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Available online at www.jpsscientificpublications.com Volume 1; Issue - 1; Year 2017; Page: 15 20 ISSN: 2456-7353 DOI: 10.22192/ijias.2017.1.1.4 I International Journal of Innovations in Agricultural Sciences (IJIAS) Journal of In EFFECT OF Glomus fasciculatum ON THE GROWTH AND YIELD OF Dolichos lablab AS INFLUENCED BY THE APPLICATION OF PHOSPHATE FERTILIZER R.T.S. Nandini*, Department of Microbiology and Biochemistry, Nadar Saraswathi College of Arts and Science, Theni, Tamil Nadu, India. Abstract Arbuscular mycorrhizal fungi (AMF) are a key component for plant survival and create an intimate link between plant roots and the soil. The major role of AMF is to supply Phosphate (P) in infected plants, because P is an extremely immobile element in soil. Several species of mycorrhizal fungi especially Glomus fasciculatum can be used for P uptake and translocation in Dolichos lablab. A pot culture experiment was conducted to study the inoculation effect of G. fasciculatum on the growth and yield of D. lablab as influenced by different levels (0 %, 25 %, 50 %, 75 % and 100 %) of P fertilizer. The maximum yield was obtained in 75 % of the P level followed by 50 % and 100 % P levels. The increasing amount of P fertilizer decreases the G. fasciculatum population. The D. lablab yield was increased at 75 % of P level followed by 50 % and 100 %. Key words: Glomus fasciculatum, Dolichos lablab, P fertilizer and AM fungi. 1. Introduction Legumes are important sources of proteins, carbohydrates, dietary fiber and minerals consumed worldwide. This is a tropical vein plant from the legume family and it is widely grown in Africa, India and Indonesia. D. lablab is one of the lesser-known legumes of arid and semiarid land. It is a well known vegetable among the Indians. It is commonly called as lablab bean, bonavist bean, hyacinth bean and Indian butter bean (Tamil Avarai). Genus Dolichos, species lablab, family Leguminaceae. This vegetable which was not so famous is good for diabetics and a good source of fiber. It is also popular as a nitrogen fixing green manure to contribute the soil nitrogen and improve soil quality. It is a garden vegetable *Corresponding author: R.T.S. Nandini Nadar Saraswathi College of Arts and Science, Theni, Tamil Nadu, India. Received: 20.04.2016; Revised: 05.05.2016; Accepted: 22.05.2016. crop and used acts an intercrop in India and Australia as a weed suppressor and soil erosion retardant. Mycorrhizae (Allen, 1992; Harrison, 1997; Smith and Read, 1997) are mutually beneficial intimate relationship between diverse fungi and crop roots. Most of the plants (more than 90 % of all know species) are infected with atleast one type of mycorrhizae, and it also plays an important and decisive role in the phosphorus uptake of plants. AMF takes the nutrients (especially phosphorus) which are in the available forms and transfer to crop plants. In 1959, Baylis from New Zealand was the first to suggest that the beneficial mycorrhizal effect was mediated by P uptake. Gerdeman also demonstrated that non-mycorrhizal plants exhibited severe phosphorus deficiency symptoms and had significantly lower P concentration and higher K and Mg concentrations than mycorrhizal plants.

Nandini /International Journal of Innovations in Agricultural Sciences (IJIAS), 1(1): 15 20 16 The organic phosphates applied to soil as a fertilizer is rapidly converted to unavailable forms with low solubility. Soluble P was released from insoluble phosphates by a variety of solubilization reaction involving rhizosphere microorganisms. Mycorrhizal plants can take up more P than nonmycorrhizal plants. Legumes are relatively high P requirement for nodule development and nitrogen fixation. Normal levels of nodulation may depend on the presence of mycorrhizal fungi. Several species of mycorrhizal fungi especially G. fasciculatum was used for P uptake and translocation in D. lablab. 2. Materials and Methods The soil used for conducting the pot culture experiment was obtained from the garden land of the Experimental Farm, Department of Microbiology and Biochemistry, Nadar Saraswathi College of Arts and Science, Theni, Tamil Nadu. D. lablab (Avarai) seeds were obtained from the Orchard, Department of Horticulture, Annamalai University, Annamalai Nagar, Tamil Nadu has been used in the present study. The P fertilizer was supplied as Single Super Phosphate (SSP) as per the recommended doses in the pot culture experiments. The dose of P fertilizer per pot was worked out on the soil weight basis keeping the base of their recommended doses viz., 50 kg P 2 O 5 /ac for D. lablab. Live arbuscular mycorrhizal spores were inoculated in root tissues of the plant. A pot culture experiment was conducted to study the inoculation effect of G. fasciculatum on the growth and yield of D. lablab as influenced by different level of P fertilizer. T 0 : Control T 1 : G. fasciculatum + 0 % P 2 O 5 T 2 : G. fasciculatum + 25 % P 2 O 5 T 3 : G. fasciculatum + 50 % P 2 O 5 T 4 : G. fasciculatum + 75 % P 2 O 5 T 5 : G. fasciculatum + 100 % P 2 O 5 The biometric observation viz., mycorrhizal spore number, root colonization per cent of AM fungi, plant height, nodule number and P content of the plant were determined on 15, 30 and 45 DAS Arbuscular mycorrhizal spores were estimated from rhizosphere soil by Wet sieving and Decanting method (Gerdemann and Nicolson, 1963). The spores were cleaned of soil particles by sucrose density gradient centrifugation method and washed with distilled water (Mertz et al., 1979). This spore suspension was counted with stereo zoom microscope during counting. Per cent root colonization by AM fungi was determined after staining the roots with tryphan blue as described by Phillips and Hayman (1970) with slight modification. The height of the D. lablab was measured from the ground level to the tip of the growing plant. The total number of matured nodules was counted from each treatment. The P content of the plant was estimated using Vanadomolybdate method (Jackson, 1973). 