Biofertilizer s Bacteria Many free-living and symbiotic bacteria are discussed in Biological Nitrogen Fixation which fixes atmospheric

N2. Therefore, certain measures are adopted to increases number of such bacteria in soil which may increase the gross yield of nitrogen. The two methods, bacterization and green-manuring are the most widely used techniques. Bacterization Bacterization is a technique of seed-dressing with bacteria (as water suspension) for example, Azotobacter, Bacillus, Rhizobium etc. It has been proved that bacteria can successfully be established in root region of plants which in turn improve the growth of hosts. Bacterial fertilizers named 'azotobakterin' (containing cells of Azotobacter chroococcum) and phosphobacterin' (containing cells of Baccilus megatehum var.phosphaticum) have been used in erstwhile U.S.S.R. and East European countries, respectively. These increased the crop yield about 10-20 per cent (Cooper, 1959). Subsequently bacterization of seeds in Russia, Czechoslovakia, Rumania, Poland, Bulgaria, Hungary, England and India has clearly demonstrated the increase in crop .yield such as wheat, barley, maize, sugarbeet, carrot, cabbage and potato. In rhizosphere, bacteria secrete growth substances and antibiotic secondary metabolites which contribute to seed germination and plant growth (Subba Rao, 1982; Dwivedi et al, 1989). Biofertilizers Mass Cultivation of Microbial Inoculants Bacteria to be inoculated in soil as biofertilizer need to be multiplied on artificial media to harvest on a large scale so that it can be supplied to farmers. Mass cultivation of rhizobia,Azotobacter and Azospirillum is discussed below : Rhizobium Species of Rhizobium are kept in different specialization groups. Inoculum of different strains are prepared separately and cultivated on large scale, as required. Strains ofRhizobium sp. are grown in Yeast Extract Mannitol (YEM) broth in a small or large container as needed. Chemical composition of YEM broth is as below : Yeast extract 1g Mannitol 10g K2HPO4 0.5 g Mg SO4 7H2O 0.2 g NaCl 0.1 g Distilled Water 1000 ml pH 6.5 - 7.0 Methods of Cultivation Following are the steps of mass cultivation of Rhizobium. (a) sterilize the growth medium and inoculate with broth of mother culture prepared in advance, (b)incubate for 3-4 days at 30 - 32C, (c) test the cultures for its purity and transfer to a large fermenter, wait for 4-9 days for bacterial growth (for good bacterial growth make the device for its aeration), (d) allow to grow the bacteria either in a large fermenter containing broth or in small flasks as per demand,(e) check the quality of broth, (f) blend the broth with sterile carrier e.g. peat, lignite, farmyard manure and charcoal powder, (g) pack the culture in polyethylene bags and keep at 25C, (h) check the quality of carrier culture, (i) store at 4C in a controlled-temperature room, and (j) supply to farmers. During the blending of broth a variety of carriers are used, for example, peat, lignite, farmyard manure, charcoal powder, etc. In India powdered farmyard manure and charcoal powder are good carrier and an alternative to peat and lignite. Good quality of carrier culture is that which contains sufficient amount of rhizobial cells i.e. 1000 x 106 to 4000 x 106 rhizobia/g carrier. Seed inoculation with aqueous suspension of carrier culture during sowing has revealed the luxuriant nodulation and good yield of crops. Methods of seed inoculation with rhizobial culture The steps of seed inoculation with rhizobial culture are given in Fig. 12.1. Dissolve 10 per cent sugar or gur (jaggery) in water by boiling it for some time. Leave the content to cool down. Gum arabic solution (10% ) may also be added to the solution. This serves as sticker for Rhizobium cells to seeds. Mix this carrier based culture of Rhizobium to form the inoculum slurry. For one hectare, 400 g charcoal based culture would be sufficient for mixing the seeds. Transfer the inoculum slurry on seeds and mix properly. The number of rhizobial cells/ seed should be between 10 5 to 106. Spread the seeds in shade for drying on cement floor or plastic sheets. While using rhizobial cultures, certain precautions are taken into account. For example, use of culture before expiry date, use of small amount of pesticides when required, immediate sowing of seeds after mixing, etc. Seeds must be stored at 4C when not used immediately to protect the rhizobial cells. Pelleting When soil has the adverse conditions such as dryness, acidity, excess fertilizers and pesticides, etc., the rhizobial cells are protected by adopting special method of inoculation. One of these methods is pelleting i.e. preparation of pelleted seeds. This method involves the procedure as described earlier. High amount of gum arabic (40%) or carboxymethylcellulose (20%) is added to the inoculum slurry before mixing with seeds. Finally, pelleting agent is mixed when inoclated seeds are moist (before seed drying ) to get the seeds evenly coated.

