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 Fathers Name Date of Birth Sex Marital status Nationality : : : : : :

Raj Kumar
Sh. Man Singh 01.12.1983 Male Single Indian

Educational qualifications: Certificate/Degree B.Sc. Forestry Division First Board/University Dr Y.S. Parmar University of Horticulture & Forestry, NauniSolan (HP) 173 230 Cenral Board of School Education HP Board of School Education, Dharamshala (HP) : No Year 2005

10+2 Matriculation

Second First

2001 1999

Whether sponsored by some state/ 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

(Sr. Scientist)

Department of Silviculture and Agroforestry 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 Dated : , 2007

__________________ Dr. R.K. Nayital 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 Students Advisory Committee after an oral examination of the same in collaboration with the external examiner. ______________________ Dr. G.S. Shamet Professor (Co-opted in place of Dr. R.K. Nayital) Chairman Advisory Committee Dept. of Silviculture & Agroforestry _____________________ External Examiner

Members, Advisory Committee

______________________ Dr. S.D. Bhardwaj Professor & Head Dept. of Silviculture & Agroforestry

_____________________ Dr. H.P. Sankhyan Scientist Dept. of Tree Improvement

______________________ Dr. Bhupender Dutt Assistant Professor Dept. of Forest Products

______________________ Deans Nominee

____________________ 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)

CONTENTS
Chapter
1 2 3 4 5 6 7

Title
INTRODUCTION REVIEW OF LITERATURE MATERIALS AND METHODS EXPERIMENTAL RESULTS DISCUSSION SUMMARY REFERENCES ABSTRACT APPENDIX

Pages
1-4 5-15 16-21 22-54 55-65 66-67 68-77 78 i-iv

LIST OF TABLES
Table
1 2

Title
Germination parameters of freshly collected seed of C. deodara Effect of temperature on per cent germination and germinative capacity of C. deodara seed during storage Effect of temperature on per cent germinative energy and germination value of C. deodara seed during storage Effect of container on per cent germination and germinative capacity of C. deodara seed during storage Effect of container on per cent germinative energy and germination value of C. deodara seed during storage Interaction effect of temperature and container (T x C) on per cent germination of C. deodara seed during storage Interaction effect of temperature and container (T x C) on per cent germinative capacity of C. deodara seed during storage Interaction effect of temperature and container (T x C) on per cent germinative energy of C. deodara seed during storage Interaction effect of temperature and container (T x C) on germination value of C. deodara seed during storage Physio-biochemical parameters of freshly collected seed of C. deodara Effect of temperature on moisture content and total sugar of C. deodara seed during storage Effect of temperature on reducing and non reducing sugar of C. deodara seed during storage Effect of temperature on starch and phenol of C.deodara seed during storage

Page(s)
22 24

25

4 5 6

28 29 30

32

33

34

10 11 12 13

35 37 38 40

Table
14 15 16. 17.

Title
Effect of container on moisture content and total sugar of C. deodara seed during storage Effect of container on reducing and non reducing sugar of C. deodara seed during storage Effect of container on starch and phenol of C. deodara seed during storage Interaction effect of temperature and container (T x C) on Moisture content (%) of C. deodara seed during storage Interaction effect of temperature and container (T x C) on total sugar (%) of C. deodara seed during storage Interaction effect of temperature and container (T x C) on reducing sugar (%) of C. deodara seed during storage Interaction effect of temperature and container (T x C) on non reducing sugar (%) of C. deodara seed during storage Interaction effect of temperature and container (T x C) on starch (%)of C. deodara seed during storage Interaction effect of temperature and container (T x C) on phenol (mg/g) of C. deodara seed during storage Correlation of per cent germination with germinable and physio-biochemical parameters of deodara seed.

Page(s)
41 43 44 46

18. 19.

47 49

20.

50

21.

52

22. 23

53 54

LIST OF PLATES Plates Title Between Page(s)

1 2

Determination of biochemicals 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 68 o 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,

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 to 13 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

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

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 1C to 5C. 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

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 (+ 4C) 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 4.4C. 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

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 5C for 2 weeks to obtain uniform germination (Rudolf, 1974 and Allen, 1995). Thapliyal and Gupta (1980) found that 9C (48.2F) was a better temperature for stratification than 3C 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 20C. International Seed Testing Association rules, however, specify diurnally alternate temperature of 20C at night and 30oC (86F) 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 4C 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 +5C, 10C and 20C, as well as in a basement at fluctuating temperature of +10C/20C. Storage in airtight boxes and at temperature range +5C to 10C, 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 51C for 15 days. The common storage conditions (10C/20C) as well as temperature lower than 10C had a negative effect on germination of both

species. The soaking of the seed in water for 3 hours and the cold stratification at 51C 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

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 31C. 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

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.

