Impact of pre-sowing treatment on seed germination and seedlings ...

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Journal of Food, Agriculture & Environment Vol.12 (3&4): 395-399. 2014

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Impact of pre-sowing treatment on seed germination and seedlings growth attributes of Calamus longisetus Griff. at nursery and field conditions M. Rafiqul Haider 1*, Md. Sah Alam 1, Md. Aktar Hossain 1

2, 3

* and Nor Aini Ab. Shukor

3, 4

Bangladesh Forest Research Institute, P. O. Box 273, Chittagong - 4000, Bangladesh. Institute of Forestry and Environmental Sciences, Chittagong University, Chittagong - 4331, Bangladesh. 3 Faculty of Forestry, 4 Institute of Tropical Forestry and Forest Products, University Putra Malaysia, 43400 UPM, Serdang, Selangor, Malaysia. *e-mail: [email protected], [email protected] 2

Received 30 May 2014, accepted 12 September 2014.

Abstract The paper describes the germination behaviour of Calamus longisetus Griff. seeds under three different pre-sowing treatments and seedlings growth performance in nursery and field condition. Whole fruits, seeds with pulp and cleaned seeds were sown in germination trays filled with soils mixed with decomposed cowdung at a ratio of 3:1. The growth performance of the seedlings were determined by transferring the young seedlings (having 2 - 3 leaves) from germination bed to the polybags filled with soil mix followed by outplanting (at 1-year old) in the field. Germination percentage was significantly (p ≤ 0.001) enhanced by the pre-sowing treatment where the highest germination percentage was in cleaned seeds followed by the seeds with pulp and lowest in whole fruits. The survival percentage was over 91% in the field after one year and the average height 126.2 cm after two years of planting. Clean seeds for nursery raising and one-year old seedlings for outplanting were found suitable for raising successful plantations for the species. Key words: Calamus longisetus, cleaned seeds, germination potential, survival percentage, pre-sowing treatment, seedling growth.

Introduction Rattan is a climbing spiny palm belonging to the family Arecaceae (Palmeae) and constitutes an integral part of the tropical forest ecosystem 1. Rattans are the most important non timber forest products (after bamboo) in the tropical and sub-tropical counties of Asia and Africa. It has gained additional interest among the people because of increased awareness as well as their vital roles in socioeconomic development and socio-ecological research issues 2. Rattans are important sources of income and employment for millions of people all over the world directly or indirectly. In India, rattan industries alone provides jobs for 200,000 people 3 and its contribution is about 25 - 35% of the total household income of the tribal communities in North Eastern India. Rattan furnitures are much valued in many countries, and are being exported abroad from the producing countries 4. Six hundred species of rattans under 13 genera were recorded in the world in early nineties 4 but recent report shows that only 558 species exist 5. Of these, 15 species were recorded in Bangladesh 6-11 but currently there are only 10 species of rattans under two genera growing in the country 12, 13. These rattan resources have depleted fast in recent years due to over exploitations and poor management 14 including almost complete removal of stems from the forest during harvesting. Consequently, steady loss of forest habitat due to urbanization and industrialization is also posing a serious threat to rattan supply. High demand for these resources, coupled with uncontrolled harvesting and deforestation, has led the resources towards the exhaustion in many rattan-producing areas 15. To cope with the increasing global demand for rattan, there is an urgent need for sustainable management of rattan resources. There are two different approaches to achieve the goals, by establishing rattan plantations and managing existing rattans

properly in their natural habitats 15. However, establishing the rattan plantation requires appropriate knowledge about the nursery development, plantations raising and stand management for sustainable development 14. Calamus longisetus locally known as udum bet, occurs naturally in the dry hill slopes, sandy loam soils of mixed evergreen forests of Chittagong, Chittagong Hill Tract and Cox’s Bazaar district in Bangladesh 7, 12. The species is found in robust clustering to form dense clumps, erect for climbing where support tree is available. Stems covered with sheath and internode ranges from 18 - 25 cm long. Leaf is ecirrate, 3 - 4 cm long with petiole. Leaf sheath is greenish-yellow, densely armed with 3 - 5 cm long, flat, erect, spreading and reflexed spines (Fig. 1). The species is dioecious; flowering occurs in October to December, fruiting in March to

Figure 1. Leading shoot and spiny stem of Calamus longisetus.

