Optimizing the Agrobacterium tumefaciens-mediated transformation conditions in Colletotrichum lindemuthianum: a step forward to unravel the functions of pathogenicity arsenals

Abstract

Significance and Impact of the Study: In the present study, we for the first time, report an efficient Agrobacterium tumefaciens-mediated transformation (ATMT) system for a hemibiotrophic fungus, Colletotrichum lindemuthianum. This optimized ATMT protocol is versatile and can be used for the functional genetic studies. Insertional mutagenesis studies produce thousands of mutant lines that need to be evaluated for pathogenicity loss and the traditional methods of pathogenicity assay in bean-anthracnose pathosystem are time consuming and laborious. Hence, we developed a quick and robust screening method that produce consistent results similar to the established bioassay methods. Our optimized ATMT and virulence analysis protocols will prove useful in understanding the bean-anthracnose interface.

Polyethylene glycol–mediated genetic transformation and fluorescent labelling of Colletotrichum sublineola for studying sorghum anthracnose pathogenicity

“…Hygromycin B is the most common selective marker used for filamentous fungal transformant screening. Previous studies reported that 200 μg mL −1 hygromycin B can completely inhibit the germination and growth of wild‐type Colletotrichum lindemuthianum race 2047 (Nabi et al., 2022), whereas 250 μg mL −1 hygromycin B can completely inhibit the growth of Colletotrichum acutatum isolate 397 (Talhinhas et al., 2008). This result indicates that the fungus is sensitive to hygromycin B.…”