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Emerging Technology Marketplace

EAFIT University’s New Fungicidal Technology

EAFIT University, located in Medellin, Colombia new fungicidal technology is based on a Bacillus species, discovered in banana plantation soil in Colombia. Laboratory, greenhouse, and field trials have all indicated that this microorganism has fungicidal properties and is effective against typical fungal pathogens of plants. EAFIT is seeking licensees to the technology.

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Tech Brief

Microbial Control of Plant Disease

Description
EAFIT University and The Banana Growers Association (AUGURA) have collaboratively developed a microbial biopesticide based on an isolated strain of Bacillus subtilis (EA-CB0015) and its metabolites for the control of phytopathogens. The technology includes an optimized production process for the bacterial strain that produces a high yield of microbial biomass, viable spore count, and metabolites (lipopeptides), and a novel formulation that allows long-term shelf life, UV protection of the bacterial strain, good adherence of the biopesticide to plant surfaces and compatibility with chemical fungicides.

The biopesticide formulation has been tested in banana crops for the control of black Sigatoka (fungal) disease in the greenhouse and field with a control efficacy equal to the fungicides chlorotalonil and mancozeb. Additionally, the strain B. subtilis EA-CB0015 has been successfully tested against the fungi Colletotrichum (tomato), and Botrytis cinerea (chrysanthemum flowers), showing high control efficacy. This bacterial strain also has high inhibition activity in vitro against other phytopathogens including the fungi Mycosphaerella fijiensis, Fusarium oxysporum, Monilia sp. Rhizoctonia solani and Fusarium solani and bacteria Ralstonia solanacearum.

Technical merits
The strain B. subtilis EA-CB0015, isolated from a leaf of a banana plant in Colombia, has been rigorously selected from 649 bacterial strains for its high antimicrobial activity, with an average inhibition of 92% against the fungus M. fijiensis. The metabolites produced by this strain, lipopeptides from the surfactin, fengycin and iturin family, are highly effective against phytopathogenic microorganisms. Research has demonstrated that this strain produces high amounts of a new fengycin isoform, “fengycin C” (781.4 mg/L) and iturins (355.4 mg/L) which generates alterations in fungal structures. The spores and vegetative cells of B. subtilis EA-CB0015 are also effective against phytopathogenic fungi with percentage of inhibitions between 40% and 92% in vitro.

The fermentation process for producing the strain B. subtilis EA-CB0015 was optimized to obtain high yields of biomass (1*10^9 CFU/mL) and lipopeptide metabolites, that can be exploited for biotechnological application.

The optimized formulation which incorporates the bacterial strains and its metabolites has a long shelf life (1 year), is stable with other chemical fungicides, has protection from UV light and has a good adherence to hydrophobic surfaces.

Screening process for strain selection
Strain B. subtilis EA-CB0015 was rigorously isolated from 649 aerobic endospore-forming bacteria (AEFB) which were obtained from different banana plant cultivars in Colombia during the years 2008 and 2009. All isolated cultures were assessed for their antagonistic activity against M. fijiensis (causal agent of Black Sigatoka disease) and also tested in vitro against other phytophatogens such as F. oxysporum, R. solani, among others. To isolate AEFB, leaf samples from banana cultivars Grand Naine, Valery and the plantain Harton were obtained from different plantations and stored in sterile bags and analyzed 24 hours after their collection. A unique protocol was designed to isolate AEFB to avoid traditional methods that require the organism be in the spore form. With these traditional methods there´s no guarantee that strain, isolated from a particular environment, will actually grow at that location. Using the unique isolation protocol, many gram positive strains, not in the spore form, and which might grow in a leaf environment, were isolated. Each isolate was cultured and the supernatant assessed in vitro for its capacity to inhibit growth of M. fijiensis. From this test, 5% of the isolates showed fungal inhibitory effects above 84% and selected for further testing. Those selected were then evaluated in two other trials to determine their effect on M. fijiensis mycelium and ascospore germination, as well as their activity against different phytopathogens. After 2 years of research, strain B. subtilis EA-CB0015 was selected for its high antimicrobial characteristics.

IP and tangible property status
A PCT patent application has been filed. The tangible property includes cultures of the strain. In addition, there are trade secrets involving the bioprocessing/cultivation of the strain and formulation of the biopesticide. The ownership of the intellectual and tangible property is equally shared between AUGURA and EAFIT University through the Biological Sciences and Bioprocess Research Group. AUGURA has given a legal power to EAFIT to lead contract negotiations and the commercialization process of this technology.

Development status
The biopesticide has been manufactured at the lab scale (14 liters) and numerous experiments carried out over several years have shown good results in greenhouse and field trials of banana crops. The production process for the banana field trials has been scaled up to 14 liters. Methods developed includes the operational units and conditions, as well as the amounts of raw materials required. Other formulations for other crops can be developed using liquid and microencapsulation techniques.

Types of deals sought
EAFIT University and AUGURA are seeking to license the IP and tangible property and will consider exclusive licenses on a country-by-country or regional basis.

Contact details
Dr. Richard S. Cahoon, President, BioProperty Strategy Group, Inc. rscahoon@biopropertystrategy.com, 607.229.0802


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