Emergence of Plant Growth Promoting Rhizobacteria for an Environmentally Sustainable Agriculture

Neha Upadhyay, Radha Rani, Shivesh Sharma* Department of Biotechnology, MNNIT Allahabad, Allahabad -211004 U.P.
Abstract
Plant growth promoting rhizobacteria (PGPR) are valuable bacteria commercialized as biofertilizer and biopesticides. Any technology that deals with natural environment is always turning to an innovative way and PGPR isolates are categorized as an eco-friendly microbial tool. PGPR inoculants are emerged as a novel technique that reduces the excessive use of chemical fertilizers and pesticide. The industrial requirement for PGPR is progressively increased, as it propounds attractive way to maintain a sustainable agriculture system. The successful commercialization of PGPR strains depend on the association between the scientific organizations and industries. A number of plant growth promoting rhizobacterial isolates has been reported that enhance plant growth either directly or indirectly by inhibiting the growth of plant pathogen. Production of phytohormones (IAA), solublizing phosphate and nitrogen fixation are some of the mechanism that directly involve in plant growth and promotion. Indirect mechanism for plant growth promotion involves production of antifungal substances, HCN production, production of siderophores and cell wall degrading enzymes. The miscellaneous and favourable interactions between plant and PGPR inoculants lead towards a hopeful solutions for environmentally sustainable agriculture. Recent progress renders the use of PGPR as biopesticides as well as biofertilizers. Its long term benefits for successful agriculture are presented here.

Introduction
Fertilizers are essential components of modern agriculture because they provide essential plant nutrients. However, overuse of fertilizers and pesticide can cause unanticipated environmental impacts such as accumulation of toxic chemical substances in the soil, depletion of organic carbon content, micro flora and fauna, soil fertility having deleterious effects on crop productivity. One potential way to decrease negative environmental impacts resulting from continued use of chemical fertilizers is inoculation with plant growth-promoting rhizobacteria (PGPR). These bacteria influencing root growth and morphology or by aiding other beneficial symbiotic relationships exert beneficial effects on plant growth and development and many different genera have been commercialized for use in agriculture. During the past couple of decades, the use of plant growth promoting rhizobacteria (PGPR) for sustainable agriculture has increased tremendously in various parts of the world. Significant increases in growth and yield of agronomically important crops in response to inoculation with PGPR have been repeatedly reported (1, 2, 5).

Siderophore Production
Iron is an essential nutrient of plants, but it is relatively insoluble in soil solutions. Siderophores are low molecular weight, extracellular compounds with a high affinity for ferric iron, that are secreted by microorganisms to take up iron from the environment and their mode of action in suppression of disease were thought to be solely based on competition for iron with the pathogen (Bakker et al., 1993; Duijff et al., 1997). Plant roots prefer to absorb iron as the more reduced ferrous (Fe2+) ion, but the ferric (Fe3+) ion is more common in well aerated soil although it is easily precipitated in iron-oxide forms (Salisbury and Ross, 1992). Plants commonly excrete a soluble organic compound (chelators and phytosiderophores) which binds Fe3+ and helps to maintain it in solution. Chelators deliver the Fe3+ to the root surface where it is reduced to Fe2+ and immediately. Some rhizospheric bacteria also produce siderophores and there is evidence that a number of plant species can absorb bacterial Fe3+-siderophore complexes (Wang et al., 1993). Majority of research on microbial siderophores in the rhizosphere is associated with their biocontrol activities due to their competitive effects with plant pathogens.

Nitrogen Fixation
Nitrogen is one Nitrogen is one of the most common nutrients required for optimal plant growth and productivity. Approx 60 % of the earths available nitrogen is fixed by the process of biological nitrogen fixation and represents an economically beneficial and environmentally sound alternative to chemical fertilizers (Ladha et al., 1997. The process of N2 fixation is carried out by nitrogenase enzyme coded by nif genes (Kim and Rees, 1994).

