World Journal of Agricultural Sciences 12 (6): 384-392, 2016 ISSN 1817-3047 © IDOSI Publications, 2016 DOI: 10.5829/idosi.wjas.2016.384.392

Effect of Different Pre-Sowing Treatments on Seed Germination of ebenum Koenig

11K. Jeyavanan, S. Sivachandiran, 1 S. Vinujan and 2 D.K.N.G. Pushpakumara

1Department of Agronomy, Faculty of Agriculture, University of Jaffna, Sri Lanka 2Department of Crop Science, Faculty of Agriculture, University of Peradeniya, Sri Lanka

Abstract: Diospyros ebenum Koenig is one of canopy dominant commonly called as Ceylon ebony and heavily extracted and threaten border line by anthropogenic activity in the dry zone forests in Sri Lanka, has led to the decrease in their population densities in the wild. Hence, conservation of the species is vital due its delayed germination and unprotected seedlings during its vegetative and reproductive periods. Mature fruits were randomly collected from and seeds were extracted from the fruits for assessment of microscopic, morphological and germination tests in the laboratory. Study was carried out at the Department of Agronomy, Faculty of Agriculture, University of Jaffna, Sri Lanka. The study aims to improve germination with the

application of pre sowing seed treatments such as different levels of H24 SO , KCl, hot water and distilled water in addition to control and morphological studies of seeds, fruits and seedlings. Experiment was conducted using a completely randomized design (CRD) with three replicates for germination test. Mean value of thickness, diameter and length of seeds were 0.64±0.02 cm, 1.19±0.02 cm and 0.79±0.02 cm, respectively. All the seed treatments were effective on germination percentage except distilled water soaked with 12 hours and highest value recorded as 63 %. Rate of root growth was high in seedling than the stem. Seedling had attained the mean height of 2.73 cm, 4.31 cm, 7.75 cm and 10.11 cm tall at 20, 30, 60 and 120 days, respectively after germination.

Key words: Germination Pre-sowing Seed treatments Sri Lankan ebony

INTRODUCTION lobed style and 8 uni-ovulate locules [5, 6, 7]. The sapwood is light yellowish grey, often streaked with Diospyros is the largest genus of the family black; heartwood very black, heavy with a specific gravity with more than 500 species, making it also one of 1.12 [8, 9, 10]. The wood is straight grained, fine and of the largest angiosperm genera. The greatest species of even-textured with a high glossy finish. The wood is diversity is in Asia and the Pacific region, the species difficult to season and work by hand. It is resistant to native to and Sri Lanka and Exotic in Malasiya [1]. insect attack and fungi and very durable [11]. It is mainly Diospyros ebenum Koenig is a ever green dry deciduous exported to China for furniture and to Europe as fancy forest species commonly called as black ebony, wood. It finds use in sports goods, musical and Ceylon ebony or “Kaluwara or Karungali” in Sri Lanka mathematical instruments, ornamental carvings and [2]. Tree has been known for its black wood since ancient turnery. Ebony requires overhead light once seedlings times. It produces one of the best commercial and most establish. The gummy astringent fruits are used as a expensive timbers in the world [2, 3, 4]. medicine and eaten in times of famine. It is a slow growing medium sized tree grown up to Anthropogenic activities such as selective logging or 30 m tall and up to 90 cm in diameter. Flowers mostly male extraction of canopy dominant timber of D. ebenum have and bisexual; male flowers in 3-16-flowered cymes, 4- led to the decrease in their population densities in the wild merous, stamens-16; female and bisexual flowers solitary, [12]. Based on the, [13], D. ebenum is identified as an 3-4 merous, calyx lobes valvate, glabrous, corolla divided endangered species. However, review of literature to about halfway, staminodes-8, ovary with a single 4-5- revealed that studies on seed, fruits, seedlings and

