CRISPR-mediated genome editing in entompathogenic fungi for improved fungal biopesticides

• Grant number: 22/00242-3
• Support Opportunities: Scholarships abroad - Research
• Effective date (Start): August 27, 2022
• Effective date (End): August 26, 2023
• Field of knowledge: Agronomical Sciences - Agronomy - Plant Health
• Principal Investigator: Gabriel Moura Mascarin
• Grantee: Gabriel Moura Mascarin
• Host Investigator: Jeffrey Coleman
• Host Institution: Embrapa Meio-Ambiente. Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA). Ministério da Agricultura, Pecuária e Abastecimento (Brasil). Jaguariúna , SP, Brazil
• Host Company: Universidade de São Paulo (USP). Escola Superior de Agricultura Luiz de Queiroz (ESALQ)
• Research place: Auburn University, United States
• Associated research grant: 18/02317-5 - Center for Excellence in Biological Control, AP.PCPE

Abstract

Beauveria bassiana is one of the most studied fungi for use in biological control of many agricultural and forest arthropod pests of economic importance worldwide. However, compared with chemical pesticides, the performance of this and other entomopathogenic fungi recurrently shows inconsistent or unsatisfactory results under field conditions due to their slow speed and high lethal dose required to kill target insects. To overcome these setbacks, a possible and feasible strategy lies in a precise and efficient CRISPR genetic engineering system enabling generation of fungal strains with improved virulence. To this end, our main goals are to develop CRISPR/Cas9 systems for B. bassiana a) to generate knockout mutants of Cry toxin-like encoding genes, which appear to be functionally analogous to the Cry toxins in Bacillus thuringiensis, allowing evaluation of their role in fungal virulence against an insect host; and b) to overexpress the two most important cuticle degrading enzymes (protease/chitinase) in order to enhance the fungal virulence. Fungal mutants generated through these aims will be evaluated in regards to their virulence on insect hosts and in vitro blastospore production in comparison to the parental (wild-type) strain. Finally, we anticipate that our results will foster the development of improved and customized fungal biopesticides based on novel strains phenotypically superior than their wild-type through the implementation of an innovative and robust CRISPR platform in Brazil. (AU)