General Description:
Introduction of high yielding varieties to meet the demand of food production ultimately rest on applying huge quantities of inorganic fertilizers to nurture the nutrient requirements of high yielding plants. This practice of applying high nutrient content inorganic fertilizers , slowly wiped out the traditional agricultural practices of applying organic source of nutrients to the soil to improve soil health and sustain soil fertility. In India 60% of the total cultivable soil has less than the minimum requirement of organic carbon and also over thirty years period of time the organic carbon content has come down to 0.60% from 1.2%. This reduction in soil health reflected the reduction in microbial population in the soil. If microbial diversity in the soil is reduced, the chances of improving the soil fertility becomes a question, because soil microbes act on various organic matter and release the all the nutrients. Because of this lack of soil organic carbon, the technologies adopted, or new varieties tested are not responding to the desired level. The problem is not with the variety north the technology. But with the soil health. A healthy soil respond very well to the new technologies adopted or bring out the maximum potential from any good seed. By taking into account all these factors, research was taken up to improve soil health as well as to establish healthy plant. Out of three years of research carried out this TNAU Bio-Capsule was successfully developed for farmers use.
Prior Art:
Biofertilizers:
Biofertilizers are of recent origins and are successfully applied to crop plants to enhance the yield. Air has 78% Nitrogen and 88,000 tons of nitrogen in every hectare of land on the world. Azospirillum form an associative symbiosis with many plants particularly with those having the C4 decarboxylic acid pathway of photosynthesis. Azospirillum applied to crops like Wheat, Rice, Barley, Sorghum showed good response under various agroclimatic conditions. ( Verma & Bhattacharya 2000)

The availability of phosphorous in Indian soil is poor. About 25 to 30% of the applied phosphorous becomes available to crops and the remaining portion gets converted into insoluble forms. These fixed forms are solubulized and mineralized by certain microorganisms such as Bacillus sp and Pseudomonas sp . ( Prabhakaran and Uthayakumar 2006)
Azospirillum:
Azospirillum is well adapted to establish in the competitive environment of the rhizosphere due to the versatility in utilizing different carbon and nitrogen sources. This makes Azospirillum as a more potent bioinoculant to improve plant growth (Steenhoudt and Vanderleyden, 2000).
Nitrogen fixation in culture media by Azospirillum was precisely assessed through acetylene reduction assay by Van Berkum and Bohlool (1980). Incorporation of atmospheric nitrogen into wetland rice was assessed through acetylene reduction assay (ARA) by Watanabe and Gin (1984). In an experiment conducted with hydroponic cultures, within 7 days after inoculation of Azospirillum lipofertim in rice, significant increase in the uptake of PO4' and NHU4 ion were observed. These observations had positive correlation with increase in biometric observations (Murty and Ladha, 1988). Auxins
The auxin, indole acetic acid is the most important plant growth stimulating substance produced by Azospirillum cultures (Kapulink et al, 1985; Okon and Kapulink, 1986). Azospirillum was found to produce lAA during stationary phase of growth. Addition of lAA precursor- tryptophan and nitrogen sources to the culture medium enhanced the lAA synthesis (Baca et al, 1994).
Many researchers have reported that plant growth hormones, predominantly indole acetic acid are produced by Azospirillump. It is mainly responsible for the

enhanced plant growth as well as yield of crop plants (Okon and Labandera- Gonzalez, 1994; Dobbelaere et aL, 1999). In culture medium, wild strain Azospirillum brasilense was able to produce indole-3-butyric acid. The compound obtained from culture filtrate when sprayed on the maize seedlings under in vitro conditions increased the growth of maize seedlings considerably (Martinez-Morales et al, 2003). Addition of tryptophan to culture media stimulated auxin biosynthesis. The strain that synthesized highest amount of auxin under in vitro conditions recorded the maximum increase in plant growth. Gibberellin
Bottini et al. (1989) identified and quantified like gibberellin substances- GAi, GA2 and IS0-GA3 in the culture media of Azospirillum lipoferum. Gibberellins normally produced in the range of 20-40 / ml of culture media by Azospirillum lipoferum was reported to be fairly sufficient to enhance root growth of field crops under in vitro conditions. Piccoli and Bottini (1994) reported the close correlation between gibberellin (GA3) production, viability and growth stage in pure culture of Azospirillum lipoferum,
Cytikinins
Cacciari et al (1989) identified and quantified cytokinin production by Azospirillum sp. under various culture conditions through high performance liquid chromatography.
Crop response to Azospirillum inoculation
Increase in total plant dry weight, plant height, leaf size, amount of N in shoots and grains, total number of tillers, fertile tillers, ears, number of spikes and grains per spike, grain weight, germination rates and earlier heading and flowering were reported by several workers due to inoculation of Azospirillum sp. in graminaceous plants (Bouton et al, 1979, Cohen et al 1980; Baldani and Dobereiner, 1980; Schank et al, 1981).