Journal of Food, Agriculture & Environment, Vol.12 (3&4), July-October 2014

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June and is pollinated mainly by insects. It is mainly used for making furniture frame, umbrella handles, walking sticks, binding materials and stem water for drinking by the tribal people. With depletion of rattan resources C. longisetus is also depleted all over the country and hardly found in the natural condition in forests. To ensure sustainable development and continuous supply of rattans, several initiatives have already been taken to increase the rattan plantations and the overall production by involving the government, non-government and other agencies in the country. For the purpose sound knowledge on nursery raising and plantation development, updated techniques and management systems are required for the species. Rattan research in Bangladesh was initiated about four decades back and considerable information relating to germination and nursery techniques for selected rattan species are available 16-21. Rattans cannot be grafted or rooted by cuttings, therefore their propagation largely depends on seed germination, wildings, suckers and tissue cultured materials for plantation. Among them, seeds are the most important propagating materials for large-scale plantations program for rattans 22, 23. There are considerable variations among the rattan species in respect to germination period and percentage. The seed viability after collection may affect the germination period and percentage. Moreover, because of hard seed coat in association with possible seed dormancy, most of the species of cane require longer period to germinate 24. Pre-sowing treatments, especially, removal of fleshy pulp or scales from the intact fruits has been reported to influence the seed germination and seedling growth performance in many tropical tree species. For instance, depulping of fruits and soaking of the seeds in water for 48 h enhanced the seed germination and seedling growth of T. belerica 25-27, T. chebula 28-30 and Grevillea robusta 31. Nainar et al. 28 showed that seed pre-treatments including mechanical scarification offered 60% germination in T. chebula seeds. Although there are some reports on pre-sowing treatment and germination of some rattan species, for example, pre-treatment of Daemonorops jenkensiana clean seeds with 10% H2SO4 and HCl enhanced seed germination (68.14% with both the treatments) 21, the information about the regeneration of Calamus longisetus with seed treatments is very scare. Further research related to the nursery raising techniques including seed germination, pre-sowing treatments and initial seedling growth performance of this species is therefore required with an ultimate objective of successful rattan plantation. Considering the facts the present study was undertaken to investigate the effects of pre-sowing treatment on seed germination and seedling growth performance of C. longisetus in nursery and field condition. Materials and Methods Present research was carried out in the nursery of Bangladesh Forest Research Institute (BFRI), Chittagong, Bangladesh, over a period of three and half years from June 2010 to November 2013. The study area was located between 22°22·27´´ and 22°29·0´´ North latitude and 91°46·30´´ and 91°46·30´´ East longitude enjoys a tropical climate, characterised by hot humid summer and cool dry winter. The maximum and minimum temperature in the area are 28.3 - 31.9°C and 15.2 - 25.2°C 32. Relative humidity is usually low in winter (November - February) and high in summer (June September). Mean annual rainfall is around 300 cm, mainly occurring from June to September. 396

Seed collection and pre-sowing treatments: Ripe fruits of Calamus longisetus were collected from natural forest of Ukhia, under Cox’s Bazaar district of Bangladesh in the first week of June 2010. To determine the effect of pre-sowing treatment on seed germination and seedling growth attributes, three different treatments were applied namely, i) whole fruit ( seed + sarcotesta + scale), ii) seed with pulp (seed + sarcotesta without scale) and iii) clean seed (seed without sarcotesta and scale) (Fig. 2). Clean seeds were obtained by removing the scale and pulp through rubbing. The numbers of whole fruits and cleaned seeds per kg were 200 - 220 and 600 - 650, respectively. Seeds of C. longisetus were sown in the germination trays filled up with soil mixed with decomposed cowdung at a ratio of 3:1 by volume. Complete Randomized Design (CRD) was adopted for the experiment with three replications. Fifty seeds were sown in each replication and total of 450 seeds were subjected to three treatments for the germination trial. The trays were kept under nursery shade for one week and then exposed to partial sunlight. A)

(A) Fi

B)

