Microbial inoculants (MI), a biofertilizer, composed of many different beneficial microorganisms has positive role on seed germination and growth of plants. In the present study, its efficacy on seed germination and seedling growth of Albizia lebbeck in the nursery was studied. The seeds were sown in polybags filled with a mixture of forest soil and cow dung (3:1) and treated with 0.1%, 0.5%, 1%, 2%, 5% and 10% concentrations of MI. Most of the parameters studied (seed germination, shoot and root lengths, dry weights of shoot and root, collar diameter, leaf number etc) were found maximum in 2% of MI . Although the highest vigor index, volume index and quality index (7053, 3738 and 1.106, respectively) were found in 2% MI, but the highest sturdiness (65.95) was found in 1% MI solution. The nodule number was higher at a very low (0.5%) concentration of MI but it normally decreased with the increase of concentration. Total pigment content in leaf was recorded highest (112.86 mg.L-1) in 2% of MI. Therefore, MI influences seed germination and seedling growth of A. lebbeck and the low concentration (2%) of the inoculant can be recommended for getting maximum seed germination and seedling growth of the species studied. K e y w or d s : Albizia lebbeck, germination, microbial inoculant, seedling growth Microbial inoculant influences the germination and growth of Albizia lebbeck seedlings in the nursery B. M. Khan1*, M. A. Kabir2, M. K. Hossain1 and M. A. U. Mridha3 Albizia lebbeck (L.) Benth [Kalo sirish in Nepali, Kala koroi in Bengali], is a moderate to large deciduous tree with a straight bole and broad crown under the family Leguminosae (Mimosoideae). The species is widely spread in the world, and is native to Asia, Africa and Northern Australia. It grows naturally in Nepal, Bangladesh, Myanmar and Pakistan and has been cultivated in tropical and subtropical regions in Northern Africa, the West Indies, South America, and south Asia. Extensive plantations had been established in Nepal and in South India (Luna, 1996; Kumar et al., 2010; Elzaki et al., 2012; Missanjo et al., 2013). The wood is excellent for furniture and timber for general uses, post, piles, fuel wood and charcoal. The tree is used as ornamental and nurse tree for tea gardens, coffee and cocoa orchards (Mishara et al., 2010; Missanjo et al., 2013; Shaikh et al., 2014). It is a soil improver because of its inherent ability to fix nitrogen (Qadri and Mahmood, 2005). The leaves are excellent fodder for livestock. The species is also very useful for the rehabilitation of degraded lands because of its rapid growth, ability to fix nitrogen and tolerance for a range of soil conditions (Luna, 1996; Qadri and Mahmood, 2005; Mishara et al., 2010; Bobby et al., 2012; Shaikh et al., 2014). The plant has already been proven successful for afforestation, reforestation, social forestry and agroforestry programs in Bangladesh (Zabala, 1990; Dey, 2006). To fulfill the high demand, many organizations are producing A. lebbeck seedlings in the nursery in Bangladesh to supply those in the plantation programs. Because the plants are grown mostly in unfavorable soil conditions, beneficial soil microorganisms can play a significant role in early establishment and better growth of the inoculated seedlings under field conditions. The microbial inoculant (MI) in this study, with the commercial name “Effective Microorganisms” or EM was developed at the University of Ryukyus, Okinawa, Japan, in the early 1980s by a distinguished professor of horticulture, Dr. Teruo Higa (Kyan et al., 1999). The main species comprising MI are lactic acid bacteria ( Rhodopseudomonas spp.), photosynthetic bacteria (Lactobacillus spp., Streptococcus 1 Institute of Forestry and Environmental Sciences, University of Chittagong, Chittagong-4331, Bangladesh. *E-mail: bmkhan2004@yahoo.com 2 Department of Agroforestry, Patuakhali Science and Technology University, Patuakhali-8602, Bangladesh. 