Construction and evaluation of a transformant library of <em>Lasiodiplodia theobromae</em> generated through Restriction Enzyme-Mediated Integration

Abstract

Grapevine dieback, caused by Lasiodiplodia theobromae, is an important trunk disease worldwide. Transformants of L. theobromae were generated in an attempt to identify potential pathogenicity-related genes. Lasiodiplodia theobromae strain JZB 0300251, a highly virulent isolate, was selected for the genetic transformation. Based on optimised conditions, the Restriction Enzyme-Mediated Integration (REMI) methodology was established in L. theobromae using pUCATPH (a plasmid carrying a hygromycin B marker). A total of 6,036 transformants were generated with four restriction enzymes, respectively and the transformant library was evaluated based on 200 randomly selected transformants. Mutants that exhibited various degrees of virulence and different growth rates were obtained. The study provides basic results that will lead to increased understanding of the role of the pathogenicity-related genes involved in the infection process of L. theobromae