2 Wh l f it (A)

d l

d

(B) d (B) f C l

i t

d f

i

Figure 2. Whole fruits (A) and cleaned seeds (B) of C. longisetus are ready for sowing in the germination trials.

i

Assessment of seed germination and seedling growth performance: The effects of pre-sowing treatments on seed germination and seedling growth were explored periodically through counting the germinated seeds and assessing initial growth performance of seedlings. Cumulative germination was recorded in every third day from the day of sowing and continued till ending the germination (106 days after sowing the seeds). Germination phase like imbibition period was determined by counting the number of days required for the commencement of germination from the day of sowing and germination period was the number of days required for completion of germination from sowing the seeds. For assessing the growth performance, all seedlings were measured for above ground height (form base to leaf tip) and number of leaves four months after sowing the seeds. Besides these ten seedlings from each replicate (30 from each treatment) were randomly uprooted and measured for total length (root length and shoot length separately) for the assessment. Seedling vigor index (VI) was calculated according to Abdul-Baki and Anderson 33 as the germination percent multiplied by total length of seedling (i.e. sum of shoot and root length). Assessment of seedling growth performance in the nursery and the field: To determine the seedling growth performance in the nursery and the field, clean seeds were first sown in the nursery bed. When the seedlings were about two months old (with 2 - 3 leaves), they were transferred in to the polybags (23 cm x 15 cm in size) filled with soil mixed with cowdung. The polybags were kept


under full shade for one week and then placed under direct sunlight where they were allowed to grow. When the seedlings were about one-year old, 1000 seedlings were out planted in five plots in the field at the beginning of the monsoon, e.g. June. Rest of the seedlings (1000), were allowed to grow in the nursery for one more year. Data on shoot and root length and leaf number of these seedlings were also recorded at three, six, twelve and twenty four months after transferring them in polybags. Seedlings in the field were planted at 2 m × 2 m spacing at the foot hills of Hinguli Research Station, Chittagong, Bangladesh. The soil was sandyloam with a pH 5.7 - 6.0. Average rainfall of the area was about 3200 mm and average maximum and minimum temperature was 34.7°C and 20.7°C, respectively, indicating the ecophysiological conditions of the planting site were ideal for the rattan plantations34. Weeding was done at every six months in the field but no fertilizer or water was added after planting. One hundred seedlings from each of the five selected plots were measured to determine the height growth of the species in the field. Data on the height of each plant were recorded at six, twelve and twenty four months after planting. Survival percentage of the planted seedling in the field was determined one year after planting the seedlings. All data were analyzed with computer software IBM SPSS ver. 21 to determine the significant (p ≤ 0.001) variations among the treatments.

400 200 00

Whole fruit

80 80

800 600

Cummulative germination(%)(%) Cummulative germination

Germination percentage

Results and Discussion Seed germination and initial growth performance of the seedlings: Pre-sowing treatments significantly affected the germination period and germination percentage of C. longisetus seeds. Seed germination started 56 days after sowing (DAS) the seeds and continued until 106 DAS. The fastest germination was noticed 56 DAS in cleaned seeds followed by seeds with pulp (62 DAS) and slowest seed germination (64 DAS) was in whole fruits. Clean seeds showed 60% germination between 70 and 80 DAS days and germination continued up to 100 days. In the seeds with pulp and whole fruits 40% and 35% germination occurred in between 70 and 80 DAS and germination continued up to 106 DAS (Fig. 2). Finally, the germination percentage reached to 73.3 in clean seeds, 45.7 in seeds with pulp and 35.7 in whole fruits (Fig. 3). Although the germination percentage in the cleaned seeds was significantly (F = 56.083; p < 0.001) higher than in the other treatments, there was no variation between whole fruits and seeds with pulp. Like other members of the family Palmeae, the species Calamus required long time to germinate. Generalao 35 reported that cane seeds take weeks to six months to germinate depending on the

(A)

60 60

Whole fruit

(B)