Azospirillum sp. inoculation in wheat was reported to bring about favorable changes in crop growth including increase in nitrogen nutrition, total nitrogen content and grain yield (Dreesen and Vlarsak, 1984). About 3 to 54 per cent yield increase in rice, wheat, maize and barley on inoculation of Azospirillum combined with reduced rates of fertilizer nitrogen was reported by Favilli et al (1987).
Pages and Arsac (1991) identified two efficient strains oi Azospirillum lipoferum through germination paper test and reported that inoculation of Azospirillum lipoferum enhanced the plant growth and yield of sunflower crop. Increase in shoot length, root length, biomass production and grain yield due to Azospirillum inoculation in rice was reported by Sivakumar (1992).
In the absence as well as in the presence of nitrogenous fertilizers, growth stimulatory effect of Azospirillum inoculation in rice was observed by Okon and Labandera-Gonzalez (1994). Azospirillum lipoferum inoculation under rice-wheat cropping system was reported to bring about significant increase in the grain yield and in soil nitrogen content upto 35 to 36 per cent (Ghosh and Puste, 1997). The ability of Azospirillum sp. to alleviate the effects of water deficit in cereal seedlings under salt and osmotic stresses was reported by Creus et at. (1997).
Singh et al (1997) reported that Azospirillum inoculation in rice increased the total biomass and grain yield up to 14.3 per cent over uninoculated control. Azospirillum inoculation in rice, irrespective of the method of application was reported to increase seedling height, shoot and root dry matter production, root volume and yield (Natarajan and Kuppusamy, 1998). Significant increase in chlorophyll content, K, Ca, soluble sugars and protein content of Zea mays when inoculated with Azospirillum lipoferum was reported by Hamdia and Elkomy (1998).

Gunarto et aL (1999) reported that Azospirillum inoculation in lowland rice enhanced the early growth and shoot weight by 12-25 per cent over the uninoculated control. Significant increase in the growth, N, P and micronutrient content of tea seedlings due to inoculation of Azospirillum amazonense was reported by Gnanachitra (2000). Higher green forage yield, dry root biomass and nitrogen uptake were noticed with the inoculation of Azospirillum lipoferum in Cenchnus ciliaris. Application of Azospirillum increased the green fodder yield by 28.9 per cent and dry matter yield by 28.0 per cent (Singh et al, 2000). Inoculation of Azospirillum to paddy seedlings significantly enhanced the shoot length, fi'esh and dry weight of the plants (Kavitha 2000).
Plant response to inoculation with Azospirillum sp., correlate with basic plant physiological response, plant respiration, mineral and water uptake changes. As an example, the plant growth promotion effect of Azospirillum occurs only in nitrogen limited conditions where as insignificant effects were documented under nitrogen abundant conditions (Dobbelaere, 2001).
Inoculation of Azospirillum in wheat crop substantially increased the total nitrogen, non-hydrolysable and hydrolysable organic nitrogen fi-actions of rhizosphere soils. Total and mineral nitrogen content increased at maximum tillering to flowering stages of the crop, followed by a decline at maturity, where as, hydrolysable organic nitrogen decreased with a concomitant increase in non-hydrolysable fraction with the age of the crop (Das and Saha, 2003).
Inoculation of foxtail millet with three strains of Azospirillum lipoferum either alone or in combination with nitrogen fertilizer (40 kg N / ha) increased the plant height, dry weight of shoot and root and total N content of shoot, root and grain. The grain yield of the inoculated plants was significantly higher compared to the control plants. Both the