The commonly used pelleting agents are calcium carbonate, rock phoshate, charcoal powder, gypsum and betonite. Effects of rhizobial inoculants on crop yield Effect of rhizobial culture on yield of different pulses, and subsequent crops grown after harvesting the pulse crops are given in Table 12.1 based on the study made under All India Coordinated Pulse Improvement Research Programme of I.C.A.R., New Delhi. Yield of pulse crops can be substantially increased by rhizobial inoculation. Legume crops get benefit from rhizobial symbiosis. In addition, certain amount of N is left over in soil which is taken by the other plants also.

Fig. 12.1. Procedure for seed inoculation with rhizobial culture. Subba Rao and Tilak (1977) studied the effect of residual nitrogen of many legumes on the yield of subsequent crops of wheat or rice. They always found more yield of subsequent crops in Rhizobium inoculated fields than in uninoculated control. Table 12.1. Response of different pulse crops to rhizobial cultures under different agroclimatic condition and residual effect on yield of subsequent crops. Crop response (range in % increase in grain yield (q/ha) over UI control)* 5 -25 2-25 9-21 13-29 3-40 4-19 25-40 18-28 24-43 33-67 20-41 8-12 4-26 No response 4-21 11-29 Yield of subsequent crops (% increase in yield over control, soil pH 7.3)** Yield % increase Crops (q/ ha) over control Wheat UI-20.75 16.4 (Triticwn RI-24.15 aestivum)

Crops

Location

Arhar (Cajanu cajan)

Chickpea (Cicer aritinum)

Lentil (Lens culinaris) Urd bean (Vigna munga)

Hisar, Haryana Pantnagar, U.P. S.K. Nagar, Gujarat Sehore, M.P. Rehari (Maharastra) Varanasi, U.P Dholi, Bihar Delhi Hisar Dohad, Gujarat Sehore Maharastra Pantnagar, U.P Ludhiana, Punjab

Rice (Oryza sativa)

UI-25.15 RI-27.15

7.9

Rice Rice Wheat RI-21.25

UI-22.57 RI-25.55 UI-20.75

13.2

Pudukkotti, T.N. Dholi, Bihar Pantnagar Pudukkotti, T.N. * Rewari (1984, 1985); ** with Rhizobium culture.

2.4

Subba

Rao

and

Jilak

(1977);

UI,

Uninoculated

control;

RI,

Inoculated

Azotobacter, Azospirillum and Phosphate Solubilizers Growth medium prepared for Rhizobium also supports the growth of A. chroococcum. Media used for mass cultivation of Azotobacter, Azospirillum and phosphate solubilizing bacteria differs in chemical composition (Table 12.2). Procedure for preparation of inoculants on large scale is similar to that of Rhizobium. Similarly, seed bacterization is also done as described earlier. The roots of transplantable crops e.g. vegetable) are dipped into aqueous suspension of carrier culture and then sown in fields. Carriers used for Azospirillum and Azotobacter are the same as for Rhizobium. Soil plus farmyard manure (1:1) gave the good results as far as survival of bacterial cells are concerned. Bacterial cells survived for about 31 weeks. Procedure for inoculation of seeds is the same as described for Rhizobium. Seed inoculation with A. brasilense increased the grain yield of sorghum, pearlmillet, fingermillet and rice over the uninoculated control. Even in these crops, this saves the nitrogen to about 20-30 Kg/ha (Tilak, 1991). Table 12.4 shows the effect of Azotobacter inoculant on yield of many crops. Tien et at (1979) have attributed that increased yield in pearlmillet obtained by inoculation of Azospirillum was due to the production of indoleacetic acid, gibberellins and cytokinin like substances by the bacterium and their subsequent effect on the plant. Gaur and Algawadi (1989) studied the interaction of N2 fixing bacteria (A. chroococcum) and phosphate solubilizing bacteria (B. megaterium and P. striata) and their effect on sorghum and rice crops in a green house experiment by dual inoculation of sorghum with A. brassilens and P. striata. They found a significant increase in root nitrogenase activity, dry matter and seed yields as compared to single inoculation of both the organisms and control. Similarly, nutrient uptake and yield of rice increased when inoculated together with A. chroococcum and P. striata as compared to uninoculated control or single inoculated experiments.

