 1800mm.
Deodar is typically a gregarious species. It is commonly associated
with conifer like, Piinus wallichiana, Picea smithiana and Abies pindrow. At
lower elevation, its association with Pinus roxburghii has been reported,
where chir pine occupies the drier ridges and deodar the moist and cooler
depressions. In some belts, it is also associated with Cupressus torulosa,
Pinus gerardiana (In the dry inner Himalaya) and Taxus baccata (In the mosit
shaddy situation). Many broad leaved species are also associated with
deodar. Quercus leucotrichophora and Quercus dilatata are frequent
companions. Apart form Oaks, Pyrus pashia, Rhododendron arboretum,
Prunus puddum, Aesculus indica, Populus ciliata, Cornus macrophylla,
Juglans regia, Ulmus wallichiana and Acer species are other broad leaved
associates. The ground flora mainly consist of species of Rosa, Rubus,
2
Lonicera, Berberis, Vibernum, Indigofera, Desmodium, Clematis, Montana,
Hedera helix etc.
Cedrus deodara is a large evergreen tree, branches horizontal or
slightly ascending or descending, not whorled (Gamble, 1881; Brandis, 1906
and Troup, 1921), leaves are acicular; 2.5 to 3.75 cm long arranged spirally
on long shoots and is pseudowhorls in short shoots, generally dark green in
colour. Bark is grayish brown with vertical and diagonal cracks.
As a rule, deodar is monoecious with male and female cones on
separate branches. Occasionally, it shows a dioecious habit. Male flowers
appear in June. There is no resting stage and entire course of pollen
development is accomplished in about three months (Johri, 1936). The female
cones appear in August. The time required from first appearance to the
ripening of cones is about 12 ½ to 13 ½ month (Raizada and Sahni, 1960).
Deodar is the strongest of the coniferous woods. It is fairly hard and very
durable (Rao and Juneja, 1971). This is commonly used in the building,
furniture and carpentry. It is also used for electrical poles and battery
separators besides, its use for making second grade pencils (Anonymous,
1950). The wood of deodar on steam distillation yields reddish brown oil,
which is used in making scents, soaps and perfumes. The wood oil of deodar
is useful for fevers, piles and urinary disorders (Anonymous, 1950). The bark
astringent is useful for fevers, diarrhea and dysentery. The oleoresin of
deodar and the dark colored oil obtained from the wood are valued as an
application for ulcers and skin diseases.
Being an important timber species, it is very important to regenerate
this species. The difficulties in getting natural regeneration of deodar is
because of low viability of seed, heavy grazing and drought sensitive nature of
seedling, necessitate collection of seed in large quantities for artificial
regeneration. Most of forest trees are irregular seed producer with long cycles
of good seed production such as 4-5 in Cedrus deodara. Due to this reason,
large quantity of high quality seed need to be collected and stored in good
3
seed year for use in intervening years to ensure a continuous supply of seed
for the sustained annual production of nursery stock to meet the exigency of
afforestation programme.
Storage may be defined as preservation of viable seed from time of
collection until they are required for sowing (Holmes and Buszewicz, 1958).
Successful seed storage of different trees requires the knowledge of seed
behaviour such as seed maturity, seed handling and processing, seed
moisture content, storage temperature and storage methods. F.A.O. (1995)
has suggested three reasons for the storage of forest tree seed: (a) to
preserve seed in the best condition to retain their germination energy during
the interval between collection and time of sowing; (b) to protect seed from
damages by pathogens, insects, rodents and birds; (c) to preserve quantities
of seed collected in good year in order to have seed in reserves to be made
available for years when little or no seed have been produced.
Roberts (1973) has classified seed into the following two categories.
Orthodox seed : seed which can withstand drying to low moisture content
around 5% and successfully stored at low or sub freezing temperature for
longer periods e.g., Acacia, Albizia, Cassia, Eucalyptus, Pinus, Picea etc.
Recalcitrant: seed are characterized by large size and high moisture content,
which cannot be dried without causing injury. Most of the species of
Diptocarpaceae, Fagaceae and Lauraceae produce recalcitrant seed. The
genera Aesculus, Castenea, Dipterocarpus, Hopea, Quercus, Shorea etc also
fall into this category.
The principle factors affecting the viability of seed in storage are
moisture content of seed, maturity, temperature and relative humidity. At the
time of harvesting the state of maturity of seed is found to be major factor
responsible for viability. Therefore, a decision as to when to harvest the seed
is of a great importance. Many seed have been reported to harbor great
variety of fungi and insect pests. Therefore, sound seed at the time of
harvesting, may be invaded by fungi that have been known to contribute
4
significantly towards reduction in the viability or death of seed during storage
(Singh and Mathur, 1993) and insects damage only those seed, which were
already infested at the time of storage (Thakur, 2000). Oxygen also affect
seed during storage and if higher the oxygen pressure, the shorter the viability
and the effects are more at higher temperature and moisture content, which
lead to loss of seed viability (Roberts 1973).
Harrington (1963) suggested that sum of the percentage of relative
humidity and temperature in degrees Fahrenheit of storage environment
should not exceed 100 for safer storage. Moisture content is also one of most
important factors in maintaining the viability of seed during storage. By
lowering the moisture, the metabolic activity is considerably reduced resulting
in reduction of respiration and consumption of reserve nutrients that is a vital
factor in the maintenance of viability.
The seed of Cedrus are oily and do not keep well under ordinary
storage conditions (Rudolf, 1974 and Allen, 1995). If Cedrus seed are dried
below a critical level, they will not imbibe water in a way that will allow the
food reserves to be used by the embryo (Macdonald, 1986). According to
Rudolf (1974) and Erkuloglu (1995), seed of Cedrus could retain viability for 3
to 6 years when dried to a moisture content of less than 10%, placed in
sealed containers, and held at temperatures of –1°C to –5°C.
Keeping in view the aforesaid problems, an investigation entitled
“Effect of storage conditions on germinability of Himalayan cedar
(Cedrus deodara Loud.) seeds” was carried out with the following
objectives:
To observe the effect of temperature on seed viability
To find out suitable storage container
Chapter-II
REVIEW OF LITERATURE
The information pertaining to the present study has been reviewed in
the light of work done on the conifer species in India and abroad. Though
sufficient work has been done on various conifer species in Europe and
United States, the work is still in its infancy in India. Literature on storage
technique of deodar is scanty, cross references on other species have been
incorporated as per following main heads:
2. Storage of seed
2.1 Effect of storage temperature and container on germinability
2.2 Physio-biochemical indices of seed
2. Storage of seed
Storage may be defined as the preservation of viable seed from the
time of seed collection, until they are required for sowing. Seed storage is
virtually a practical necessity associated with artificial regeneration
programmes in many tree species for regular and sustained supply of seed
(Holmes and Buszewicz, 1958; Magini, 1962; Vlase, 1974 and Sharma et al.,
2004). Better storage technique could have significant effect on afforestation
programmes and therefore to reverse deforestation and at the same time
preserve gene pool in many forest species.
2.1 Effect of storage temperature and container
The storage container is a prerequisite for seed storage to facilitate
handling of individual seed lot while the choice of storage temperatures varies
considerably according to species and period for which the seed is to be
stored. Moisture proof container and controlled temperature provide maximum
6
protection against mechanical damage to the seed and are equally suitable
for storage and shipment (Robbins, 1984).
According to many references, fresh seed of Cedrus are not normally
dormant and thus do not require treatment in order to germinate (Dirr and
Heuser, 1987; Takos and Merou, 1995 and Hartmann et al., 1997). However,
it is possible for dormancy to develop in some seed lots whose germination,
without treatment, can be irregular. In such cases, if the seed are treated with
cold stratified (+ 4°C) for 2 months, then they germinate readily in 4 to 7 days
(Fodham and Spraker, 1977 and Dirr and Heuser, 1987). The main factors
that affect seed viability during the storage are moisture content and
temperature (Bradbeer, 1988; Bonner, 1990; Gordon, 1992 and Takos, 1999).
Storage of many species seems to induce dormancy so that further treatment
is necessary (Willemsen, 1975).
Chandra and Ram (1980) referred to dormancy in stored seed of C.
deodara, which was broken after stratification for 15 or 30 days at 4.4°C. The
resulting germination percentages were 16% and 45%, respectively, whereas
the control (untreated) germination was 11 per cent. On the contrary, Fodham
and Spraker (1977) did not find any improvement in the germination
percentage of Cedrus seed after cold stratification. Struck and Whitcomb
(1977) proposed the soaking of Cedrus seed for 2 or 3 hours as an alternative
method for breaking of dormancy. Therefore, although Cedrus seed do not
possess primary dormancy, it is later induced, especially during the long-term
storage of 3 to 6 years (Young and Young, 1992). Even short-term storage,
from the time of the collection in the autumn till the sowing in the spring, can
affect germination negatively. In many species a decrease in germination
capacity, appears during the first few months after collection, due to
inappropriate storage conditions (Romanas, 1991). Krussmann (1981)
suggested that Cedrus seed should remain in the cones during winter,
because their germination percentage was better. The same storage method
was proposed by Young and Young (1992) for C. atlantica, C. libani and C.
brevifolia, however C. deodara was not studied. Cedrus seed exhibit little or
7
no dormancy and will germinate without pretreatment. However, variable
degrees of dormancy may be observed within a single lot of seed (Dirr and
Heuser, 1987). Seed should be stratified at 3 to 5°C for 2 weeks to obtain
uniform germination (Rudolf, 1974 and Allen, 1995).
Thapliyal and Gupta (1980) found that 9°C (48.2°F) was a better
temperature for stratification than 3°C for Cedrus deodara and Cedrus libani.
Seed are prone to damping-off disease caused by Fusarium, Rhizoctonia and
Pythium species. Therefore, an appropriate fungicide should be used (Mittal,
1983 and Tewari, 1994). The Association of Official Seed Analysts rules for
Cedrus (Rudolf, 1974) specified germination tests of stratified seed on top of
blotters for 3 weeks at 20°C. International Seed Testing Association rules,
however, specify diurnally alternate temperature of 20°C at night and 30oC
(86°F) during the day for a period of 4 weeks (Rudolf, 1974). Light apparently
is not required and tests may also be made in sand flats (Rudolf, 1974).
Deodar seed stratified at 4°C in moist sand for 30 days germinated 45%
versus 11% without stratification (Dirr and Heuser, 1987). Thapliyal and Gupta
(1980) also found percentage of germination without stratification to vary from
16 to 69 per cent. Singh et al. (1992) found that seed from larger-sized cones
exhibited higher seed germination (66%) in Himalayan cedar. Singh et al.
(1997) also found significant differences between tree diameter classes in
fresh and dry weight of seed, and germination in the laboratory and in the
nursery.
The seed of Cedrus deodara and C. libani species were stored during
the winter at various temperatures. The storage of the seed was made in
airtight PVC boxes at temperature of +5°C, –10°C and –20°C, as well as in a
basement at fluctuating temperature of +10°C/20°C. Storage in airtight boxes
and at temperature range +5°C to –10°C, were effective short-term storage
methods for both of the species. It must be pointed out that, during storage,
the seed became dormant that was successfully broken by cold stratification
at 5±1°C for 15 days. The common storage conditions (10°C/20°C) as well as
temperature lower than –10°C had a negative effect on germination of both
8
species. The soaking of the seed in water for 3 hours and the cold
stratification at 5±1°C for 15 or 30 days resulted in a higher seed germination
value (Takos and Merou, 2001).
Mughal and Thapliyal (2006) stored the seed of Cedrus deodara at four
different temperature namely 30oC, 15oC, -5oC and ambient room temperature
(which varied between 10oC to 17oC) with three different moisture level of 18,
14 and 10 per cent in sealed poly bag. It was found that seed stored at a
temperature of -5oC with a moisture content of 10 percent retained viability
even after 650 days from start of storage. At increased temperature and
moisture content, the viability period is negatively influenced. Viability
recorded at room temperature and at 15oC was very close. However, storage
at -5oC with a moisture content of 10 per cent significantly eclipsed other
combinations, thereby advocating storage of deodara seed at lower
temperature (-5oC) with low moisture content (10%) and simultaneously
providing it to be sub orthodox nature.
Seed of Cedrus libani species could be stored at moisture content of
7.9 per cent for three year at -5oC with 15-30 per cent reduction in
germination, whereas there was no significant difference between containers
(Erkuloglu, 1995). Cedrus atlantica seed stored for 3 year at temperature of
3oC in container and germination was found around 50 per cent (Piotto and
Gradi, 1998). Normal germination of Cedrus deodara is 70-80 per cent. Seed
stored in gunny bags in cool and dry place for one year gave about half of this
germination percentage (Kaushik et al., 1967). Seed of Abies and Cedrus can
be stored at moderate moisture content and low temperature. The moisture
and temperature during storage were (1) for period of 1-3 years, moisture and
temperature should be 12-13 per cent and - 4 to - 5oC (2) for period over 3
years, moisture and temperature should be 7-9 per cent and - 10 to – 20oC.
Khan et al. (2007) conducted nursery trial of Cedrus deodara and
Pinus helepensis with nine different dates spread over autumn, spring and
winter was conducted during 2002-2003. Germination of Cedrus was better
9
when seed were sown in the month of February. But in Pinus sowing dates
did not affect the germination upto first fortnight of February. Time taken to
completion of germination decreased as the sowing proceeded from autumn
to spring. The viability of seed was recorded 98 per cent for Cedrus and 91
per cent for Pinus. Seed during the intervening period of sowing were stored
in airtight poly bag in a refrigerator at a temperature of 3±1°C.
