Supplementary material from "Chitinase genes from Metarhizium anisopliae for the control of whitefly in cotton"

Entomopathogenic fungi produces endochitianses, involved in the degradation of insect chitin to facilitate the infection process. Endochitinases (Chit1) gene of family 18 glycosyl hydrolyses were amplified, cloned and characterized from genomic DNA of two isolates of Metarhizium anisopliae. Catalytic motif of family 18 glycosyl hydrolyses was found in Chit1 of M. anisopliae, while no signal peptide was found in any isolate, whereas substrate-binding motif was found in Chit1 of both isolates. Phylogenetic analysis revealed the evolutionary relationship among the fungal chitinases of Metarhizium. The Chit1 amplified were closely related to the family 18 glycosyl hydrolyses. Transient expressions of Chit1 in cotton plants using Geminivirus-mediated gene silencing vector of Cotton Leaf Crumple Virus (CLCrV) revealed the chitinase activity of Chit1 genes amplified from both of the isolates of M. anisopliae when compared with the control. Transformed cotton plants were virulent against fourth instar nymphal and adult stages of Bemisia tabaci which resulted in the mortality of both fourth instar nymphal and adult B. tabaci. Thus, the fungal chitinases expressed in cotton plants played a vital role in plant defence against B. tabaci. However, further studies are required to explore the comparative effectiveness of chitinases from different fungal strains against economically important insect pests.