title: Molecular cloning and characterization of gloverin from the diamondback moth, Plutella xylostella L. and its interaction with bacterial membrane.
abstract: Gloverin restricted to Lepidoptera is known to be a glycine-rich and heat stable antimicrobial protein. The current research reports a 650 bp full-length cDNA encoding gloverin from Plutella xylostella (PxGlo) by reverse transcription polymerase chain reaction and rapid amplification of cDNA ends. PxGlo transcript was detected in both developmental stages and several tissues of 4th instar naïve larvae of P. xylostella with higher levels in the fat bodies. The mRNA levels of PxGlo increased appreciably in fat bodies after injection of Escherichia coli K12. The recombinant PxGlo expressed in S2 cells was purified by Anti-V5 M2 agarose beads which showed high activity against E. coli K12, while low activity against Bacillus thuringiensis, Staphylococcus aureus and E. coli D31. The analysis of transmission electron microscope and scan electron microscope showed PxGlo to cause significant morphological alteration in the E. coli K12 cell surface. Knockdown of PxGlo expression by RNAi increased the larval susceptibility towards the pathogenic bacteria i.e., Serratia marcescens and B. thuringiensis. Our results showed that PxGlo is an inducible antibacterial peptide which exhibits high activity mainly against E. coli K12, and PxGlo performs vital roles against the infection of pathogenic bacteria.
authors.affiliation: College of Natural Resources and Environment, Engineering Research Centre of Biological Control, Ministry of Education, South China Agricultural University Guangzhou, Guangzhou, 510642, China.
authors.forename: X X
authors.initials: XX
authors.lastname: Xu
authors.affiliation: College of Natural Resources and Environment, Engineering Research Centre of Biological Control, Ministry of Education, South China Agricultural University Guangzhou, Guangzhou, 510642, China. jflbang@scau.edu.cn.
authors.forename: F L
authors.initials: FL
authors.lastname: Jin
authors.affiliation: State Key Laboratory of Respiratory Disease, Guangzhou Institute of Respiratory Disease, The First Affiliated Hospital of Guangzhou Medical College, Guangzhou, 510230, China.
authors.forename: Y S
authors.initials: YS
authors.lastname: Wang
authors.affiliation: Department of Entomology, Faculty of Agricultural Sciences and Technology, Bahauddin Zakariya University, Multan, 60800, Pakistan. shbfkh@gmail.com.
authors.forename: Shoaib
authors.initials: S
authors.lastname: Freed
authors.affiliation: College of Natural Resources and Environment, Engineering Research Centre of Biological Control, Ministry of Education, South China Agricultural University Guangzhou, Guangzhou, 510642, China.
authors.forename: Q B
authors.initials: QB
authors.lastname: Hu
authors.affiliation: College of Natural Resources and Environment, Engineering Research Centre of Biological Control, Ministry of Education, South China Agricultural University Guangzhou, Guangzhou, 510642, China.
authors.forename: S X
authors.initials: SX
authors.lastname: Ren
mesh_terms: D000818:Animals; D023181:Antimicrobial Cationic Peptides / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D001424:Bacterial Infections / Q000276:immunology; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects*; D020869:Gene Expression Profiling; D020868:Gene Silencing; D006090:Gram-Negative Bacteria / Q000187:drug effects* / Q000648:ultrastructure; D006094:Gram-Positive Bacteria / Q000187:drug effects*; D036341:Intercellular Signaling Peptides and Proteins; D007915:Lepidoptera / Q000235:genetics / Q000254:growth & development / Q000276:immunology; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D023181:Antimicrobial Cationic Peptides; D036341:Intercellular Signaling Peptides and Proteins; D011506:Proteins; D011994:Recombinant Proteins; C107617:gloverin
keywords: Antibacterial mechanism; Diamondback moth; Gloverin; RNA interference; RT-PCR