Corresponding Author: K. Jeyavanan, Department of Agronomy, Faculty of Agriculture, University of Jaffna, Sri Lanka. Tel: +94776381736, +94212060175. 384 World J. Agric. Sci., 12 (6): 384-392, 2016 germination of seeds of this species is lacking or floating them in distilled water prior to the experiment and nonexistence in global scale as well as local scale. those that floated were removed as they may not have Therefore, the objective of this study is to identify been viable whereas, those that sank and settled at the mechanisms for improving its seed germination by bottom were used in the experiment. Three glass petri applying different pre-sowing seed treatments. dishes each with a germination paper and allocated 10 seeds were used for each replicate. Germination papers MATERIALS AND METHODS were kept moistened and distilled water was added whenever necessary throughout the duration of the Experiments were conducted on seeds and fruits of experiments. Anon. (undated) reported that seeds are very D. ebenum in the laboratory at the Department of sensitive to desiccation during germination and early Agronomy, Faculty of Agriculture, University of Jaffna, growth, so must be regularly watered at this time. Seeds Sri Lanka during December to February in 2016. The were placed on moistened filter papers in 14.8 cm diameter

Ariviyal Nagar in Kilinocchchi District falls under DL3 and Petri dishes under laboratory condition for germination.

DL4 Agro-ecological regions of Sri Lanka located on Three hundred sixty seeds of D. ebenum were selected for latitude 23°34’S and longitude 25°57’E with an altitude of germination test and 10 seeds per Petri dish with three 36 m above mean sea level, located at 1.9 km from A-9 replicates were used for the experiment. Forest soil where highway [14]. the seedlings grown in natural place was taken as medium in poly bags only for control experiment. About 10 seeds Laboratory Environment: The environment of the were planted per poly bag with 3 replicates as the control. laboratory is medium shade with relatively high relative Three levels of H24 SO , KCl, Hot water, Distil water humidity than the outside environment. Average humidity were treated on ebony seeds with control. Total of twelve and light intensity were 78.68±2.52 % and 59.22±3.44 lux, treatments were compared with control (Table 5). Different respectively. Germination test was performed at natural concentrations of H24 SO were prepared by standard altering day and night room temperature varied from procedure in 250 mL volumetric flask. Seeds were divided 25±0.23°C to 28.52±0.35°C. and put into nine 50 mL heat resistant non-corrosive glass

beaker and H24 SO with different concentration (98%, 60% Seed Collection: Seeds of D. ebenum were collected from and 3%) was poured separately slowly on the side of the seed bearing tree in the Faculty during the period of beaker to a level where all the seeds were covered (25 mL). research was carried out [15]. Seeds were then stored in The seeds in the nine beakers with different levels or room temperature ranges from 25 to 30°C for three months. concentration were left for different times; 6 minutes, 30 Germination test was conducted using standard methods. minutes and 24 hours, respectively, after which the seeds were removed and the acid drained off into another beaker Measurement: Length, thickness and diameter of fruit and [16]. Seeds were thoroughly washed and rinsed to remove seeds were also measured by using a Vernier caliper and all the acid in tap water and distilled water, respectively. about 50 seeds and fruits were used for these Seeds were then put into petri dishes for germination measurements. Number of seeds or fruits per kg and 1, 000 examination. Seeds were divided and put into four 100 mL seeds or fruits weight were measured in the basis of dry heat resistant glass beaker for hot water treatments. The weight by using a digital weighing balance. Root, stem seeds in the nine beakers were soaked in hot water and and crown length was measured by using a calibrated left for different times with different temperature, after ruler. Stem of the seedlings was measured from embryonic which the seeds were removed from the hot water bath axis to bottom of leaf. Root of seedlings was measured and cool in room temperature and put in the Petri dishes from the embryonic axis. Crown length was taken from the for experiment. Different concentration of KCl was tip of leaf to bottom of the leaf. About 10 seedlings were prepared by measuring the weight of KCl and then, put randomly selected for the measurement in each stage such into volumetric flask 250 mL, then, slowly it was as 2 leaves younger stage (20 days age old), 2 leaves solubilized and make up to the mark [17]. About nine non mature stage (300 days age old), 5 leaves stage (60 days corrosive beakers were used and different concentrations age old) and 10 leaves stage (120 days age old). of KCl were left in different time. For water soaking, distilled water was used in nine beakers left in different Germination Test: Seeds were randomly selected among time [18] (Table 1). Germination test for all treatments the selected uniform seeds and were tested for viability by performed between moistened layers of filter paper.

