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Biocontrol Agents of Crops Diseases

#KR-1881


Scale up of Technology for Production and Application of Rhizobacteria-Based Preparations as Biocontrol Agents of Crop Diseases

Tech Area / Field

  • AGR-DIS/Disease Surveillance/Agriculture
  • AGR-PPR/Plant Protection/Agriculture
  • BIO-IND/Industrial Biotechnology/Biotechnology

Status
3 Approved without Funding

Registration date
01.09.2010

Leading Institute
Innovative Phytotechnology Center, Kyrgyzstan, Bishkek

Collaborators

  • Washington State University, USA, WA, Pullman\nUniversity of Saskatchewan, Canada, SK, Saskatoon

Project summary

Project is focused on the scale-up technology and establishment of pilot production of plant growth promoting biological control agents as alternative strategy for agrochemical substances (pesticides, mineral fertilizers). Development of Pseudomonas rhizobacteria is one of perspective agricultural field for plants rhizobacteria research, which is potential agrobiotechnology objects for development of biological control agents of crops phytopathogens, and also plant growth-promoting biopreparations (Plant Growth-Promoting Rhizobacteria - PGPR).

Annually, bacterial and fungal diseases cause millions of dollars damage to Kyrgyz crops and billions of dollars damage to crops worldwide. In the Kyrgyz republic, for example, there are many economically important plant diseases of commercial crops including seedling blight, root rot, blackleg and stem rot, which cause serious damage to wheat, barley, maize, potato, melon, sugar beet and oilseed crops, specially in the irrigated areas. Although chemical pesticides are used to control these diseases, development of natural humic substances and/or antagonistic rhizosphere microorganisms for biological control may prove more successful. Biocontrol systems using antagonistic bacteria have been considered as an alternative strategy to agrochemicals that are harmful the environment and human health.

In course of three years of the ISTC project KR-993.2 development the project participants have developed very promising technologies for production of biocontrol agent formulations from perspective plant rhizobacteria that can be scaled up to industrial production. These technologies have been described in detail in the final report of project KR-993.2. Project collaborators - Prof. Renato de Freitas and Dr. Lynne Carpenter-Boggs strongly recommended scale-up and thus, demonstrating the newly developed technology in field scale trials. The new technology originally hypothesized in project KR-993.2 has been tested and backed-up by extensive laboratory microbiological data including biological tests on product quality of the extracts obtained under laboratory conditions.

The following highlights have been established during project KR-993.2 implementation and provide a solid background for scaling-up a production line from the lab to a pilot plant:

  • Establishment of a stock culture collection comprising of rhizobacteria isolated from soils and rhizosphere of the weed couch grass (Elytrigia repens L. Nevski) and commercial crops of Kyrgyzstan including tomato and corn. A total of 132 bacteria isolated from various crop’s rhizosphere were assessed for biocontrol activity in vitro against phytopathogenic fungi including Fusarium culmorum, Fusarium heterosporum, Fusarium oxysporum, Drechslera teres, Bipolaris sorokiniana, Piricularia oryzae, Botrytis cinerea, Colletothrichum atramentarium, Cladosporium sp. and Stagonospora nodorum. A culture collection comprising of 64 potential biocontrol agents (BCA) exhibiting activity against wheat and barley root diseases has been established.
  • Superior rhizobacteria strains have been studied extensively on in vitro bioassays, in vivo in laboratory and greenhouse studies and in micro-plot field trials using crops. These studies confirmed not only the antagonistic potential of these rhizobacteria against pathogenic fungi but also their plant growth-promoting activities. As a result, four bacterial strains exhibiting high fungicidal activity were selected from rhizosphere of weed and used as pure cultures on plant growth-stimulating studies. Microbial classification using nucleotide sequence of 16S ribosomes indicated that Pseudomonas and Bacillus species were the most dominant bacteria exhibiting biocontrol activity.
  • Technology to establishing liquid and dry microbiological formulations using beneficial rhizobacteria has been developed in the laboratory. Two dry bio-formulations have been tested including: (i) spray drying and (ii) low-temperature convective-contact drying. Our results demonstrated that low-temperature drying had higher beneficial effect in terms of high yield as compared to spray drying.
  • Active metabolites (antibiotics) have been isolated and purified from mutants of rhizobacteria resistant to antibiotics including ten spontaneous rifampicin- and streptomycin-resistant Bacillus subtilis, strain BSRB-43 and rifampicin-resistant Pseudomonas, strain PsRB-7. Rifampicin-resistant Bacillus subtilis, strain BSRB-43 is a strong antagonist against the plant pathogen Bipolaris sorokiniana.