Table 12.2. Composition of media for mass cultivation of bacteria (g/l). Chemical Azotobacter Azospirillum Bacillus composition chroococcum megaterium, Pseudomonas striata KH2 PO4 1.0 0.50 MgSO4 7H2O 0.50 0.10 0.10 NaCl 0.50 0.02 KCL 0.20 MnSO4 .H20 0.01 CaCO2 2.00 KOH 4.00 (NH4)2 . SO4 0.50 MnSO4 0.002 FeSO4 0.10 0.002 FeSO47. H2O 0.05 5.00 Ca3(Po4)2 Na2MoO4 0.002 5.00 CaCl2 0.01 Sucrose 20 Glucose 10 Malic acid 5.00 Yeast extract 0.5 pH 6.0-7.0 Field experiments conducted at IARI, New Delhi, have shown that upon seed inoculation of sorghum and cotton by A. chroococcum the yield was increased by 38 pet cent and 27 per cent respectively, whereas increase in yield of wheat was 10 per cent at the same conditions. These experiments suggest that small and marginal farmers can follow the seed bacterization and increase crop yield. Effects of these bacteria on yield of crops are given in Tables 12.3. and 12.4. Table 12.3. Effect of seed inoculation with Azospirillum brasilense on grain yield (q/ha). Plant species * Treatments References Control A. Urea Urea + (N-free) brasilense A. brasilense 1. Eleusine coracana 16.54 19.47 23.07 28.09 Subba Roa et (fingermillet) al. (1985) 2. Pennisetum 12.4 14.3 20.4 Tilak and americanum Subba Rao (pearlmillet) (1987) 3. Sorghum biocolor ** 19.23 22.78 30.48 31.05 Tilak (1991) (sorghum var. CSH 5) * Values of 1, 2 and 3 are all India mean of 4.4 and 9 centres respectively, ** Average of 4 years' results; a, 40 Kg N/ha. Table 12.4. Effect of Azotobacter inoculant on crop yield. Crops % increase in yield over Azotobacter uninoculated control chroococcum ** Azotobakterin* Cotton 6.7 -26.6 Barley 9.0 Maize 8.0 36.5 71.7 Oat 12.0 Potato 8.0 Sorghum 9.3 38.1 Sugar beat 7.6 * Mishustin and Shilnikova(1969)j ** Shende and Apte (1982). Green Manuring Ghai and Thomas (1989) have defined green manuring as "a farming practice where a leguminous plant which has derived enough benefits from its association with appropriate species of Rhizobium is ploughed into the soil and then a non legume is grown and allowed to take the benefits of the already fixed nitrogen". The practice of green manuring was started from several century B.C. in India and China. During the course of time availability of chemical fertilizers decreased the significance of green manuring. In recent years, due to hike in price of chemical fertilizers, the practice of green manuring is reemphasized. Ghai and Thomas (1989) have given a list of various leguminous plants to be used as green manure. Some of them are: cultivated annual legumes (e.g. Crotalaria juncea, C. striata, Cassia mimosoides, Cyamopsis pamas, Glycine wightii, Indigofera linifolia, Sesbania rostrata, Vigna radiata), perennial legumes (e.g. Acacia nilotica, Cassia hirsuta, Sesbania aegyptica, Leucaena Jeucocephala), and wild annual legumes (e.g. Cassia cobanensis, Lathyrus sativus, Mimosa invisa, Mucuna bracteata). In a report of International Rice Research Institute (Philippines) it has been suggested that fast growing tropical legumes can accumulate more than 80 Kg N/ha when grown as green manures.