385 World J. Agric. Sci., 12 (6): 384-392, 2016

Table 1: Different seed treatments used in the experiment Treatments Level 1 Level 2 Level 3

H24 SO 98% 6 minutes 60% 30 minutes 3% 24 hours Hot water 100°C 5 minutes 80°C 10 minutes 60°C 15 minutes KCl 15 mmol 12 hours 10 mmol 24 hours 5 mmol 72 hours Distilled water Soaking 72 hours Soaking 24 hours Soaking 12 hours Control Poly bags

Table 2: The formula used to calculate the index of germination Germination indices Unit The formula used References Final Germination Percentage (FGP) % Ng [22-24]. [17, 25, 26]. FGP = ∗100 Nt Germination Mean Time (GMT) Day NiTi [27-32]. GMT =∑ ×100 ∑ Ni

Germination Rate Index (GRI) %/day NN12 Ni [22, 30]. GRI = + ++... 12 i

Germination Value (GV) ΣDGS FGP [33]. GV = × N 10 Germination Energy (GE) %/day FGP [20]. GE = FGD

Coefficient Velocity Germination (CVG) ΣNi [29, 34]. CVG = ×100 ΣNiTi Imbibition Period (IP) day Days on which seeds start to germinated [20].

Days lapsed for 50 % Germination (T50) day (N + 1) [35-38]. − ni T50 = ti +−2 ()tj ti nj− ni   Final Germination Day (FGD) day Days on which seeds finally germinated [20].

Time Spread for Germination (TSG) day TSG = FGD – IP [21]. Note: DGS- Daily germination speed, obtained by dividing the cumulative germination percent by the number of days since sowing; ?DGS –sum of DGS, N- Total number of germinated seeds, starting from the date of first germination; Where Ni is the germination day of Ti represents the corresponding day of germination; 10 = is a constant; ni and nj are total number of seeds germinated by adjacent counts at time ti and tj, where ni, < (N +1)/2 < nj., if not, negative value is multiply with negative sign to get the germination days. If 50 % germination recorded in particular day not in the range, days on which is taken for 50 % germination; Ng-number of germinated seeds; Nt-total number of seeds in the trial

The Indices of the Germination: Germination value (GV) and increases when number of germinated seeds an expression of germination energy (GE) or speed of increases and time required for germination decreases. germination. Germination Value, as an integrated measure The germination rate index (GRI) gives an indication of the of seed quality [19]. Germination energy (GE) defined as percentage of seeds germinating per day of the the germination percentages when the mean daily experimental period. Time spread of germination (TSG) is germination (cumulative germination percent divided by the times elapsing between the first day of germination the time germination period) reached its peak. Germination (IP) and final germination day (FGD) [21]. energy is also treatments significantly affected the The tree is rarely raised artificially [39]. Chacko et al. germination period measure of the speed of germination [15] reported that fresh seeds are sown in germination and measure of the vigor of seedling [20]. trays and the seedlings are pricked out to polythene The germination mean time (GMT) represents the containers. A seed was considered germinated when the mean time a seed lot requires to initiate and end tip of the radical had grown free of the seed coat [40-42] germination. The coefficient of velocity of germination and Protrusion of radicle was considered as a marker for (CVG) gives an indication of the rapidity of germination germination as identified by Nasim et al. [43].