Seed with pulp

Seed with pulp

species and method of treatment. Sumantakul 36 reported that C. longisetus seed in different media starts to germinate from 30 days and continues till 60 days. Banik and Nabi 16 mentioned that the seeds sown with intact sarcotesta require two or three months to start germination and give poor germination percentage (10 - 26 only). Manokaran and Wong 37 reported that removal of the sarcotesta is a necessary pre-treatment for cane to shorten the germination period and obtain good levels of success. The findings of the present research are also unidirectional to those reported in the previous studies mentioned here. The initial growth performance of the C. longisetus seedlings was also significantly affected by the pre-sowing treatment of the seeds. The shoot length (from base to leaf tip), root length, leaf number and vigor index are shown in Table 1. The highest length of shoot and root and vigor index was noticed in the cleaned seeds which values were much higher than those in the whole fruits and seeds with pulp. However, there was no significant variation in the leaf number among the treatments. Similar results were reported by several authors and mentioned that depulping or removing the fleshy outer parts of the fruits enhance the seed germination and seedling growth performance in the nursery conditions. Hossain et al. 29, 30 mentioned that the seedling growth including root, shoot and total length of T. chebula was significantly increased with pre-sowing treatment specially by depulping the fruits. Again, in separate experiments, Hossain et al. 27, 29 showed maximum growth including shoot length, root length, total length and vigor index in T. belerica seedlings when fruits were depulped and soaked in cold water. However, similar to this experiment, there was no remarkable difference in average number of leaves per seedling of T. belerica seedlings 26. Table 1. Initial growth performance of the seedlings germinated from various treatments one month after germination. Growth parameter Shoot length (cm) Root length (cm) Leaf number Vigor index

Cleaned seeds 12.53 ± 1.6 a 6.7 ± 0.87 a 2.1 ± 0.52 a 1409.56 a

The vigor index of the seedlings in this experiment was dramatically increased from 489.8 in the whole fruits to 1409.56 in the cleaned seeds (Table 2). The vigor index usually depends on the germination percentage and the seedling length. In this study we noticed that the seedling length was marginally significantly different among the treatments but the germination percentage in the cleaned seeds was almost doubled of that in the whole seeds and the seeds with pulp for the Clean seed Clean seed species which led the vigor index substantially higher in the cleaned seeds than in the other treatments.

20 20

00

Seeds with pulp 9.04 ± 1.3 b 5.32 ± 0.36 ab 1.4 ± 0.51 a 647.11 b

Note: The same different letter(s) in each row are significantly different at p ≤ 0.05, according to Duncan’s Multiple Range Test (DMRT). ± indicates the standard error of mean.

40 40

Whole fruit Seed with pulp Clean seed Treatment

Whole fruits 9.1 ± 1.03 b 4.62 ± 0.42 b 1.3 ± 0.3 a 489.80 b

60 64 67 70 73 76 79 82 85 88 91 94 97 100 103 106 109 Day after sowing

Figure 3. Germination percentage (A) and the germination pattern (B) of C. longisetus for whole fruits, seeds with pulp and cleaned seeds over time. Journal of Food, Agriculture & Environment, Vol.12 (3&4), July-October 2014

Seedlings growth performance in nursery condition: In our preliminary experiment (Fig. 3), it was noticed that the germination percentage of the cleaned seed was almost double of that in the whole fruits and seeds with pulp. The initial growth performance of these seedlings including seedling length and vigor index were also 397

Table 2. Seedling growth of C. longisetus at different age (up to 24 months) in the nursery. Age of seedlings (month) 3 6 12 24

Average height (cm) 17.5 ± 2.5 22.8 ± 2.8 30.2 ± 3.7 82.7 ± 6.2

Average length of Roots (cm) 10.1 ± 2.2 13.4 ± 2.8 16.6 ± 1.7 27.2 ± 3.8

No. of leaves Per seedlings 3.1 ± 0.9 5.0 ± 1.2 7.6 ± 1.4 11.3 ± 2.1

The figure in each column mean followed by standard error (SE) of means.