10

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.

11

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 51oC 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

12

total sugar, reducing sugar, total nitrogenous and phosphorus compounds increased in the embryos of Pinus taiwanensis and Cumninghamia lanceolata seed during germination. Quantitative changes in sugars, -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 -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,

13

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 embryos 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 0 C 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
o

14

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.

15

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.

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 T2 T3 T4 B) Storage container The following types of storage containers were used: C1 C2 C3 C) Storage duration Total storage duration was ten months and its effect on seed viability observed at bimonthly intervals. D1 : Two months of storage : : : Poly bag Canvas bag Plastic container : : : : Room temperature 51oC 01oC -51oC

17

D2 D3 D4 D5

: : : :

Four months of storage Six months of storage Eight months of storage Ten months of storage : : : Split plot design Temperature Container

Experimental Design Main plot factor Sub plot factor 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

18

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 (%) = Total number of seed taken 3.2.1.4 Germination value Germination value (GV) is the index combining speed and x 100

completeness of seed germination. Daily germination counts were recorded and calculated as per Czabator (1962). GV Where, PV = Peak value of germination Mean daily germination = PV x MDG

MDG =

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). 19

Original weight Oven dry weight Moisture content (%) = 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. x 100

20

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).

21

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 4.2 Germination parameters 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 (%) 70.00 Germinative capacity (%) 94.00 Germinative energy (%) 45.00 Germination value 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.

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), 51oC (T2), 01oC (T3) and -51oC (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 -51oC (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 -51oC (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 -51oC (T4) gave the highest germinative energy of 46.33 per cent after ten months

23

24

25

(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 -51oC (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 at51oC (T2), 01oC (T3) and -51oC (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).

26

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), 51oC (T2), 01oC (T3) and -51oC (T4) and in three type of container namely, poly bag (C 1), 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 -51oC in plastic container (T4C3) which was closely followed by T3C3 (73.07 %) at the end of

27

28

29

Table 6:

Interaction effect of temperature and container (T x C) on per cent germination of Cedrus deodara seed during storage Duration (month)

Treatment T1C1 T1C2 T1C3 T2C1 T2C2 T2C3 T3C1 T3C2 T3C3 T4C1 T4C2 T4C3 Mean SEm + CD0.05

D1 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

D2 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

D3 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

D4 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

D5 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

Mean 54.13 50.40 56.00 62.73 60.47 66.27 69.87 64.27 73.07 70.40 67.93 75.20

30

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 -51oC (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 -51oC (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 -51oC (T4C3). Whereas, the lowest mean germination value (6.56) was registered in seed stored in canvas bag at room temperature (T1C2).

31

Table 7:

Interaction effect of temperature and container (T x C) on per cent germinative capacity of Cedrus deodara seed during storage Duration (month)

Treatment T1C1 T1C2 T1C3 T2C1 T2C2 T2C3 T3C1 T3C2 T3C3 T4C1 T4C2 T4C3 Mean SEm + CD0.05

D1 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

D2 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

D3 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

D4 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

D5 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

Mean 68.60 66.13 68.97 73.27 71.60 75.33 77.60 74.67 79.93 78.47 77.00 82.73

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Table 8:

Interaction effect of temperature and container (T x C) on per cent germinative energy of Cedrus deodara seed during storage Duration (month) D3 D4 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

Treatment T1C1 T1C2 T1C3 T2C T2C2 T2C3 T3C1 T3C2 T3C3 T4C1 T4C2 T4C3 Mean SEm + CD0.05

D1 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

D2 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

D5 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

Mean 33.60 31.33 35.07 37.00 36.13 41.33 43.93 39.00 47.53 46.07 42.27 50.67

33

Table 9:

Interaction effect of temperature and container (T x C) on germination value of Cedrus deodara seed during storage Duration (month)

Treatment T1C1 T1C2 T1C3 T2C1 T2C2 T2C3 T3C1 T3C2 T3C3 T4C1 T4C2 T4C3 Mean SEm + CD0.05

D1 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

D2 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

D3 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

D4 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

D5 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

Mean 7.29 6.56 7.45 8.71 8.45 10.05 10.81 9.40 11.53 11.03 10.39 12.69

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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 (%) Nonreducing sugar (%) 22.00 8.50 1.68 6.60 11.70 39.50 Starch (%) Phenol (mg/g)

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), 51oC (T2), 01oC (T3) and -51oC (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

35

seed stored at -51oC (T4), followed by 01oC (T3), 51oC (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 -51oC (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 -51oC (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 -51oC (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).