Seed dormancy in many conifers such as Abies alba, Abies densifolia,
Picea smithiana and Pinus densifolia can be overcome by cold stratification
and overwintering over periods of time from 21 to 90 days (Barton, 1930;
Edwards, 1962 and Singh and Singh, 1984). On the other hand, Nikolaeva
(1969) reported that conifer seed have generally intermediate physiological
dormancy and this is overcome by cold stratification for 14-21 days depending
upon species.
Bhardwaj et al. (2001) observed that seed of Ulmus leavigata kept in
polyethylene bag and stored at 5oC maintained higher viability with 62%
germination after 4 months of storage. Barner (1975) observed that the seed
of Alnus, Betula, Cupressus, Picea, Pinus and Thuja can be stored at
moisture content of 6-8.5% and temperature 2 to 4oC for 3-5 years.
Barton (1954) revealed that seed of Pinus ponderosa, Pseudostuga
menziessii and Tsuga heterophylla could be stored successfully in canvas
bag at sub-freezing temperature of -4oC, -11oC and -18oC for three years. The
experiment conducted by Gordon et al. (1972) indicated that Pinus merkusii
responded well to storage at low temperature. The storage temperatures of
2oC produced maximum germination of 80 per cent after three years of
storage while room temperature storage showed significant loss of
germination after 3-4 months of storage.
Similarly, Zlobin (1973) studied the effect of seed storage in Pinus
sylvestris, Picea abies and Larix sibirica in sealed metal container at different
temperatures viz., 18oC, 2oC, -3oC and natural temperature regime. The result
revealed best germination and germination energy in P. sylvestris after six
10
months storage at -3oC, followed by +2oC. However, L. sibirica was nearly
good at -3oC while the temperature regime of 18oC was found to be least
favourable in all the species.
Baldwin (1955) has recommended different levels for cold storage, e.g.
Pine (7.9%), Abies (6-7%), Picea (6-7%), Ulmus (3-7%), Thuja (8.0%) and
Betula (1-5%). Beside moisture, another important factor that determines the
longevity of seed is the temperature. Barner (1975) recommends moisture
content of 12-13 per cent in Abies for 1-3 years of storage. Similarly, for
satisfactory long term storage of Juniperus scopulorum a moisture content of
10-12 per cent was desirable (Strachan, 1990).
The experiment conducted by Vlase (1974) indicated that storage
temperature of 4.4oC for 4 years has no adverse affect on seed germination in
Pinus sylvestrris. According to Robbins (1983), Pinus oocarpa could retain
viability upto four years when seed were stored in sealed container at 0 to
5oC.
Barnett and Vozzo (1985) on the other hand observed 66 per cent
germination, when seed of Pinus elliottii were stored at 4oC for 50 years.
However, there was a total loss of germination in Shorea roxburghii seed after
10 days of storage at 20oC (Corbineau and Come, 1986). Similarly, Danielson
and Tanaka (1987) studied the effect of drying in Ponderosa pine and
Douglas-fir seed to three moisture levels and storing at 2oC. The air dried
seed were found to prolong the storage life of both species and resulting in
higher germination than oven dried and non dried seed.
On the other hand, Napier and Robbins (1987) studied the effect of
temperature and container type on germinability of Pinus roxburghii seed by
storing them in: a) sealed glass, b) thick polythene bag and c) cloth bag. They
found best results by storing the seed in a), followed by b) and c) conditions.
According to Haverbeka and Peterson (1989), viability was found to be
maximum in Pinus ponderosa seed stored in glass jar at -16oC (34-84%),
followed by 5oC (22-80%) for all the seed sources.
11
Donald and Jacobs (1990) studied the effect of storage in Pinus elliottii,
P. patula, P. radiata and P. taeda seed in linen bag and PVC container at
room temperature, 2-3oC and-16oC for 25 years. It was concluded that while
germination and germination capacity decreased rapidly at room temperature,
the cold storage of 2-3oC and -16oC maintained the germination and
germination capacity for 20 years in both the containers. The PVC container
was however found to be more efficient than linen bag in this regard.
The experiment conducted by Singh et al. (1992) in Chilgoza pine
revealed that germinability was greatly reduced with the reduction in the
biochemical properties when seed were stored in gunny bag at room
temperature. As the storage period increased, the biochemical and
germinability of seed were found to decrease accordingly. Similarly, Effendi
and Sinaga (1996) studied the effect of storage duration on redwood seed for
0, 2, 4, 6, 8, 10, 12, 14, 20, 22 and 24 months at 4oC. They recommended
four month storage for maximum germinability as germination capacity,
germination value and mean daily germination decreased rapidly after 4
month of storage.
Singh (1989) found that Picea smithiana cones collected from 2400
and 2700m elevation and stored for four weeks gave 49.4 and 43.4 per cent
germination, respectively as compared to seed extracted from fresh cones,
there germination was found 30.9 and 30.4 per cent, respectively.
Muller et al. (1999) on the other hand, indicated that seed of Douglas fir
and Pseudotsuga menziessii pre-treated and dried to 6.7 per cent moisture,
stored appreciably (15oC) upto 6 months without any detrimental effect on
germination. However, in another experiment, the seed pre-chilled for 18
weeks and stored at three different moisture levels of 6.7, 7.2 and 8.1 per cent
over a period of 17 months revealed that the seed stored at the lowest
moisture content germinated faster and to the highest percentage as
compared to other treatments. The similar results were recorded under
nursery conditions.
12
Phartyal et al. (2001) on the other hand, studied the effect of different
temperature regimes from 20 to 50oC and three relative humidity levels viz.,
11.2, 51.4 and 85.3 per cent in Ulmus wallichana seed. Their results indicated
that 16.2 per cent RH and 20oC temperature treatment resulted in good
viability for a longer period with higher vigour in the species. Similarly, Hilli et
al. (2003) studied the change in germination behaviour of pretreated Pinus
sylvestris seed at different incubation temperature for long term storage of
upto 10 years. The study revealed that germination indices of forest tree seed
whether pre-treated or not were preserved equally well in cold storage (2oC)
than frozen storage (-18oC).
Similarly, Gautam et al. (2005) stated that germination of chir pine seed
was significantly affected by various storage temperatures. Seed stored at
5±1oC resulted in maximum germinability than other temperature. Shiva et al.
(2006) indicated that seed of Aegle marmelos stored well in closed plastic
containers under ambient and 10-12oC temperatures revealing higher
germinability even 12 months after storage. The seed stored at low
temperature (0-5oC) were however found to be deleterious.
2.2 Physio-biochemical indices of seed
Study of physio-biochemical indices is a reliable and biologically sound
technique to understand the various internal changes in seed during storage
and stratification. Although the determination of these indices is a time
consuming proposition requiring special laboratory technique yet they have
marked effect on seed germination and consequent growth and development
of seedlings in forest tree species (Nancy et al., 2000).
Temayching (1966) studied the changes in weight, water content,
nucleic acid, nucleotides, carbohydrate, lipid, nitrogenous and phosphorus
compounds in embryo and gametophyte of Psudotsuga menziesii during
germination process and reported that lipids, proteins and phosphorus
compound were utilized for the synthesis of carbohydrate and other soluble
compounds in the seedling. Ching (1973) on the other hand, observed that
13
total sugar, reducing sugar, total nitrogenous and phosphorus compounds
increased in the embryo’s of Pinus taiwanensis and Cumninghamia lanceolata
seed during germination. Quantitative changes in sugars, α-keto organic acids
and amino acids were also observed in the seed of both the species.
Murphy (1985) studied the production of acetones at various stages of
seed germination in Pinus edulis, P. lambertiana and P. pinea and revealed
that acetone productions occurred predominantly in the embryo and embryo
axes of the seed. Similarly, Singh and Puri (1987) examined the seed of Pinus
roxburghii (eight seed stands of Himachal Pradesh) for starch, sugar, protein
and α-amylase contents and reported increase of starch content with the
increase in altitude. There was however negative but significant correlation in
protein levels with regards to altitude while the sugar content was also more
in seed stands of higher altitude.
Sehgal et al. (1989) conducted experiment on oil characteristics of
composite seed samples of eight seed stands of Pinus roxburghii. They
observed maximum oil and saponification value in Dhar-Chaubutra seed
stand situated at lower altitude. Schmeidar and Gifford (1995) on the other
hand, reported that moist stratification of 35 days at 2oC increased seed
germination from 19-76 per cent in Pinus taeda as the total lipid did not
change whereas, the protein content of both megagametophyte and embryo
was more variable. The rate of synthesis of buffer soluble proteins in these
two tissues, however, increased with the increase in stratification period.
Working with seed of Alnus firmifolia, Abies religiosa, Cedrela odorata,
Swietenia microphylla and Cordia eleagnoides storage at room temperature
and 5oC, Carritto et al. (1994) reported that except Abies religiosa, the per
cent seed oil declined during three months storage with greater decline at
room temperature than 5oC.
Stolyhwo and Janson (1999) working with Norway spruce observed a
small decrease in fat content over extended storage from initial 36.6 per cent
of dry mass to 33.5 per cent and 32.5 per cent after 15 and 20 years,
14
respectively. Similarly, germination capacity decreased from 90 per cent in
freshly collected seed to 70 per cent and 30 per cent after 15 to 20 years,
respectively. Janice et al. (2002) reported that moist chilling of Pinus taeda
seed did not affect the embryo’s ability to mobilize storage protein.
Similarly, Gautam (2005) on the other hand studied the biochemical
contents of different seed stands of Pinus roxburghii in Himachal Pradesh and
concluded that oil content; acid value, saponification value, total sugar, total
phenol and soluble proteins were different among different seed stands.
Shivani (2003) while working with Abies pindrow and Picea smithiana seed
reported that total sugar, reducing sugar and soluble proteins increased
steadily upto 60 days of wet or dry stratification and thereafter all the
biochemical contents showed a declining trend.
Kao and Rowan (2005) indicated that stratification of Pinus radiata at
0oC accelerated subsequent germination with increase in organic acids,
sucrose and organic phosphate while, lipase and invertase had low activities
and did not increase during stratification treatment. Sofi and Bhardwaj (2007)
while subjecting the seed of Cedrus deodara to different stratification periods
of 0, 15, 30, 45, 60 and 90 days revealed an increase in the soluble proteins
and total sugar content over that of control upto 60 days while, the starch
content showed a continuous decline with successive stratification.
Moisture content is probably the most important single factor in
determining seed longevity, and almost all seed that remain viable for more
than one year can withstand considerable drying without injury. Maintenance
of a constant low moisture content secured by preliminary drying is the most
suitable method for prolonged storage of most tree seed. Various worker have
established critical moisture content for seed above which viability is rapidly
lost and below which it is retained for considerable period (Holmes and
Buszewiz,1958) within a few per cent below the critical level, the actual
moisture content has little effect on keeping seed quality. However any
15
increase above this level greatly accelerates respiration and the rate of seed
deterioration (Barton, 1941).
Baldwin (1955) has recommended different moisture levels for cold
storage, e.g. pine (7.9%), Abies (6-7%), Picea (6-7%), Ulmus (3-7%), Thuja
(8.0%) and Betula (1-5%). High moisture content at the time of dispersal
affect storage under normal storage conditions (Khan et al., 2007).
Khan et al. (2007) stored the seed of Cedrus at three different moisture
contents viz., 10, 14, and 18 per cent and maximum germination was
recorded in those seed having moisture content of 10 per cent. At higher
moisture content seed (18%) seed lost its viability.
Barner (1975) recommended moisture content of 12-13 per cent in
Abies for 1-3 years of storage; similarly, for satisfactory long term storage of
Juniperous scopuloruma moisture content of 10-12 per cent was desirable.
If cedar seed dried below a critical level, they will not imbibe water in a
way that will allow the food reserve to be used by the embryo (Macdonald,
1986). Matziris (1998) noted that moisture content ranging between 12-13 per
cent is better for storage of Cedrus seed for 1-5 years. Cedar seed retained
viability for 3-6 years when dried to a moisture content of less than 10 per
cent (Rudolf, 1974 and Erkuloglu, 1995).
Muller et al. (1999) indicated that seed of Douglus fir pretreated and
dried to 6.7 per cent moisture and stored upto six months without any
detrimental effect on germination.
Chapter-III
MATERIALS AND METHODS
The present investigation entitled “Effect of storage conditions on
germinability of Himalayan cedar (Cedrus deodara Loud.) seeds” was
conducted in the laboratory of the department of Silviculture and Agroforestry,
Dr Y.S. Parmar University of Horticulture & Forestry, Nauni, Solan, Himachal
Pradesh during the year 2006-2007. The details about the experimental site,
materials used and methodology adopted are given in this chapter.
3.1 EXPERIMENTAL SITE
3.1.1 Location
The experiments were conducted in departmental laboratories of the Dr
Y.S. Parmar University of Horticulture & Forestry, Nauni, Solan located at
30o51' N latitude and 76o11' E longitude at an altitude of 1250 m above mean
sea level. The place lies 14km south east of Solan town of Himachal Pradesh
on Solan-Rajgarh road.
The climate of the area ranges from sub-tropical to sub-temperate and
experience a mean precipitation of 928.4 mm per annum during the study
period. The major part of rain is received during July and August (Monsoon
period) months. Winter showers though common, frost occurs recurrently from
December to February. Snow fall is also experienced in alternate year. In
general, May and June are the hottest, while December and January form the
coldest months.