Biofertilizers

The Blue-Green Algae (Cyanobacteria) Role of the blue-green algae (e.g. Aulosira, Anabaena, Cylindrospermum, Nostoc, Plectonema, Tolypothrix) in the paddy fields was realized much earlier (Singh, 1961). In water-logging condition, the cyanobacteria multiply, fix atmospheric N2 and release it into the surroundings in the form of amino acids, proteins and other growth promoting substances (Sewart, 1970). Recent works done at Central Rice Research Institute, (Cuttack), Indian Council of Agricultural Research (New Delhi) and other centres, e.g. Centre of Advanced Study in Botany, Banaras Hindu University, Varanasi are of much importance. Algalization It were Japanese workers (Watanabe and coworkers) who developed techniques for mass cultivation of blue-green algae to be used as biofertilizer in paddy fields. Venkataraman (1961) coined the term 'algalization' to denote the process of application of blue-green algal culture in field as biofertilizer. He initiated algalization technology in India and demonstrated the way how this technology could be transferred to farmer level who hold small lands (Vekataraman, 1972). At present, algalization is being followed in Tamil Nadu and Uttar Pradesh, and tried in Jammu and Kashmir, Andhra Pradesh, Karnataka, Maharashtra and Haryana. Algalization is also being practiced in China, Egypt, Philippines and erstwhile U.S.S.R. In 1990, Department of Biotechnology, Government of India, New Delhi establishment four major centres in different paddy growing areas of the country for acceleration and extension of works on algal biofertilizers. The programme was launched under 'Technology Development and Demonstration Projects on Cyanobacterial Biofertilizers' in U.P. (Lucknow), Tamil Nadu (Madurai), West Bengal (Calcutta) and New Delhi. The main objectives of the programme were (i)to develop low cost indigenous technology for mass production of cyano bacteria, (ii) to isolate regional specific fast growing and better N2 - fixing strains, (iii)to develop starter inoculum, (iv) to demonstrate the farmers in field, and (v) to study the benefits on both economy and ecology. As a result of these studies cyanobacterial biofertilizer was found very useful, especially for small and marginal farmers of the country with the view point of both economy and ecology (Venkataraman, 1993). Table 12.5. Contribution of nitrogen by some of the nodulated legumes when used as green manure/cover crop. N2 fixing system Amount of N contributed (q/ha) 1. Green manure legumes : Sesbania aculeata Rhizobium 70-120 Leucaena leucocephala - Rhizobium 500-600 Beans (broad beans / lupines /soybean / lentil, 60-210 etc.) - Rhizobium Fodders (Trifolium / Medicagol Melilotus, etc. ) - 100-300 Rhizobium 2. Cover crop legumes : Lablab purpureus - Rhizobium 240 Glycine jawanica - Rhizobium 210 3. Non-legumes : Casuarina equisitifolia - Frankia 100 Alnus - Frankia 30-300 4. Others : Azolla - Anabaena 25-190 Grasses - Azospirillium 15-100 Source : Ghai and Thomas (1989). D.B.T. Centre of U.P. (Lucknow) has reported the increase in yield of paddy (about 12.5 q/ha) to be due to cyanobacterial biofertilizer. Moreover, its use continued with chemical fertilizer gave the best results. Because the cyanobacterial biofertilizer not only increase the productivity and quality of paddy, but also minimize the harmful effects of the chemical fertilizers.

