Phrase ABSTRACT Vesicular - arbuscular mycorrhiza fungi are associated with the majority of the terrestrial plants. Their function ranges from stress alleviation to bioremediation in soils polluted with heavy metals. However, our knowledge about this symbiosis is still limited. The study was done to wind up best kind growth requirements for VAM growth including pot composition and host plant. The investigation was done to identify maize and sorghum as host and 4 divers substratum composition with materials like vermicompost, ecru, soil and cocopeat which are beneficial and eco - friendly compared to chemical fertilizers. Desertification and aforestation is the elder problem nowadays so there is a need to use the VAM fungi as biofertilizers. So cool of the experiment is to find out the better host and lining for VAM and effect of VAM on plant growth. KEY WORDS: Vesicular Arbuscular mycorrhiza, Biofertilizer, Fungi, Soil 1. INTRODUCTION
Rapid desertification and erosion is a problem in tropical region due to adverse soil and climatic genius. Sustainable agriculture and forestry have in consequence taken prime cynosure for researcher, public and policy makers. Revegetation and afforestation of barren lands through vesicular arbuscular mycorrhizal technology is an immediate necessity. The present paper emphasizes on beneficial factors like VAM and its application as biofertilizer. Scientists are currently curious in developing alternative technology to minimize the dependence on chemical fertilizer to refresh the use of biofertilizer. ( Robert, J. and D. G. Pallid House, 1976 ) VAM fungi are an integral component of sustainable agriculture and tropical forestry with symbiosis between fungus and tree host for common functioning and survival with wide host scope. ( more fitting, 1991, Dixon et al. 1997 ). - The mutualistic nature of the interaction is a critical disposition that differentiates mycorrhiza from other plant - fungus associations. The relationship is primarily characterized by the flow of inorganic components from fungus to plant and organic components from plant to fungus - ( Varma 1995 ). VAM are immensely common symbiont consisting aseptate hyphae desired to the order Glomales of class Zygomycetes. ( Morten and Benny, 1990; Mukharji, 1996 ). VAM fungi are create in assosiation with plant roots of arctic, temperate and tropical region and are distributed in all family of angiosperms. ( Kendric and Berch, 1985 ). They are beneficial to plant by providing nutrients especially phosphorus and zinc, copper, potassium, calcium. ( Cupper and Tinker, 1978 a, b ). VAM structure helps the plan to uptake nutrients from low concentration segment around roots and uptake of water ( Bhandari and Mukerji, 1993 ). The present study was planned to scale up VAM culture and its effect on Various crops. The close obtain from the proposed studies will help in its use as biofertilizer and application for crops production. 2. MATERIALS AND METHODS
Procurement of VAM culture: VAM culture was procured from Indian Council of Agricultural Research, New Delhi. Identification of procured culture: The culture was identified by spore stain method. Large production of VAM: For mammoth scale production of VAM two experiment were performed: 1. Composition Variable: 2. Host Variable: Composition variable: For collar preparation buff and soil sieve with opposed sieves to empty immense gravels. Ergo soil and buff mixed in equal proportion and fill it in flasks. Same procedure followed in other 3 composition: - Neutral: Perlite, Sand: Soil: Vermicompost, Cocopeat. Autoclave it and neighboring autoclave let it cool. Fill this alloy in transparent cups and in each cup add VAM soil culture.. Unaccompanied one cup taken as a control. In each cup make little hole at bottom to withdraw inessential water. Since dissemble 4 wells for seeds inoculation in mix and inoculate seeds in it consequent pretreatment of IPA. Since for growth of plant fit out proper environmental peculiarity approximative as temperature, moisture. Observe the growth of plant intermittently till 60 days. Harvest the spore after 30, 60, 90 days and calculate the spore count. Host variable: In this experiment mixture of Sand: Soil: Serlite: Vermicompost: Cocopeat were used and other procedure same as upper method. But in this two type of seed used 1. Maize and 2. Sorghum. Observe the growth of plant intermittently till 60 days and harvest the spore after 30, 45, 60 days and calculate the spore count. Spore harvesting: Soil sample from Rhizosphere of plant ( Approx 7 cm diameter surrounding the roots ) was collected. From this 1 g of soil sample was taken and sieved it with different sieve ( 650, 150, 45 m ) by simultaneously washing with water. Roots and large soil particles were retained on 650 m and 150 m while fine soil particles and spores were obtained on 45 m sieve. Content collected from 45 m sieve was collected in centrifugal tube and suspended in 27 ml distilled water. Then centrifuged at 2000 rpm for 5 minutes and the supernatant was discarded. Then the pellet was resuspended in 50 % sucrose solution and centrifuged at 2000 rpm for 1 minute. Spores were found to be in the supernatant after centrifugation. Then supernatant was filtered through vacuum filtration apparatus. The spores obtained on filter paper was collected and calculated with the help of dissecting microscope.