36

37

38

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 -51oC (T4), followed by 01oC (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 -51oC (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 -51oC (C3), followed by poly bag

39

40

41

(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

42

43

44

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), 51oC (T2), 01oC (T3) and -51oC (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 51oC 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 (T 1C2) 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 -51oC in plastic container (T4C3) throughout the storage period. It was closely followed by T4C1 and T3C3 treatment combinations. The lowest mean sugar contents of

45

Table 17:

Interaction effect of temperature and container (T x C) on Moisture content (%) of Cedrus deodara seed during storage Duration

Treatment T1C1 T1C2 T1C3 T2C1 T2C2 T2C3 T3C1 T3C2 T3C3 T4C1 T4C2 T4C3 Mean SEm + CD0.05

D1 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

D2 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

D3 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

D4 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

D5 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

Mean 9.26 8.10 9.62 11.80 9.76 12.12 11.86 11.24 12.20 12.56 11.84 12.92

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Table 18:

Interaction effect of temperature and container (TxC) on total sugar (%) of Cedrus deodara seed during storage Duration (month)

Treatment T1C1 T1C2 T1C3 T2C1 T2C2 T2C3 T3C1 T3C2 T3C3 T4C1 T4C2 T4C3 Mean SEm + CD0.05

D1 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

D2 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

D3 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

D4 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

D5 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

Mean 6.42 5.82 6.42 7.56 7.15 7.88 8.14 7.69 8.52 8.24 8.15 8.94

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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 -51oC (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 -51oC 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 (T 1C2) registered the minimum value of 4.50 per cent.

48

Table 19:

Interaction effect of temperature and container (T x C) on reducing sugar (%) of Cedrus deodara seed during storage

Duration (month) Treatment T1C1 T1C2 T1C3 T2C1 T2C2 T2C3 T3C1 T3C2 T3C3 T4C1 T4C2 T4C3 Mean SEm + CD0.05 D1 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 D2 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 D3 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 D4 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 D5 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 Mean 1.13 1.07 1.20 1.43 1.42 1.52 1.60 1.55 1.64 1.63 1.57 1.72

49

Table 20:

Interaction effect of temperature and container (T x C) on non reducing sugar (%) of Cedrus deodara seed during storage Duration (month)

Treatment T1C1 T1C2 T1C3 T2C1 T2C2 T2C3 T3C1 T3C2 T3C3 T4C1 T4C2 T4C3 Mean SEm + CD0.05

D1 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

D2 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

D3 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

D4 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

D5 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

Mean 5.02 4.50 5.16 5.82 5.44 6.04 6.21 5.87 6.52 6.27 6.30 6.85

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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 -51oC (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 -51oC (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 physiobiochemical 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.

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Table 21:

Interaction effect of temperature and container (T x C) on starch (%) of Cedrus deodara seed during storage Duration (month)

Treatment T1C1 T1C2 T1C3 T2C1 T2C2 T2C3 T3C1 T3C2 T3C3 T4C1 T4C2 T4C3 Mean SEm + CD0.05

D1 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

D2 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

D3 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

D4 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

D5 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

Mean 8.66 8.28 8.76 10.03 9.59 10.28 10.76 10.27 11.02 10.99 10.62 11.12

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Table 22:

Interaction effect of temperature and container (T x C) on phenol (mg/g) of Cedrus deodara seed during storage Duration (month)

Treatment T1C1 T1C2 T1C3 T2C1 T2C2 T2C3 T3C1 T3C2 T3C3 T4C1 T4C2 T4C3 Mean SEm + CD0.05

D1 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

D2 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

D3 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

D4 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

D5 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

Mean 28.57 27.81 28.94 30.50 30.17 32.05 30.04 31.66 32.35 32.62 32.35 33.44

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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 5.2 Storage of Cedrus seed Germination and physio-biochemical parameters of freshly collected seed 5.3 5.4 5.5 5.6 Effect of temperature on germination parameters of seed during storage Effect of container on germination parameters of seed during storage Biochemical parameter of stored seed 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

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 collected seed physio-biochemical parameters of freshly

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 -51oC (T4) excelled all other storage temperature witnessing the maximum germinability of seed. All germination parameters such as

56

germination per cent, germinative capacity, germinative

energy and

germination value were recorded to be maximum from seed stored at -51oC (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 51oC (T4) storage temperature. The results are in agreement with Matziris (1995), who noted that temperatures between 4C and 15C, 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 (+5C,5C). 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 (T 1) 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 -51oC (T4). The seed of C. deodara were totally frozen below 5C temperature, a result that agrees with Matziris (1995), who reported

57

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 51oC (T2) and 01oC (T3) were found best after 51oC (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 4 oC 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).

58

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 whos 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).

59

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

60

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

61

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

62

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

63

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 31oC and thereafter declined. The better germinability at

64

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.