Mass cultivation of cyanobacterial biofertilizers For outdoor mass cultivation of cyanobacterial biofertilizers, the regional specific strains should be used. However, many germplasm collection laboratories have been established by the D.B.T. in different parts of the country for the development of starter inoculum. Mixture of 5 or 6 regional acclimatized strains of cyanobacteria, e.g. species of Anabaena, Aulosira, Cylindrospermum, Gloeotrichia, Nostoc, Plectonema, Tolypothrix are generally used for starter inoculum. The following four methods are used for mass cultivation : (i) cemented tank method., (ii) shallow metal troughs method, (iii) polythene lined pit method, and (iv)field method. The polythene lined pit method is most suitable for small and marginal farmers to prepared algal biofertilizer. In this method, small pits are prepared in field and lined with thick polythene sheets. Mass cultivation of cyanobacteria is done by using any of the four methods under the following steps: (i) Prepare the cemented tanks, shallow trays of iron sheets or polythene lined pits in an open area. Width of tanks or p facilitate the proper handling of culture. (ii) Transfer 2 -3 Kg soil (collected from open place for lm 2 area of the tank) and add 100 g of superphosphate. Water adjust the pH 7. Add 2 ml of insecticide e.g. malathion to protect the culture from mosquitoes. Mix well and allow to se (iii) When water becomes clear, sprinkle 100 g of starter inoculum on the surface of water. (iv) When temperature remains between 35-40 during summer, optimum growth of cyanobacteria is achieved. Alway during this period, (v) After drying, the algal mat will get separated from the soil and forms flakes. During summer about 1 kg pure alg collected, powdered, kept in sealed polythene bags and supplied to the farmers. (vi) The algal flakes can be used as starter inoculum if the same process is repeated. Azolla Azolla is an aquatic heterosporous fern which contains an endophytic cyanobacterium,Anabaena azollae, in its leaf cavity. The significance of Azolla as biofertilizer in rice field was realized in Vietnam. Recently, it has become very popular in China, Indonesia, Philippines, India and Bangladesh. A total of six species of Azolla are known so far viz., A. caroliniana, A. filiculoides, A. mexicana, A. microphylla, A. nilotica, A. pinnata and A. rubra. Out of these A. pinnate is commonly found in India. The global collections of several

species of Azolla are maintained at CRRI (Cuttack). Within the leaf cavity filaments of Anabaena azollae are present. Dr. P.K. Singh, at CRRI has done an outstanding work on mass cultivation of Azolla and its use as biofertilizer in rice and other crop fields. Mass cultivation of Azolla Microplots (20m2) are prepared in nurseries in which sufficient water (5-10 cm) is added. For good growth of Azolla, 4-20 Kg P2O5/ha is also amended. Optimum pH (8.0) and temperature (14-30C) should be maintained. Finally, microplots are inoculated with freshAzolla (0.5 to 0.4 Kg/ m2). An insecticide (furadon) is used to check the attack of insects. After three, week of growth mat formed by Azolla is harvested and the same microplot is inoculated with fresh Azolla to repeat the cultivation. Azolla mat is harvested and dried to use as green manure. There are two methods for its application in field: (a) incorporation of Azolla in soil prior to rice plantation, and (b)transplantation of rice followed by water draining and incorporation of Azolla (Singh, 1977, 1979, 1980). However, reports from the IRRI (Philippines) reveal that growing of Azolla in rice field before rice transplantation increased the yield equivalent to that obtained from 30Kg N/ha as urea or ammonium phosphate. Moreover, Azolla shows tolerance against heavy metals viz. As, Hg, Pb, Cu, Cd, Cr, etc. It tolerates low concentration but at high levels a setback in biochemical pathways is caused.A. pinnata absorbs heavy metals into cell walls and vacuoles through evolution of specific metal resistant enzymes. Therefore, heavy metal resistant species such as A. pinnata can also be incorporated as green manure in rice field near the polluted areas where heavy metal concentration is between 0.01 and 1.5 mg/liter. Due to development of chemical industries and discharge of effluents into water bodies, heavy metal concentration is gradually increasing day by day. Industries where work of electroplanting, fertilizers, tanning, etc. are done, they act as a chief source for soil and water pollution. For example, disturbed vegetation in aquatic system around Damodar river valley in India has received a great attention.

Mycorrhiza as Biofertilizers Mycorrhiza (fungus roots) is a distinct morphological structure which develops as a result of mutualistic symbiosis between some specific root - inhabiting fungi and plant roots. Plants which suffer from nutrient scarcity, especially P and N, develop mycorrhiza i.e. the plants belong to all groups e.g. herbs, shrubs, trees, aquatic, xerophytes, epiphytes, hydrophytes or terrestrial ones. In most of the cases plant seedling fails to grow if the soil does not contain inoculum of mycorrhizal fungi. In recent years, use of artificially produced inoculum of mycorrhizal fungi has increased its significance due to its multifarous role in plant growth and yield, and resistance against climatic and edaphic stresses, pathogens and pests. Mechanism of Symbiosis The mechanism of symbiosis is not fully understood. Biorkman (1949) postulated the carbohydrate theory and explained the development of mycorrhizas in soils deficient in available P and N, and high light intensity. Slankis (1961) found that at high light intensity, surplus carbohydrates are formed which are exuded from roots. This in turn induces the mycorrhizal fungi of soil to infect the roots. At low light intensity, carbohydrates are not produced in surplus, therefore, plant roots fail to develop mycorrhizas.