17
3.2 DETAILS OF EXPERIMENT
3.2.1 Seed collection and extraction
Seed of Cedrus were collected from Salooni forests in Chamba district
of Himachal Pradesh during October, 2006. Cones were collected and seed
were extracted. Thereafter seed were packed in polythene bag and
transported to the laboratory of department of Silviculture and Agroforestry, Dr
Y.S. Parmar University of Horticulture & Forestry, Nauni, Solan for
undertaking germination test and physio-biochemical analysis. Total 2.0 kg of
seed was collected and 1.4 kg was utilized for the research purpose.
3.2.2 Technical programme of work
A) Storage temperature
Storage was done at four different temperatures given below:
T1 : Room temperature
T2 : 5±1oC
T3 : 0±1oC
T4 : -5±1oC
B) Storage container
The following types of storage containers were used:
C1 : Poly bag
C2 : Canvas bag
C3 : Plastic container
C) Storage duration
Total storage duration was ten months and its effect on seed viability
observed at bimonthly intervals.
D1 : Two months of storage
18
D2 : Four months of storage D3 : Six months of storage D4 : Eight months of storage D5 : Ten months of storage
Experimental Design : Split plot design
Main plot factor : Temperature
Sub plot factor : Container
3.2.3 Germination Test
The germination test was carried out by placing the seed on moist filter
paper in germination tray to be kept in germinator at 30oC. For that a sample
of 300 seed divided into three replications of 100 seed each was used to run
the germination tests. Germinator was fitted with specially designed chamber
with the control of temperature, humidity and light. Germination test was
conducted at bimonthly interval. The seed were counted as germinated when
radical emerged about 2mm. The testing period was 21 days.
3.2.4 Viability test
The viability of seed was determined by using Tetrazolium test (Tz) by
taking 100 seed in each replication (Bonner, 1974). The seed with completely
stained (red) embryos and other tissues were considered viable
3.3 OBSERVATIONS RECORDED
3.3.1 Germination studies
The following germination parameters were recorded under laboratory
conditions.
3.3.1.1 Germination per cent
19
Germination per cent was calculated as the number of seed taken and
number of seed germinated, expressed in percentage.
3.3.1.2 Germinative capacity
The cumulative number of seed that germinated during the 21 days of
test period plus the number of ungerminated viable seed at the end of the test
expressed in percentage.
3.3.1.3 Germinative energy
Germinative energy (GE) was calculated on the basis of the
percentage of the total number of seed that had germinated when the
germination reached its peak generally taken as the highest number of
germination in 24 hours period.
Number of seed germinated upto time of peak germination GE (%) = x 100
Total number of seed taken 3.2.1.4 Germination value
Germination value (GV) is the index combining speed and
completeness of seed germination. Daily germination counts were recorded
and calculated as per Czabator (1962).
GV = PV x MDG
Where,
PV = Peak value of germination
MDG = Mean daily germination
3.3.2 Physio-biochemical studies
3.3.2.1 Moisture content
The moisture content expressed in percentage on fresh weight basis was
determined by the following formula (low constant temperature oven
methodgivenbyWillian,1985).
20
Original weight – Oven dry weight
Moisture content (%) = x 100
Original weight
3.3.2.2 Total sugar and starch
3.3.2.3 Extraction of total sugar and starch
One gram of dried sample were filtered placed in 20-25 ml of boiling
ethanol (80%) for 10 minute and decanted. Another 10-15 ml of boiling
ethanol was added to the residue. Thereafter, the two extracted samples were
filtered and combined. The final volume was made to 50 ml. The alcoholic
extract was then used for the estimation of total sugar while the residue for
the determination of starch.
3.3.2.4 Total sugar
Total sugar was estimated by phenol-sulphuric acid method given by
Dubois et al. (1951).
3.3.2.5 Reducing sugar
Reducing sugar was established by di-nitrosalicylic acid method
developed by Miller (1972).
3.3.2.6 Non reducing sugar
The content of non reducing sugar was calculated by deducting the
quantity of reducing sugar from that of the total sugar and then multiplied by
the factor 0.95.
3.3.2.7 Starch
Glucose in the sample was determined by phenol-sulphuric acid
method of Dubois et al. (1951) and then starch content was calculated by
multiplying the glucose value with conversion factor of 0.90.
21
3.3.2.8 Total Phenol
Total phenol was estimated with Folin phenol reagent method of Bray
and Thorpe (1954)
3.4 STATISTICAL ANALYSIS
The entire data generated from the present investigation were
subjected to statistical analysis as per methods described by Gomez and
Gomez (1984).
Chapter-IV
EXPERIMENTAL RESULTS
The present investigation entitled “Effect of storage conditions on
germinability of Himalayan cedar (Cedrus deodara Loud.) seeds” were
carried out during the year 2006-07 to determine the longevity of seed under
different storage conditions. The results obtained on different aspects are
presented as under:
4.0 Storage of Cedrus deodara seed
4.1 Germination parameters
4.2 Physio-biochemical parameters
4.0 STORAGE OF Cedrus deodara SEED
4.1 Germination parameters
4.1.1 Determination of germination parameters of freshly collected seed of Cedrus deodara
Data in Table 1 reveal that fresh Cedrus seed registered germination
success of 70 per cent, germinative capacity of 94 per cent, germinative
energy of 45 per cent and germination value of 10.50.
Table 1: Germination parameters of freshly collected seed of Cedrus
deodara
Germination (%)
Germinative capacity (%)
Germinative energy (%)
Germination value
70.00
94.00 45.00 10.50
After the initial study, the germination parameters of the seed were
examined at bimonthly interval viz., two (D1), four (D2), six (D3), eight (D4)
and ten month (D5) of storage period.
23
4.1.2 Effect of temperature on per cent germination and germinative capacity of Cedrus deodara seed during storage
Seed of Cedrus deodara were stored under four different temperature
namely room temperature (T1), 5±1oC (T2), 0±1oC (T3) and -5±1oC (T4). Data
pertaining to germination parameters viz., germination per cent, germinative
capacity of stored seed are presented in Table 2. It is quite evident that
germination parameters manifested significant effect under different storage
temperature.
A perusal of data in Table 2 indicates that per cent germination
increased upto four months’ (D2) storage under all temperature except for
room temperature (T1), thereafter a steep decline in germination values
occurred till the termination of experiment after ten months’ (D5) storage
under all temperature. However seed stored at -5±1oC (T4) gave the highest
mean germination of 71.18 per cent after ten months’ (D5) storage. On the
other hand, the lowest germination of 53.51 per cent was observed after ten
months’ (D5) storage at room temperature (T1).
The maximum germinative capacity of 94.00 per cent was observed
immediately after harvest of seed (Table 1). However the data in the Table 2
indicate that there was a decline in values till the end of storage period under
all temperatures. The maximum mean germinative capacity of 79.40 per cent
was registered under temperature -5±1oC (T4) and the minimum (68.20 %) at
room temperature (T1) after ten months’ (D5) of storage.
4.1.3 Effect of temperature on per cent germinative energy and
germination value of Cedrus deodara seed during storage Data on per cent germinative energy (Table 3) indicate that per cent
germinative energy increased upto four months’ (D2) of storage under all
temperatures except for room temperature (T1), thereafter a steep decline in
germinative energy occurred till the termination of experiment after ten
months’ (D5) storage under all temperatures. However seed stored at -5±1oC
(T4) gave the highest germinative energy of 46.33 per cent after ten months’
24
25
26
(D2) storage. Whereas the lowest mean value of 33.33 per cent was observed
after ten months’ (D5) storage at room temperature (T1).
Data on germination value (Table 3) which also shows an identical
pattern of increase and decrease in their values like per cent germination,
capacity and energy. Peak germination value of 15.77 was recorded at the
end of four months’ (D2) storage of seed at -5±1oC (T4) temperature. After
four months’ (D2) storage, with the advancement of storage period there was
a corresponding decline of germination value. However, the germination value
declined faster at room temperature (T1) in comparison to those stored at-
5±1oC (T2), 0±1oC (T3) and -5±1oC (T4).
4.1.4 Effect of container on per cent germination and germinative
capacity of Cedrus deodara seed during storage
Three type of container namely poly bag (C1), canvas bag (C2), and
plastic container (C3) were assessed for their influence on germination
parameters of Cedrus deodara seed in storage. Data pertaining to
germination parameters viz., germination per cent and germinative capacity of
stored seed are represented in Table 4. The germination parameters were
significantly affected by different storage containers.
All type of containers significantly influenced the per cent germination
during storage. At the end of four months’ (D2) storage the highest per cent
germination of 80.08 was recorded in plastic container (C3) and the lowest of
73.92 per cent in seed stored in canvas bag (C2). Plastic container (C3)
maintained its superiority till the end of ten months (D5) when seed drawn
from it exhibited 49.67 per cent germination which was significantly higher
than poly bag (C1). On the other hand, the lowest per cent germination of
39.83 per cent was recorded in seed taken from canvas bag (C2).
It is evident from Table 4 that the highest mean per cent germinative
capacity of 76.92 per cent was determined in seed drawn from plastic
container (C3) and the lowest of 72.25 per cent from canvas bag (C2).
27
Per cent germinative capacity decreased till the termination of experiment
after ten months’ (D5) storage under all containers.
4.1.5 Effect of container on per cent germinative energy and
germination value of Cedrus deodara seed during storage
A perusal of values in Table 5 indicates that the seed germinative
energy showed a pattern of increase upto four months’ (D2) storage in seed
stored in different container except canvas bag (C2). Thereafter germinative
energy sharply declined till ten months’ storage (D5) in seed stored in different
containers. However, the highest mean germinative energy of 43.65 per cent
was exhibited by the seed stored in plastic container (C3) and the lowest of
37.18 per cent was registered by the seed stored in canvas bag (C2) at the
end of experiment.
An inquisition of data in Table 5 reveals that different containers
significantly influenced the germination value during storage. With the
advancement of storage, there was an increase in germination value upto four
month (D2) and thereafter a continuous decrease upto ten month (D5).
However the seed drawn from plastic container (C3) reflected continuous
higher germination value as compared to poly bag (C1) and canvas bag (C2).
4.1.6 Interaction effect of temperature and container (TxC) on the
germination parameters of Cedrus deodara seed during storage
Seed of Cedrus deodara were stored under four different temperature
namely, room temperature (T1), 5±1oC (T2), 0±1oC (T3) and -5±1oC (T4) and in
three type of container namely, poly bag (C1), canvas bag (C2), and plastic
container (C3) were assessed for their influence on germination parameters of
Cedrus deodara seed in storage. Interaction due to temperature and container
(TxC) was found to be significant except for two months of storage (D1).
An overview of Table 6 reveals that the maximum mean per cent
germination of 75.20 resulted when seed were stored at -5±1oC in plastic
container (T4C3) which was closely followed by T3C3 (73.07 %) at the end of
28
29
30
Table 6: Interaction effect of temperature and container (T x C) on per cent germination of Cedrus deodara seed during storage
Treatment
Duration (month)
D1 D2 D3 D4 D5 Mean
T1C1
T1C2
T1C3
T2C1
T2C2
T2C3
T3C1
T3C2
T3C3
T4C1
T4C2
T4C3
Mean
SEm +
CD0.05
66.67
62.00
68.67
76.67
75.33
78.67
82.33
77.67
83.33
82.33
82.33
83.33
77.31
-
NS
64.00
61.00
65.67
78.33
77.67
81.67
84.33
79.33
85.67
85.00
81.67
87.33
76.61
0.40
0.97
55.00
52.33
57.67
63.67
62.33
66.33
70.33
65.67
72.67
70.67
69.67
76.67
65.25
0.18
0.43
48.67
45.00
49.67
53.33
51.33
60.00
32.67
56.33
67.33
64.33
60.33
69.33
57.36
0.72
1.76
36.33
31.67
38.33
41.67
39.67
44.67
49.67
42.33
56.33
49.67
45.67
59.33
44.61
0.90
2.10
54.13
50.40
56.00
62.73
60.47
66.27
69.87
64.27
73.07
70.40
67.93
75.20
31
ten months’ (D5) storage. On the other hand, minimum mean value of 50.40
per cent was found in seed stored in canvas bag at room temperature (T1C2).
Data on interaction effects of temperature and container (TxC) on per
cent germination capacity are given in Table 7. It is seen that there was a
clear cut pattern of decrease in values as the seed coursed through storage
period. The seed stored in plastic container at -5±1oC (T4C3) recorded the
highest mean germinative capacity of 82.73 per cent at the end of ten months’
(D5) storage. Whereas the lowest mean value of 66.13 per cent was recorded
in seed stored in canvas bag at room temperature (T1C2) after ten months’
storage (D5).
Comparison of storage temperature with individual container level
(TxC) for seed germinative energy (Table 8) reveals that in general there was
an increase in germinative energy upto four month’ (D2) storage except in
seed stored in different container at room temperature and thereafter, a
continuous decrease in energy was observed upto ten months’ (D5) storage
irrespective of temperature and container. However, seed stored in plastic
container at -5±1oC (T4C3) registered the maximum mean germinative energy
of 50.67 per cent. The minimum mean germinative energy (31.33 %) on the
other hand was recorded from seed stored in canvas bag at room
temperature (T1C2).
Data on interaction effect of temperature and container (TxC) on
germination value are presented in Table 9. Data reveal a gradual increase in
germination value upto four months’ (D2) storage irrespective of temperature
and container. Past four months’ (D2) storage, there was a constant decrease
in values upto ten months’ (D5) storage. The highest mean germination value
of 12.69 was recorded from seed stored in plastic container at -5±1oC (T4C3).