Fig.4 Function of siderophores

Production of Phytohormones
Plant Growth Promoting Rhizobacteria (PGPR) PGPR (Biofertilizers & Biofungicides) are Plant Growth-Promoting Rhizobacteria defined as rootcolonizing bacteria that exert beneficial traits on plant growth and development. Root colonization comprises the ability of PGPR to establish on or in the root or rhizosphere to multiply, survive and colonize along the growing root in the presence of the indigenous microflora. Genera of PGPR generally include Acinobacter, Agrobacter, Arthrobacter, Azospirillum, Bacillus, Bradyrhizobium, Frankia, Pseudomonas, Rhizobium, Serratia, Thiobacillus, and others. In addition to plant growth promotion, PGPR are also used for controlling several plant pathogens, enhancement of nutrient up-take and in rhizomediation. .
PGPR facilitates plant growth by the production of plant growth regulators or phytohormones like IAA, gibberllic acid and cytokinins (5). Tryptophan has been identified as main precursor molecule for biosynthesis of IAA in bacteria. IAA controls a diverse array of functions in plant growth and development and acts as a key component in shaping plant root architecture such as root vascular tissue differentiation, regulation of lateral root initiation, polar root hair positioning, and root gravitropism (4).

Development of PGPR inoculant

Selection and characterization of PGPR strains

PGPR strains have diverse applications in agriculture, horticulture and forestry. Specific PGPR strains are initially selected from several hundreds of root-colonizing bacteria isolated from excised roots of field grown plants. During the experiment, those PGPRs that consistently caused statistically significant increases in root or shoot development or both are selected for further testing in agricultural field. Recently, selections of efficacious PGPR strains have been made by mass screening technique (4). Primary screenings of new isolates are done based on physiological, nutritional and biochemical characteristics as in Bergeys Manual of Determinative Bacteriology. While DNA and RNA homology tests are also considered as most reliable tools for the characterization of potent PGPR strains (4).

Table: Examples of plant growth promoting rhizobacteria tested for various crop types

Fig.1 Schematic representation of PGPR

Mechanism of action
PGPR has a significant impact on plant growth and development in both indirect and direct ways. the direct promotion of plant growth by PGPR generally entails providing the plant with compound that is synthesized by the bacterium or facilitating the uptake of nutrients from the environment On the other hand, indirect promotion of plant growth occurs when bacteria prevent some of the deleterious effects of a phytopathogenic organism by one or more mechanisms. (Glick, 1995; Glick et al.1999).
Fig.2 mechanism of Plant growth promotion by PGPR

Challenges in Field Application of PGPR

The application of PGPR for control of fungal pathogens in greenhouse systems shows considerable promise (30), due in part to the consistent environmental conditions and high incidence of fungal disease in greenhouses. Achieving consistent performance in the field where there is heterogeneity of abiotic and biotic factors and competition with indigenous organisms is more difficult. Knowledge of these factors can aid in determination of optimal concentration, timing and placement of inoculant, and of soil and crop management strategies to enhance survival and proliferation of the inoculant. The concept of engineering or managing the rhizosphere to enhance PGPR function by manipulation of the host plant, substrates for PGPR, or through agronomic practices, is gaining increasing attention (8 ). Development of better formulations to ensure survival and activity in the field and compatibility with chemical and biological seed treatments is another area of focus; approaches include optimization of growth conditions prior to formulation and development of improved carriers and application technology (8,3).

Conclusion
As our understanding of the complex environment of the rhizosphere, of the mechanisms of action of PGPR, and of the practical aspects of inoculant formulation and delivery increases, we can expect to see new PGPR products becoming available. The success of these products will depend on our ability to manage the rhizosphere to enhance survival and competitiveness of these beneficial microorganisms (4). Rhizosphere management will require consideration of soil and crop cultural practices as well as inoculant formulation and delivery (3,8). The use of multi-strain inocula of PGPR with known functions is of interest as these formulations may increase consistency in the field (8). They offer the potential to address multiple modes of action, multiple pathogens, and temporal or spatial variability . PGPR offer an environmentally sustainable approach to increase crop production and health. The application of molecular tools is enhancing our ability to understand and manage the rhizosphere and will lead to new products with improved effectiveness.

Phosphate solublization
PGPR stimulate plant growth directly through increase in nutrition acquisition, such as phosphate solubilization, or by rendering the inaccessible nutrients available to the plants. Phosphorous is taken up from soil solution as phosphate (Pi, H2PO4_). Although soils generally contain a large amount of total P but only a small proportion is available for uptake by the plants. Several phosphate solubilizing microorganisms (PSMs) are now recorded to convert the insoluble form of phosphorus to soluble form through acidification, secretion of organic acids or protons (7) and chelation and exchange reactions. Phosphate-solubilizing microorganisms (PSM) include a wide range of symbiotic and non symbiotic organisms, such as Pseudomonas, Bacillus, and Rhizobium species; actinomycetes; and various fungi-like Aspergillus and Penicillium species (7).

References
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Fig.3 Mechanism of P solublization