3 Plant Production Department, King Saud University, Kingdom of Saudi Arabia. 82 Banko Janakari, Vol. 26, No. 1 83 spp.), yeast (Saccharomyces spp., Candida spp.), Actinomycetes (Streptomyces spp.) and beneficial fungi (Aspergillus spp., Penicillium spp.). The microorganisms are added into the inoculant in the manufacturing process and can survive in the inoculant liquid at pH 3.5 or below. The density of most of the above mentioned microbes is in the range of 1 × 106 to 1 × 108 mL-1 (Xu, 2000). MI can be applied as inoculant to increase the microbial diversity of soils. It has been used with considerable success to improve soil quality and yield of crops, particularly in nature farming and organic farming systems (Xu, 2000). Inoculation of MI culture can improve photosynthesis and fruit yield (Xu, 2000; Wang et al., 2000). Although A. lebbeck is used for wide range of purposes and even planted intensively in the field, the initial growth potential under the influences of MI was not studied. Therefore, the aim of this study was to observe the effectiveness of MI on germination of seed and the growth of seedlings of A. lebbeck and also to find out the best concentration of MI solution for ensuring maximum seedling development in the nursery. Materials and methods Collection of seeds and soils The experiment was carried out in the nursery of the Institute of Forestry and Environmental Sciences; University of Chittagong, Bangladesh (lies approximately at the intersection of 91o50´E and 22o30´N) (Fig. 1). The seeds of A. lebbeck were collected from the seed orchard division of Bangladesh Forest Research Institute (BFRI) where the source of seed was a mother tree of 25 years old. The soils collected from the degraded hills of the University Campus was sieved well (<3 mm) and mixed thoroughly with decomposed cow dung in a ratio of 3:1. The brown hill soils (Rashid, 1991), are sandy loam to sandy clay loam, moderately to strongly acid and poorly fertile with pH <5.5, organic matter <2.0%, CEC <10 me/100 g, BSP <40% (Osman et al., 2001). The white polybags of 15 cm x 10 cm in size were filled with the prepared mixture and a thin layer of coconut husk was added to each bag as top layer to reduce evaporation and to supply organic matter. Fig. 1: Map showing the location of the nursery of IFESCU (Institute of Forestry and Environmental Sciences, University of Chittagong) in Bangladesh where the experiment was conducted. Treatment design The experiment was conducted in 03 March to 02 August 2015. There were seven treatments including control and 25 replications for each treatment. Seeds were sown in polybags (filled with soil and cow dung) with no added MI but water only for control treatment. Other treatments included; sowing the seeds in polybags with the concentrations of 0.1%, 0.5%, 1%, 2%, 5% and 10% of MI, respectively. For preparing 0.1% of MI solution, 0.1 mL stock solution of MI was added with 99.9 mL of water while for preparing 0.5% of MI solution, 0.5 mL stock solution of MI was added with 99.5 mL of water and the same formula was used for preparing the other solutions. For each treatment, 50 mL of MI of required concentration was poured in each polybag (filled with soil and cow dung) before a week of sowing the seeds while another 50 mL was poured after a week of sowing the seeds. Five seeds were sown in each polybag to observe the influence of MI on germination in the nursery conditions (temperature, 28oC; humidity, 75%). After completion of germination, only one seedling (the best one) per polybag was managed to observe growth performance and nodulation status of the seedlings. Partial shade and cover was ensured using polythene sheet on the nursery roof to protect the seedlings from strong sunlight and rains. Khan et al. Banko Janakari, Vol. 26, No. 1 84 Khan et al. Growth measurement Germination was recorded daily from the date of seed sowing to the last of germination. The seedlings were allowed to grow altogether for five months from the time of seed