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The seed germination was recorded for 34 days. shaped seeds, 24.0% of small size shaped less seeds and The germination parameters of final germination 44.9% of very small seeds (Table 3). Orwa et al. [1] percentage (FGP); germination mean time (GMT); reported that fruit depressed globose to subglobose, up germination rate index (GRI); germination value (GV); to 1.5 cm across and glabrous and there are about 9, 000 germination energy (GE); coefficient of velocity of seeds/kg. In Sri Lanka, ebony occasionally seeds twice germination CVG; imbibition period (IP); time lapsed for per year. It occurs naturally in comparatively dry mixed 50% of germination (T50); final germination day (FGD); evergreen forests usually as an understory tree with other time spread for germination (TSG) were calculated by Diospyros species, Vitex altissima and Albizia using the standard formula (Table 2). odoratissima [1]. Flowering and fruiting period of the species were Data Analysis: The data collected was subjected to observed during the month of Februray and April 2015. analysis of variance (ANOVA) using the SAS program Fortunately, seedlings growth was observed during using Excel spread sheet. Treatment means were November 2015. This observation resulted that seeds of separated using Duncan's multiple range tests. Means the species had delayed germination account 7 months within columns followed by the same letters are not after fruit fall in the ground floor and growth rate of the significantly different. seedlings is very low (Table 3). Elias et al. [28] reported that Germination mean time GMT does not show the time from the start of imbibition Seed Germination Process: Fruits and seeds were oblong to a specific germination percentage. GMT has been used to ovo-oblong shape (Table 3). Micropyle was found to to compare specific pairs or groups of means and to opposite site of the fruit peduncle attachment specifically evaluate seed vigour. However, it is not the real time to in dorsal end of the seed. Embryo was found in the center mean germination but just an index of germination speed. of the seed and extended up to dorsal end (Fig. 1). Using GMT was not correct for ANOVA, post-ANOVA or Cotyledons were developed on the tip of the axis and later the other comparison tests, because it does not show time axis developed radicle for root development (Fig. 2). to a specific germination percentage. Elias et al. [28] recommend that T50 be used instead of GMT. The T50 Seedlings Growth and Development: Seedlings growth has all benefits of GMT, but it does not have the problems was epigeal type [15, 44, 45]. Root growth showed in of GMT in treatment comparisons. Hence, in this increasing trend compared with stem growth at 2, 5 and 10 experiment, time elapsed for 50% germination (T50) was leaves stages (Figs. 4 and 5). also calculated for treatment comparison with other Crown was initiated when seedling grown from two germination parameters. leaves mature stage to five leaves stage (Fig. 4). Growth rate of root, stem and crown had shown decreasing trend RESULTS AND DISCUSSION from two leaves younger stage to ten leaves stage. Root growth rate of seedling had decreased from 0.25 cm per Structure of Seeds and Fruits of Ebony: Seed structure day to 0.14 cm per day from two leaves stage to ten leaves and morphological charterer were examined by both stage. Similarly stem growth had decreased in trend from observation and microscopic studies in the laboratory. 0.14 cm per day to 0.04 cm per day. In each stage of Colour of the fruit and seed were shiny olive green seedlings, rate of growth high in root than stem and and dark brown to browinish black, respectively. Shape of crown (Table 4). the fruit and seed were globose to sub globose and Troup [48] reported that the seedlings develop long oblong to globose, respectively. Mean length of the fruit taproots at an early stage, often before any appreciable and seed were 1.74±0.05 and 1.68±0.03, respectively. elongation of the shoot takes place and the growth of the Mean diameter of the fruit and seed were 1.19±0.02 and seedling is decidedly slow [15, 39]. 0.79±0.02, respectively. 1, 000 fruits and seeds dry weight Colour of root was black in all stages of seedlings. were 565.40 g and 111.36 g, respectively (Table 3). However, stem colour was light green in younger 2 leaves Larger, small and very small seeds were recoded per stage and up to 5 leaves stage showed dark green in fruit. Total number of seed per fruit was 6.90±0.31. Out of colour. But, in 10 leaves stages colour turn in to black total number of seeds, fruit had 31.01% of large well (Table 4).