65

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), 51oC (T2), 01oC (T3) and -51oC (T4) reflected significant differences. The storage temperature of -51oC (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.

6.5

Interaction study reveals that the seed stored in plastic container at -51oC (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), 51oC (T2), 01oC (T3) and -51oC (T4) reflected significant differences. The storage temperature -51oC (T4) proved effective in increasing the biochemical attributes of seed followed by 01oC (T3), 51oC (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 physiobiochemical contents. Like germinability, maximum physio-biochemical, were estimated from the seed stored in plastic container at a temperature of -51oC (T4C3)

CONCLUSION
Cedrus deodara seed should be stored in plastic container (airtight) at a temperature of -51oC for prolonging their viability.

67

Chapter-VII

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Dr. Y. S. Parmar University of Horticulture and Forestry, Nauni, Solan 173 230 (HP) India

Department of Silviculture and Agroforestry Effect of storage conditions on germinability of Himalayan cedar (Cedrus deodara Loud.) seeds Raj Kumar F-2005-16-M Dr. R.K. Nayital Silviculture i) Forest Product ii) Tree Improvement M.Sc. Forestry (Silviculture) 2008 77+ IV 148

Title of Thesis Name of the Student Admission Number Major Advisor Major Field Minor Field(s) Degree Awarded Year of Award of Degree No. of Pages in thesis No. of words in Abstract

: : : : : : : : : :

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), 51oC (T2), 01oC (T3), -51oC (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 -51oC (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)

77

Table 2:

Effect of temperature on per cent germination and germinative capacity of Cedrus deodara seed during storage Germination (%) Duration (Month) Germinative capacity (%) Duration (Month) D5 35.44 42.00 49.44 51.56 44.61 0.31 0.76 Mean 53.51 63.16 69.07 71.18 D1 81.78 85.00 87.00 89.00 85.69 0.94 2.30 D2 76.78 80.44 84.67 86.67 82.14 0.42 1.04 D3 70.78 76.44 79.78 81.89 77.22 0.16 0.28 D4 61.78 67.56 71.44 73.44 68.56 0.16 0.24 D5 49.89 57.56 63.44 66.00 59.22 0.20 0.50 Mean 68.20 73.40 77.27 79.40

Treatment T1 T2 T3 T4 Mean SEm + CD0.05

D1 65.78 76.89 81.11 82.67 76.61 0.50 1.23

D2 63.56 77.87 83.11 84.67 77.31 0.24 0.60

D3 55.00 64.11 69.56 72.33 65.25 0.20 0.50

D4 47.78 54.89 62.11 64.67 57.36 0.63 1.50

Table 3:

Effect of temperature on per cent germinative energy and germination value of Cedrus deodara seed during storage Germinative energy (%) Duration (Month) Germination value Duration (Month) D5 22.22 27.00 32.44 35.00 29.17 0.27 6.57 Mean 33.33 38.16 43.49 46.33 D1 9.55 11.75 13.06 14.10 12.12 NS D2 10.37 12.58 15.05 15.77 13.44 0.04 0.10 D3 7.98 9.43 10.69 11.76 9.96 0.07 0.18 D4 4.84 6.78 8.25 9.06 7.22 0.05 0.12 D5 2.88 4.78 5.86 6.16 4.92 0.07 0.17 Mean 7.12 9.07 10.58 11.36

Treatment T1 T2 T3 T4 Mean SEm+ CD0.05

D1 46.67 45.44 48.44 50.33 46.53 0.72 1.77

D2 40.33 44.78 52.22 56.22 48.39 0.87 2.13

D3 34.44 38.78 45.3 49.11 42.17 0.65 1.58

D4 27.78 33.78 39.00 41.00 35.39 0.52 1.28

Table 4:

Effect of container on per cent germination and per cent germinative capacity of Cedrus deodara seed during storage Germination (%) Duration (Month) Germinative capacity (%) Duration (Month) D5 44.33 39.83 49.67 44.61 Mean 64.28 60.77 67.63 D1 85.67 84.08 87.33 85.69 0.70 0.262 D2 82.08 79.75 84.58 82.14 0.402 2.07 D3 76.92 74.83 79.92 77.22 1.60 2.13 D4 68.33 66.25 71.08 68.56 0.14 2.84 D5 59.42 56.33 61.92 59.22 0.19 1.92 Mean 74.48 72.25 76.92

Treatment C1 C2 C3 Mean SEm+ CD0.05

D1 77.00 74.33 78.50 76.61 0.85 1.82

D2 77.92 73.92 80.08 77.30 0.29 0.61

D3 64.92 62.50 68.33 65.25 0.12 0.30

D4 57.25 53.25 61.58 57.36 0.51 1.00

Table 5:

