Secretome Analysis of Metarhizium anisopliae Under Submerged Conditions Using Bombyx mori Chrysalis to Induce Expression of Virulence-Related Proteins

Abstract

The entomopathogenic fungus Metarhizium anisopliae is used to control insect pests. This species is specialized for the secretion of an enzymatic complex consisting of proteases, lipases, and chitinases related to pathogenicity and virulence. In this context, the secretomes of strains IBCB 167 and IBCB 384 of M. anisopliae var. anisopliae, grown under submerged fermentation in the presence of chrysalis as an inducer, were analyzed. Analysis of two-dimensional gels showed qualitative and quantitative differences between secreted proteins in both isolates. Around 102 protein spots were analyzed, and 76 % of the corresponding proteins identified by mass spectrometry were grouped into different classes (hydrolases, oxidases, reductases, isomerases, kinases, WSC domains, and hypothetical proteins). Thirty-three per cent of all the proteins analyzed were found to be common in both strains. Several virulence-related proteins were identified as proteases and mannosidases. Endo-N-acetyl-β-D-glucosaminidase expression was observed to be 10.14-fold higher for strain IBCB 384 than for strain IBCB 167, which may be an important contributor to the high virulence of IBCB 384 in Diatraea ssaccharalis. These results are important for elucidation of the host-pathogen relationship and the differences in virulence observed between the two strains.

Identification of Pathogenicity-Related Effector Proteins and the Role of Piwsc1 in the Virulence of Penicillium italicum on Citrus Fruits

“…Forty-seven secreted proteins have functional annotations or belong to known protein families. Among them, 11 secreted proteins belong to known effector families or contain domains that have been found in fungal effectors, which include cell wall integrity and the stress response component (WSC) domain (PF01822) [ 47 ], concanavalin A-like lectin (PF00457) [ 46 ], cerato-platanin (PF07249) [ 11 , 48 , 49 ], necrosis-inducing protein (NIP) (PF05630) [ 16 ], pectin lyase fold/virulence factor (PF00544) [ 50 ], ribonuclease (RNase) (PF00545, PF00445) [ 51 , 52 ], and peptidase family (PF00082, PF09286, PF05922, PF05577, PF00450) [ 24 , 28 , 53 , 54 ] ( Table S7 ).…”

“…A core candidate effector, Piwsc1, was selected to verify its role in P. italicum infection in this study. WSC proteins were regarded as candidate effector proteins in P. expansum colonization on apples and identified as secreted proteins of entomopathogenic fungus Metarhizium anisopliae when grown under submerged fermentation in the presence of chrysalis as an inducer [ 24 , 47 ]. Similar to homologs in other fungi, the RT-qPCR results exhibited that Piwsc1 was highly induced during the P. italicum infection of citrus fruits.…”

MrSVP, a secreted virulence-associated protein, contributes to thermotolerance and virulence of the entomopathogenic fungus Metarhizium robertsii

“…Bioinformatics analyses revealed that there is a signal peptide sequence (from aa position 18 to 43), and conserved functional domains were not identified. According to the experimental results that the gene affects virulence [13], it was assumed to be a secreted virulence-associated protein and named as MrSVP.…”

Evaluación in vitro del potencial acaricida de Beauveria bassiana DS3.17 sobre la garrapata común (Rhipicephalus microplus) en Oaxaca, México

“…Broglio-Forti et al (21) observaron el 100 % de mortalidad de garrapatas con concentraciones de 1x10 8 y 1x10 9 conidios en un periodo de 14 días. De igual manera, Del Pozo et al (22) reportaron valores de mortalidad de 87. procesos fisiológicos y enzimáticos de los hongos entomopatógenos, que están influenciados por factores como la variación en la excreción enzimática durante el proceso de infección, así como la variabilidad genética entre las cepas del mismo género, que favorece la expresión diferencial de genes de virulencia o específicos (23,24,25) .…”