panicle and the seed weight increased due to inoculation (Bhaskara Rao and Charyulu, 2005).
Organic manures:
Compost comparable to natural humus preserves the soil at minimum cost. Scientific tests showed that compost can be effective manure even though the nutritive value of the compost was less in comparison with mineral fertilizers. The organic components, presence of macro and micronutrient add to the value (Parr et al., 1994). It has been shown that the use of compost can improve the physical, chemical and microbiological characteristics of cultivates soil, reduces the water requirement for plant growth by increasing the availability of moisture and increase the crop production (Gaur and Singh, 1995).
The addition of organic manures improves soil texture, structure and tilth, sandy soils become more compact and clayey soil become more arable ( Gaur, 1982). Various organic acids are released during the decay of organics that act as strong binding agents in the formation of large and stable soil aggregates ( Larson and Clapp, 1984). Composted coir pith being the source of organic matter has extremely high water holding capacity ( more than five times its dry weight) contributing towards increased soil moisture ( Biswas and Khosla,1971). The bulk density of both the surface (10-15 cm) and sub surface (15-30cm) soil reduced to considerable extent in organic manure applied soils. (Appavu and Saravanan, 1999).
As compost is prepared from plant residues and by products, it contains all elements of which the plant is made up of Its addition therefore increases the total stock of these elements in the soil. ( Gaur, 1982). In light textured soils, coir pith applied plots recorded highest Cation Exchange Capacity ( CEC) and organic carbon content of the soil ( Jaganathan et al 1993). The availability of zinc and iron was increased by the incorporation of organic manures ( Appavu et al., 2000).

Compost carries with it a very large population of actinomycetes, fungi and bacteria and by their incorporation into soil not only are millions of microorganisms added, but those already present in the soils are stimulated by the fresh supply of humic materials. Compost also stimulates the mycorrhizae, which live in symbiotic association with the roots of plants and trees and play an important role in transferring certain nutrients from soil to plant. ( Gaur,1982). Soil microbes are sole agents responsible for all of the biological transformations in the soil. These are carried out through a variety of biochemical reactions carried out or catalysed by group of enzymes( De and De, 1988).
All crops responded to organic manuring and the extent of response depends on several factors such as degree of humification, maturity of the compost, its C/N ratio, the time and method of its application and on soil type, agro climatic conditions and moisture regime of soil during the growth of the crop (Gaur, 1982).

Claims:
Biocapsule development
Empty water dissolvable capsule available in the market, which is being used for filling ayurvedic medicine is used for preparing Biocapsule. These capsules are different from capsule which is being used in allopathic medicine, which gets dissolved only in mild acid. These capsules are available in different capacities of 0.5g, 0.75g and 1.0 g capacity. All the three capacities were taken for preparing biocapsule. For smaller seeds 0.5 g capacity was used. For medium size seeds 0.75g and for larger seeds 1.0 g was used. The capsule are having two parts. One base and one cap fixing over the base. The materials were put only in the base of the capsule. The cap fixed over the base will have air space and the organisms filled at the bottom will have sufficient air for survival. Ingredients preparation for filling in the Biocapsule :

Azospirillum:'
The Azospirillum lipoferum pure culture was mass multiplied in Nitrogen free malic acid medixmi. After one week of growth the cultures were mixed with sterilized lignite carrier material to the moisture level of 40%, After mixing the Azospirilum population was enumerated through Most probable number method. The Azospirillum population was 1x10 CFU per gram of carrier based inoculant. It was kept separately in cold storage to maintain the population,
Phosphobacteria:
The phosphobacteria Bacillus megaterium var phosphaticum pure culture was mass multiplied in nutrient broth. After 3 days, maximum growth was obtained and the grown culture was mixed with sterilized lignite carrier material to the moisture level of 40%. The phosphobacteria population in the carrier based inoculum was enumerated through serial dilution plate count method. The phosphobactial population in the lignite based material was Ix 10^^ CFU per gram of material. The cultures were kept in cold storage to maintain the population as well as for future use.
Vermicompost:
Vermicompost was obtained from vermicompost production unit of Department of Environmental Science, and the nutritive value was assessed. The nutrient content of vermicompost was nitrogen 1.75%, phosphorous 1.0% and potash was 1.2%. The vermicompost was sieved thoroughly through 4 mm sieve and removed all debris. The sieved material was properly stored for preparing biocapsule.
Coir dust compost:
Coir dust compost was prepared by composting coir dust. For composting one ton of Coir dust, poultry litter 200 kg, Pleurotus 1 kg, and rock phosphate 20 kg were mixed and composted for 120 days. After composting compost maturity was assessed and the nutritive value was assessed. The nutrient content in the