3. Result and Discussion A preliminary survey was taken upto study the mycotrophy of D. lablab. The mycorrhizal status of rhizosphere soil sample was studied by estimating the per cent root colonization and AM fungal spore number after inoculate the G. fasciculatum on 15, 30 and 40 DAS. The AM spore population increased with the increase in the age of the plant. The maximum was observed at 45 DAS. Application of P at different levels viz., 0, 25, 50, 75 and 100 per cent recorded the AM spore number (163.89, 168.0, 159.0, 148.0, 101.3 and 85.66 per 100 g of rhizosphere soil). The lowest spore number in 100 per cent P 2 O 5 level (85.66 per 100 g of soil). The colonization of G. fasciculatum in D. lablab root at 45 DAS in control shows highest (38.16) followed by 25 % P 2 O 5 (35.20). The per cent root colonization reduced with the increasing level of P and the per cent gradually decreased of root

Nandini /International Journal of Innovations in Agricultural Sciences (IJIAS), 1(1): 15 20 17 colonization at 50 %, 75 % and 100 % of P 2 O 5 level were respectively are presented in Table - 1. Inoculation of G. fasciculatum at 50 % significantly increased the plant height than the control. At different levels of P application the maximum plant height was observed at 75 % P application (36.46 cm) and increased plant height was noticed with increased in the levels of P application. The plant height observed at 0, 25, 50, 75 and 100 % of P 2 O 5 were 27.16, 30.56, 35.76, 36.46 and 34.33cm respectively under G. fasciculatum inoculated D. lablab plants. The data are presented in Table - 2. The nodule number of D. lablab increased with the increased levels of P fertilizer. At all the three intervals of observations the P content at different levels viz., 0, 25, 50, 75 and 100 % recorded the increase in nodule number of 20.44, 22.88, 25.21,27.22 and 30.66 in the rhizosphere region of plant root respectively. Nodule number was more in 100 % P 2 O 5 (30.66 %) and the lowest in the control (18.29 %) are shown in Table - 3. At 60 DAS, the uptake of P was the maximum. The maximum level was observed in the 75 % P (5.19 %) and followed by 50 % and 100 % of P level were 4.63 % and 4.49 % respectively. The minimum level of P uptake was by control plant (2.11 %). The P uptake was increased at the increased level of P 2 O 5 (Table - 4). At different levels of P application, the minimum yield was obtained at 75 % (497 kg ac -1 ) followed by 100 % (495 kg ac -1 ) and 50 % (493 kg ac -1 ) P levels of the yield of D. lablab was more (or) less similar. The results revealed that the inoculation effect of G. fasciculatum was influenced by the application of graded levels of P fertilizer, to develop a technology for treatment for AM fungi in D. lablab are shown in Table - 5. The inoculation of VAM fungi significantly increased the nodulations, nitrogenase activity, per cent root colonization and spore number in cluster bean, mung bean and moth bean (Tarafdar and Rao, 1997). Inoculation of plants with AM fungi can stimulate nodulation and nitrogen fixation by legumes (Mosse, 1981). Inoculation of G. fasciculatum increased the nodulation, shoot N and P concentration, shoot dry matter and seed yield in soy bean plants Srinivasan et al. (1995). However, the effect appears to operate indirectly through increased P uptake because the response to inoculation can be completely overcome by increasing P supply (Abort and Robson, 1977; 1982). AM fungi which constitute important group of microorganism are ubiquitous throughout the world and are known to improve the plant growth through better uptake of P and other nutrients. It was generally accepted that the plant growth was enhanced because the fungal hyphae explore the soil outside nutrition depletion zones and absorb ions especially P from soil solution and around roots and transfer them to plants (Hayman and Mosse, 1972). Higher levels of soluble P have negative effect on VA mycorrhizal colonization in wheat (Ryan, 1994). On this basis 75 and 100 % of P 2 O 5 decrease the fungal spore and the root colonization. In the 50 % P 2 O 5 level the fungal colonize are survived and the plant uptake the phosphorus content in from the soil. But, both 100 and 50 per cent level yield are similar moderate increase P availability can maintain or elevate VA mycorrhizal colonization (Habte and Fox, 1993; Lue et al., 1994). The increasing amount of P fertilizer (100 %) decreases the G. fasciculatum population. The D. lablab yield was increased at 75 % of P level followed by 50 % and 100 %. They are more or less similar to each other. The result revealed that 50 % of P fertilizer can be saved by G. fasciculatum inoculation.