sowing. After five months, five representative seedlings from each treatment were selected for measuring growth parameters. The recorded parameters were shoot and root lengths, collar diameter, leaf number, fresh shoot and root weights, dry shoot and root weights, and nodulation status. For recording dry weights, shoots and roots were oven dried at 80oC for 48 hr. To assess the seedling vigor, total height (from the soil surface to seedling tip) of each seedling in each sub-plot was measured using a ruler to the nearest 0.1 cm. Vigor index was calculated according to Abdulbaki and Anderson (1973) as germination percent x seedling total length i.e. total shoot and root length. Volume index was obtained by multiplying shoot height or shoot length (cm) with the square of collar diameter (mm)2 of the seedling. Quality index was developed following Dickson et al. (1960) to quantify seedlings morphological quality. The formula for calculating quality index is as follow: H S dw QI = T dw / ( + ) D c R dw where, QI is quality index, T dw is total dry weight (g), H is seedling height or shoot length (cm), D c is collar diameter (mm), S dw is shoot dry weight (g), R dw is root dry weight (g). Sturdiness was obtained by dividing shoot height or shoot length (cm) with collar diameter (cm) of the seedling. Measurement of pigment contents The pigment contents (chlorophyll-a, chlorophyll-b, and carotenoid) were determined from the fresh leaves of seedlings in different treatments (Wettstein, 1957; Khan, 2012). Ten leaf discs were cut with a cork borer (inside diameter of 5 mm), weighed immediately after cutting, and dipped in 100% acetone in 5 mL in test tube with stopper. After 24 hr of incubation, the supernatant-colored solution from the top was decanted carefully into a 25 mL volumetric flask. The leaf discs were then crushed with a blunt glass rod gently and 5 mL fresh acetone was added to the test tube and left for 15 min. Then the supernatant-colored solution from the top was again decanted to the same volumetric flask very carefully, avoiding the fragmented plant tissues. The process was repeated until the leaf fragments became colorless. Finally, the volume was made up to 25 mL with fresh acetone and the measurement was taken immediately after preparation of the solution. The measurement of chlorophyll-a, chlorophyll-b, and carotenoid were made at 662 nm, 644 nm, and 440.5 nm respectively, with a spectrophotometer (Spectronic-20). The pigment contents in the extract were calculated following the formula of Wettstein (1957). Ca = (9.784E662 – 0.99E644) x V x d (1) 1000 Fw Cb = (21.426E644 – 4.650E662) x V x d (2) 1000 Fw Cc = [4.695E440.5 – 0.268(Ca+CbE662)] x V x d (3) 1000 Fw where, Ca is the chlorophyll-a (mg.L-1); Cb the chlorophyll-b (mg.L-1); Cc the carotenoid (mg.L-1); V the total volume (25 mL); d the dilution factor; FW the fresh weight of leaf disc (g); and E is the absorbance at a particular wavelength (440.5, 644 and 662 nm). Statistical analysis All data were analyzed statistically using computer software SPSS (Version 20, SPSS Incorporation, Chicago, USA). Possible significant variations among the treatments were explored by Duncan’s Multiple Range Test (DMRT). Prior to statistical analysis, the normality of each data set was tested using an Anderson-Darling test, in case a transformation was necessary. Log base 10 transformation was done where required. Means were separated by MS Excel (MS Office 2010). Results and discussion Germination and seedling growth The highest (77%) seed germination was recorded in 2% of MI, while the lowest (61%) was recorded in control treatment (without inoculation). Shoot (54.0 cm) and root (37.6 cm) lengths were also highest in 2% of MI (Table 1). With the application of 2% of MI, shoot length increment was 33% compared to control (Fig. 2). Collar diameter was maximum (8.32 mm) in 2% of MI and was significantly (P=0.015) different Banko Janakari, Vol. 26, No. 1 85 from the control. Seedlings