3. RESULTS 3. 1 Quality check of VAM culture: VAM culture procured from Indian Council of Agricultural Research was check through spore stain method. The result for staining is shown in fig. 1 ( Fig. 1: Identification of procured VAM culture ) 3. 2 Large scale production of VAM: Composition Variable: In the composition variable experiment 4 type of composition were used as medium in which Sand: Soil: Vermicompost mixture was better growth medium than the other three medium. This result was shown in fig. 2 and Table 1. Fig. 2 shows the growth of plant and Table 1 shows the plant growth and VAM growth. Sand: Perlite composition shows 30 % better growth than Sand: Soil composition. Sand: Soil: Vermicompost composition shows 44 % better growth than Sand: Soil composition. Cocopeat shows 34 % lesser growth than Sand: Soil. Table 1: Observation of plant growth and VAM count at different interval time Incubation time Parameters Soil bed composition
Sand: Soil Sand: perlite Sand: Soil: Vermicompost Cocopeat 30 days Plant length 25. 34 21 22 10
Spore count ( Spore / g ) 40 32 36 25 45 days Plant length 28 31 35 15
Spore count 145 184 190 37 60 days Plant length 30 39 43 20
Spore count 678 814 890 45
Sand: Soil: Vermicompost ( 1: 1: 1 ) Sand: Perlite ( 1: 1 )
Sand: Soil ( 1: 1 ) Cocopeat ( Fig. 2: Observation of plant growth in different bed composition used as growth medium )
- Host Variable: In the host variable experiment 2 type of seeds were used as host: Maize and Sorghum. In which Maize shows better growth of VAM which was shown in Table 2 and Fig. 3. Table 2 shows spore count at different harvest time. Because of the better growth of VAM in Maize, Maize grows at high rate than the Sorghum. Table 2: Effect of plant host on VAM spores at different harvest time
Sr. No. Incubation time Spore count ( Spores / gm )
Maize Sorghum 1 30 days 40 31 2 45 days 183 101 3 60 days 884 746 From this table we conclude that when maize seeds were used as a host, Spore count seen to be 18 % higher than the sorghum which indicate the better growth of VAM culture in Maize. So, as compare to sorghum Maize provide beneficial condition for spore generation in VAM culture.
( Fig. 3: Comparative study of spore count in Maize and Sorghum at different harvest time )
( Fig. 4 Growth of Sorghum when it used as host for VAM )
( Fig. 5 Growth of Maize when it used as host for VAM )
- Fig. 4 and 5 shows the effect of VAM on Maize and Sorghum. In Maize VAM spore count is high as compare to Sorghum therefore because of the better growth of VAM in Maize Plant growth is high as compare to Sorghum. 4. CONCLUSION There are three main components involved in VAM association and plant growth with soil fertility1 ) the soil, 2 ) the fungus and 3 ) the plant. The fungal component involves, the fungal structure within the cell of the root and the extraradical mycelium in the soil. The last may be not extensive under some conditions, but does not form any vegetative structures ( Smith and Read, 1997 ). Its primary function is the absorption of resources from the soil. The increased efficiency of mycorrhizal roots versus non - mycorrhizal roots is caused by the active uptake and transport of nutrients by mycorrhizae. AMF have been shown to improve productivity in soils of low fertility ( Jeffries, 1987 ) and are particularly important for increasing the uptake of slowly diffusing ions such as PO4 ( Jacobsen et al., 1992 ), immobile nutrients such as P, Zn and Cu ( Lambert et al., 1979; George et al., 1994; George et al., 1996; Ortas et al., 1996; Liu et al., 2002 ) and other nutrients such as Cadmium ( Guo et al., 1996 ). In the present investigation Sand: Perlite, Sand: Soil: Vermicompost, Cocopeat and studied for number of AM spores and root colonization compared to Sand: Soil. So results of the experimental study for the analysis of AM growth in four different bed composition shows that Vermicompost in combination of sand and soil gives batter growth for VAM spores. The results are supported by the views of Nence ( 1987 ) who stated the superiority of perlite: sand mix over only sand. Douds and Reider ( 2003 ) inoculated AM in soil amended with composted dairy cow manure and reported enhanced sporulation. Muthukumar and Udaiyan ( 2002 ) reported increased spore - population of G. aggregatum in cowpea in response to organic manure. Lins et al. ( 1999 ) reported better growth of Carica papaya even after 30 days with the application of cattle manure to soil. Similarly Tarkalson et al. ( 1998 ) experimented on dry bean and reported higher VAM root colonization in soil amended with manure or compost. Sharma et al. ( 2005 ) also recommended farmyard manure to be best for culture of G. mosseae spores with maize. Bajwa and Javaid ( 2002 ), Srivastava et al. ( 2001 ) observed enhanced colonization in maize on application of farmyard compost. Ellis ( 1998 ) also showed that the absence of host plants could drastically reduce AM fungal populations in soil. Live roots present in the soil produce volatile organic compounds and carbon dioxide, which are believed to stimulate spore germination ( Becard and Piche, 1989; Carpenter - Boggs et al., 1995 ). In the absence of active roots, the likelihood that spores will stay dormant. Delayed germination could be a survival mechanism utilized by sporulating fungi ( Abbott andRobson, 1982 ). However, in the present investigation, the four substrates and 2 hosts used were able to maintain the growth of the hyphae and the development of VAM fungal spores but vermicompost found to be the best tool for the production of VAM fungi in a fertilizer form. One of the reasons for this may be the supply of minearals by vermi - compost. A stimulation of AM sporulation by organic substances has already been described by Gryndler et al. ( 2003 ). Presumably, soil amended with substrates and VAM fungal spores can be a good ameliorative solution for reclaiming acidic soils. AM fungi are and will be a major component in any sustainable plant productive systems, which can be explicitly used for the maintenance of species diversity and productivity.
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