Whereas, the lowest mean germination value (6.56) was registered in seed
stored in canvas bag at room temperature (T1C2).
32
Table 7: Interaction effect of temperature and container (T x C) on per cent germinative capacity of Cedrus deodara seed during storage
Treatment
Duration (month)
D1 D2 D3 D4 D5 Mean
T1C1
T1C2
T1C3
T2C1
T2C2
T2C3
T3C1
T3C2
T3C3
T4C1
T4C2
T4C3
Mean
SEm +
CD0.05
82.33
80.67
82.33
84.67
84.33
86.00
87.67
85.00
88.33
88.00
86.33
92.67
85.69
-
NS
77.60
74.67
78.00
80.00
78.00
83.33
84.33
82.33
87.33
86.33
84.00
89.67
82.14
-
NS
71.00
68.33
73.00
76.33
74.33
78.67
79.67
77.33
82.33
80.67
79.33
85.67
77.22
0.23
0.56
62.00
59.33
64.00
68.00
65.67
69.00
71.33
68.67
74.33
72.00
71.33
77.00
68.56
0.18
0.46
50.00
47.67
52.00
57.33
55.67
59.67
65.00
58.00
67.33
65.33
64.00
68.67
59.22
0.26
0.63
68.60
66.13
68.97
73.27
71.60
75.33
77.60
74.67
79.93
78.47
77.00
82.73
33
Table 8: Interaction effect of temperature and container (T x C) on per cent germinative energy of Cedrus deodara seed during storage
Treatment
Duration (month)
D1 D2 D3 D4 D5 Mean
T1C1
T1C2
T1C3
T2C
T2C2
T2C3
T3C1
T3C2
T3C3
T4C1
T4C2
T4C3
Mean
SEm +
CD0.05
42.33
41.00
42.33
45.33
43.33
47.67
49.67
46.33
43.33
49.67
46.33
52.00
48.39
-
NS
40.33
38.33
40.33
49.67
47.67
49.33
53.33
48.67
57.67
55.00
50.33
63.33
43.53
1.17
2.85
34.33
32.67
36.67
38.67
37.33
42.67
46.33
39.33
50.33
48.67
44.00
54.67
42.17
1.13
2.75
28.33
24.33
30.67
33.33
31.33
35.67
38.00
36.33
43.67
42.67
37.67
45.00
35.39
0.623
1.52
22.67
20.33
23.67
27.33
25.33
31.33
32.33
29.33
36.67
37.33
33.67
39.67
39.17
1.36
3.31
33.60
31.33
35.07
37.00
36.13
41.33
43.93
39.00
47.53
46.07
42.27
50.67
34
Table 9: Interaction effect of temperature and container (T x C) on germination value of Cedrus deodara seed during storage
Treatment
Duration (month)
D1 D2 D3 D4 D5 Mean
T1C1
T1C2
T1C3
T2C1
T2C2
T2C3
T3C1
T3C2
T3C3
T4C1
T4C2
T4C3
Mean
SEm +
CD0.05
9.76
9.14
9.77
11.32
11.02
12.97
13.44
11.32
14.40
13.64
13.29
15.37
13.44
-
NS
10.35
10.16
10.60
11.80
11.51
14.41
15.34
13.40
16.40
15.98
14.64
16.67
12.12
0.12
0.29
7.26
7.00
8.89
9.20
9.17
9.92
10.95
9.79
10.33
11.19
10.32
13.75
9.96
0.06
0.14
5.17
4.11
5.23
6.50
6.16
7.69
8.36
7.40
8.89
8.39
8.32
10.35
7.22
0.12
0.29
2.89
2.36
3.39
4.72
4.38
5.26
5.64
5.11
6.54
5.94
5.34
7.21
4.92
0.06
0.15
7.29
6.56
7.45
8.71
8.45
10.05
10.81
9.40
11.53
11.03
10.39
12.69
35
4.2 Physio-biochemical parameters
4.2.1 Estimation of physical and biochemical parameters of freshly
collected seed of Cedrus deodara The results of physical and biochemical attributes of C. deodara seed
after harvest are presented here under:
Table 10: Physio-biochemical parameters of freshly collected seed of
Cedrus deodara
Moisture content (%)
Total sugar (%)
Reducing
sugar (%)
Non-
reducing
sugar (%)
Starch
(%)
Phenol
(mg/g)
22.00 8.50 1.68 6.60 11.70 39.50
The appraisal of the physio-biochemical attributes of the seed
immediately after harvest (Table 10) reveals that the moisture content and
total sugar was to the tune of 22.00 and 8.50 per cent, respectively. The
reducing and non reducing sugars were 1.68 and 6.60 per cent, respectively.
The starch per cent estimated was 11.70 and phenol to the level of 39.50
mg/g.
4.2.2 Effect of temperature on moisture content and total sugar of
Cedrus deodara seed during storage
Seed of Cedrus deodara were stored under four different temperatures
namely room temperature (T1), 5±1oC (T2), 0±1oC (T3) and -5±1oC (T4). Data
pertaining to physio-biochemical parameters viz., moisture content and total
sugar of stored seed are presented in Table 11. A critical review of data in
Table 11 reveals that storage temperature exerted significant effect on
moisture content and total sugar in Cedrus seed.
It is clear from data in Table 11 that moisture content of seed was
significantly affected by different temperature. There was continuous
decrease in moisture content from first month till the termination of
experiment. However, the highest mean value of 12.43 per cent was found in
36
seed stored at -5±1oC (T4), followed by 0±1oC (T3), 5±1oC (T2) and the lowest
content of 8.99 per cent at room temperature (T1) at the termination of
experiment.
A scrutiny of data in Table 11 reflects that total sugar content of seed
was significantly affected by temperature. There was an increasing trend in
total sugar after harvest of seed upto four months’ (D2) storage and
thereafter it declined till the termination of the experiment under all
temperature except for room temperature (T1) where seed showed a
continuous decrease in total sugar right from seed harvest upto ten months’
(D5) storage. However, seed stored at -5±1oC (T4) registered the highest
mean sugar content of 8.44 per cent. The minimum mean sugar content of
6.29 per cent was recorded at room temperature (T1).
4.2.3 Effect of temperature on reducing sugar and non reducing sugar
of Cedrus deodara seed during storage
A perusal of data in Table 12 indicates that reducing sugar increased
upto four months’ (D2) storage under all temperature except for room
temperature (T1), thereafter a decline in reducing sugar values occurred till the
end of experiment after ten months’ (D5) storage under all temperature.
However seed stored at -5±1oC (T4) gave the maximum mean reducing sugar
of 1.64 per cent. On the other hand, the lowest mean reducing sugar of 1.13
per cent was observed after ten months’ (D5) storage at room temperature
(T1).
A scrutiny of data in Table 12 reflects that non reducing sugar contents
of seed were significantly affected by temperature. There was an increasing
trend in non reducing sugar from seed harvest to four months’ storage (D2)
after that it decreased till the termination of experiment. However seed stored
at -5±1oC (T4) registered the highest mean sugar content of 6.47 per cent.
The lowest mean non reducing sugar contents of 4.89 per cent were observed
at room temperature (T1).
37
38
39
4.2.4 Effect of temperature on starch and phenol of Cedrus deodara seed during storage
Data in respect of starch are given in Table 13. The highest starch
content (10.91%) was estimated in seed kept at -5±1oC (T4), followed by
0±1oC (T3). The lowest contents (8.56%) on the other hand, were found in
seed kept at room temperature (T3). The starch content decreased with the
corresponding increase of the storage period from two (D2) to ten months’
(D5) under all temperature conditions.
It is seen from Table 13 that phenol contents estimated during storage
differ significantly under all temperature. A comprehensive view of Table
reveals that the highest mean total phenol of 32.80 mg/g were found in seed
stored at -5±1oC (T4). The lowest values of 28.44 mg/g were obtained under
room temperature (T1). The phenol content decreased with the advancement
of storage period under all storage conditions.
4.2.5 Effect of container on moisture content and total sugar of
Cedrus deodara seed during storage The seed were stored in three different containers and their impact on
moisture content and total sugar was assessed.
It is crystal clear from Table 14 that moisture content differs
significantly in seed stored in different containers. There was a declining trend
for moisture content throughout the storage period in all containers. It is clear
from mean values that the highest mean moisture content of 11.70 per cent
was estimated in seed stored in plastic container (C3), followed by poly bag
(C1) and minimum (10.23%) in canvas bag (C2).
An appraisal of data in Table 14 reflects that total sugar contents of
seed were significantly affected by containers. There was an increasing trend
in total sugar from first month to four months’ (D2) storage except for canvas
bag after that there was decrease in total sugar till the termination of
experiment in all containers. The highest mean sugar content of 7.99 per cent
was obtained when seed were stored at -5±1oC (C3), followed by poly bag
40
41
42
(C1) and canvas bag (C2). The lowest mean sugar contents of 7.20 per cent
were observed in canvas bag (C1).
4.2.6 Effect of container on reducing sugar and non reducing sugar of
Cedrus deodara seed during storage
Going through the values of reducing sugar in Table 15 it becomes
clear that the highest value of 1.53 per cent was determined in seed drawn
from plastic container and the lowest of 1.40 per cent in seed taken from
canvas bag after four months’ (D2). The same trend continued ten months’
(D5) storage. Inspite of increase in value upto four months’ (D2) storage
irrespective of container type and then declined till the termination of
experiment. But after ten months’ storage, the mean reducing sugar of 1.53
per cent was registered in seed drawn from plastic container (C3) which in turn
was the highest value. On the other hand, seed taken from canvas bag (C2)
gave the lowest reducing sugar value of 1.40 per cent.
It is clear from data in Table 15 that non reducing sugar contents of
seed were significantly affected by container. There was an increase in non
reducing sugar upto two months’ (D1) storage and thereafter it exhibited a
continuous decrease with the advancement of storage period upto termination
of experiment. However, seed stored in plastic container (C3) registered
higher non reducing sugar than those stored in poly bag (C1) and canvas bag
(C2). The highest mean values of 6.14 per cent were found in seed stored in
plastic container (C3) followed by poly bag (C1) and the lowest contents of
5.52 per cent in seed drawn from canvas bag (C2).
4.2.7 Effect of container on starch and phenol contents of Cedrus
deodara seed during storage
Data in the Table 16 indicate that storage container exhibited a
significant effect on starch and phenol content of seed.
A perusal of data in the Table 16 indicates that seed exhibited the
maximum mean value of 10.30 per cent when seed stored in plastic container
(C3) followed by poly bag (C1) and the minimum mean value of 9.69 per cent
43
44
45
in canvas bag (C1). The starch contents were found to decrease continuously
from the day of seed harvest till the termination of experiment after ten
months’ (D5) storage.
Data regarding phenol reveal a gradual decrease in value till the
termination of experiment (Table 16). The highest mean phenol contents of
31.19 mg/g and the lowest of 30.50 mg/g were estimated in seed drawn from
plastic container (C3) and canvas bag (C2), respectively.
4.2.8 Interaction effect of temperature and container (TxC) on
moisture content and total sugar contents of Cedrus deodara seed during storage
Seed of Cedrus deodara were stored under four different temperature
namely, room temperature (T1), 5±1oC (T2), 0±1oC (T3) and -5±1oC (T4) and in
three type of container namely, poly bag (C1), canvas bag (C2), and plastic
container (C3) were assessed for their influence on moisture contents and
total sugar of Cedrus deodara seed in storage.
A perusal of data in Table 17 reveals that the interactions between
temperature and container were found to be significant. The highest mean
moisture content of 12.92 per cent was estimated when seed were stored at -
5±1oC in plastic container (T4C3) throughout the storage period. It was closely
followed by T4C1 (12.56%) and T3C3 (12.20%) treatment combinations.
Whereas, the seed stored in canvas bag at room temperature (T1C2)
registered the lowest mean moisture content of 8.10 per cent. Data in the
Table 17 reveal that moisture content declined with the corresponding
increase in storage period.
A comprehensive overview of the data in Table 18 indicates that the
total sugars were low in the beginning, increased during the early period of
storage to peak at D2, and there after declined towards the end of storage.