treated with MI had more leaf compared to the control (Table 1). Fig. 2: Influence of microbial inoculant on shoot length increased to decreased (%) with respect to control in Albizia lebbeck. Cont. = Control. Both fresh (21.4 g) and dry (6.95 g) shoot weights were maximum in 2% of MI. Both fresh and dry root weights were also maximum (9.1 g and 3.01 g, respectively) in 2% of MI and were significantly ( P<0.001 and P=0.026, respectively) varied from control (Table 2). Though maximum vigor index, volume index and quality index (7053, 3738 and 1.106, respectively) were found in 2% of MI (Table 2 and Fig. 3), the maximum sturdiness (65.95) was found in 1% of MI treatment (Fig. 4). The total dry biomass increased gradually with the increase of the concentrations of MI up to 2% while decreased gradually from its maximum point as the concentrations of MI increased above 2%. Increased biomass production might be due to the better root development in the treated seedlings (Khan et al., 2011, 2014). Such promotion might also be due to the biological active substances in the inoculant (Lim et al., 1999), such as indole acetic acid (IAA) and gibberellins produced by Lactobacillus spp., Rhodopseudomonas spp., Aspergillus spp. and Saccharomyces spp. which enhance plant growth (Chowdhury et al., 1994). However, germination rate was below 80% which was probably due to the quality of seeds as well Khan et al. Table 1: Influence of microbial inoculant on germination, shoot and root lengths, collar diameter and leaf number of Albizia lebbeck in the nursery Concentration of MI (%) Germination (%) Length (cm) Collar dia. (mm) Number of leafShoot Root Total Control 61c 40.6b 27.8b 68.4b 6.41b 16.8b 0.1 65b 45.3ab 31.2ab 76.5ab 6.95b 17.6b 0.5 72a 48.4a 33.5a 81.9a 7.42a 18.0a 1 74a 52.3a 35.7a 88.0a 7.93a 19.6a 2 77a 54.0a 37.6a 91.6a 8.32a 20.2a 5 68b 47.1ab 32.3ab 79.4ab 7.47a 18.4a 10 65b 42.9b 29.4b 72.3b 6.94b 17.2b P value <0.001 0.002 <0.001 <0.001 0.015 0.006 F value 137.18 16.37 74.82 104.51 6.35 9.21 Note: a-c = Mean values with different lowercase superscripts in a column are significantly different at P<0.05, according to Duncan’s Multiple Range Test (DMRT). Table 2: Influence of microbial inoculant on fresh and dry weights of shoot and root, vigor index and volume index of Albizia lebbeck Concentra- tion of MI (%) Fresh weight (g) Dry weight (g) Total dry biomass increment (%) Index Shoot Root Total Shoot Root Total Vigor Volume Control 14.9b 5.9b 20.8b 4.48b 1.89c 6.37b 00.00 4172c 1668c 0.1 15.7b 6.7b 22.4b 4.72b 2.25b 6.97b +9.42 4973b 2188b 0.5 16.5b 7.3a 23.8ab 4.65b 2.37b 7.02b +10.20 5897a 2665a 1 19.3a 8.0a 27.3a 5.88a 2.58a 8.46a +32.81 6512a 3289a 2 21.4a 9.1a 30.5a 6.59a 3.01a 9.60a +50.71 7053a 3738a 5 20.7a 8.8a 29.5a 6.29a 2.92a 9.21a +44.58 5399b 2628a 10 17.6b 7.8a 25.4ab 5.16a 2.57a 7.73a +21.35 4700b 2066b P value <0.001 <0.001 <0.001 0.017 0.026 0.021 -- <0.001 <0.001 F value 146.12 114.09 127.65 6.19 4.83 5.27 -- 57.65 88.34 a-c = Mean values with different lowercase superscripts in a column are significantly different at P<0.05, according to Duncan’s Multiple Range Test (DMRT). Banko Janakari, Vol. 26, No. 1 86 Khan et al. as the environmental factors regulating seed germination. Fig. 3: Influence of microbial inoculant on quality index of Albizia lebbeck. Cont. = Control. Fig. 4: Influence of microbial inoculant on sturdiness of Albizia lebbeck. Cont. = Control. Microbial inoculants are being applied in Japan, China, Thailand, United States, France, Brazil and many other countries of the world. Application of MI can play a role in enhancing germination, growth, and yield of various agricultural crops and vegetables (Vongprachanch, 1995; Zacharia, 1995; Iwaishi, 2000; Chowdhury et al., 2002). MI with organic fertilizer and other chemicals is also reported to enhance germination, growth, and yield of different grains (Ahmed et al., 1995; Anuar et al., 1995; Xu, 2000). But the influence of MI on forest crops has not been