coir dust compost was 1.8% Nitrogen, 1.4% Phosphorous and 1.5% potash. The coir dust compost was sieved through 4 mm sieve and the sieved material was stored properly for preparing biocapsule.
Poultry litter compost.
Poultry litter compost was prepared by composting fresh poultry litter. Moisture was maintained 60% through out the composting period. Periodical turning was given and after 90 days compost maturity index was assessed to confirm the maturity of the compost and the nutritive value was determined in the laboratory. The nutrient content in the poultry litter compost was 2.2% nitrogen, 0.8% phosphorous and 1.2 % potassium. The composted material was sieved through 4 mm sieve and the sieved material was stored safely for future use.
Preparation of Biocapsule
The organic manures vermicompost, coir dust compost and poultry litter compost were mixed in equal quantities at 30% moisture level. The biofertilizer inoculum Azospirillum and phosphobacteria were taken equal quantities and mixed thoroughly. The mixed inoculum was added to the organic manure blend at 2% level. The final moisture content will be 40%. Now the mixture is ready for filling. Biocapsule with different capacity was taken. The cap was removed from the capsule and the bottom portion was filled to the capacity manually and it was covered with the cap. Now the biocapsule is ready for use.
Advantage of Biocapsule:
Farmers are advised with many practices to follow. But shortage of labor and too many materials to handle for a single crop, indirectly put heaviness in the minds of the farmer and finally farmer skip some of beneficial material for adoption. To avoid this many benefits are packed in a single capsule, so that at one stroke of application farmers will deliver important beneficial material in to their field.

The second advantage in this Bio-Capsule is precise placing of beneficial organisms like Azospirillum and Phosphobacteria in the vicinity of the root, so that these organism colonize on the roots and multiply there itself in large numbers over the period of time. These organism will not be moved away from the root, because they take root exudates as source of nutrients and multiply very well. There is no necessity for search of nutrients other than the nutrients available in the root zone and the root exudates oozing out from the roots. Once they multiply in the root zone, the benefits given by the microbes are absorbed by the root system as plant nutrient. Azospirillum fixes atmospheric nitrogen and Phosphobacteria solubulizes unavailable form of phosphorous into available form. The quantum benefits delivered by the microbes increased multifold when these microbes increased their numbers in the root zone.
The third advantage is organic manures contained in the capsule. The organic manures contain all macro and micro nutrients that a crop require. The quantity available in the capsule is sufficient for the young seedling emerges from the seed. At the developmental stage, all the required nutrients by the seedlings are given by the organic manure. Some times the soil may deficit with selective micro nutrients which are very essential for good development of the plant. Apart from these the organic manure especially vermicompost contain plant growth promoting substances like Indole acetic acid and Gibbrelic acid. These plant growth promoting substances induce the plant growth by profusely increase the root growth and developing more leaves. The profuse development of root penetrates deep into the soil and absorb plant nutrients. More leaves fix more carbon through photo synthesis.
Fourth advantage is improvement of soil characters. The physical character of soil is improved by increasing in soil porosity and water holding capacity. The chemical character of the soil is improved by increased the cation exchange capacity of the soil and improving the nutrient status of soil. In the biological character of soil microbial diversity in increased in the soil because of addition of beneficial organism as well as organic manure addition. Once microbial diversity is increased other life forms

like. Protozoa and earthworms, are developed on its own. This is good trend for a healthy soil.
Utility of Biocapsule:
Biocapsule can be used for all crops which are raised by sowing seeds by dibbling in the soil. Seed can be sown along with a Biocapsule. The irrigation
water dissolve the outer coating of the capsule. The contents in the capsule available near
f
the seed. When the root emerges out of the seed it contact with the microbes. The microbes colonize over the roots.
When stem cuttings are used for establishing new plant the biocapsule can be put under the cuttings. In direct seeding in main field these capsules can be sown through seed drill machine,
In forest nursery where thousands of tree seedlings are rearing in poly pack, the Bio-Capsules are sown along with tree soeds in poly bags. In the three month period microorganism colonize in the root and healthy seedlings will develop. When these seedlings are taken from poly bag and planted in forest soil the microorganism colonize in the root region also transfer to the main field and multiplies there.
In Horticulture precision farming is followed to raise vegetables in large areas. The vegetable seedlings are raised in trays filled with coir dust and Farm yard manure. Bio-Capsules are suited for this purpose.