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Table 3: Morphological characters of seeds and fruits Category Characters Observation or Measurement Flowering and fruiting Flowering period February-April Fruiting period April- January Fruit Calyx Valvate Colour Shiny olive green colour Shape Berry, Globose to sub globose with lobes Length 1.740±0.049 cm Diameter 1.684±0.029 cm Number per 1, 000g (Dry weight basis) 1768.66 g 1, 000 fruits weight (Dry weight) 565.40 g Seeds Colour Dark brown to brownish black Shape Oblong globose Thickness 0.640±0.020 cm Length 1.193±0.022 cm Diameter 0.789±0.019 cm Number per 1000g (Dry weight basis) 8979.68 g 1000 seeds weight (Dry weight) 111.36 g Total number of seeds in fruit 6.900±0.312 Large size-well shaped 2.140±0.204 Small-Shapeless seeds 1.660±0.201 Very Small 3.100±0.225 Note: values are given with mean ± standard error

Table 4: Length, colour and growth rate of root, stem and crown of D. ebenum seedlings at 2, 5 and 10 leaves Root Stem Crown ------Stages Colour Length±SE (cm) Growth rate (cm/day) Colour Length±SE(cm) Growth rate (cm/day) Colour Length±SE(cm) Growth rate (cm/day) 2 leaves younger stage (20 days) Black 5.04±0.11 0.25 Light green 2.73±0.05 0.14 Light green ….. 2 leaves mature stage (30 days) Black 6.63±0.07 0.22 Dark green 4.31±0.03 0.14 Dark green ….. 5 leaves stage(60 days) Black 8.04±0.23 0.13 Dark green 4.98±0.06 0.08 Dark green 2.77±0.14 0.09 10 leaves stage (120 days) Black 16.79±0.77 0.14 Black 5.13±0.22 0.04 Dark green 4.98±0.08 0.08 Note: values are given with mean ± standard error (SE)

Figs. 1, 2 and 3: 1-longitudional cut of the seed before germination; 2-longitudition cut of the seed after seed germination; 3- Cotyledons (Magnification: 40x)

Figs. 4, 5 and 6: 4-different growth stages of D.ebenum seedlings; 5-Length of crown, stem and root; 6-leves number in different growth stages; A-2 leaves younger stage (20 days age old) B-2 leaves mature stage (30 days old), C-5 leaves stage (60 days age old), D-10 leaves stage (120 days age old)

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Fig. 7: Cumulative germination percentage of seeds during the experimental period

Seedling had attained the height of 2.73 cm, 4.31 cm, germination (T50), final germination day (FGD) and time 7.75 cm and 10.11 cm tall at 20, 30, 60 and 120 days, spread of germination (TSG), were 1-5 days (control: 23 respectively after germination. Lemmens et al. [45] days), 1-5 days (control: 17 days), 1-6 days (control: 14 reported that seedlings reach about 10 cm tall 75 days days), 2-7 days (control: 28 days) and 3-5 days (control: after germination and 20-30 cm tall 180 days after 12 days), respectively (Table 5). These result shows that germination. This results shown that tree has slow growth control had delayed germination compared to other seed rate. treatments. Anon. (undated) reported that normally the seed will Germination Test: Final germination percentage was not germinate after one week. In one trial, fresh seed, sown significantly differed among the treatments and its value one day after collection, showed 85% germination rate ranges from 63 % to 33 %. Treatment of distilled water within 17 - 65 days. Lemmens et al. [45] also reported that with 12 hours had lowest germination percentage account 80 % of germination was observed with simple water 17 %. soaking. Chacko et al. [15] and FRI [39] reported that However, in comparison with other parameters, germination of seeds were recorded up to 81 %. germination rate index (GRI), germination value (GV), Germination value is as defined by Czabator [19], aims germination energy (GV) and coefficient of velocity of to combine in a single figure an expression of total germination (CVG) should have higher value for the good germination at the end of the test period with an germination ability and lowest value for germination mean expression of germination energy or speed of germination. time (GMT), imbibition period (IP), times taken for 50 % of Germination value, as an integrated measure of seed germination (T50), final germination day (FGD), times quality, has been used by several tropical seed workers spread of germination (TSG) [21]. e.g. for Terminalia ivorensis [46]. and for Pinus kesiya Comparatively, all the seed treatments had higher [47]. Number of days from sowing to commencement of value for germination rate index (GRI), germination value germination is called as imbibition period of the seeds. (GV), germination energy (GV) and coefficient of velocity Germination energy is closely related to speed of the of germination (CVG). The control had significantly higher germination and vigour of the seedlings. value for germination mean time (GMT), imbibition period Troup [48] reported that as a rule fresh seeds have a (IP), times taken for 50 % of germination (T50), final high percentage of fertility. Chacko et al. [15] reported germination day (FGD), times spread of germination (TSG) that viability period of the seeds is two months. Seeds shows that control had delayed germination compared to were collected and stored at room temperature in the other seed treatments. laboratory about three months. The germination test was