Effect of container on per cent germinative energy and germination value of Cedrus deodara seed during storage Germinative energy (%) Duration (Month) Germination value Duration (Month) D5 28.92 26.08 32.50 29.17 0.097 0.203 Mean 40.15 37.18 43.65 D1 12.04 11.19 13.13 12.12 NS D2 13.37 12.43 14.52 13.44 0.08 1.74 D3 9.90 9.07 10.92 9.96 0.04 0.09 D4 7.10 6.50 8.06 7.72 0.08 0.17 D5 4.87 4.30 5.60 4.92 0.04 0.08 Mean 9.45 8.70 10.45

Treatment C1 C2 C3 Mean SEm+ CD0.05

D1 46.67 45.00 47.92 46.53 0.77 1.63

D2 47.83 4.17 53.17 48.39 0.83 1.77

D3 42.00 38.58 45.92 42.17 0.93 1.98

D4 35.33 32.08 38.75 35.39 0.44 0.93

Table 11:

Effect of temperature on moisture content and total sugar of Cedrus deodara seed during storage Moisture content (%) Duration (Month) Total sugar (%) Duration (Month) D5 6.17 9.45 10.00 11.13 9.19 0.13 0.32 Mean 8.99 11.22 11.75 12.43 D1 8.33 8.76 9.12 9.36 8.89 0.18 0.44 D2 7.57 8.89 9.51 9.90 8.97 0.16 0.41 D3 6.89 7.90 8.55 8.82 8.04 0.004 0.011 D4 5.14 6.91 7.40 7.75 6.80 0.15 0.37 D5 3.53 5.18 5.98 6.38 5.26 0.08 0.20 Mean 6.29 7.53 8.11 8.44

Treatment T1 T2 T3 T4 Mean SEm+ CD0.05

D1 10.85 12.47 13.00 13.43 12.44 0.06 0.14

D2 10.35 11.95 12.50 12.93 11.93 0.04 0.10

D3 9.48 11.58 12.05 12.63 11.44 0.02 0.05

D4 8.11 10.68 11.22 12.06 10.52 0.24 0.59

Table 12:

Effect of temperature on reducing and non reducing sugar of Cedrus deodara seed during storage Reducing sugar (%) Duration (Month) Non reducing sugar (%) Duration (Month) D5 0.60 0.91 1.15 1.16 0.95 0.02 0.05 Mean 1.13 1.45 1.59 1.64 D1 6.42 6.69 6.95 7.34 6.85 0.040 0.099 D2 5.87 6.66 7.07 7.35 6.73 0.055 0.11 D3 5.41 6.05 5.58 6.79 6.21 0.02 0.05 D4 4.00 5.37 5.75 5.92 5.26 0.14 0.36 D5 2.78 4.04 4.63 4.95 4.10 0.05 0.12 Mean 4.89 5.76 6.20 6.47

Treatment T1 T2 T3 T4 Mean SEm+ CD0.05

D1 1.56 1.71 1.79 1.84 1.72 0.06 0.13

D2 1.40 1.87 2.06 2.15 1.87 0.038 0.092

D3 1.148 1.53 1.61 1.66 1.49 0.04 0.010

D4 0.92 1.25 1.34 1.39 1.22 0.007 0.002

Table 13:

Effect of temperature on starch and phenol of Cedrus deodara seed during storage Starch (%) Duration (Month) Phenol (mg/g) Duration(Month) D5 5.50 7.29 8.02 8.29 7.28 0.023 0.005 Mean 8.56 9.96 10.68 10.91 D1 36.73 37.71 38.13 38.57 37.78 0.11 0.29 D2 32.98 33.87 35.72 36.83 34.85 0.92 2.26 D3 29.27 31.83 33.25 33.59 31.98 0.64 1.59 D4 24.53 26.51 29.41 29.94 27.60 0.005 0.01 D5 18.71 21.76 23.59 25.08 22.16 0.026 0.061 Mean 28.44 30.24 32.03 32.80

Treatment T1 T2 T3 T4 Mean SEm+ CD0.05

D1 11.14 11.98 12.49 12.69 12.14 0.12 0.31

D2 9.90 11.03 11.70 11.99 11.18 0.19 0.47

D3 8.70 10.43 11.29 11.49 10.48 0.007 0.018

D4 7.30 9.09 9.83 10.09 9.08 0.005 0.012

Table 14:

