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.
Cell size : 2-3 µm in diameter in length and shape Curved rods .
Gram reaction: Negative

Development of white pellicles 2-4 mm below the surface of NFB medium

PHYSIOLOGY
CLASSIFICATION

Family –
Spirillaceae
1. A. brasilance
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 " .
MICROSCOPHIC OBSERVATION
ORIGIN AND CHARACTERISTIC OF AZOSPIRILLUM

ORGIN AND
CHARACTERISTIC
ASOPIRILLUM SPP
 PRODUCTION

① Isolation of Azospirillum
② Confirmation of Azospirillum
③ Making the starter culture
④ Mass culture
⑤ Carrier based inoculant
ISOLATION OF AZOSPIRILLUM
 RESUL
T

Pellicle formation in semisolid media

Figure 1
Pure colonies in solid NFB media

Figure 2

Antibiotic resistance against common

antibiotics

Figure 3
 MASS PRODUCTION

Mother culture Growth of Azospirillum in broth

Mixed with sterlized carrier materials

Packaging and sealing
 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.
AZOSPIRILLUM MEDIUM

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.
MODE OF ACTION
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..
 APPLICATIONS

①
Seed treatment
②
Soil treatment
③
Root dip treatment
 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.
 ROOT DIP TREATMENT
INTRODUCTION

• 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.
 SOIL TREATMENT
INTRODUCTION
• 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.
CROP APPLICATION BIOFERTILIZER
 REFERENCE

• Text book of ENVIRONMENTAL AND AGRICULTURAL MICROBIOLOGY Author name :

Ragur Tim bhatt, pg.no : 342 -351.

• https://pubmed.ncbi.nlm.nih.gov/10978548/

• https://www.sciencedirect.com/science/article/pii/B9780128214060000187
 Confidence level
Communication
skill

SKILLS
GAINED BY
presentatio
SEMINAR
n skil

Innovative skills

Gained subject
knowledge
 THANKS TO

THE RESPECTED CHAIRMAN ,

THE RESPECTED SECRETARY,
THE RESPECTED PRINCIPAL ,
THE HEAD OF THE DEPARTMENT - DEPARTMENT OF
MICROBIOLOGY ,
THE STAFF MEMBERS - DEPARTMENT OF
MICROBIOLOGY.