Abstract
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.
Highlights
The whitefly (Bemisia tabaci) is recognized as an economically important insect pest of crop plants causing damage in both field and greenhouse through feeding on phloem sap, excretion royalsocietypublishing.org/journal/rsos R
The chitinase gene was isolated from the genomic DNA of M. aniopliae and PCR reaction produced an expected size bands for Chit1 from M. anisopliae
The partial endochitinase Chit1 recombinant clones were confirmed through restriction digestion and sequenced using M13 primers employing the primer walking technique
Summary
The whitefly (Bemisia tabaci) is recognized as an economically important insect pest of crop plants causing damage in both field and greenhouse through feeding on phloem sap, excretion royalsocietypublishing.org/journal/rsos R. Of honey dews and by transmitting viruses [1,2,3,4]. Major damage caused by B. tabaci is associated with 2 its notable property of transmitting numerous viruses. Brown [5] reported 1100 whitefly species worldwide; only three among them, named as B. tabaci, Trialeurodes vaporariorum and T. abutiloneus, were known as the vector of plant viruses [5]. B. tabaci (Gennadius) is considered the most significant of the whitefly vectors with the ability to transmit geminiviruses [6]. There are about 288 species of begomoviruses (family: Geminiviridae) which are known to be transmitted by B. tabaci [7]
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