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Researcheres: Abolfazl Moghadam,Shirin Amirtiemoori Instructor: Dr.Rokhbakhsh

 Rhizosphere : the rhizospher is the volume of soil

surrounding and under the influence of plant roots, and the rhizoplane is the plant root surfaces and strongly adhering soil particles ( Kennedy,2005) Rhizobacteria : Rhizobacteria are rhizospher competent bacteria that aggressively colonize plant roots ; they are able to multiply and colonize all the ecological niches found on the root at all stages of plant growth , in the presense of a competing microflora ( Antoun and Kloepper , 2001)
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Neutral bacteria

Pathogenic bacteria
Beneficial bacteria ( PGPR )
Suslow, Kloepper, Schroth, Burr, 1979.

Figure 1 Nodules of bacteria soybean root

 PGPR (Plant growth promoting rhizobacteria) A group of bacteria that can actively colonize plant

roots and improve plant grows and yield by direct and indirect mechanisms (Noel et al.,1996)
(A) Soybean plant infected with Bradyrhizobium japonicim left, uninfected control plant (right).

(B) Legume root with nodules induced by B.japonicum

Figure 2

Figure 3

Figure 4

(MS medium)
Effective Two strain of PGPR icluding B.subtilis GB03 , B.amyloliquefaciens IN937a & the nongrowth-promotin strain E.coli DH5 on growth of A.thaliana.
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DH5

Refrence ; chhong-min Ryu,mohamed A.Farag,,2003.,Bacteria volatiles promote growth in Arabidopsis

These mechanisms Plays a vital role in the management of plant diseases to increase crop productivity via various mechanisms.
N2 - fixation

Phosphate solublization

Direct
Siderophore

Phytohormones

Siderophore

Indirect
Antibiotic & Antifungal

Figure 4 N2 Fixation cycle

 Using growth medium with Tricalcium phosphate (NBRIP). Bacterial mechanisms: phosphatase enzymes or production of acids

Figure 5

Figure 6

Solubilization of P

No solubilization
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Figure 7 Siderophore
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Figure 8 type of phytohormone produced by PGPR

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 Antibiotic encompass a chemically heterogeneous

group of organic,low-molecular weight compounds produced by microorganisms at low concentrations that are deleterious to the growth of metabolic activities of other microorganisms (Fravel,1988;Thomashow et al.,1997). Antibiotics produced by different PGPR have a broad-spectrum activity.

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PGPR Acinetobacter Agrobacterium Arthrobacter Azospirillum Bacillus Bradyrhizobium

Detailes
Gram-negative bacilli Gram-negative bacilli
Gram-positive , rods during exponential growth and cocci in their stationary phase.

Gram-negative bacilli Gram-Positive , rod-shaped Gram-negative bacilli

Frankia
Pseudomonas Rhizobium Serratia Thiobacillus

Gram-Positive , filamentous bacteria

Gram-negative bacilli Gram-negative bacilli Gram-negative , rod-shaped Gram-negative , rod-shaped 13

Figure 9

PGPR

Antibiotic

structure

Fungus
Gaeumannomyces graminis var.tritici

Disease
Strains of P.fluorescens may be involved in the suppression of G.graminis var. tritici. In pythium contaminated sites, and signficant increases were observed in plant height,number of heads and grain yield of winter wheat.

Reference
Raaijmakers & Weller , 2000

P.fluorescens

Phenazines

P.fluorescens

Pyoluteorin

Pythium sp.

Weller and Cook,1986

P.fluorescens

2,4-Diacetyl phloroglucinol

Rhizoctonia solani

Out of 40 strains, 18 strains showed strong antifungal activity.

Kumar et al.,2002

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 One of the most effective mechanisms that PGPR

employ to prevent proliferation of phytopathogens is the systhesis of antibiotic. some well know PGPR strains are pseudomonas,Bacillus, Azospirillum,Rhizobium,and serratia species.the primary mechanism of biocontrol by PGPR involves the production of antibiotics such as phenazine-1-carboxyclic acid, 2,4-diacetyl phloroglucinol, oomycin, pyoluteorin, pyrrolnitrin, kanosamine, zwittermycin-A, and pantocin.
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Antibiotics
DAPG Aerugine Phenazine PCA Pyrrolnitrin Viscosinam ide Pyoluteorin

PGPR
Pseudomonas sp. P,fluorescens Pseudomonas sp. P.fluorescens Burkholderia cepacia P.Fluorescens P.fluorescens

Pathogen
Pythium ultimum Phytophthor C.orbiculare Fusarium oxysporwn
G.g.Var.tritici

Crop
sugar Pepper Cucumber tomato Wheat Potato Sugar beet Cotton Sugarbeet

Reference
Shanahan et al.(1992b)
Lee et al.(2003) Lee et at.(2003)

Chin-A-Woeng et al.(1998) Thomashow and Weller(1988) Burkhead et al.(1994) Nielsen et al.(1998) Howell and Stipanovic(1980)
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Fusarium sambucinum
Rhizoctonia Solani Pythium ultimum

Pythium spp Pythium spp

Gaeumannomyces graminis Var. tritici

 Primary site of Action pyrrolnitrin is terminal electron

transport system.