The highest total sugar was estimated when seed were stored at -5±1oC in
plastic container (T4C3) throughout the storage period. It was closely followed
by T4C1 and T3C3 treatment combinations. The lowest mean sugar contents of
46
Table 17: Interaction effect of temperature and container (T x C) on Moisture content (%) of Cedrus deodara seed during storage
Treatment
Duration
D1 D2 D3 D4 D5 Mean
T1C1
T1C2
T1C3
T2C1
T2C2
T2C3
T3C1
T3C2
T3C3
T4C1
T4C2
T4C3
Mean
SEm +
CD0.05
11.10
10.06
11.40
13.10
11.01
13.30
13.20
12.50
13.50
13.60
12.90
13.80
12.44
0.07
0.17
10.60
9.55
10.90
12.50
10.55
12.80
12.60
12.00
12.90
13.10
12.40
13.30
11.93
0.06
0.14
9.70
8.65
10.10
12.50
10.15
12.40
12.05
11.65
12.45
12.80
12.00
13.10
11.44
0.10
0.24
8.40
7.15
8.80
11.30
9.25
11.50
12.35
10.75
11.55
12.20
11.40
12.60
10.52
0.002
0.005
6.50
5.10
6.90
9.90
7.85
10.60
10.10
9.30
10.60
11.10
10.50
11.80
9.19
0.0022
0.0054
9.26
8.10
9.62
11.80
9.76
12.12
11.86
11.24
12.20
12.56
11.84
12.92
47
Table 18: Interaction effect of temperature and container (TxC) on total sugar (%) of Cedrus deodara seed during storage
Treatment
Duration (month)
D1 D2 D3 D4 D5 Mean
T1C1
T1C2
T1C3
T2C1
T2C2
T2C3
T3C1
T3C2
T3C3
T4C1
T4C2
T4C3
Mean
SEm +
CD0.05
8.43
8.04
8.50
8.78
8.56
8.96
9.04
8.90
9.82
9.20
9.02
9.88
8.97
-
NS
7.54
7.14
8.05
8.88
8.68
9.12
9.57
9.10
9.87
9.68
9.48
10.54
8.89
0.072
0.176
7.10
6.26
7.13
7.98
7.37
8.36
8.46
8.10
9.12
8.74
8.42
9.30
8.04
0.009
0.021
5.42
4.56
5.45
6.95
6.68
7.12
7.56
7.10
7.56
7.12
7.94
8.10
6.80
0.14
0.34
3.60
3.10
3.90
5.23
4.45
5.86
6.10
5.25
6.40
5.35
5.90
6.89
5.27
0.09
0.22
6.42
5.82
6.42
7.56
7.15
7.88
8.14
7.69
8.52
8.24
8.15
8.94
48
5.82 per cent were estimated in seed drawn from canvas bag at room
temperature (T1C2).
4.2.9 Interaction effect of temperature and container (TxC) on reducing
sugar and non reducing sugar contents of Cedrus deodara seed during storage
From Tables 19 and 20 it appears that storage temperature and
storage container exerted significant effect on reducing sugar and non
reducing sugar of C. deodara seed except for two (D2) and four months’ (D5)
of storage.
Reducing sugars demonstrated an increasing and decreasing pattern
similar to total sugars, irrespective of temperature and container (Table 19). It
is seen from data that the highest reducing sugar contents were estimated in
seed stored in plastic container at -5±1oC (T4C3), followed by (T3C3), (T4C1)
and (T3C1) in descending order of their values. The lowest mean value of 1.07
per cent was recorded by T1C2 and the highest mean contents of 1.72 per
cent in seed stored under T4C3 at the end of storage period.
It is evident from Table 20 that there was an increase in non reducing
sugar irrespective of container and temperature upto to four months’ (D2)
storage and a decline occurred thereafter till the termination of storage (D5).
The seed stored at -5±1oC and in plastic container (T4C3) continued their
supremacy in maintaining the highest mean non reducing sugar content of
6.85 per cent right from two months (D1) upto ten months’ (D5) storage, which
was followed by T3C3 than the remaining treatment combinations. The seed
stored in canvas bag and at room temperature (T1C2) registered the minimum
value of 4.50 per cent.
49
Table 19: Interaction effect of temperature and container (T x C) on reducing sugar (%) of Cedrus deodara seed during storage
Treatment
Duration (month)
D1 D2 D3 D4 D5 Mean
T1C1
T1C2
T1C3
T2C1
T2C2
T2C3
T3C1
T3C2
T3C3
T4C1
T4C2
T4C3
Mean
SEm +
CD0.05
1.55
1.50
1.63
1.70
1.68
1.75
1.82
1.72
1.85
1.84
1.78
1.90
1.87
-
NS
1.42
1.32
1.48
1.86
1.78
1.90
2.08
1.92
2.18
2.14
2.02
2.30
1.73
-
NS
1.20
1.10
1.25
1.52
1.50
1.57
1.64
1.54
1.67
1.66
1.60
1.72
1.50
0.004
0.010
0.94
0.85
0.98
1.24
1.20
1.32
1.36
1.28
1.40
1.38
1.32
1.48
1.23
0.0043
0.010
0.52
0.60
0.85
0.68
0.92
0.98
1.08
1.26
1.12
1.12
1.15
1.22
0.10
0.30
0.07
1.13
1.07
1.20
1.43
1.42
1.52
1.60
1.55
1.64
1.63
1.57
1.72
50
Table 20: Interaction effect of temperature and container (T x C) on non reducing sugar (%) of Cedrus deodara seed during storage
Treatment
Duration (month)
D1 D2 D3 D4 D5 Mean
T1C1
T1C2
T1C3
T2C1
T2C2
T2C3
T3C1
T3C2
T3C3
T4C1
T4C2
T4C3
Mean
SEm +
CD0.05
6.53
6.21
6.52
6.72
6.53
6.84
6.85
6.82
7.19
6.99
7.50
7.58
6.86
-
NS
5.80
5.52
6.24
6.66
6.55
6.77
7.11
6.82
7.30
7.16
7.08
7.82
6.74
-
NS
5.60
4.90
5.75
6.13
5.57
6.45
6.46
6.23
7.07
6.72
6.47
7.20
6.21
0.03
0.073
4.25
3.52
4.24
5.42
5.20
5.51
5.89
5.52
2.85
5.53
5.95
6.28
5.26
0.11
0.27
2.92
2.37
3.05
4.16
3.35
4.63
4.76
3.94
5.19
4.96
4.51
5.38
4.10
0.12
0.30
5.02
4.50
5.16
5.82
5.44
6.04
6.21
5.87
6.52
6.27
6.30
6.85
51
4.2.10 Interaction effect of temperature and container (TxC) on starch and phenol contents of Cedrus deodara seed during storage
The comprehensive overview of Table 21 and 22 show that starch and
phenol contents declined with the advancement of storage period under all
treatment combinations.
Data in the Table 21 indicate that seed starch contents were found to
be significant except for two and four months’ storage. The best results were
obtained in seed stored at T4C3. In other words the maximum mean starch of
11.12 per cent was found in seed stored in plastic container at -5±1oC (T4C3).
Whereas, the minimum mean starch contents of 8.28 per cent were registered
from seed stored at T1C2.
It is clear from the data in Table 22 that total phenol contents of 33.44
mg/g (mean) were found to be the highest in plastic container at -5±1oC
(T4C3), closely followed by T3C3 (32.35 mg/g) and T4C1 (32.62 mg/g). The
lowest mean phenol of 27.81 mg/g was found at room temperature in canvas
bag (T1C2).
4.2.11 Correlation of per cent germination with germinable and physio-
biochemical parameters of deodara seed.
From the matrix correlation as exhibited in Table 23, it is clear that the
per cent germination showed a negative correlation with starch content.
However, it was observed to have positive correlation with rest of the
germinable and physio-biochemical attributes.
52
Table 21: Interaction effect of temperature and container (T x C) on starch (%) of Cedrus deodara seed during storage
Treatment
Duration (month)
D1 D2 D3 D4 D5 Mean
T1C1
T1C2
T1C3
T2C1
T2C2
T2C3
T3C1
T3C2
T3C3
T4C1
T4C2
T4C3
Mean
SEm +
CD0.05
11.50
11.12
11.60
12.01
11.75
12.19
12.54
12.12
12.80
12.77
12.40
12.90
12.14
-
NS
10.00
9.62
10.10
11.06
10.80
11.24
11.84
11.24
12.10
12.07
11.70
12.20
11.80
-
NS
8.80
8.42
8.90
10.56
10.00
10.74
11.34
10.92
11.60
11.50
11.20
11.70
10.48
0.008
0.020
7.40
7.02
7.50
9.16
8.60
9.52
9.94
9.34
10.20
10.17
9.80
10.30
9.08
0.006
0.015
5.60
5.22
5.70
7.36
6.80
7.72
8.14
7.54
8.40
8.37
8.02
8.50
7.28
0.006
0.014
8.66
8.28
8.76
10.03
9.59
10.28
10.76
10.27
11.02
10.99
10.62
11.12
53
Table 22: Interaction effect of temperature and container (T x C) on phenol (mg/g) of Cedrus deodara seed during storage
Treatment
Duration (month)
D1 D2 D3 D4 D5 Mean
T1C1
T1C2
T1C3
T2C1
T2C2
T2C3
T3C1
T3C2
T3C3
T4C1
T4C2
T4C3
Mean
SEm +
CD0.05
36.44
36.82
36.92
37.50
37.76
37.87
37.94
38.15
38.29
38.52
38.55
38.65
37.78
-
NS
33.41
32.03
33.15
35.01
34.74
31.86
35.80
35.19
36.17
36.05
35.79
37.45
34.85
-
NS
29.37
28.98
29.46
31.95
31.40
32.14
33.30
32.88
33.56
33.67
33.30
33.80
31.98
-
NS
24.83
23.82
24.93
26.56
26.03
26.95
29.50
28.96
29.76
30.02
29.65
30.15
27.60
0.002
0.005
18.80
17.42
19.90
21.48
20.92
21.38
23.70
23.10
23.96
24.82
24.47
25.95
22.16
0.0022
0.0054
28.57
27.81
28.94
30.50
30.17
32.05
30.04
31.66
32.35
32.62
32.35
33.44
54
Chapter-V
DISCUSSION
The results obtained from the present investigation “Effect of storage
conditions on the germinability of Himalayan cedar (Cedrus deodara Loud.)
seeds” have been discussed in this chapter, establishing a cause and effect
relationship wherever necessary or feasible, in the light of the available literature,
under the following headings:
5.1 Storage of Cedrus seed
5.2 Germination and physio-biochemical parameters of freshly collected
seed
5.3 Effect of temperature on germination parameters of seed during storage
5.4 Effect of container on germination parameters of seed during storage
5.5 Biochemical parameter of stored seed
5.6 Interaction effect of temperature and container (TxC) on germination and
physio-biochemical parameters of seed during storage
5.1 Storage of Cedrus seed
Seed storage is an important aspect of any sound seed management
programme especially in conifer species where storage is a common rather
necessary practice due to short supply and viability problems. Cedar seed are
orthodox in storage behaviour, they are very oily and do not keep well under
many storage conditions (Allen, 1995). Cedrus has erratic and infrequent seed
years and being severely infested by various fungi as has been reported by Mittal
(1983) that cedar seed are prone to damping off disease. Temperature
determines the respiration rate and if higher the temperature higher will be the
rate of respiration (Barton, 1941) which will consume all the reserve food material
56
of the seed and consequently, the seed will loose its viability. Container on the
other hand controls the relative humidity and seed moisture content and also
determines the viability of seed (Toole et al., 1948). Therefore, various storage
techniques like regulation of storage temperature and use of storage devices
were tested to preserve and enhance viability of Cedrus deodara seed – An
important species of North -West Himalaya.
5.2 Germination and physio-biochemical parameters of freshly
collected seed
The high germinability and viability is an imperative criterion of seed that
are subjected to long storage. Keeping this in mind the freshly collected
Cedrus deodara seed were put to test for confirming their germinability and
physio-biochemical status under laboratory conditions. It is evident from the
results in Table 1 that freshly collected Cedrus seed possessed high viability
but moderate germinability as compared to two and four months’ storage
which may be attributed to inherent dormancy prevalent in the seed. The
biochemical parameters like total sugar, reducing sugar and non reducing
sugar also exhibited low values as compared to two and four months of
storage. The problem of moderate germinability but high viability indicates that
Cedrus exhibit dormancy in fresh seed. This has also been demonstrated by
Takos (1999) for Cedrus species and suggested cold period of stratification
before sowing. The present study corroborates the findings of Rudolf (1974),
Dirr and Heuser (1987), Young and Young (1992) and Hartman et al. (1997) in
Cedrus.
5.3 Effect of temperature on germination parameters of seed during
storage
The present investigation reveals that storage temperatures exerted
significant influence on germination parameters of Cedrus seed. The seed
stored at -5±1oC (T4) excelled all other storage temperature witnessing the
maximum germinability of seed. All germination parameters such as
57
germination per cent, germinative capacity, germinative energy and
germination value were recorded to be maximum from seed stored at -5±1oC
(T4), whereas room temperature (T1) proved to be the least effective
registering the minimum values of all the above mentioned parameters.
In the present study, it is pertinent to mention that the seed were stored at
their initial moisture content of 14 per cent and storage temperature has
significantly affected the seed germination parameters. The enhanced longevity
of C. deodara seed may be ascribed to the slow rate of biological processes at -
5±1oC (T4) storage temperature. The results are in agreement with Matziris
(1995), who noted that temperatures between –4°C and –15°C, and moisture
content ranging between 12 to 13 per cent were observed to be better for the
storage of Cedrus seed for 1 to 5 years. Erkuloglu (1995) observed best
temperature of -5oC at 7-9% of moisture content for 3 years in Cedrus libani.
Almost similar findings were reported by the findings of Takos and Merou (2001)
for Cedrus deodara. The result gets further supported by the findings of Kaushik
et al. (1967) in Cedrus, Barton (1954) in Pinus ponderosa and Zlobin (1973) in
Pinus palustris.
According to the present study, storage of Cedrus seed can be made by
reducing the initial seed moisture content when using the range of temperature
(+5°C,–5°C). Higher temperatures and specifically the ones of the room
temperature (T1) reduced the seed germination. Young and Young (1992) also
reported the unsuitability of this storage method because they pointed out that
the Cedrus seed were not well suited for storage under these conditions (T1)
because of their high oil content. In the present study the room temperature (T1)
was found to be unsuitable. At this temperature the mean germination
percentage of seed of C. deodara was approximately 17.67 per cent lower as
compared to that of -5±1oC (T4). The seed of C. deodara were totally frozen
below –5°C temperature, a result that agrees with Matziris (1995), who reported
58
that the Cedrus seed could maintain their viability at such low temperatures only
if their moisture content was very low.