Types of Mycorrhizas By earlier mycologists the mycorrhizas were divided into the following three groups : (i) Ectomycorrhiza. It is found among gymnosperms and angiosperms. In short roots of higher plants generally root hairs are absent. Therefore, the roots are infected by mycorrhizal fungi which, in turn, replace the root hairs (if present) and form a mantle. The hyphae grow intercellularly and develop Hartig net in cortex. Thus, a bridge is established between the soil and root through the mycelia. (ii) Endomycorrhiza. The morphology of endomycorrhizal roots, after infection and establishment, remain unchanged. Root hairs develop in a normal way. The fungi are present on root surface individually. They also penetrate the cortical cells and get established intracellulary by secreting extracellular enzymes. Endomycorrhizas are found in all groups of plant kingdom. (iii) Ectendomycorrhiza. In the roots of some of the gymnosperms and angiosperms, ectotrophic fungal infection occur. Hyphae are established intracellularly in cortical cells. Thus, symbiotic relation develops similar to ecto- and endo-mycorrhizas. Marks (1991) classified the mycorrhizas into seven types on the basis of types of relationships with the hosts (i) vesicular-arbuscular (VA) mycorrhizas (coiled, intracellular hyphae, vesicle and arbuscules present), (ii) ectomycprrhizas (sheath and inter-cellular hyphae present), (iii) ectendomycorrhizas (sheath optional, inter and intra-cellular hyphae present), (iv) arbutoid mycorrhizas (seath, inter-and coiled intracellular hyphae present), (v) monotropoid mycorrhizas (sheath, inter- and intra- cellular hyphae and peg like haustoria present), (vi) ericoid mycorrhizas (only coiled intracellular hyphae, long coiled hyphae present), and (viii) orchidaceous mycorrhizas (only coiled intracellujlar hyphae present). Type (i) is present in all groups of plant kingdom; Types (ii) and (iii)are found in gymnosperms and angiosperms. Types (iv), (v) and (vi) are restricted to Ericales, Monotropaceae and Ericales respectively. Types (vii) is restricted to Orchidaceous only. Types (iv) and (v) were previously grouped under ericoid mycorrhizaes.

Methods of Inoculum Production and Inoculation Methods of inoculum production of VAM fungi differ; however, some of these two are briefly described here. (a) Ectomycorrhizal fungi: The basidiospores, chopped sporocarp, sclerotia, pure mycelia culture, fragmented mycorrhizal roots or soil from mycorrhizosphere region can be used as inoculum. The inoculum is mixed with nursery soils and seeds are sown. Institute for Mycorrhizal Research and Development, U.S.A., Athens and Abbort Laboratories (U.S.A) have developed a mycelial inoculum of Pisolithus tinctorius in a vermiculite-peat moss substrate with a trade name Myco-Rhiz which is now commercially available on large quantities. In 1982, about 1.5 million pine seedlings were produced with MycoRhiz in the U.S.A. (Marx and Schenck, 1983). (b) VA mycorrhizal fungi : VA mycorrhizas can be produced on a large scale by pot culture technique. This requires the host plants, Fig. 12.5. Method of production of VAM (VA mycorrhizal) inoculum for mycorrhizal fungi and natural soil. application in fields. The host plants which support large scale production of inoculum are sudan grass, strawberry, sorghum, maize, onion, citrus, etc. The starter inoculum (spores) of VA mycorrhizal fungi can be isolated from soil by wet sieving and decantation technique (Gerdeman and Nicolson, 1963). VA mycorrhizal spores are surface sterilized and brought to the pot culture. Commonly used pot substrates are sand: soil (1:1, w/w) with a little amount of moisture. An out line for inoculum production is given in Fig. 12.5. There are two methods of using the inoculum : (i) using a dried spore-root- soil to plants by placing the inoculum several centimeters below the seeds or seedlings, (ii) using a mixture of soil-roots, and spores in soil pellets and spores adhered to seeds with adhesives. Commercially available pot culture of VA mycorrhizal hosts grown under aseptic conditions can provide effective inoculum. Various types of VA mycorrhizal inocula are currently produced by Native Plants, Inc (NPI), Salt Lake City. In India, Forest Research Institute, Dehra Dun has established mycorrhizal bank in different states of the country. Inocula of these can be procured as needed and used in horticulture and forestry programmes. Benefits from Mycorrhizas to Plants (i) They increase the longevity of feeder roots, surface area of roots by forming mantle and spreading mycelia into soil and, in turn, the rate of absorption of major and minor nutrients from soil resulting in enhanced plant growth. (ii) They play a key role for selective absorption of immobile (P, Zn and Cu) and mobile (S, Ca, K, Fe, Mn, Cl, Br, and N) elements to plants. These are available to plants in less amount (Tinker, 1984). (iii) Some of the trees like pines cannot grow in new areas unless soil has mycorrhizal inocula because of limited or coarse root hairs. (iv) VA mycorrhizal fungi enhance water uptake in plants, (v) VA mycorrhizal fungi reduce plant response to soil stress such as high salt levels, toxicity associated with heavy metals, mine spoils, drought and minor element (e.g. Mn) imbalance. (vi) VA mycorrhizal fungi decrease transplant socks to seedlings. They produce organic 'glues' which bind soil particles into semistable in aggregates. Thus, they play a significant role in augmenting soil fertility and plant nutrition. (vii) Some of them produce metabolites which change the ability of plants to induce roots from woody plant cuttings and increase root development during vegetative propagation. (viii) They increase resistance in plants and with their presence reduce the effects of pathogens and pests on plant health. Benefits from Biofertilizers Following are benefits from the biofertilizers : (i) It is a low cost and easy technique, and can be used by small and marginal farmers. (ii) It is free from pollution hazards and increase soil fertility. (iii) On application of algal biofertilizers increase in rice yields ranges between 10-45 per cent and about 40-50 Kg N is left over in the soil which in turn is used for the subsequent crops (Venkataraman, 1972). Moreover, benefits from algalization is about 25-30 Kg N/ ha/cropping season in rice fields (Subba Rao, 1992). After 3-