Effect of container on moisture content and total sugar of Cedrus deodara seed during storage Moisture content (%) Duration (Month) Total sugar (%) Duration (Month) D5 9.470 8.18 9.98 9.18 0.12 0.26 Mean 11.37 10.23 11.70 D1 8.86 8.63 9.19 8.89 0.10 0.21 D2 8.91 8.60 9.39 8.97 0.05 0.10 D3 8.06 7.53 8.52 8.04 0.006 0.01 D4 6.78 6.57 7.05 6.80 0.10 0.36 D5 5.32 4.67 5.81 5.26 0.06 0.13 Mean 7.59 7.20 7.99

Treatment C1 C2 C3 Mean SEm+ CD0.05

D1 12.75 1.62 12.93 12.45 0.05 0.11

D2 12.20 11.12 12.47 11.93 0.03 0.10

D3 11.69 10.61 12.01 11.43 0.70 0.15

D4 10.81 9.63 11.11 10.81 0.23 0.49

Table 15:

Effect of container on reducing and non reducing sugar of Cedrus deodara seed during storage Reducing sugar (%) Duration (Month) Non reducing sugar (%) Duration (Month) D5 0.89 0.98 1.00 0.95 0.02 0.05 Mean 1.44 1.40 1.53 D1 6.77 6.76 7.03 6.85 0.07 0.15 D2 6.68 6.49 7.03 6.73 0.12 0.26 D3 6.22 5.79 6.61 6.21 0.01 0.04 D4 5.27 5.04 5.47 5.26 0.078 0.16 D5 4.20 3.54 4.56 4.10 0.08 0.17 Mean 5.83 5.52 6.14

Treatment C1 C2 C3 Mean SEm+ CD0.05

D1 1.72 1.67 1.78 1.723 0.014 0.08

D2 1.87 1.75 1.98 1.87 0.018 0.039

D3 1.50 1.43 1.55 1.49 0.0030 0.006

D4 1.23 1.16 1.29 1.22 0.003 0.006

Table 16:

Effect of container on starch and phenol of Cedrus deodara seed during storage Starch (%) Duration (Month) Phenol (mg/g) Duration (Month) D5 7.36 9.68 7.58 7.28 0.003 0.008 Mean 10.11 9.69 10.30 D1 37.60 37.82 37.92 37.78 0.07 0.016 D2 35.07 34.44 35.05 34.85 0.95 2.01 D3 32.07 31.64 32.24 31.98 0.50 1.00 D4 27.73 27.11 27.95 27.60 0.01 0.003 D5 22.20 21.48 22.80 22.16 0.055 0.11 Mean 30.93 30.50 31.19

Treatment C1 C2 C3 Mean SEm+ CD0.05

D1 12.20 11.85 12.37 12.14 0.10 0.21

D2 11.24 10.88 11.41 11.81 0.14 0.30

D3 10.57 10.13 10.73 10.48 0.07 0.016

D4 9.17 8.69 9.38 9.08 0.003 0.008

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 Germinative capacity Germinative energy Germination value Moisture content Total sugar Reducing sugar Non reducing sugar Starch Phenol

1.0000 0.9265 0.9582 0.3966 0.0246 0.9411 0.8928 0.9184 1.0000 0.9182 0.3664 0.0401 0.8930 0.8499 0.8973 1.0000 0.3899 0.0201 0.9390 0.8432 0.9353 1.0000 -0.0016 0.3632 0.3675 0.3623 1.0000 0.0171 0.0237 0.0219 1.0000 0.8749 1.0000 0.9562 0.8388 1.0000

-0.1145 0.8681

-0.1003 0.7967

-0.1291 0.8996

-0.0307 0.3672

-0.0170 -0.0254

0.1648 -0.1161 0.8590 0.7509

-0.1302 0.8446

1.0000 -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

DURATION (MONTH) S.V D.F SS T (A) R (B) A*B C (C) A*C A*B*C 3 2 6 2 6 16 1566.3 32.05 6.83 106.89 24.67 69.78 D1 MS 522.11 16.02 1.34 53.44 4.11 4.36 SS 2495.6 22.39 1.61 234.89 11.11 8.00 D2 MS 831.88 11.19 0.27 117.44 1.85 0.50 SS 1575.6 24.67 1.11 206.17 23.61 1.56 D3 MS 525.21 12.33 0.19 103.08 3.93 0.097 SS 1574.60 70.72 10.61 408.72 44.61 25.33 D4 MS 524.85 35.36 1.77 204.36 7.43 1.58 SS 367.64 341.06 14.28 51.38 13.28 57.33 D5 MS 122.55 170.53 2.38 25.69 2.21 3.58

SV D.F SS

= Source of variation = Degree of freedom = Sum of square

R (B) = Replication A*B = Temperature * Replication

C (C) = Container A*C = Temperature * Container (Interaction)