The present study showed that after short-term storage and depending
upon the storage conditions, the seed acquire some degree of dormancy. Rudolf
(1974), Dirr and Heuser (1987), Young and Young (1992), Hartmann et al. (1997)
and Takos (1999) also referred to this and this was the reason why they
suggested a short period of cold stratification before the spring sowing.
Storage temperatures 5±1oC (T2) and 0±1oC (T3) were found best after -
5±1oC (T4) for all germination and physio-biochemical parameters. Piotto and
Gradi (1998) stored Cedrus atlantica seed at temperature of 3oC for 3 years and
germination was found to be 50 per cent. Zlobin (1973) found -3oC as best
followed by 2oC, Gordon et al. (1972) in Pinus merkussii observed 2oC as best
temperature. Similar findings observed by Barnett and Vozzo (1985) in Pinus
elliottio at 4oC temperature. The result gets further supported by the findings of
Havarbeka and Peterson (1989) in pinus ponderosa, Vlase (1974) in pinus
sylvestris, Robbins (1983) in pinus oocarpa. Barton (1954) in Thuja, Annon
(1948) in liriodendron, Johannsen (1921) in Quercus, Rehmedar (1953) in Abies
alba and Isaac (1934) in Abies nibilis also found similar results.
The above observations are in line with the findings of Gordon et al.
(1972) who reported that Pinus merkusii seed responded well to storage
temperature of 2oC giving maximum germination of 80 per cent after 3 years of
storage while room temperature showed significantly loss of germination after 3-4
months of storage. On the other hand, Effendi and Sinaga (1996) experimenting
with red wood reported that 4 months of storage at 4oC proved best resulting in
maximum germinability in the species. He further concluded that germination
capacity, germination value and mean daily germination decrease rapidly after 4
months of storage. The results are in harmony with the findings of Yap and Wang
(1983), Sharma and Bhardwaj (1999) and Manisha (2000).
59
The loss of germinability during storage at room temperature was probably
due to maximum loss of moisture content and biochemical parameters viz. total
sugar at 10 months’ storage. This might also be due to wide temperature
fluctuation, the high relative humidity and high activity of mycoflora present in
seed which become active under optimum environmental conditions and start
feeding on endosperm renderous the seed non-viable. The results are also in
agreement with the findings of Gupta and Raturi (1975) who while conducting
viability test of forest tree seed have reported that those insect infected seed
who’s embryos were eaten failed to germinate.
5.4 Effect of container on germination parameters of seed during
storage
The present investigation in Cedrus deodara reveals that various
germination parameters were significantly affected by different storage container
viz poly bag (T1), canvas bag (T2) and plastic container (T3). It is evident from the
data presented in tables 4 to 6 that plastic container (C3) proved superior as
compared to other container by registering maximum values in respect of
germination per cent, germinative capacity, germinative energy and germination
value. After ten months’ (D5) storage there was reduction in germination
parameters. The minimum germinability parameters were observed when canvas
bag (C2) was used as the storage container.
The better performance of plastic container (C3) in storage might be due to
the less reduction in moisture. Similar result was found by (Manisha, 2000;
Napier and Robbins, 1987; Chadurvedi and Das, 2004). The result also gets
supported by Takos and Merou (2001) in Cedrus deodara. The better
performance of plastic container (C3) may be due to proper preservance of
moisture and prevent contamination by fungi and micro organism. The results are
in harmony with the findings of Barnet and Maclemore (1970) and Athaya (1985).
60
The germination percentage of C. libani was very high in the seed stored
inside the cones through winter, as noted by Krussmann (1981) and Young and
Young (1992). On the contrary, in C. deodara the results of this storage method
were disappointingly low. These results contradict Krussmann (1981) who
proposed that, for all the Cedrus species, the seed should remain in the cones till
the sowing.
The intermediate performance of poly bag (C1) may be ascribed to
exchange of gases and moisture because they are not completely impermeable
to moisture and gases. Similar results, with poly bag were found by Brydrum
(1971) in Khasi pine, Maithani et al (1989) in Azadirachta indica, Gurdev (1994)
in Toona ciliata and Sharma (1996) in Q. leucotricophora. The results are also in
harmony with the findings of Khan et al. (2007) for Cedrus deodara.
In canvas bag (C2) the least germination was observed as compared to
other containers. Almost similar results were obtained by Napier and Robbin
(1987). Further, almost - identical trend was mentioned by Donald and Jacob
(1990) who studying the effect of storage in pinus species seed in linen bag and
PVC container. The PVC container was found to be more efficient than linen bag
in maintaining the germination parameters. The poor germinability of seed stored
in canvas bag (C2) may be attributed to the reduction in moisture content during
storage which reduces seed longevity. Almost identical findings were also
reported by Mehta (1999) and Manisha (2000). The deterioration of seed
longevity in storage may be attributed to the changes in the physiological stage
of seed particularly the respiratory metabolism. The changes in respiratory
metabolism are reported as the major factor responsible for seed deterioration
and hence fall in germinability (Abdul Baki, 1980). Other possible cause for the
loss of viability with time can be depletion of food reserve of seed during storage.
The results also get support from the work of Bhardwaj and Gupta (1998)
who reported that cloth bags storage of Pinus gerardiana seed exhibited
61
minimum germination per cent because of the incidence of fungi during storage
of nine months. These results are also in line with those of Barton (1954) in Pinus
pseudostuga, Pinus ponderosa and Tsuga heterophylla and Robbins (1983) in
Pinus elliottii, P. patula, P. radiata and P. taeda.
5.5 Biochemical parameters associated with seed storage
Biochemicals namely total sugar, reducing sugar, non-reducing sugar,
phenol and starch contents were estimated at bimonthly interval. Like
germination, the highest total sugar as well as reducing and non-reducing sugars
were found more in seed kept at -5 ± 1oC (T4) storage temperature as compared
to 0 ± 1oC (T3), 5 ± 1oC (T2) and the least were obtained from seed kept under
room temperature (T1). Similar results were obtained for starch and phenols
where maximum contents were obtained under -5 ± 1oC (T4) temperature.
The month wise comparison of these parameters indicates that there was
an increasing trend in sugars upto four months’ (D2) storage except in different
containers at room temperature and after that it declined continuously till the
termination of experiment. These observations are in accordance with the
findings of many other workers who also found an initial increase in these
biochemical parameters followed by a decrease in the later stages of storage in
seed of various tree species (Singh et al., 1992; Blanche et al., 1990 and
Mellareddy and Sharma 1983). The initial increase in sugars in seed during
storage may possibly be attributed to numerous catabolic processes taking place
in the seed, preparing for senescence. Other reason may be that seed gets
stratified during and some chemical changes occur in this period those are
required for seed germination. The decline in starch was not reflected in changes
in the level of sugar.
The lack of stoichiometry between starch and sugar levels may be
ascribed to rapid utilization of sugars as a result of elevated metabolism during
aging process with the advance of storage period. Also, we did not measure the
62
leakage of soluble sugars that is known to occur in greater amount in aged seed
(Abdul-Baki and Anderson, 1970; Bonner, 1970 and Ghosh et al., 1981). As
result of increased metabolism during senescence and may possibly be the
insect damage which has been reported in Cedrus deodara and Pinus
helepensis by Khan et al. (2007), the food reserves that would otherwise be
available for germination may no longer be adequate and thus a decline in seed
vigour results.
There are two school of thoughts that a number of seed have been
described to contain phenolics having both inhibitory (Lalmen and Misra, 1980;
Mellarreddy and Sharma, 1983 and Enu and Dumsoff, 1990) as well as
promoting effect on germination. There is still another observation that phenolics
do not permanently inhibit germination but only delay the event until the inhibitors
are apparently metabolized by the germinating seed (Kosuge and Conn, 1959;
Haskins and Gorz, 1963 and Sivan et al., 1965).
The phenol and starch decreased continuously with increase in storage,
but germination increased upto four months’ storage which may be ascribed to
the presence of higher levels of sugars and may be higher endogenous growth
promoters, but as the storage period progressed, there was corresponding
decrease in phenols as well as in germination. It is not possible to establish
relation between phenols and germination, since no attempt has been made in
the present investigation to identity the specific phenolic compound responsible
either for inhibiting or for promoting germination of Cedrus deorara seed.
A critical review of data in the tables 11 to 13 reveals that storage
temperature exerted significant effect on different biochemical parameters. It is
observed that there was increasing trend in total, reducing and non reducing
sugars from seed harvest to four months’ (D2) storage period and thereafter they
decreased till the termination of experiment. But on the other hand, there was a
continuous decrease in moisture content, starch and phenol from first month till
63
the end of experiment. However, seed stored at -5 ± 1oC (T4) have been found to
maintain steady decrease in all biochemical parameters as compared to room
temperature (T1).
The findings are in agreement with the work of Singh et al. (1992) in
Chilgoza in which they revealed that germ inability was greatly reduced with the
reduction in the biochemical properties when seed were stored in gunny bags at
room temperature. As the storage period was increased, the biochemical and
germinability of seed were correspondly found to decrease accordingly. Almost
identical results pertaining to beech seed below freezing temperature were
obtained by Muller et al. (1999). The other reason for decreasing viability with
storage period could be attributed to the depletion of food reserve of seed during
storage and change in respiratory metabolism (Abdul Baki, 1980). The findings
are thus in accordance with the results of Gordon et al. (1972) in Pinus merkusii;
Donald and Jacobs (1990) in Pinus elliottii, P. patula, P. radiate and P. taeda;,
Stoyhwo and Janson (1990) in Norway spruce and Gautam (2005) in Pinus
roxburghii seed.
5.6 Interaction effect of temperatures and containers (TxC) on
germination and biochemical parameters of Cedrus deodara seed during storage
In the present study it was observed that the maximum mean per cent
germination, total sugar, reducing sugar, non-reducing sugar, total phenol and
starch were registered in seed stored in plastic container at -5 ± 1oC (T4C3) which
was closely followed by T3C3 at the end of ten months’ (D5) storage. The
increased germination and other biochemical attributes of seed in plastic
container may be attributed to relatively slow biological process of seed at -5 ±
1oC in comparison to other combinations of temperature and container causing
comparatively less physiological disturbance to seed longevity. Although poly
bag is not suitable for long term storage of orthodox seed for genetic
conservation, it is very suitable for short term or medium term storage and given
64
excellent results upto 5 years storage of Pinus caribaea and P. oocarpa seed
with no significant change in moisture content (Robbins, 1983). Plastic container
has advantage of restricting moisture passage, exchange of oxygen and carbon
dioxide in contrast to poly bag. Seed stored at room temperature in canvas bag,
on the other hand resulted the lowest value for both attributes which may
possibly be ascribed to wide fluctuation of physiological process of seed by wide
variation in room temperature along with the loss of moisture in canvas bag
(T1C2). These observations are in accordance with the findings of Kramer and
Kozlowski (1960) who found that storing possibilities of seed could be best in
sealed container and under low temperature and humidity conditions so as to
keep the respiration at the lowest rate.
The appraisal of present data (Tables 6 to 9 and 17 to 22) reveals that
treatment interaction (TxC) has significant influence on germination and
biochemical parameters in Cedrus deodara seed. Data presented in above given
tables reveal that seed stored at -5 ± 1oC in plastic container (T4C3) excelled all
other treatment combinations in germination and biochemical parameters. The
significantly highest mean germination, mean germinative capacity, mean
germinative energy and mean germination value were observed when seed were
stored at -5 ± 1oC in plastic container (T4C3). This represents an increase in
germination per cent, germinative capacity over that of T1C2 which resulted
significantly least values for all the parameters. Data are supported by the fact
that treatments T4C3 excelled all other treatments in having higher total, reducing
and non reducing sugar during storage. There was decrease in germination with
corresponding decrease in biochemical parameters. Singh et al. (1992) who
stated that germinability was considerably reduced with the reduction of
biochemical parameters in Pinus gerardiana seed. Khan et al. (2007) also
reported that germination per cent of Cedrus deodara after four month of storage
was maximum (98.00%) when seed were stored in poly bag and at a
temperature of 3±1oC and thereafter declined. The better germinability at
65
treatment combination T4C3 might also be due to relatively better biological
process or less physiological deterioration of biochemical parameters as
compared to other treatment combinations. This obviously resulted in maintaining
the seed longevity in the species. The least values of above parameters found in
treatment combination T1C2 (Room temperature x Canvas bag) might be
ascribed to faster physiological deterioration which therefore consequently
reduced the germ inability performance in the seed. The results are in harmony
with the findings of Bhardwaj and Gupta (1998) in Pinus gerardiana, Barton
(1954) in Pinus ponderosa, Pseudostuga menziessii and Tsuga heterophylla,
Donald and Jacobs (1990) in Pinus elliotii, P. patula, P. radiata and P. taeda,
Gautam (2005) in Pinus roxburghii and Stolyhwo and Janson (1990) in Norway
spruce. Almost similar studies were conducted by Young and Young (1992) for
C. deodara.
Chapter-VI
SUMMARY AND CONCLUSION
The investigation “Effect of storage conditions on germinability of
Himalayan cedar (Cedrus deodara Loud.) seeds” was conducted in the department
of Silviculture and Agroforestry, Dr. Y.S. Parmar University of Horticulture and Forestry,
Nauni-173230, Solan (HP) during the year 2005-06. The results of the same are
summarized below:
6.1 Germination of freshly collected seed was found to be 70.00, whereas
germinative capacity and germinative energy 94.00 and 45.00 per cent,
respectively. The germination value was 10.50.