studied widely (Khan et al., 2006, 2011). From this study, it has been observed that soil amended with different concentrations of MI can also improve the seedling growth of forest crops. Mridha (2005), Khan (2012) and Khan et al. (2011, 2014) also reported enhanced seed germination rate and seedling growth with the application of low concentrations of MI. Specially, applying 2% of MI enhanced seed germination and seedling growth at an optimum level which might be due to the presence of microbial population in such a density within this concentration which was cable to produce optimum level of growth enhancing hormones and other chemicals. However, as the concentration of MI increased, seed germination and seedling growth was depressed, which may be due to the perniciousness exerted by the higher concentrations of the inoculant (Mridha, 2005; Khan et al., 2014). Nodulation status Although most of the parameters were highest in 2% of MI, the highest (134) nodule number was in 0.5% of MI while the lowest (95) was in 10% of MI (Table 3). Both fresh and dry nodule weights were maximum (2.07 g and 0.67 g, Table 3: Influence of microbial inoculant on nodule number and their fresh and dry weights of Albizia lebbeck Concentration of MI (%) Nodule Number Weight (g) Weight increased or decreased (%) Fresh Dry Fresh Dry Control 127a 1.77a 0.56ab 0.00 0.00 0.1 131a 2.01a 0.65a +13.56 +16.07 0.5 134a 2.07a 0.67a +16.95 +19.64 1 121a 1.89a 0.61a +6.78 +8.93 2 116b 1.75a 0.54ab -1.13 -3.57 5 104b 1.57b 0.51ab -11.30 -8.93 10 95c 1.42b 0.44b -19.77 -21.43 P value <0.001 0.013 0.019 -- -- F value 51.42 6.68 5.3 -- -- a-c = Mean values with different lowercase superscripts in a column are significantly different at P<0.05, according to Duncan’s Multiple Range Test (DMRT) Banko Janakari, Vol. 26, No. 1 87 respectively) at 0.5% of MI and lowest (1.42 g and 0.44 g, respectively) were in 10% of MI. The rate of nodule increment was positive in case of 0.1% and 0.5% of MI, while negative for all other treatments compared to control (Fig. 5). The nodule dry weight increment rate was positive for 0.1%, 0.5%, and 1% of MI while negative for all other treatments, with respect to control (Table 3). The number of nodule in soybean root was also not changed significantly due to application of low concentrations of MI (Thach et al., 1999) while decreased in higher concentrations in Dalbergia sissoo and Acacia auriculiformis (Khan et al., 2011, 2014). The higher concentrations of MI solution in the substratum may affect the growth of plants because of toxic effect of secretion of toxic metabolites. Fig. 5: Influence of microbial inoculant on nodule number increased to decreased (%) with respect to control in Albizia lebbeck. Cont. = Control. Content of leaf pigments Effects of MI on the content of leaf pigments (Chlorophyll-a, chlorophyll-b, and carotenoid) were also determined (Table 4). Chlorophyll-a was highest (56.12 mg.L-1), in 2% of MI and lowest (37.39 mg.L-1) in the control treatment. Chlorophyll-b was highest (15.67 mg.L-1) in 2% of MI, followed by 14.98 mg.L-1 in 5% and 13.59 mg.L-1 in 1% of MI, and was significantly ( P<0.001) varied from control. Carotenoid was maximum (41.07 mg.L-1) in 2% of MI. Total pigment was recorded highest (112.86 mg.L-1) in 2% of MI and was significantly (P<0.001) different from control. The results are in agreement with the findings of Xu (2000), Wang et al. (2000), Mridha et al. (2002), Khan et al. (2006, 2011) and Khan (2012) that low concentrations of MI with organic fertilizer promote root growth and enhance photosynthetic efficiency and yield of seedling. Conclusion Microbial inoculant influences seed germination and seedling growth of A. lebbeck and the low concentration (2%) of the inoculant can be recommended for getting maximum seed germination and seedling growth of the species in the nursery conditions that may also be effective for seedling development in the field. References Abdul-Baki, A. and Anderson, J. D. 