In Vitro Induction of Infection-Related Hyphal Structures in Plant Pathogenic Fungi

Abstract

In recent years, a voluminous amount of genomic data has been generated for several plant pathogenic fungi. Multiple studies have utilized these genomic data to advance our knowledge about the molecular mechanisms of plant pathogenesis. However, not all plant pathogenic fungi share the same infection strategies, and several genes have been identified that are crucial for plant pathogenesis in one fungus, but dispensable in others. In order for data on biological relevance to keep pace with accumulating genomic data, new biological assays need to be developed for several pathogenic fungi. Accordingly, we have developed an in vitro assay that allows us to monitor morphological changes in hyphal development as the head blight pathogen Fusarium graminearum infects wheat. Using previously frozen detached wheat glumes, we are able to monitor both subcuticular and intercellular hyphal development of F. graminearum. The method described takes only 3-5 days from inoculation to microscopic observation (depending on time point) and does not require any elaborate laboratory equipment or supplies. This method could be adapted for different necrotrophic or hemibiotrophic pathogens, on their host tissue types, in order to characterize their hyphal differentiation in vitro.

Aspergillus flavus virulence in pods and seeds of peanut with different drought responsive genotypes related to water status

“…Fungal stain was firstly made overnight with 5% lactophenol cotton blue dye solution. Removal of residual dye was modified from the previous procedure [24] by subsequent washings for 1 min twice in distilled water, 5 min in 20% glycerol, and 1 min in distilled water. Secondly, plant cells were counterstained for 20 s by 1% safranin O [25], and the sections were then washed as aforementioned.…”