Nandini /International Journal of Innovations in Agricultural Sciences (IJIAS), 1(1): 15 20 18 Table - 1: Effect of G. fasciculatum inoculation on the AMF spore number and per cent root colonization of D. lablab Number of AM fungal spore (per 100g of soil) Root colonization 15 DAS 30 DAS 45 DAS 15 DAS 30 DAS 45 DAS T 0 Control 70.57 110.68 163.89 23.43 29.52 35.73 T 1 G. fasciculatum + 0 % P 2 O 5 78.33 116.0 168.0 25.9 31.66 38.16 T 2 G. fasciculatum + 25 % P 2 O 5 73.67 109.6 159.0 23.66 28.30 35.20 T 3 G. fasciculatum + 50 % P 2 O 5 69.00 99.67 148.0 20.53 25.16 32.30 T 4 G. fasciculatum + 75 % P 2 O 5 62.00 89.00 101.3 18.43 23.36 29.50 T 5 G. fasciculatum + 100 % P 2 O 5 57.00 73.30 85.66 15.13 20.33 27.66 SED 3.76 4.39 2.54 1.40 1.26 2.92 CD (p = 0.05) 7.55 8.83 5.11 2.82 2.53 5.88 Table - 2: Effect of G. fasciculatum inoculation on the plant height of D. lablab Plant height (cm) 30 DAS 45 DAS 60 DAS T 0 Control 16.75 20.98 25.34 T 1 G. fasciculatum + 0 % P 2 O 5 18.83 23.83 27.16 T 2 G. fasciculatum + 25 % P 2 O 5 19.66 26.66 30.50 T 3 G. fasciculatum + 50 % P 2 O 5 20.00 30.16 35.76 T 4 G. fasciculatum + 75 % P 2 O 5 22.66 31.10 36.46 T 5 G. fasciculatum + 100 % P 2 O 5 29.90 27.66 34.33 SED 3.38 1.38 1.38 CD (p=0.05) 6.80 2.77 2.97 Table - 3: Enumeration of number of root nodules in the rhizospheric region of D. lablab Number of nodules 30 DAS 45 DAS 60 DAS T 0 Control 10.89 14.78 18.29 T 1 G. fasciculatum + 0 % P 2 O 5 12.53 16.55 20.44 T 2 G. fasciculatum + 25 % P 2 O 5 15.98 18.99 22.88 T 3 G. fasciculatum + 50 % P 2 O 5 18.55 21.71 25.21 T 4 G. fasciculatum + 75 % P 2 O 5 19.44 23.11 27.22 T 5 G. fasciculatum + 100 % P 2 O 5 20.44 23.22 30.66 SED 1.63 1.39 4.29 CD (p = 0.05) 3.28 2.81 8.63

Nandini /International Journal of Innovations in Agricultural Sciences (IJIAS), 1(1): 15 20 19 Table - 4: Effect of G. fasciculatum on phosphorus content of the plant D. lablab Phosphorus content (%) 30 DAS 45 DAS 60 DAS T 0 Control 0.62 1.26 2.11 T 1 G. fasciculatum + 0 % P 2 O 5 0.92 1.82 2.72 T 2 G. fasciculatum + 25 % P 2 O 5 1.51 2.95 3.98 T 3 G. fasciculatum + 50 % P 2 O 5 1.96 3.61 4.63 T 4 G. fasciculatum + 75 % P 2 O 5 2.33 3.76 5.19 T 5 G. fasciculatum + 100 % P 2 O 5 2.41 3.40 4.49 SED 0.23 0.37 0.29 CD (p = 0.05) 0.47 0.75 0.59 Table - 5: Effect of G. fasciculatum on the yield of D. lablab Yield (kg /ac -1 ) T 0 Control 378 T 1 G. fasciculatum + 0 % P 2 O 5 413 T 2 G. fasciculatum + 25 % P 2 O 5 461 T 3 G. fasciculatum + 50 % P 2 O 5 493 T 4 G. fasciculatum + 75 % P 2 O 5 497 T 5 G. fasciculatum + 100 % P 2 O 5 495 SED 2.92 CD (p=0.05) 5.88 4. References 1) Abort, L. K and A. D. Robsen. 1977. Growth stimulation of subterranean cloves with vesicular arbuscular mycorrhizae. Aus. J. Agri. Res, 28: 639-649. 2) Abort, L. K and A. D. Robsen. 1982. The role of vesicular arbuscular mycorrhizal fungi in agricultural and the selection of fungi for inoculum. Aus. J. Agri. Res, 3: 389-405. 3) Allen, M. F. 1992. Mycorrhizal functioning. An Integrative plant fungal process. Chapman and Hall, New York, P: 534 [ISNO 412-01891-8]. 