1973. Vigor determination in Soybean seed by multiple criteria. Crop Science 13: 630–633. Ahmed, R., Hussain, T., Jilani, G., Shahid, S. A., Akhtar, S. N. and Abbas, M. A. 1995. Use Khan et al. Table 4: Influence of microbial inoculant on pigment contents in fresh leaves of Albizia lebbeck Concentration of MI (%) Pigments concentration of leaf (mg.L-1) Total pigment increment (%)Chlorophyll-a Chlorophyll-b Carotenoid Total Control 37.39c 10.34b 28.41c 76.14c 0 0.1 41.51b 11.75ab 32.97b 86.23b +13.25 0.5 43.34b 12.06ab 34.02b 89.42b +17.44 1 50.02a 13.59a 37.18a 100.79a +32.37 2 56.12a 15.67a 41.07a 112.86a +48.23 5 54.17a 14.98a 40.84a 109.99a +44.46 10 47.49b 12.32ab 31.49b 91.30b +19.91 P value <0.001 <0.001 <0.001 <0.001 -- F value 38.26 49.58 91.63 61.47 -- a-c = Mean values with different lowercase superscripts in a column are significantly different at P<0.05, according to Duncan’s Multiple Range Test (DMRT) Banko Janakari, Vol. 26, No. 1 88 of effective microorganisms for sustainable crop production in Pakistan. In Effective Microorganisms (EM) (eds) Sharifuddin, H. A. H., Anuar, A. R. and Shahbudin, M. F. Proceedings of the 2nd International Conference on Effective Microorganisms (EM). Kyusei Nature Farming Center, Saraburi, Thailand, 15–27. Anuar, A. R., Sharifuddin, H. A. H., Shahbudin, M. F. and Zaharah, A. R. 1995. Effectiveness of effective microorganisms (EM) on maize grown on sandy tin tailings. In Effective Microorganisms (EM) (eds) Sharifuddin, H. A. H., Anuar A. R. and Shahbudin, M. F. Proceedings of the 2nd International Conference on Effective Microorganisms (EM). Kyusei Nature Farming Center, Saraburi, Thailand, 42–54. Bobby, N., Wesely, E. G. and Johnson, M. 2012. HPTLC profile studies on the alkaloids of Albizia lebbeck. Asian Pacific Journal of Tropical Biomedicine 1–3:1–6. Chowdhury, A. R, Hussain, M. M., Mia, M. S., Karim, A. M. M. S., Haider, J., Bhuyan N. I. and Shifuddin, K. 1994. Effect of organic amendments and EM on crop production in Bangladesh. In Kyusei Nature Farming (eds) Parr, J. F., Hornick S. B. and Simpson, M. E. Proceedings of the 2nd International Conference on Kyusei Nature Farming, Piracicaba, Brazil, 155–163. Chowdhury, M. H. U, Mridha, M. A. U., Khan, B. M. and Xu, H. L. 2002. Effects of effective microorganisms on seed germination and seedling growth of Oryza sativa L. Nature Farming and Environment 3 (1): 23–30. Dey, T. K. 2006. Useful Plants of Bangladesh. The Ad. Communication, Anderkilla, Chittagong, Bangladesh. Dickson, A., Leaf, A. L. and Hosner, J. F. 1960. Quality appraisal of white spruce and white pine seedling stock in nurseries. Forestry Chronicle 36: 10–13. Elzaki, O. T., Khider, T. O., Omer, S. H. and Shomeina, S. K. 2012. Environment friendly alkaline pulping of Albizia lebbeck from Sudan. Nature and Science 10 (4): 76–82. Iwaishi, S. 2000. Effect of organic fertilizer and effective microorganisms on growth, yield and quality of paddy-rice varieties. Journal of Crop Production 3 (1): 269–273. Khan, B. M., Hossain, M. K. and Mridha, M. A. U. 2014. Improving Acacia auriculiformis seedlings using microbial inoculant (Beneficial Microorganisms). Journal of Forestry Research 25 (2): 359–364. Khan, B. M., Hossain, M. K. and Mridha, M. A. U. 2006. Effect of microbial inoculants on Albizia saman germination and seedling growth. Journal of Forestry Research 17 (2): 99–102. Khan, B. M., Hossain, M. K. and Mridha, M. A. U. 2011. Nursery practice on seed germination and seedling growth of Dalbergia sissoo using beneficial microbial inoculants. Journal of Forestry Research 22 (2): 189–192. Khan, B. M. 2012. Environmental Biology: Growth Improvement of Forest Seedlings using Beneficial Microbial Inoculant (Effective Microorganisms). Lap lambert academic publishing, Germany. 