Handling of concentrated H24 SO and KCl are harmful also delayed after seed collection. This may lead to lower to public and it is a costliest method than the treatment of germination in the experiment. Due to this, research is hot water with all levels and distilled water. However, needed after immediate collection of the seeds. distilled water treatment soaked with 12 hours had lower Germination period will takes about 226 days [15, 39]. germination percentage. Hence, application of seed treatments improve For all seed treatments, germination mean time (GMT), germination and break the dormancy of the seeds of the imbibition period (IP) and time lapsed for 50 % ebony.

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Table 5: Comparison of different levels of treatments for germination parameters Treatments Levels FGP GMT GRI GV GE CVG IP T50 FGD TSG

H24 SO 98% 5min 60.00±5.77a 3.59±0.23 bced 2.01±0.20 bced 15.20±1.41 ab 1.00±0.14a 28.08±1.81bcd 2.00±0.00 b 3.56±0.44 b 6.33±1.20 Bc 4.33±1.20b 60% 30 min 63.33±6.67a 4.53±0.29 bced 1.67±0.23 bcdef 14.26±1.66 ab 0.90±0.10 a 22.25±1.47cd 2.00±0.00 b 5.22±0.91 b 7.00±0.00 bc 5.00±0.00b 3% 24 hr 63.33±3.33a 4.43±0.21 bced 1.70±0.17 bcdef 14.40±1.02 ab 0.94±0.13 a 22.67±1.01 cd 2.00±0.00 b 4.39±0.31 b 7.00±1.00 bc 5.00±1.00b KCl 15 mmol 12 hr 36.67±12.02ab 3.39±0.56bced 1.20±0.31 cdef 9.20±2.57 bc 0.81±0.10 b 31.02±4.56 b 2.33±0.33 b 1.42±1.42 b 4.67±1.67 bc 2.33±1.86b 10 mmol 24hr 46.67±16.6ab 2.47±8.18 ced 2.69±0.17 bc 14.55±4.97 ab 1.72±0.49 ab 40.86±2.82ab 1.33±0.33b 0.67±0.67b 2.67±0.33Bc 1.33±0.33b 5 mmol 72 hr 50.00±15.2ab 1.38±0.38 ed 4.46±1.01 a 19.16±4.90 a 2.71±0.84 a 82.35±17.6a 1.00±0.00 b 0.25±0.25 b 3.00±2.00 bc 2.00±2.00b Hot water 100°C 5 min 43.33±12.0ab 6.46±0.27 b 0.73±0.23 def 7.85±2.33 bc 0.58±0.15 a 15.55±0.63d 5.00±1.00 b 4.67±2.33 b 7.33±0.33 b 2.33±1.20b 80°C10 min 43.33±8.82ab 5.44±0.29 bc 0.85±0.21 def 8.59±1.94 bc 0.67±0.17 a 18.47±0.97 cd 4.33±0.33 b 1.44±1.44 b 6.67±0.67 bc 2.33±0.88b 60°C 15 min 63.33±8.82a 5.04±0.23 bcd 1.29±0.12 cdef 12.89±1.45 ab 0.95±0.13 a 19.90±0.86cd 4.00±0.00 b 5.81±0.6 b 6.67±0.33 bc 2.67±0.33b Distilled water 72 hr 33.33±8.82 ab 1.27±0.27a 3.07±0.64ab 13.03±2.98 ab 2.00±0.58a 85.19±14.8a 1.00±0.00b 1.67±1.67b 2.33±1.33C 1.33±1.33b 24 hr 43.33±12.02ab 2.27±0.37 ced 2.22±0.78 bcd 13.51±4.18 ab 2.06±0.71 a 46.30±6.68 b 2.00±0.58 b 0.53±0.53 b 2.33±0.33C 0.33±0.33b 12 hr 16.67±8.82 b 4.25±2.16 bced 0.41±0.27 ef 3.26±1.86 c 0.24±0.13 a 10.56±5.36 d 3.00±2.08b 0.00±0.00 b 4.67±2.33 bc 1.67±1.67b Control 56.67±8.82 a 22.37±3.27 a 0.29±0.07f 3.04±1.11 c 0.21±0.04a 11.87±7.56 d 16.67±9.18 a 14.00±7.02 a 28.00±3.00a 11.33±3.94a Note: Values were mean ± standard error. Similar letters in the column were statistically not significant at á 0.05. DW: Distilled water; Con.: control; each treatment level was tested for Duncan’s means with separately. FGB: final germination percentage; GMT: germination mean time; GRI: germination rate index; GV: germination value; GE: germination energy; CVG; coefficient of velocity of germination; IP: imbibition period; T50: time lapsed for 50% of germination; FGD: final germination day; TSG: time spread for germination