4) Baylis, G. T. S. 1959. Effect of Vesicular arbuscular mycorrhizas on growth of Griselinia littoralis (Cornaceae). New Phytology, 58: 274. 5) Fedderman, N. R., T. Finally, F. Boller and Eifstr. 2010. Functional diversity in arbuscular mycorrhizae the role of green expression phosphorus nutrition and symbiotic efficiency. Fungal Ecology, 3: 1-8. 6) Gray, L. E. 1964. Endotrophic mycorrhizae on trees and field crops. M.Sc Thesis, University of Illinosis, Urbana. III. 7) Habte, M and R. L. Fox. 1993. Effectiveness of VA-mycorrhizal fungi in non-sterile soils before and after optimization of P in soil solution. Plant and Soil, 151: 219-226. 8) Harrison, M. J. 1997. The arbuscular mycorrhizal symbiosis: An underground association. Trends in Plant Science, 2: 54-60.

Nandini /International Journal of Innovations in Agricultural Sciences (IJIAS), 1(1): 15 20 20 9) Haymen, D. S and B. Mosse. 1972. Plant growth responses to vesicular arbuscular mycorrhizae. Increased uptake of labile P from soil. New Phytology, 71: 41-47. 10) Hector Valenzuela and Jody Smith. 2002. Lablab, Sustainable Agri Green manure Crops. 11) Jackson, M. L. 1973. Soil chemical analysis, Prentice Hall of India Ltd., New Delhi. 12) Lu, S. L., P. G. Braunberger and M. H. Miller. 1994. Response of vesicular arbuscular mycorrhizae of maize to various rates of P addition to different rooting zones, Plant and Soil, 158: 119-128. 13) Magdi A. Osman. 2007. Effect of different processing methods on nutrient composition, anti-nutritional factors and in vitro protein digestibility of Dolichos lablab bean (Lablab purpuresus L.). Pak. J. Nutri., 6 (4): 299-303. 14) Mosse, B., D. P. Stribey and F. Letacon. 1981. Ecology of mycorrhizae and mycorrhizal fungi. Adv. Microbiol. Ecol,, 5: 137-210. 15) Muchovej, R. M. 2002. Importance of Mycorrhizae Agricultural crops. University of Florida, 1-5. 16) Myrene, R. D Souza and V. R. Devaraj. 2011. Specific and nonspecific responses of Hyacinth bean (Dolichos lablab) to drought stress, Ind. J. Biotech., 10: 130-139. 17) Ryan, M. H., G. A. Chilvers and D. C. Dumaresq. 1994. Colonization of wheat by VA-mycorrhizal fungi was found to be higher on a farm managed in an organic matters than on a conventional neighbor. Plant and Soil, 160: 33-40. 18) Sevaraj, T and P. Chellappan. 2006. Arbuscular mycorrhizae: A diverse personality. Review Paper, 2: 349-358. 19) Smith, S. E and D. J. Read. 1997. Mycorrhizal symbiosis. Second edition. Academic press, New York, P: 605 [ISBNO 12-652840-3]. 20) Srinivasa, M. N., G. T. Basavaraja and J. H. Kulkarni. 1995. Vesicular arbuscular mycorrhizae assist in nodulation and N 2 - fixation in soybean. Maharastra J. Agric, 20: 292-293. 21) Tarafder, J. C and A. V. Rao. 1997. Response of aid Legumes to VAM fungal inoculation. Symbiosis, 22(3) :265-274. 22) Tufenkci, S. Feritsonmez and Gazioguz Sensoy, R. I. 2005. Effects of Arbuscular Mycorrhizae fungus inoculation and phosphorus and nitrogen fertilizations on some plant growth parameters and nutrient content of Chickpea. J. Biol. Sci., 5 (6): 738-743.