160 pp. Kumar, S., Bansal, P., Gupta, V., Sannd, R. and Rao, M. 2010. The clinical effect of Albizia lebbeck stem bark decoction on Bronchial Asthma. International Journal of Pharmaceutical Sciences and Drug Research 2 (1): 48–50. Kyan, T., Shintani, M., Kanda, S., Sakuria, M., Ohashi, H., Fujisawa, A. and Pongdit, S. 1999. Kyusei Farming and the Technology of Effective Microorganisms. International Nature Farming, Bangkok, Thailand and Research Center, Atami, Japan and Asia Pacific Natural Agriculture Network. Lim, T. D., Pak, T. W. and Jong, C. B. 1999. Yields of rice and maize as affected by effective microorganisms. In Kyusei Nature Farming and Effective Microorganisms (eds) Senanayake, Y. D. A. and Sangakkara, U. R. Proceedings of the 5th International Conference on Kyusei Nature Farming and Effective Microorganisms for Agricultural and Environmental Sustainability. Bangkok, Thailand, 92–98. Luna, R. K. 1996. Plantation Trees. International Book Distributors. Dehra Dun, India. Mishara, S. S., Gothecha, V. K. and Sharma, A. 2010. Albizia lebbeck: a short review. Khan et al. Banko Janakari, Vol. 26, No. 1 89 Journal of Herbal Medicine and Toxicology 4 (2): 9–15. Missanjo, E., Maya, C., Kapira, D., Banda, H. and Kamanga-Thole, G. 2013. Effect of seed size and pretreatment methods on germination of Albizia lebbeck. ISRN Botany 1: 1–4. Mridha, M. A. U, Khan, B. M., Hossain, M. K., Tasnim, H. and Rahman, M. F. 2002. Effective microorganisms (EM) for neem tree. APANews 20: 5–6. Mridha, M. A. U. 2005. Growth improvement of seedlings of a tropical forest species, Khair (Acacia catechu Willd.) using beneficial microbial inoculant (effective microorganisms). Saudi Journal of Biological Sciences 12 (1): 67–73. Osman, K. T., Rahman, M. M. and Barua, P. 2001. Effects of some forest tree species on soil properties in Chittagong University Campus, Bangladesh. Indian Forester 127 (4): 431– 442. Qadri, R. and Mahmood, A. 2005. Ultrastructural studies on root nodules of Albizia lebbeck (L.). Pakistan Journal of Botany 37 (4): 815– 821. Rashid H. 1991. Geography of Bangladesh. University Press Ltd. Dhaka, Bangladesh. Shaikh, F. K., Gadge, P. P., Shinde, A. A., Padul, M. V. and Kachole, M. S. 2014. Characterization of the A1T113 protein from Indian siris ( Albizia lebbeck) that inhibits the growth of cotton bollworm (Helicoverpa armigera). Journal of Asia-Pacific Entomology 17 (3): 319–325. Thach, N. Q., Long, C. A., Liet, V. V., Trung, N. V., Thanh, N. X., Dich, T. V., Duong, N., Tuna, N. K., Xuan L. T. H. and Dan, P. V. 1999. Preliminary results of effective microorganisms (EM) application in Vietnam. In Kyusei Nature Farming and Effective Microorganisms (eds) Senanayake, Y. D. A. and Sangakkara, U. R. Proceedings of the 5th International Conference on Kyusei Nature Farming and Effective Microorganisms for Agricultural and Environmental Sustainability. Bangkok, Thailand, 254–260. Vongprachanch, B. 1995. The use of EM fermented materials in farm trials. In Effective Microorganisms (EM) (eds) Sharifuddin, H. A. H., Anuar A. R. and Shahbudin M. F. Proceedings of the 2nd International Conference on Effective Microorganisms (EM). Saraburi, Thailand, 90–94. Wang, R., Xu, H. L. and Mridha M. A. U. 2000. Effect of organic fertilizer and EM inoculation on leaf photosynthesis and fruit yield and quality of tomato plants. Journal of Crop Production 3 (1): 173–182. Wettstein, D. 1957. Formula of chlorophyll determination. Exp. Cell Research 12 (3): 427–489. Xu, H. L. 2000. Effect of a microbial inoculants and organic fertilizer on the growth, photosynthesis and yield of sweet corn. Journal of Crop Production 3 (1): 183–214. Zabala, N. Q. 1990. Silviculture of Species. Development of the Professional Education in the Forestry Sector, Bangladesh. UNDP/ FAO BGD/85/011, Field Document No. 14. 174 pp. Zacharia, P. P. 1995. Studies on the application of effective microorganisms (EM) in paddy, sugarcane and vegetables in India. In Effective Microorganisms (EM) (eds) Sharifuddin, H. A. H., Anuar, A. R. and Shahbudin, M. F. Proceedings of the 2nd International Conference on Effective Microorganisms (EM). Kyusei Nature Farming Center, Saraburi, Thailand, 31–41. Khan et al.