azo

1. INTRODUCTION
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. Martinus Willem Beijernick
• Dutch microbiologist & botanist.
• Founder of Environmental microbiology and virology.
• German chemist
• Coined the term " Associative symbiosis".
Johann Wolfgang Döbereiner

3. 1.Cell size : 1mm in
diameter - size and
shape Curved rods or
may be vary.
2.Accumulate :
PHB 3.Gram
reaction:
Negative
4.Development
of white pellicles
: 2-4 mm below
the surface of
NFB medium
MORPHOLOGY
PHYSIOLOGY

4. CLASSIFICATION
Family – Spirillaceae
1. A. brasilense
2. A. lipoferum
3. A. amazonense
4. A. halopraeferens
5. A. irkense
6. A. dobereinerae
7. A. largimobilis
" 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 " .

5. MICROSCOPHIC OBSERVATION

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

7. See
d
EFFECTS OF AZOSPIRILLUM

8. PRODUCTION
① Isolation of Azospirillum
② Confirmation ofAzospirillum
③ Making the starter culture
④ Mass culture
⑤ Carrier based inoculant

9. ISOLATION OF AZOSPIRILLUM

11. Pellicle formation in semisolid media
Pure colonies in solid NFB media
Antibiotic resistance against common
antibiotics
RESULT
Figure 1
Figure 2
Figure 3

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

13. 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.

14. 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

15. MODE OF ACTION

16. 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..

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

18. SEED TREATMENT
• 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.

19. INTRODUCTION
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, 500gmAzospirillum 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.

20. INTRODUCTION
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.

21. CROP APPLICATION BIOFERTILIZER