6.2 Three different temperatures namely room temperature (T1), 5±1oC (T2),
0±1oC (T3) and -5±1oC (T4) reflected significant differences. The storage
temperature of -5±1oC (T4) out classed for all temperature registering the
maximum values for germination parameters. On the other hand minimum
values were exhibited by seed stored at room temperature (T1).
6.3 The different container viz. poly bag (C1), canvas bag (C2) and plastic
container (C3) demonstrated a mark bearing on germinability. The plastic
container (C3) excelled over the other two registering maximum mean
germination success of 67.33 per cent. On the other hand, minimum mean
values were exhibited by the seed stored in canvas bag (C2).
6.4 For different duration, four month of storage registered maximum values for
different germination parameters except for germination capacity and after
this all germination parameters started declining till the termination of
experiment.
67
6.5 Interaction study reveals that the seed stored in plastic container at -5±1oC
(T4C3) excelled all other combinations of storage temperature (T) and
container(C).
6.6 Physio-biochemical contents namely moisture content, total sugar,
reducing sugar, non reducing sugar, starch and phenol of fresh seed
immediately after harvest were observed to be 22.00, 8.50, 1.68, 6.60,
11.70 per cent, and 39.50 mg/g, respectively.
6.7 Among three different temperature namely room temperature (T1), 5±1oC
(T2), 0±1oC (T3) and -5±1oC (T4) reflected significant differences. The
storage temperature -5±1oC (T4) proved effective in increasing the
biochemical attributes of seed followed by 0±1oC (T3), 5±1oC (T2) and room
temperature (T1).
6.8 Among the three different container viz., poly bag (C1), canvas bag (C2)
and plastic container (C3), the plastic container (C3) excelled over the other
two registering maximum contents of all biochemical attributes.
6.9 For different duration, seed storage upto four month’ storage registered
maximum values for total sugar, reducing sugar and non reducing sugar.
6.10 Interaction of temperature and container (TxC) effected the various physio-
biochemical contents. Like germinability, maximum physio-biochemical,
were estimated from the seed stored in plastic container at a temperature
of -5±1oC (T4C3)
CONCLUSION
Cedrus deodara seed should be stored in plastic container (airtight) at a
temperature of -5±1oC for prolonging their viability.
Chapter-VII
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77
Dr. Y. S. Parmar University of Horticulture and Forestry, Nauni, Solan 173 230 (HP) India
Department of Silviculture and Agroforestry
Title of Thesis : “Effect of storage conditions on germinability of Himalayan cedar (Cedrus deodara Loud.) seeds” Name of the Student : Raj Kumar Admission Number : F-2005-16-M Major Advisor : Dr. R.K. Nayital Major Field : Silviculture Minor Field(s) : i) Forest Product ii) Tree Improvement Degree Awarded : M.Sc. Forestry (Silviculture) Year of Award of Degree : 2008 No. of Pages in thesis : 77+ IV No. of words in Abstract : 148
ABSTRACT
The present study on the “Effect of storage conditions on germinability of Himalayan cedar (Cedrus deodara Loud.) seeds” was carried out in the laboratory of department of Silviculture and Agroforestry, Dr. Y.S. Parmar University of Horticulture and Forestry, Nauni-Solan during the year 2006-2007. Seeds were stored at four different temperature namely, room temperature (T1), 5±1oC (T2), 0±1oC (T3), -5±1oC (T4) and in three different container viz.; poly bag (C1), canvas bag (C2) and plastic container (C3). Observation on germination and physio-biochemical parameters were observed at bimonthly for storage duration of ten month. It was observed that seed stored at temperature of -5±1oC (T4) in plastic container (C3) singly or in combination gave overall best result in term of germination parameters (per cent germination, germinative capacity, germinative energy, germination value) and physio-biochemical parameters (moisture content, total sugar, reducing sugar, non-reducing sugar, starch and phenol) after ten months of storage.
Signature of Major Advisor Signature of the Student
Countersigned
Professor and Head
Department of Silviculture and Agroforestry
Dr. Y. S. Parmar University of Horticulture and Forestry,
Nauni, Solan-173 230 (HP)
Table 2: Effect of temperature on per cent germination and germinative capacity of Cedrus deodara seed during storage
Treatment
Germination (%) Germinative capacity (%)
Duration (Month) Duration (Month)
D1 D2 D3 D4 D5 Mean D1 D2 D3 D4 D5 Mean
T1
T2
T3
T4
Mean
SEm +
CD0.05
65.78
76.89
81.11
82.67
76.61
0.50
1.23
63.56
77.87
83.11
84.67
77.31
0.24
0.60
55.00
64.11
69.56
72.33
65.25
0.20
0.50
47.78
54.89
62.11
64.67
57.36
0.63
1.50
35.44
42.00
49.44
51.56
44.61
0.31
0.76
53.51
63.16
69.07
71.18
81.78
85.00
87.00
89.00
85.69
0.94
2.30
76.78
80.44
84.67
86.67
82.14
0.42
1.04
70.78
76.44
79.78
81.89
77.22
0.16
0.28
61.78
67.56
71.44
73.44
68.56
0.16
0.24
49.89
57.56
63.44
66.00
59.22
0.20
0.50
68.20
73.40
77.27
79.40
Table 3: Effect of temperature on per cent germinative energy and germination value of Cedrus deodara seed during storage
Treatment
Germinative energy (%) Germination value
Duration (Month) Duration (Month)
D1 D2 D3 D4 D5 Mean D1 D2 D3 D4 D5 Mean
T1
T2
T3
T4
Mean
SEm+
CD0.05
46.67
45.44
48.44
50.33
46.53
0.72
1.77
40.33
44.78
52.22
56.22
48.39
0.87
2.13
34.44
38.78
45.3
49.11
42.17
0.65
1.58
27.78
33.78
39.00
41.00
35.39
0.52
1.28
22.22
27.00
32.44
35.00
29.17
0.27
6.57
33.33
38.16
43.49
46.33
9.55
11.75
13.06
14.10
12.12
-
NS
10.37
12.58
15.05
15.77
13.44
0.04
0.10
7.98
9.43
10.69
11.76
9.96
0.07
0.18
4.84
6.78
8.25
9.06
7.22
0.05
0.12
2.88
4.78
5.86
6.16
4.92
0.07
0.17
7.12
9.07
10.58
11.36
Table 4: Effect of container on per cent germination and per cent germinative capacity of Cedrus deodara seed during storage
Treatment
Germination (%) Germinative capacity (%)
Duration (Month) Duration (Month)
D1 D2 D3 D4 D5 Mean D1 D2 D3 D4 D5 Mean
C1
C2
C3
Mean
SEm+
CD0.05
77.00
74.33
78.50
76.61
0.85
1.82
77.92
73.92
80.08
77.30
0.29
0.61
64.92
62.50
68.33
65.25
0.12
0.30
57.25
53.25
61.58
57.36
0.51
1.00
44.33
39.83
49.67
44.61
64.28
60.77
67.63
85.67
84.08
87.33
85.69
0.70
0.262
82.08
79.75
84.58
82.14
0.402
2.07
76.92
74.83
79.92
77.22
1.60
2.13
68.33
66.25
71.08
68.56
0.14
2.84
59.42
56.33
61.92
59.22
0.19
1.92
74.48
72.25
76.92
Table 5: Effect of container on per cent germinative energy and germination value of Cedrus deodara seed during storage
Treatment
Germinative energy (%) Germination value
Duration (Month) Duration (Month)
D1 D2 D3 D4 D5 Mean D1 D2 D3 D4 D5 Mean
C1
C2
C3
Mean
SEm+
CD0.05
46.67
45.00
47.92
46.53
0.77
1.63
47.83
4.17
53.17
48.39
0.83
1.77
42.00
38.58
45.92
42.17
0.93
1.98
35.33
32.08
38.75
35.39
0.44
0.93
28.92
26.08
32.50
29.17
0.097
0.203
40.15
37.18
43.65
12.04
11.19
13.13
12.12
-
NS
13.37
12.43
14.52
13.44
0.08
1.74
9.90
9.07
10.92
9.96
0.04
0.09
7.10
6.50
8.06
7.72
0.08
0.17
4.87
4.30
5.60
4.92
0.04
0.08
9.45
8.70
10.45
Table 11: Effect of temperature on moisture content and total sugar of Cedrus deodara seed during storage
Treatment
Moisture content (%) Total sugar (%)
Duration (Month) Duration (Month)
D1 D2 D3 D4 D5 Mean D1 D2 D3 D4 D5 Mean
T1
T2
T3
T4
Mean
SEm+
CD0.05
10.85
12.47
13.00
13.43
12.44
0.06
0.14
10.35
11.95
12.50
12.93
11.93
0.04
0.10
9.48
11.58
12.05
12.63
11.44
0.02
0.05
8.11
10.68
11.22
12.06
10.52
0.24
0.59
6.17
9.45
10.00
11.13
9.19
0.13
0.32
8.99
11.22
11.75
12.43
8.33
8.76
9.12
9.36
8.89
0.18
0.44
7.57
8.89
9.51
9.90
8.97
0.16
0.41
6.89
7.90
8.55
8.82
8.04
0.004
0.011
5.14
6.91
7.40
7.75
6.80
0.15
0.37
3.53
5.18
5.98
6.38
5.26
0.08
0.20
6.29
7.53
8.11
8.44
Table 12: Effect of temperature on reducing and non reducing sugar of Cedrus deodara seed during storage
Treatment
Reducing sugar (%) Non reducing sugar (%)
Duration (Month) Duration (Month)
D1 D2 D3 D4 D5 Mean D1 D2 D3 D4 D5 Mean
T1
T2
T3
T4
Mean
SEm+
CD0.05
1.56
1.71
1.79
1.84
1.72
0.06
0.13
1.40
1.87
2.06
2.15
1.87
0.038
0.092
1.148
1.53
1.61
1.66
1.49
0.04
0.010
0.92
1.25
1.34
1.39
1.22
0.007
0.002
0.60
0.91
1.15
1.16
0.95
0.02
0.05
1.13
1.45
1.59
1.64
6.42
6.69
6.95
7.34
6.85
0.040
0.099
5.87
6.66
7.07
7.35
6.73
0.055
0.11
5.41
6.05
5.58
6.79
6.21
0.02
0.05
4.00
5.37
5.75
5.92
5.26
0.14
0.36
2.78
4.04
4.63
4.95
4.10
0.05
0.12
4.89
5.76
6.20
6.47
Table 13: Effect of temperature on starch and phenol of Cedrus deodara seed during storage
Treatment
Starch (%) Phenol (mg/g)
Duration (Month) Duration(Month)
D1 D2 D3 D4 D5 Mean D1 D2 D3 D4 D5 Mean
T1
T2
T3
T4
Mean
SEm+
CD0.05
11.14
11.98
12.49
12.69
12.14
0.12
0.31
9.90
11.03
11.70
11.99
11.18
0.19
0.47
8.70
10.43
11.29
11.49
10.48
0.007
0.018
7.30
9.09
9.83
10.09
9.08
0.005
0.012
5.50
7.29
8.02
8.29
7.28
0.023
0.005
8.56
9.96
10.68
10.91
36.73
37.71
38.13
38.57
37.78
0.11
0.29
32.98
33.87
35.72
36.83
34.85
0.92
2.26
29.27
31.83
33.25
33.59
31.98
0.64
1.59
24.53
26.51
29.41
29.94
27.60
0.005
0.01
18.71
21.76
23.59
25.08
22.16
0.026
0.061
28.44
30.24
32.03
32.80
