Expression analysis of Crz1 and Rho-GDP of Aspergillus flavus during Interaction with Triticum aestivum

Abstract

Fungus is a group of eukaryotic organism which include yeasts, moulds also most famous mushrooms. Majorly yeast and moulds are responsible for the spoilage of food .due to the large amount of spores produced by fungus, these spores when land into some food, it starts reproducing .beside causing spoilage and economic loss some fungal strain also produce mycotoxins .major strain of fungus which infect foods are Aspergillus, Penicillium. In this study the objective is to find out genes responsible for the attachment and growth of fungus on wheat. Aspergillus flavus was taken for this study. Pure colony of Aspergillus flavus was formed using sub-culturing and conidia harvesting. Microscopic and molecular analysis was made for the spores of Aspergillus flavus was stained with lacto-phenol cotton blue and observed under compound microscope. Further for the molecular analysis wheat was infected with the spores of A.flavus. After the 3rd and 5th day of infection, RNA was isolated and cDNA from the best purified RNA was made. RT-PCR was carried out to check the quantification of gene. Genes selected for quantifying expression analysis during interacting with host are Crz1 and Rho-GDP. Tubulin was used as housekeeping gene. Crz1 is a transcriptional factor regulates morphogenesis via calcium signalling pathway. It helps fungal cell to promote growth and their by assist in virulence during infection on suitable host. Rho-GDP is a transcriptional inhibitor. Expression of these two genes will reveal the morphogenesis of fungus. There is high level of expression of Crz1 gene at 5 th day of infection as compared to 3rd day and the expression of Rho-GDP was higher on 3rd day as compared to 5th day. The expression levels of gene were quantified by RT-PCR and normalized using tubulin gene. The expression analysis of transcriptional factor may help to add in our understanding on how A. flavus colonizes on wheat