4 consequent years, the algal effects become consistent and there is no need of using this practice. The parental inoculum is sufficient for growth and multiplication. (iv) Cyanobacteria secrete growth promoting substances like IAA, IBA, NAA, aminoacids, proteins, vitamins, etc. They add sufficient amount of organic matter in soil. (v) Cyanobacteria can grow and multiply under wide pH range of 6.5-8.5. Therefore, they can be used as the possible tool to reclaim saline or alkaline soil because of their ameliorating effect on the physico-chemical properties of the soil. (vi) Rhizobial biofertilizer can fix 50-150 kg N/ha/annum. (vii) Azotobacter and Azospirillum, besides supplying N to soil, secrete antibiotics which act as pesticides (viii) Azolla supplies N, increases organic matter and fertility in soil and shows tolerance against heavy metals. (ix) The bioferilizers increase physico-chemical properties of soils such as soil structure, texture, water holding capacity, cation exchange capacity and pH by providing several nutrients and sufficient organic matter. (x) The mycorrhizal biofertilizers make the host plants available with certain elements, increase longevity and surface area of roots, reduce plant response to soil stresses, and increase resistance in plants. In general, plant growth, survival and yield are increased. Commercial Producers of Biofertilizers Following are some of the companies producing microbial inoculants (the addresses are subject to change): A. Indian Companies: Trade name 1. Bacfil, 25 N.K. Road, Lucknow 'Rhizoteeka' 2. Microbes India, 87 Lenin Sarani, Calcutta-13 3. Rallis India Ltd, 87 Richmond, Road, Bangalore-25 4. Indian Organic Chemicals Ltd, 15 'Nodin', Mathew Road, Mumbai - 4 'Natrin B. Foreign Companies: 1. Union Chemique S.A., Belgium 'Nodosit' 2. Phyluxia Allami, Hungary 'Rhizonit' 3. Laboratorie de Microbiologie, 'N-germ' France 4. Root Nodue Pvt. Ltd., Australia 'Nitrogerm' 5. Agricultural Laboratories, Austria. 'Nodulud' 6. Radicin Institute, Germany. 'Radicin Impfsfoff' 7. Abbot Laboratories , U.S.A. and 'MycoRhiz Institute for Mycorrhizal Research and Development, U.S.D.A., Athens.

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