Advantages of the technology developed by the participants of Project KR-993.2 are:
  • availability of their own collection of high-productivity strains;
  • developed technologies for formulation of biopreparations; their toxicological tests showing biological safety;
  • possibility of formulation of biological products in the liquid, concentrated and dry standardised commodity forms using natural fertilizers – humates as filling agent, which are possessed protection at concentration, drying and long-term storage, and are provided prolonged action in field conditions;
  • integrated approach to the solution of the scientific and technical problem – foundation of a new branch of production of biopreparations with consumer-oriented qualities and high export potential; a ready team of researchers and specialists in various fields (microbiology, biotechnology, chemistry, engineering, and agriculture).

Thus, results of the project will allow to provide the researches at innovative level meaning scaling up a newly developed technology including a pilot-plant production and field-testing settings for the new PGPR-based products – biocompatible, non-toxic and effective. The results listed above clearly qualify our research team to elaborate a new project proposal, which uses the products and technologies that have identified on project KR-993.2. Thus, it is only logical that we demonstrate these results in large field scale trials. The objective of the proposed project is to scale-up technology and to establish a pilot plant for the production of plant growth promoting biological control agents (bio-formulations).

The following objectives have been established in order to reach this goal:

  • Formulate nutrient media for cultivation of biocontrol agent and plant growth stimulating Pseudomonas. Commercial sources of carbon and nitrogen will be assessed and selected for industrial production.
  • Optimize fermentation conditions for cultivation of biocontrol agents and plant growth stimulating rhizobacteria. Rhizobacteria strains obtained during project KR-993.2 will be assessed for growth efficiency using different technological strategies.
  • Establish and validate technology for production of various bio-formulations using medium-size (16 L) fermenters. Evaluation of biocontrol activity of novel bio-formulations using plants in laboratory experiments.
  • Establish and validate technology for production of various bio-formulations using large-size (100 L) fermenters. Evaluation of biocontrol activity of novel bio-formulations using crops in micro-scale field experiments.
  • Pilot production of product bio-formulations (powders, concentrates, solutions) and assessment of biocontrol activity/efficiency of bio-formulations using crops in large-scale field experiments.
  • Development of protocols and practical recommendations on the production and use of Pseudomonas-based formulations as biocontrol agents.

The implementation of the proposed work plan will allow us to transfer state-of-the-art technology from a lab bench top to applied research in the field. The main goal to be achieved include the development of technology for pilot production of Pseudomonas-based formulations that actively promote plant growth by killing or inhibiting fungal diseases of commercial crops. Kyrgyz biotechnology companies will market these microbial formulations as commercial biocontrol products. Such products are a viable first line for controlling crop diseases and thus have a great potential to be registered for release. Control of these diseases will lead to increased crop yields that will be worth hundreds of millions of dollars to Kyrgyz producers.

Benefits to the Industry. The current proliferation of biotechnology companies worldwide underscores the importance and economic potential of biological inoculants in agriculture. Such inoculants have a potential for commercialization of biocontrol agents. In addition to the enhanced commercial feasibility of utilizing natural humic substances and bacteria for biological control, these inoculants will:

  • Contribute to technologies and management strategies for production of healthy crops in an environmentally acceptable manner i.e., reduced dependence on agrochemicals;
  • Help to conserve soil resources by promoting soil-plant-microbe interactions which sustain and enhance soil fertility
  • Enhance the cost-effectiveness of crop production by increasing fertilizer use efficiency.


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The International Science and Technology Center (ISTC) is an intergovernmental organization connecting scientists from Kazakhstan, Armenia, Tajikistan, Kyrgyzstan, and Georgia with their peers and research organizations in the EU, Japan, Republic of Korea, Norway and the United States.

 

ISTC facilitates international science projects and assists the global scientific and business community to source and engage with CIS and Georgian institutes that develop or possess an excellence of scientific know-how.

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