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& Department of Microbiology Institute of Bioinformatics and Biotechnology
Agricultural Biotechnology ))Biofertilizers
Biological, Chemical and Physical features to long term, sustainable agricultural productivity with minimal environmental impact .)Arias, 2005(
Are: soil organic matter )including microbial Biomass(, Soil texture, soil structure, soil depth, content of nutrients, storage capacity adsorption capacity(, soil reactions( & absence of toxic elements
2. Soils may be naturally low in nutrients 3. Deficient due to nutrient removal by crops 4. When high yielding varieties are grown (In order to obtain high yields, Fertilizers are needed).
Chemical Fertilizers )Conventional Farming(
Biological Fertilizers )Organic Farming(
The threat of chemical Fertilizers:
1. Threaten Human Health. 2. Threaten Agricultural soils, Food safety and Waterways. • Soil quality • Plant uptake • Water quality
The threat of chemical pesticides:
*Health & Environmental Problems. WHO-3million acute sever cases of poisoning
Although, during last 50 years, farmers have dramatically increased crop yields through the use of chemical fertilizers & pesticides, and improved varieties, today, the rising costs of chemical inputs and a host environmental concerns have caused farmers to consider alternative agri-industrial managements (e.g.Organic Farming) to reduce costs, protect human health, and conserve the resource base. (Kritcher, 1993)
Organic Farming by Bioorganic fertilizers:
• If continuous exploitation of land for cultivation has caused a progressive decline in soil health, it can be restored and maintained to a greater extend by the use of organic manures. • Further improvement in this regard was observed by incorporating microorganisms in organic manures to develop Bioorganic Fertilizers (Chakradhar, 2004).
• So, Utilization of microbial inoculants specially PGPR for sustainable agri-industrial applications has been subjected of a number of recent reviews to manipulate rhizosphere conditions by innovative techniques for a better plant growth and plant health (Bloemberg, 2001).
Any of large number of natural & synthetic material like manure, N, P, K compounds, spread on or worked into soil to increase its fertility.
6. Living fertilizers compounds of Microbial inoculants: eg,Plant growth promoting substances like Hormones and Auxins. 7. Group of microorganisms which are able to fix atmospheric nitrogen or solubilize Phosphorus, decompose organic material or oxidize sulfur in soil.
• Importance of Biofertilizers:
• Eco friendly. • In addition to N2 ,Provide certain PGP substances like hormones , vitamins,… . • Supplying N2 , continuously throughout the entire period of crop growth in the field under favorable conditions. • Without toxic effects. • When applied to soil improve the soil structure. • Low production cost.
Plant Growth Rhizobacteria Endophytic .1 Bacteria Exophytic .2 Bacteria
Bacterial Endophytes: Why Are They There?
• Opportunists? –Some have no apparent effect on plant performance • Mutualists? –Evidence is accumulating to support this possibility
• Benefit to microorganism:
Bacterial Endophytes: Another Mutualistic Symbiosis
– Provides an environment buffered from external stresses – Steady source of nutrients and water
• Benefit to the plant host:
– Nitrogen fixation – Biological control of plant pathogens and pests – Enhanced uptake of nutrients and water
Bacterial Genera With Endophytes:
Acinetobacter Actinomyces Alcaligenes Arthrobacter
Agrobacterium Azoarcus Bacillus
Azorhizobium Azospirillum Bordetella Comamonas
Chryseobacter Clavibacter ium
Deleya Erwinia Kingella Leuconostoc Moraxella
Escherichia Herbaspirillum Lactobacillus Micrococcus Pasteurella Providencia Serratia
Pseudomonas Psychrobacter Rahnella Rhizobium Shewanella Vibrio Rhodococcus
PGPR affect plant growth
* Production of Plant hormones*Antibiosis * Phosphorous solubilization *Induced resistance * Enhanced iron availability *Iron scavenging * Nitrogen Fixation *Competition for nutrients/niche * Etcetera * Parasitism & Predation *Etcetera
• Arthrobacter • Acetobacter • Azotobacter • Azosperillum • Bacillus • Enterobacteria • Klebsiella • Proteus
Well known PGPRs:
• Pseudomonas • Rhizobium
Pseudomonas spp. & related genera:
• Although a range of different bacterial genera
and species have been studied, the overwhelming number of papers have involved the use of Pseudomonas species. It’s so because Pseudomonas and related genera are characteristically:
• Fast growing • Easy to culture • Manipulate genetically in the laboratory • Able to utilize a range of organic
• Sin ce o th er rhizobacte ria a re a lso found i n t he rh iz osphere o f many c rop p la nts L ik e w heat and th ere were l it tle d etail ed stu dies o n t hem fr om p lant r hizosphere: • It becomes in teresting t o f in d o ut th e probable role of oth er s i n r hizosphere of wheat & o ther c rop p la nts.
Other rh izobacteria
Di r ect Pla nt Gr owt h Pr omotio n
1.Microbial Production of Plant Hormones:
• Plants themselves synthesize Auxin, Gibberellins, Cytokinins, Ethylene, and Abscisic acid, but under less than ideal climatic and environmental conditions, Plants may not synthesize sufficient endogenous concentrations to sustain optimal growth and development.
Scientists have shown recently That Plant Growth can be improved when specific microbial strains are used to • ‘ inoculate’ seeds or roots of agricultural crops due to microbe’s production of Plant Growth Hormones (Regulators). • Exogenous Supplementation of PGPHs to plant roots is reletively new approach to maximize crop yield.