Figure 11Structure of pyrrolnitrin Figure 10 Mechanism of Antibiotic Pyrrolnitrin

Refrence : R. K. TRIPATHI AND DAVID GOTTLIEB , July 1969, Mechanism of Action of the Antifungal Antibiotic Pyrrolnitrin

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Figure 11 Mechanism of action of Antibiotic Phenazines

Refrence: Latifi, A. M., Winson, K., Foglino, M., Bycroft, B. W., Stewart, G. S. A. B., Lazdunski, A. and Williams, P., Multiple homologues of LuxR and LuxI control expression of virulence determinants and secondary metabolites through quorum sensing in Pseudomonas aeruginosa PA01. Mol. Microbiol., 1995, 17, 333343.

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Biofertilizer : living organism which increase the

nutrient status of the host plant through their ongoing existence in associantion with the plant.( J. Kevin Vessey, 2003)

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 Puts acid in the soil Strengthens pestecides(they become stronger and more resistant to

chemicals that should keep them away) It has very little carbon which is a key element in plants Chemical fertilizer also get a lot of water out of the soil which it contaminated and therfor pollute water It degrades ecosytems It releases a green house gas called nitrous oxide

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 Increase soil fertility

Product increased to 20-30%

Stimulate plant growth Stimulate soil biological activity

Protection against dryness and some pathogenic

factors of soil Coping with Nature Reduce costs chemical fertilizers, especially nitrogen and phosphorus fertilizer. Improve soil structure
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Figure 12 difference effect chemical fertilizer with biofertilizer in plant growth

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 Bacterial biofertilizer (Azospirillum , ) Fungi biofertilizer ( Mycorhiza,) Algae biofertilizer ( blue-green algae,.) Actinomycets (frankia,)

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 Pseudomonas fluorescence bacteria effectively control wilts and root rot diseases of

Groundnut, Cotton, Banana, Soybean, Tomato, Pigeon pea etc., It also controls the rice blast and sheath blight of Paddy. This bacterium enters the plant system and act as a systemic biocontrol agent against diseases.
Suitable for: It is suitable for all kinds of Crops. Mode of action: By secreting an enzyme, it has the capability to destroy the cell wall of the fungal

pathogens and annihilate them.

It secrets Hydrogen cyanide and antibiotics such as Pycocyanin and Phenazine,

which inhibit the growth of disease causing pathogens.

Advantage: It also produces Siderospores which chelate with iron in the soil, and make it

difficult for the pathogens to proliferate.

Further, it secretes several plant growth substances, and these gibberellins like

compounds contribute to vigorous crop growth.

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 Bacillus subtilis is an antagonistic bacterial Biocontrol agent, which controls many soil

and air borne diseases of Paddy, Groundnut, Cotton, Vegetables, Soybean etc., Foliar application of Bacillus subtilis with Pseudomonas fluorescence control leaf diseases of many crops.
Suitable for: Paddy, Millets, Oilseeds, Fruits & Vegetables, Sugarcane, Banana, Coconut,

Oil palm, Cotton, Chilly, Lime, Coffee & Tea, Areca nut & Rubber, Flower, Spices, & Contiments, Herbs, Lawns & Ornaments, trees etc.
Mode of action: The bacterium colonizes the developing leaf & root system of the plant and thus

competes with & thereby suppresses plant diseases.

Advantage: The Plant Growth Promoting Rhizobacteria (PGPR) having an antagonistic

interaction with various soil borne plant pathogens.

It protects plants against seed and root diseases.

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PGPR are indigenouse to soil and plant rhizosphere that improve plant growth and yeild. one of the important applications of PGPR to use in biological combating with pathogenic agents . Some of important PGPR Rhizobacteria is Pseudomonas , Bacillus , Rhizobium , Azospirilium and Important Antibiotic produced by PGPR include Pyrrolnitrin , Phenanzine , 2,4Diacetyl phloroglucinol , oomycin and That PGPR have important role in biocontrol of plant pathogens and are used in biofertilizers.
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