Biocontrol Agents for Plants
Scale-up Technology and Establishment of Pilot Production of Plant Growth Promoting Biological Control Agents
Tech Area / Field
- AGR-DIS/Disease Surveillance/Agriculture
- AGR-PPR/Plant Protection/Agriculture
- BIO-IND/Industrial Biotechnology/Biotechnology
3 Approved without Funding
National Academy of Sciences of Kyrgyzstan / Institute of Chemistry and Chemical Technology, Kyrgyzstan, Bishkek
- JSC Biochimmash, Russia, Moscow
- University of Saskatchewan, Canada, SK, Saskatoon\nWashington State University, USA, WA, Pullman
Project summaryAnnually, 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.
It should be mentioned herein that the three years of the project KR-993.2 development have been very successful – project participants have developed very promising technologies for production of biocontrol agent formulations that can be scaled up to industrial production. These technologies have been described in detail in the final report of project KR-993.2, which has been enthusiastically and critically evaluated by our American and Canadian collaborators. In fact, Prof. Renato de Freitas and Dr. Lynne Carpenter-Boggs have sent their reviews on the final technical report to Dr. H. Visser where they expressed full satisfaction with the results of the project. Additionally, they strongly recommended scale-up and thus, demonstrating the newly developed technology in field scale trials. Our scientific collaborators have no doubt that the scientific merit of the results obtained during project KR-993.2 provides unequivocal arguments for the success of a new proposal. 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. These tests clearly indicated that the project is more than ready for the next step. This includes launching another innovative research that consists of scaling up a newly developed technology including a pilot-plant production and field-testing settings for the new products. Full-scale establishment of the project is a prerequisite for further commercialization of the developed technology. In addition to provide project sustainability, this objective constitutes a final and fundamental goal of ISTC.
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 RB-43 is a strong antagonist against the plant pathogen Bipolaris sorokiniana.
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 the ISTC 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. Systems that demonstrate superior capabilities will be selected for scaling up pilot-production.
- 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 large-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.
Expected income. Development of rhizobacteria-based preparations to enhance crop production through biocontrol of phytopathogens provides an alternative to the use of expensive fungicides. Immediate economic benefits to the producer include improved crop growth and reduced pesticide costs. Biological control agents could be expected to increase yields by approximately 5 to 15% by suppressing diseases. Hence, the potential economic benefits to crop producers and agro-industry is in the millions of dollars.
Although it is difficult to predict the actual cost of microbial inoculants, one could estimate the cost to be 25 to 50% of that of currently used chemicals. In addition to being an economical alternative to chemicals, the use of biological agents should be a more environmentally acceptable technology.