CONCLUSION 2. Purkayastha, S.K., 1996. A Manual of Indian Timbers. Sri Bhumi Publishing Company, Calcutta. From this experiment, it can be concluded that all the 3. CSIR, 1976. The Wealth of India: A dictionary seed treatments were effective on germination percentage of Indian Raw Materials and Industrial except distilled water soaked for 12 hours. In comparison Products, Raw Materials, Rh-So, Publication of all germination parameters, control was less effective, and Information Directorate. Council of even it had highest germination percentage. Handling of Science and Industrial Research, New Delhi, India, concentrated acid H24 SO and KCl harmful to researchers 9: 591. and it is a costliest method. Hence, Hot water treatment 4. Turner, B., J. Munzinger, S. Duangjai, and distilled water soaked with 72 and 24 hours was better E.M. Temsch, R. Stockenhuber, M.H.J. Barfuss, seed treatments for germination of species D. ebenum. M.W. Chase and R. Samuel, 2013. Molecular Conservation of the precious resources are crucial in the phylogenetics of New Caledonian Diospyros wild as well as in In-Situ by understanding of germination (Ebenaceae) using plastid and nuclear markers. and other related parameters. Further studies are needed Molecular Phylogenetics and Evolution, on growth and development of seedlings with these 69(3): 740-763. pre-sowing seed treatments. 5. Bose, T.K., P. Das and G.G. Maiti, 1998. Trees of the World, Regional Resource Centre, Orissa. India, ACKNOWLEDGEMENT 1: 173. 6. Brandis, D., 1978. Indian Trees: An Account I would like to express my sincere thanks to Staff and of Trees, Shrubs, Woody Climbers, Bamboos Technical officers of the Department of Agronomy, and Palms Indigenous or Commonly Faculty of Agriculture, University of Jaffna and Dr. (Mrs). Cultivated in the British India Empire. London, N. Gnanavelrajah, Head, Department of Agricultural pp: 767. Chemistry and Mr. M. Prabhakaran, Head, Agricultural 7. Sahni, K.C., 2000. The Book of Indian Trees. BNHS, Engineering for providing equipment and necessary Mumbai. support for the experiment . 8. Sankaram, A.V.B. and R.V.V. Natayana, 1984. Structure of ebenone, a possible biogenetic precursor REFERENCES of elliptinone, from Diospyros ebenum. Phytochemistry, 23(9): 2039-2042. 1. Orwa, C., A. Mutua, R. Kindt, R. Jamnadass and 9. Santhakumaran, L.N. and V.V. Srinivasan, 1993. S. Anthony, 2009. Agroforestry Database: a tree Studies on the natural bioresistance of some Indian reference and selection guide version 4.0, Date of timbers in Goa waters. Proceedings of the National accesses: 25/02/2016 (http://www.worldagroforestry. Academy of Sciences India Section B Biological org/sites/treedbs/treedatabases.asp). Sciences, 63(3): 337-340.

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