MS = Mean sum of square 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

DURATION (MONTH) SV D.F SS T (A) R (B) A*B C (C) A*C A*B*C 3 2 6 2 6 16 367.64 341.06 14.27 51.39 13.28 57.33 D1 MS 122.55 170.53 2.38 25.69 2.21 3.58 SS 1385.9 260.06 20.61 491.56 107.78 66.67 D2 MS 461.96 130.03 3.44 245.78 17.96 4.16 SS 977.64 306.06 11.28 300.39 228.94 83.33 D3 MS 325.88 153.03 1.88 150.19 38.15 5.21 SS 956.75 62.00 7.33 266.67 37.33 18.67 D4 MS 318.92 31.00 1.22 133.33 6.22 1.17 SS 879.22 15.16 1.94 248.17 33.61 0.88 D5 MS 293.07 7.58 0.32 124.08 5.60 0.05

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
DURATION (MONTH)

SV

D.F SS

D1 MS 85.36 8.44 4.0 31.69 5.02 2.94 SS 526.53 54.89 4.89 140.22 10.22 15.56

D2 MS 175.51 27.44 0.81 70.11 1.70 0.97 SS 634.00 11.56 0.67 156.72 8.83 2.44

D3 MS 211.33 5.78 0.11 78.36 1.47 0.15 SS 712.67 16.72 1.50 141.06 15.16 1.78

D4 MS 237.56 8.36 0.25 70.52 2.53 0.11 SS 1382.9 16.22 1.11 187.72 40.94 3.33

D5 MS 460.96 8.11 0.18 93.86 6.82 0.20

T (A) R (B) A*B C (C) A*C A*B*C

3 2 6 2 6 16

256.08 16.89 24.00 63.38 30.16 47.11

4:

EFFECT OF TEMPERATURE (TABLE 3), CONTAINER (TABLE 5) AND THEIR INTERACTION (TABLE 9) ON GERMINATION VALUE OF Cedrus deodara SEED DURING STORAGE
DURATION (MONTH)

SV

D.F SS

D1 MS 705.12 344.59 781.87 233.36 802.64 670.14 SS 163.46 0.11 0.47 26.38 9.49 0.65

D2 MS 54.48 0.056 0.0078 13.19 1.58 0.040 SS 84.10 0.041 0.15 21.57 7.42 0.16

D3 MS 28.03 0.020 0.025 10.78 1.23 0.010 SS 91.45 0.18 6.86 14.65 3.08 0.64

D4 MS 30.48 0.091 0.01 7.47 0.51 0.040 SS 58.80 0.041 0.12 10.18 1.23 0.14

D5 MS 19.60 0.020 0.020 5.09 0.20 0.0093

T (A) R (B) A*B C (C) A*C A*B*C

3 2 6 2 6 16

2115.40 689.18 4691.2 466.71 4815.90 1072.20

5:

EFFECT OF TEMPERATURE (TABLE 11), CONTAINER (TABLE 14) AND THEIR INTERACTION (TABLE 17) ON MOISTURE CONTENT OF Cedrus deodara SEED DURING STORAGE
DURATION (MONTH)

SV

D.F SS

D1 MS 4.44 0.13 0.016 0.87 0.089 0.016 SS 68.14 0.58 0.050 25.59 2.50 0.19

D2 MS 22.71 0.29 0.0083 12.79 0.41 0.012 SS 45.55 0.028 0.010 19.91 7.17 0.46

D3 MS 15.18 0.014 0.0017 9.59 1.19 0.029 SS 122.62 0.33 1.60 25.58 6.05 5.20

D4 MS 40.87 0.16 0.26 12.79 1.00 0.32 SS 167.02 3.56 0.345 74.53 3.64 1.40

D5 MS 55.67 1.78 0.075 37.27 0.60 8.75

T (A) R (B) A*B C (C) A*C A*B*C

3 2 6 2 6 16

13.32 0.28 0.096 1.75 0.54 0.27

ii

6:

EFFECT OF TEMPERATURE (TABLE 11), CONTAINER (TABLE 14) AND THEIR INTERACTION (TABLE, 18) ON TOTAL SUGAR OF Cedrus deodara SEED DURING STORAGE
DURATION (MONTH)

SV

D.F SS

D1 MS 1.84 0.96 0.14 0.94 0.063 0.062 SS 28.05 0.80 0.77 3.84 0.50 0.24

D2 MS 9.35 0.40 0.12 1.92 0.083 0.015 SS 19.97 0.0013 0.0005 5.83 0.31 0.0039

D3 MS 6.56 0.0006 0.00009 2.91 0.051 0.0002 SS 36.28 1.53 0.61 1.43 2.17 0.97

D4 MS 12.09 0.76 0.10 0.71 0.36 0.060 SS 42.84 0.020 0.17 7.77 0.42 0.38

D5 MS 14.28 0.010 0.029 3.88 0.071 0.023

T (A) R (B) A*B C (C) A*C A*B*C

3 2 6 2 6 16

5.55 1.93 0.89 1.89 0.37 0.99

7:

EFFECT OF TEMPERATURE (TABLE 12), CONTAINER (TABLE 15) AND THEIR INTERACTION (TABLE 19) ON REDUCING SUGAR OF Cedrus deodara SEED DURING STORAGE
DURATION (MONTH)

SV

D.F SS

D1 MS 0.13 0.011 0.013 0.037 0.0011 0.0094 SS 2.97 0.017 0.038 0.31 0.017 0.032

D2 MS 0.99 0.0089 0.0064 0.15 0.029 0.0020 SS 1.25 0.0010 0.00051 0.083 0.0075 0.00086

D3 MS 0.41 0.00050 0.000086 0.041 0.0012 0.000054 SS 0.60 0.00067 0.00021 0.18 0.062 0.00091

D4 MS 0.20 0.00033 0.000036 0.090 0.010 0.000056 SS 1.09 0.0073 0.012 0.21 0.077 0.071

D5 MS 0.36 0.0036 0.0020 0.10 0.012 0.0044

T (A) R (B) A*B C (C) A*C A*B*C

3 2 6 2 6 16

0.41 0.022 0.068 0.075 0.0070 0.15

8:

EFFECT OF TEMPERATURE (TABLE 12), CONTAINER (TABLE 15) AND THEIR INTERACTION (TABLE 20) ON NON REDUCING SUGAR OF Cedrus deodara SEED DURING STORAGE
DURATION (MONTH)

SV

D.F SS

D1 MS 1.42 0.13 0.15 0.27 0.10 0.031 SS 11.53 0.057 0.11 1.80 0.40 1.46

D2 MS 3.84 0.028 0.019 0.90 0.067 0.091 SS 10.26 0.0013 0.013 4.08 0.29 0.032

D3 MS 3.42 0.000069 0.0023 2.04 0.049 0.0020 SS 20.49 0.29 0.59 1.06 1.22 0.58

D4 MS 6.83 0.14 0.09 0.53 0.20 0.036 SS 25.61 0.54 0.065 5.56 0.76 0.63

D5 MS 8.53 0.37 0.010 2.789 0.12 0.039

T (A) R (B) A*B C (C) A*C A*B*C

3 2 6 2 6 16

4.28 0.26 0.094 01.55 0.65 0.49

iii

9:

EFFECT OF TEMPERATURE (TABLE 13), CONTAINER (TABLE 16) AND THEIR INTERACTION (TABLE 21) ON STARCH OF Cedrus deodara SEED DURING STORAGE
DURATION (MONTH)

SV

D.F SS

D1 MS 2.95 0.42 0.074 0.86 0.010 0.061 SS 21.90 21.40 1.01 1.9 0.76 1.98

D2 MS 7.30 1.07 0.68 0.84 0.12 0.12 SS 48.94

D3 MS 16.31 0.0001 0.0002 0.038 0.24 0.0003 SS 42.57 0.00002 0.0007 3.01 0.244 0.0015

D4 MS 14.19 0.00001 0.0001 1.50 0.040 0.00009 SS 42.60 0.00007 0.0001 2.94 0.25 0.0014

D5 MS 14.20 0.00003 0.00002 1.47 0.043 0.00009

T (A) R (B) A*B C (C) A*C A*B*C

3 2 6 2 6 16

8.86 0.85 0.44 1.72 0.062 0.97

0.00035 0.0016 0.76 1.49 0.0060

10:

EFFECT OF TEMPERATURE (TABLE 13), CONTAINER (TABLE 16) AND THEIR INTERACTION (TABLE 22) ON PHENOL OF Cedrus deodara SEED DURING STORAGE
DURATION (MONTH)

SV

D.F SS

D1 MS 6.01 0.084 0.058 0.54 0.05 0.037 SS 82.06 19.60 23.17 3.04 35.91 87.08

D2 MS 27.35 9.80 3.86 1.52 5.98 5.44 SS 104.12 38.18 11.38 2.29 0.086 21.15

D3 MS 13.70 19.09 1.89 1.14 0.01 1.32 SS 181.53 0.0006 0.0009 4.18 2.48 0.0002

D4 MS 60.51 0.0003 0.0001 2.09 0.41 0.00001 SS 209.14 0.27 0..16 10.87 3.77 0.29

D5 MS 69.71 0.13 0.002 5.43 0.62 0.018

T (A) R (B) A*B C (C) A*C A*B*C

3 2 6 2 6 16

18.04 0.16 0.35 1.08 0.33 0.59

iv