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1. INTRO D UCTION INTRODUCTION Azospirillum species are Free living in soil & plant-associated diazotrophs Association with the roots of cereal crops, grasses and tuber plants. subclass of Alphaproteobacteria. " Azo " comes from French word , Means " Nitrogen " Spirillum - shape of bacteria. Azospirillum was First isolated by Beijerinck (1922) in Brazil from the roots of Paspalum and named it as Azotobacter paspali and later named as Spirillum lipoferum. Dobereiner and Day (1976) reported the nitrogen fixing potential of some age grasses due to the activity of S. lipoferum in their roots. Taxonomy was re-examined and Tarrand et al. (1978) designated this organism as Azospirillum. An aerobic or micro aerophilic , Gram negative rod. Non sporeformer and inhibiting the plant roots both externally and internally. Being a micro aerophilic organism, it can be isolated on a semi solid malate medium by enrichment procedures. It is used as BIOFERTILIZER. Azospirillum fixes nitrogen from 10 to 40 kg/ha.

2. CLASSIFICATION F a mil y – S p i r ill acea e A . b r a s ile n s e A . li po f eru m A . a m a z onen s e A . ha l op r ae f e r en s A . i r k en s e A . dobe r e i n e r a e A . la r gimobili s " Tilak and subbarao 1987 have reported highest increase grain yield (33.67%) and plant biomass yield as( 32.71%) by application of inoculants azospirillum brasilance and glomus fasiculatum " .

3. MICROSCOPHIC OBSERVATION

4. ORGIN AND CHARACTERISTIC ASOPIRILLUM SPP ORIGIN AND CHARACTERISTIC OF AZOSPIRILLUM

5. Se e d EFFECTS OF AZOSPIRILLUM

6. PRODUCTION ① Isolation of Azo s pirillum ② Confirmation of Azo s pirillum ③ Making the starter culture ④ Mass culture ⑤ Carrier based inoculant

7. ISOLATION OF AZOSPIRILLUM

8. Pellicle formation in semisolid media Pure colonies in solid NFB media Antibiotic resistance against common antibiotics R E S U L T Figure 1 Figure 2 Figure 3

9. Mother culture Mixed with sterlized carrier materials Growth of Azospirillum in broth Packaging and sealing MASS PRODUCTION

10. Why we use NFB Media? ① It is used to isolate diazotrophs. ② Nitrogen fixation by diazotroph bacteria is a H+ consumer process and also ammonia is subsequently produced, the color of the NFB medium should be changed from green to blue (due to the presence of Bromothymol biue pH indicator). ③ The production of yellow color by bacteria in this medium Indicates a decrease in pH, It indicate ability of nitrogen fixation.

11. ① Azospirillum Medium with 0.17% Agar is used for cultivation of Azospirillum species. ② Malic acid is used as the carbon source. ③ Azospirillum species grow well in presence of Malic acid and are not overgrown by other nitrogen fixers. ④ Dipotassium phosphate provides buffering effect and other inorganic salt ingredients provide necessary growth nutrients. ⑤ Agar at 0.17% concentrations provides microaerophillic conditions necessary for nitrogen fixation by Azospirillum species. AZOSPIRILLUM MEDIUM

12. The Benefits ① Promotes plant growth. ② Increased mineral and water uptake, root development, vegetative growth and crop yield. ③ Inoculation reduced the use of chemical fertilizers (20- 50%, 20-40 kg N/ha). ④ Increases cost benefit ratio. ⑤ Reduces pathogen damage. ⑥ Inhibit germination of parasitic weeds. ⑦ Restoration of arid zone, margine mangrove ecosystem. ⑧ Reduces humic acid toxicity in compost. ⑨ - Recommended for rice, millets, maize, wheat, sorghum, sugarcane and co-inoculant for legumes..

13. APPLICATIONS ① Seed treatment ② Soil treatment ③ Root dip treatment

14. SEED TR E A TMENT In this method 400gm of Azospirillum biofertilizer is suspended in 500mL of water to make slurry and mixed with 10 -12kg of seed until seeds are uniformly coated. The treated seeds are dried in shade and sown immediately.

15. INTRO D UCTION ROOT DIP TREATMENT In transplanted rice, Azospirillum is generally used as root dip treatment. In this method a small size bed is prepared in a corner of the paddy field. To transplant one bigha of land, 500gm Azospirillum biofertilizer is mixed with 10-15kg compost and just sufficient quantity of water in the bed. Uprooted rice seedlings are dipped in the bed for 8-12 hours before transplanting.

16. INTRO D UCTION SOIL TREATMENT For soil treatment depending upon the crops the required quantity of biofertilizers is mixed with compost and broadcast evenly in the field. In case of integrated nutrient management (INM), to supplement phosphatic and potassic fertilizers, phosphate solubilizing bacteria and potash solubilizing bacteria can be used along with Azospirillum.

17. CROP APPLICATION BIOFERTILIZER