Signals from under ground: Bacterial volatiles promote plant growth
Examples of IAA production: • Some strains of Acinetobacter isolated and characterized
Some strains of Acinetobacter isolated and characterized from rhizosphere of wheat were showed indole-3-acetic acid (IAA) production. Pot experiments showed significant increase in plant growth inoculated with eight Acinetobacter genospecies as compared to control plants. IAA production was found to be encoded by plasmid PUP1126 and this is the first report of plasmid-encoded IAA production in the genus Acinetobacter.
• The rhizobacterium Pseudomonas putida GR 12-2 is a
strong candidate for development as a soil inoculant to enhance crop yields. Inoculation of canola, tomato and other agriculturally important plants with this strain results in substantial promotion of seedling root growth. Characteristics that may contribute to the ability of P .Putida to enhance plant growth include the capacity to synthesize siderophores and thereby provide iron for the plant, the capacity to lower growth inhibiting levels of
1.2.Gibberellic acids: Azospirillum spp. are considered to be
important plant growth promoting rhizobacteria that can improve the growth and yield of at least several plant species (Labandera-Gonza´lez, 1994). Phytohormone production, including gibberellins (Bottini et al., 1989), is one mechanism that has been proposed ( Cassan et al., 2001). Other Gibberellin producing bacteria in rhizosphere are as follows:
The effects of C2H4 have been observed in practically all aspects of plant growth and development, including seed germination (Ketring et al., 1972), seedling growth (Burg et al., 1968), root growth (Chadwick et al., 1970), growth of leaves (Primrose, 1979), and ripening, aging (Biale, 1960). Agronomically, microbial production of C2H4 could have an impact on crop production under certain management conditions. Ethylene concentrations as low as 10 ,ug liter-' can evoke plant responses, and concentrations of 25 pug liter-' result in decreased fruit and flower development (Primrose et al., 1979).
the presence of micro-organisms capable of producing cytokinins, can be expected to raise the amounts of cytokinins in both the soil solution and in plants growing there. In turn, this may have an impact on the growth of these plants. In support of this there are numerous reports that certain micro-organisms affect plant growth through their ability to produce phytohormones )Arshad and frankenberger, 1991, 1998; Steenhoudt and Vanderleyden, 2000(.
The ability of microorganisms to solubilize and mineralize p in soils is vital. Phosphate availability in soil is greatly enhanced through microbial production of metabolites leading to lowering of PH and release of phosphate from organic and inorganic complexes.The species of Pseudomonas,Micrococcus, Bacillus, Aerobacter, Xanthomonas, brevibacterium, Alcaligenes, Rhizobium have been reported to be active in phosphate solubilization (Srivastav, 2004). Although these PGPRs occur in soil, usually their numbers are not high enough to compete with other bacteria commonly established in the rhizosphere. So, for agronomic utility, inoculation of plants by target with such microorganisms at higher concentration than those normally found in
for example in this subject, the solubilization of phosphatic compounds, one of the important mechanisms of plant growth promotion shown by PGPR Acinetobacter, increases its potential in the development of future bioinoculum for crop plants. In this investigation the phosphate solubilization by Acinetobacter spp. was also compared with other rhizosphere isolates like Moraxella sp., Pesudomonas sp., Serratia sp., and Pseudomonas putida NCIM1313, Escherichia coli NCIM2810. All the phosphate solubilizing Acinetobacter strains had zone diameter of dissolution in the range 1-5cm while as control P putida had . average zone diameter in the range 1-3.5cm. Solubilization of insoluble phosphates started along with the growth of strains and maximum solubilization was achieved at logarithmic to late stationary phase. Some cultures showed reprecipitation of solubilized phosphate after
3. N 2 Fix ation:
Asymbiotic Azotobacter Azosperillum Bacillus Klebsiella Clostridium P.vulgaris
Symbiotic Rhizobium Bradyrhizobium Cyanobacteria Anabaena
In Di r ect Pla nt Gr owt h Pr omotion
• According to the United States Department of Agriculture, biological control of plant disease is defined as " the involvement of the use of beneficial microorganisms, such as specialized fungi and bacteria, to attack and control plant pathogens and the diseases they cause. • These "specialized" fungi and bacteria are microorganisms that normally inhabit most soils.
Direct Competition with the Target Organism.
In this case the biocontrol agent out competes the target organisms for nutrients and space. Example:
• Iron competition in Pseudomonads has been intensively studied and the role of the pyoverdine siderophore has been intensively studied and the role of the pyoverdine siderophore produced by many pseudomonas species has been clearly demonstrated in control of Pythium and fusarium species.
The biocontrol agent produces an chemical compound such as an antibiotic or some type of toxin that kills or has some sort of detrimental effect on the target organism. Example: phenyazine-1-carboxylic acid (PCA) from Pseudomonas aureofaciens kuyver tx-1 has even been used as a direct field treatment of the control of dollar spot on creeping bent grass (Powell et al., 2000).
3. Induced Resistance of the Host Plant.
It has been know for decades that once a plant is infected with a pathogen, that infection triggers some sort of reaction in the infected host plant that helps keep it from being infected with other pathogens. The infected plant becomes more "resistant" to other infections.
Changing that have been observed in plant roots exhibiting Induced systemic resistance (ISR) include: 1. strengthening of epidermal and cortical cell walls and deposition of newly formed barriers beyond infection sites including callose, lignin and phenolics. 2. increased levels of enzymes such as chitinase, peroxidase, polyphenol oxidase, and phenylalanine ammonia lyase. 3. enhanced phytoalexin production. 4. enhanced expression of stress- related genes. However, not all of these biochemical changes found in all bacterial-plant combinations.
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