Table 14: Effect of container on moisture content and total sugar of Cedrus deodara seed during storage
Treatment
Moisture content (%) Total sugar (%)
Duration (Month) Duration (Month)
D1 D2 D3 D4 D5 Mean D1 D2 D3 D4 D5 Mean
C1
C2
C3
Mean
SEm+
CD0.05
12.75
1.62
12.93
12.45
0.05
0.11
12.20
11.12
12.47
11.93
0.03
0.10
11.69
10.61
12.01
11.43
0.70
0.15
10.81
9.63
11.11
10.81
0.23
0.49
9.470
8.18
9.98
9.18
0.12
0.26
11.37
10.23
11.70
8.86
8.63
9.19
8.89
0.10
0.21
8.91
8.60
9.39
8.97
0.05
0.10
8.06
7.53
8.52
8.04
0.006
0.01
6.78
6.57
7.05
6.80
0.10
0.36
5.32
4.67
5.81
5.26
0.06
0.13
7.59
7.20
7.99
Table 15: Effect of container on reducing and non reducing sugar of Cedrus deodara seed during storage
Treatment
Reducing sugar (%) Non reducing sugar (%)
Duration (Month) Duration (Month)
D1 D2 D3 D4 D5 Mean D1 D2 D3 D4 D5 Mean
C1
C2
C3
Mean
SEm+
CD0.05
1.72
1.67
1.78
1.723
0.014
0.08
1.87
1.75
1.98
1.87
0.018
0.039
1.50
1.43
1.55
1.49
0.0030
0.006
1.23
1.16
1.29
1.22
0.003
0.006
0.89
0.98
1.00
0.95
0.02
0.05
1.44
1.40
1.53
6.77
6.76
7.03
6.85
0.07
0.15
6.68
6.49
7.03
6.73
0.12
0.26
6.22
5.79
6.61
6.21
0.01
0.04
5.27
5.04
5.47
5.26
0.078
0.16
4.20
3.54
4.56
4.10
0.08
0.17
5.83
5.52
6.14
Table 16: Effect of container on starch and phenol of Cedrus deodara seed during storage
Treatment
Starch (%) Phenol (mg/g)
Duration (Month) Duration (Month)
D1 D2 D3 D4 D5 Mean D1 D2 D3 D4 D5 Mean
C1
C2
C3
Mean
SEm+
CD0.05
12.20
11.85
12.37
12.14
0.10
0.21
11.24
10.88
11.41
11.81
0.14
0.30
10.57
10.13
10.73
10.48
0.07
0.016
9.17
8.69
9.38
9.08
0.003
0.008
7.36
9.68
7.58
7.28
0.003
0.008
10.11
9.69
10.30
37.60
37.82
37.92
37.78
0.07
0.016
35.07
34.44
35.05
34.85
0.95
2.01
32.07
31.64
32.24
31.98
0.50
1.00
27.73
27.11
27.95
27.60
0.01
0.003
22.20
21.48
22.80
22.16
0.055
0.11
30.93
30.50
31.19
Table 23: Correlation of per cent germination with germinable attributes and physio-biochemical parameters of
deodar seed
Germination
per cent
Germinativ
e capacity
Germinative
energy
Germination
value
Moisture
content
Total
sugar
Reducing
sugar
Non
reducing
sugar
Starch Phenol
Germination
per cent
1.0000
Germinative
capacity
0.9265 1.0000
Germinative
energy
0.9582 0.9182 1.0000
Germination
value
0.3966 0.3664 0.3899 1.0000
Moisture
content
0.0246 0.0401 0.0201 -0.0016 1.0000
Total sugar 0.9411 0.8930 0.9390 0.3632 0.0171 1.0000
Reducing
sugar
0.8928 0.8499 0.8432 0.3675 0.0237 0.8749 1.0000
Non
reducing
sugar
0.9184 0.8973 0.9353 0.3623 0.0219 0.9562 0.8388 1.0000
Starch -0.1145 -0.1003 -0.1291 -0.0307 -0.0170 0.1648 -0.1161 -0.1302 1.0000
Phenol 0.8681 0.7967 0.8996 0.3672 -0.0254 0.8590 0.7509 0.8446 -0.117 1.0000
APPENDIX
ANALYSIS OF VARIANCE
1: EFFECT OF TEMPERATURE (TABLE 2), CONTAINER (TABLE 4) AND THEIR
INTERACTION (TABLE 6) ON PER CENT GERMINATION OF Cedrus deodara SEED DURING STORAGE
S.V
D.F
DURATION (MONTH)
D1 D2 D3 D4 D5
SS MS SS MS SS MS SS MS SS MS
T (A) 3 1566.3 522.11 2495.6 831.88 1575.6 525.21 1574.60 524.85 367.64 122.55
R (B) 2 32.05 16.02 22.39 11.19 24.67 12.33 70.72 35.36 341.06 170.53
A*B 6 6.83 1.34 1.61 0.27 1.11 0.19 10.61 1.77 14.28 2.38
C (C) 2 106.89 53.44 234.89 117.44 206.17 103.08 408.72 204.36 51.38 25.69
A*C 6 24.67 4.11 11.11 1.85 23.61 3.93 44.61 7.43 13.28 2.21
A*B*C 16 69.78 4.36 8.00 0.50 1.56 0.097 25.33 1.58 57.33 3.58
SV = Source of variation R (B) = Replication
D.F = Degree of freedom A*B = Temperature * Replication
SS = Sum of square C (C) = Container
MS = Mean sum of square A*C = Temperature * Container (Interaction)
T (A) = Temperature A*B*C = Temperature * Container * Replication
2: EFFECT OF TEMPERATURE (TABLE 2), CONTAINER (TABLE 4), AND THEIR
INTERACTION (TABLE 7), ON PER CENT GERMINATIVE CAPACITY OF Cedrus deodara SEED DURING STORAGE
SV
D.F
DURATION (MONTH)
D1 D2 D3 D4 D5
SS MS SS MS SS MS SS MS SS MS
T (A) 3 367.64 122.55 1385.9 461.96 977.64 325.88 956.75 318.92 879.22 293.07
R (B) 2 341.06 170.53 260.06 130.03 306.06 153.03 62.00 31.00 15.16 7.58
A*B 6 14.27 2.38 20.61 3.44 11.28 1.88 7.33 1.22 1.94 0.32
C (C) 2 51.39 25.69 491.56 245.78 300.39 150.19 266.67 133.33 248.17 124.08
A*C 6 13.28 2.21 107.78 17.96 228.94 38.15 37.33 6.22 33.61 5.60
A*B*C 16 57.33 3.58 66.67 4.16 83.33 5.21 18.67 1.17 0.88 0.05
ii
3: EFFECT OF TEMPERATURE (TABLE 3), CONTAINER (TABLE 5), AND THEIR
INTERACTION (TABLE 8), ON PER CENT GERMINATIVE ENERGY OF Cedrus deodara SEED DURING STORAGE
SV
D.F
DURATION (MONTH)
D1 D2 D3 D4 D5
SS MS SS MS SS MS SS MS SS MS
T (A) 3 256.08 85.36 526.53 175.51 634.00 211.33 712.67 237.56 1382.9 460.96
R (B) 2 16.89 8.44 54.89 27.44 11.56 5.78 16.72 8.36 16.22 8.11
A*B 6 24.00 4.0 4.89 0.81 0.67 0.11 1.50 0.25 1.11 0.18
C (C) 2 63.38 31.69 140.22 70.11 156.72 78.36 141.06 70.52 187.72 93.86
A*C 6 30.16 5.02 10.22 1.70 8.83 1.47 15.16 2.53 40.94 6.82
A*B*C 16 47.11 2.94 15.56 0.97 2.44 0.15 1.78 0.11 3.33 0.20
4: EFFECT OF TEMPERATURE (TABLE 3), CONTAINER (TABLE 5) AND THEIR
INTERACTION (TABLE 9) ON GERMINATION VALUE OF Cedrus deodara SEED DURING STORAGE
SV
D.F
DURATION (MONTH)
D1 D2 D3 D4 D5
SS MS SS MS SS MS SS MS SS MS
T (A) 3 2115.40 705.12 163.46 54.48 84.10 28.03 91.45 30.48 58.80 19.60
R (B) 2 689.18 344.59 0.11 0.056 0.041 0.020 0.18 0.091 0.041 0.020
A*B 6 4691.2 781.87 0.47 0.0078 0.15 0.025 6.86 0.01 0.12 0.020
C (C) 2 466.71 233.36 26.38 13.19 21.57 10.78 14.65 7.47 10.18 5.09
A*C 6 4815.90 802.64 9.49 1.58 7.42 1.23 3.08 0.51 1.23 0.20
A*B*C 16 1072.20 670.14 0.65 0.040 0.16 0.010 0.64 0.040 0.14 0.0093
5: EFFECT OF TEMPERATURE (TABLE 11), CONTAINER (TABLE 14) AND THEIR
INTERACTION (TABLE 17) ON MOISTURE CONTENT OF Cedrus deodara SEED DURING STORAGE
SV
D.F
DURATION (MONTH)
D1 D2 D3 D4 D5
SS MS SS MS SS MS SS MS SS MS
T (A) 3 13.32 4.44 68.14 22.71 45.55 15.18 122.62 40.87 167.02 55.67
R (B) 2 0.28 0.13 0.58 0.29 0.028 0.014 0.33 0.16 3.56 1.78
A*B 6 0.096 0.016 0.050 0.0083 0.010 0.0017 1.60 0.26 0.345 0.075
C (C) 2 1.75 0.87 25.59 12.79 19.91 9.59 25.58 12.79 74.53 37.27
A*C 6 0.54 0.089 2.50 0.41 7.17 1.19 6.05 1.00 3.64 0.60
A*B*C 16 0.27 0.016 0.19 0.012 0.46 0.029 5.20 0.32 1.40 8.75
iii
6: EFFECT OF TEMPERATURE (TABLE 11), CONTAINER (TABLE 14) AND THEIR
INTERACTION (TABLE, 18) ON TOTAL SUGAR OF Cedrus deodara SEED DURING STORAGE
SV
D.F
DURATION (MONTH)
D1 D2 D3 D4 D5
SS MS SS MS SS MS SS MS SS MS
T (A) 3 5.55 1.84 28.05 9.35 19.97 6.56 36.28 12.09 42.84 14.28
R (B) 2 1.93 0.96 0.80 0.40 0.0013 0.0006 1.53 0.76 0.020 0.010
A*B 6 0.89 0.14 0.77 0.12 0.0005 0.00009 0.61 0.10 0.17 0.029
C (C) 2 1.89 0.94 3.84 1.92 5.83 2.91 1.43 0.71 7.77 3.88
A*C 6 0.37 0.063 0.50 0.083 0.31 0.051 2.17 0.36 0.42 0.071
A*B*C 16 0.99 0.062 0.24 0.015 0.0039 0.0002 0.97 0.060 0.38 0.023
7: EFFECT OF TEMPERATURE (TABLE 12), CONTAINER (TABLE 15) AND THEIR
INTERACTION (TABLE 19) ON REDUCING SUGAR OF Cedrus deodara SEED DURING STORAGE
SV
D.F
DURATION (MONTH)
D1 D2 D3 D4 D5
SS MS SS MS SS MS SS MS SS MS
T (A) 3 0.41 0.13 2.97 0.99 1.25 0.41 0.60 0.20 1.09 0.36
R (B) 2 0.022 0.011 0.017 0.0089 0.0010 0.00050 0.00067 0.00033 0.0073 0.0036
A*B 6 0.068 0.013 0.038 0.0064 0.00051 0.000086 0.00021 0.000036 0.012 0.0020
C (C) 2 0.075 0.037 0.31 0.15 0.083 0.041 0.18 0.090 0.21 0.10
A*C 6 0.0070 0.0011 0.017 0.029 0.0075 0.0012 0.062 0.010 0.077 0.012
A*B*C 16 0.15 0.0094 0.032 0.0020 0.00086 0.000054 0.00091 0.000056 0.071 0.0044
8: EFFECT OF TEMPERATURE (TABLE 12), CONTAINER (TABLE 15) AND THEIR
INTERACTION (TABLE 20) ON NON REDUCING SUGAR OF Cedrus deodara SEED DURING STORAGE
SV
D.F
DURATION (MONTH)
D1 D2 D3 D4 D5
SS MS SS MS SS MS SS MS SS MS
T (A) 3 4.28 1.42 11.53 3.84 10.26 3.42 20.49 6.83 25.61 8.53
R (B) 2 0.26 0.13 0.057 0.028 0.0013 0.000069 0.29 0.14 0.54 0.37
A*B 6 0.094 0.15 0.11 0.019 0.013 0.0023 0.59 0.09 0.065 0.010
C (C) 2 01.55 0.27 1.80 0.90 4.08 2.04 1.06 0.53 5.56 2.789
A*C 6 0.65 0.10 0.40 0.067 0.29 0.049 1.22 0.20 0.76 0.12
A*B*C 16 0.49 0.031 1.46 0.091 0.032 0.0020 0.58 0.036 0.63 0.039
iv
9: EFFECT OF TEMPERATURE (TABLE 13), CONTAINER (TABLE 16) AND THEIR
INTERACTION (TABLE 21) ON STARCH OF Cedrus deodara SEED DURING STORAGE
SV
D.F
DURATION (MONTH)
D1 D2 D3 D4 D5
SS MS SS MS SS MS SS MS SS MS
T (A) 3 8.86 2.95 21.90 7.30 48.94 16.31 42.57 14.19 42.60 14.20
R (B) 2 0.85 0.42 21.40 1.07 0.00035 0.0001 0.00002 0.00001 0.00007 0.00003
A*B 6 0.44 0.074 1.01 0.68 0.0016 0.0002 0.0007 0.0001 0.0001 0.00002
C (C) 2 1.72 0.86 1.9 0.84 0.76 0.038 3.01 1.50 2.94 1.47
A*C 6 0.062 0.010 0.76 0.12 1.49 0.24 0.244 0.040 0.25 0.043
A*B*C 16 0.97 0.061 1.98 0.12 0.0060 0.0003 0.0015 0.00009 0.0014 0.00009
10: EFFECT OF TEMPERATURE (TABLE 13), CONTAINER (TABLE 16) AND THEIR
INTERACTION (TABLE 22) ON PHENOL OF Cedrus deodara SEED DURING STORAGE
SV
D.F
DURATION (MONTH)
D1 D2 D3 D4 D5
SS MS SS MS SS MS SS MS SS MS
T (A) 3 18.04 6.01 82.06 27.35 104.12 13.70 181.53 60.51 209.14 69.71
R (B) 2 0.16 0.084 19.60 9.80 38.18 19.09 0.0006 0.0003 0.27 0.13
A*B 6 0.35 0.058 23.17 3.86 11.38 1.89 0.0009 0.0001 0..16 0.002
C (C) 2 1.08 0.54 3.04 1.52 2.29 1.14 4.18 2.09 10.87 5.43
A*C 6 0.33 0.05 35.91 5.98 0.086 0.01 2.48 0.41 3.77 0.62
A*B*C 16 0.59 0.037 87.08 5.44 21.15 1.32 0.0002 0.00001 0.29 0.018