significantly higher than those in the other treatments (Table 1). We, therefore, sowed only the cleaned seeds in the seed beds for assessing the seedling growth performances in the nursery and the field. Three thousand cleaned seeds were sown in three blocks (considered as replications) of the nursery bed for the purpose. The germination percentage (71.6 ± 2.3) was almost similar of the previous experiment (Fig. 3). Two-months old (after germination) seedlings having 2 - 3 leaves were transferred in to the polybags filled with soil mixed with cowdung and allowed them for growing there. After one year of transferring the seedlings in the polybags, 1000 seedlings were outplanted in the field. Rest of the seedlings (1000) were grown in the nursery for one more year. The seedling mortality in the nursery bed (less than three percent) and during and after transplanting the seedlings to the polybags (around 1%) was very negligible. Growth performance of seedlings in the nursery determined at different age is shown in Table 2. The seedlings attained 17.5 cm height (above ground) with average length of root 10.1 cm and 3.1 leaves in three months. The seedlings became quite tough and attained a height of 22.8 cm with 13.4 cm root and 5.0 leaves at six months. The average height 30.2 cm with 16.6 cm root and 7.6 leaves was recorded at twelve months. The seedlings attained a height of 82.7 cm with 27.2 cm long root and 11.3 leaves at twenty four month (Table 3 and Fig. 3).

Table 3. Survival percentage and seedling growth of Calamus longisetus after out planting. Plots Plot - 1 Plot - 2 Plot - 3 Plot - 4 Plot - 5

Survival (%) at 12 months 90.0 88.5 92.5 91.0 94.5

Average height (cm) at 6 months 12 months 24 months 52.8 ± 2.9 c 70.9 ± 2.4.7 b 117.3 ± 3.8 b 67.6 ± 3.7 a 82.4 ± 3.7 ab 128.5 ± 2.8 ab 61.7 ± 2.2 ab 88.6 ± 4.4 a 134.0 ± 6.6 a 69.4 ± 1.8 b 122.9 ± 3.8 b 58.8 ± 2.0 b 63.5 ± 1.0 ab 80.2 ± 3.5 ab 128.3 ± 4.7 ab

Means followed by the same letter (s) are not significantly different at p ≤ 0.05, according to Duncan’s Multiple Range Test (DMRT). ± indicates the standard error of the mean.

(A)A)

(B) B)

Seedling survival and growth performance in the field: One year old seedlings of C. longisetus developed from cleaned seeds grown in polybags were planted in the field. Survival and seedlings growth performances were determined at 6, 12 and 24 months after planting in the field (Table 3). Survival percentage varied from 88.5 to 94.5 with an average of 91.3 among the plots. The seedling height varied from 52.8 to 67.6 cm at six months, from 69.4 to 88.6 cm in one year and from 117.3 to 134.0 cm in two years of planting (Figs 4 and 5).

Figure 5. Growth performance of C. longisetus 18 months after transplanting in the field.

The height growth of seedlings was always significantly higher in plot 3 than in the other plots and lowest in plot 1. This variation of the height growth in the seedlings was probably due to the microclimate of the plots or some other unknown reasons. Since the survival percentage of the seedlings in the field was quite satisfactory, out planting of one-year old seedlings instead of older seedlings may be considered for the planting the species in the field. Similar report was also made by the Kerala Forest Research Institute, India, and mentioned that rattan seedlings were out planted at one-year old 38. Another interesting finding in this study was that the plant height in the nursery condition at two years (82.7) after transferring them into polybags was almost similar to that in the field (69.4 - 88.6 cm) after one year of planting (two years after transferring in to the polybags). However, it was not possible to maintain the seedlings in the nursery after two years of transferring them in polybags due to some unavoidable circumstances.

(C) C)

Figure 4. Different stages of seedlings of C. longisetus. One-month old seedlings of C. longisetus in nursery bed (A), an individual seedling prior to transfer into the polybag (B) and two-year old seedlings in the nursery (C). 398

Conclusions Germination behaviour of C. longisetus is similar to other members of the family Palmeae and needs longer time to germinate. C. longisetus seeds start germination after 56 days of sowing and complete within 106 days. Maximum seed germination and highest initial growth performance of the seedlings was observed in clean seeds which was much higher than those in the other two treatments. Delayed pricking at 100 - 120 days after sowing the seeds from nursery bed to polybags ensures least or no mortality of the seedling. Survival of seedlings (91%) and growth performance in the field was satisfactory when one year old seedlings were outplanted. Therefore, clean seeds for the nursery


raising and one-year old seedlings for outplanting the species may be recommended for large-scale plantation programs. Acknowledgements We are thankful to Mr. S. M. Zahirul Islam, Research Officer for his support in data analysis. Thanks are due to Mr. Abul Bashar, Forest Ranger, Mr. Abdus Salam, Nursery Supervisor and other staff members of Minor Forest Products Division, BFRI for their help in the execution of the study. References Moore, H. E. Jr. 1973. The major groups of palms and their distribution. Gentes Herbarum 11(2):27-141. 2 Nagabhatla, N., Roy, P. S. and Jagdale, R. 2007. Monitoring spatial distribution of commercial rattans and palms in the tropical forest of Baratang Islands (Andaman and Nicobar Islands). Indian Journal Traditional Knowledge 6(4):630-635. 3 Manokaran, N. 1990. The State of Bamboo and Rattan Trade. Rattan Information Centre Occasional Paper No. 7. RIC, Forest Research Institute, Kepong, Malaysia, 39 p. 4 Renuka, C., Indira, E. P. and Muralidharan, E. M. 1998. Genetic Diversity and Conservation of Certain Species of Rattan in Andaman and Nicobar Islands and Southern India. KFRI Research Report 157, Kerala Forest Research Institute, Peechi, Thrissur, 38 p. 5 Dransfield, J., Uhl, N. W., Asmussen-Lange, C. B., Baker, W. J., Harley, M. M. and Lewis, C. E. 2008. Genera Palmarum: The Evolution and Classification of Palms. Kew Publishing, UK, 732 p. 6 Wong, K. M. 1984. On the Feasibility of an Export Oriented Rattan Furniture Industry in Bangladesh. Occasional Paper No. 1, Rattan Information Centre, Forest Research Institute, Kepong, Malaysia, 15 p. 7 Alam, M. K. 1990. Rattans of Bangladesh. Bulletin 7. Plant Taxonomy Series. Bangladesh Forest Research Institute, 33 p. 8 Alam, M. K. 1991. Rattan resources of Bangladesh and their status. RIC Bulletin 10(1):2-5. 9 Basu, S. K. 1991. Living rattan collection in the Forest Research Institute, Chittagong. RIC Bulletin 10(2):5-6. 10 Basu, S. K. 1992. Rattans (Canes) in India: A Monographic Revision. Rattan Information Centre, Forest Research Institute, Keopong, Malaysia, 141 p. 11 Ali, S. S. 2003. Forest and Forestry. In Islam, S. and Miah, S. (eds). Banglapedia. Asiatic Society of Bangladesh, Dhaka, 4:248-250. 12 Ara, R. 2008. An Overview of Rattan Resources in Bangladesh. Document no.2. Market Development of Bamboo and Rattan Products with Potential. CFC INBAR-01 Project, 9 p. 13 Ara, R. 2011. Assessment of reproductive features and propagation potential in rattans of Bangladesh. Indian Forester 137(12):1445-1450. 14 Siddiqi, N. A. 1995. Rattan research in Bangladesh. INBAR Newsletter (India) 3(2):8. 15 Supardi, M. N. N., Khali Aziz, H. and Wan Razali, M. 1999. Considerations in rattan inventory practices in the tropics. INBAR Technical Report 14. International Network for Bamboo and Rattan, Beijing, China, 57 p. 16 Banik, R. L. and Nabi, M. N. 1979. A note on the flowering periodicity and the seed germination in bara bet. Bano Biggyan Patrika 8(1&2):52-56. 17 Alim, A. and Kalimuddin, M. 1985. Development of nursery procedure of three cane species. Bano Biggyan Patrika 14(1&2):26-29. 18 Mohiuddin, M., Rashid, M. H. and Rahman, M. A. 1986. Seed germination and optimal time of transfer of seedlings of Calamus sp. from seed bed to polythene bag. Bano Biggyan Patrika 15(1&2):21-24. 19 Mohiuddin, M., Rashid, M. H. and Akhter, R. 1988. Easy identification method for different rattan species of Bangladesh. Minor Forest Products Series. Bangladesh Forest Research Institute Bulletin 2:15 (in Bengali). 20 Rashid, M. H., Merry, S. R., Ara, R., Mohiuddin, M. and Alam, M. J. 1993. How to Cultivate Rattan and Patipata. Bulletin 6. Minor Forest 1

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