title: Detection of carboxylesterase and esterase activity in culturable gut bacterial flora isolated from diamondback moth, Plutella xylostella (Linnaeus), from India and its possible role in indoxacarb degradation.
abstract: Diamondback moth (DBM), Plutella xylostella (Linnaeus), is a notorious pest of brassica crops worldwide and is resistant to all groups of insecticides. The insect system harbors diverse groups of microbiota, which in turn helps in enzymatic degradation of xenobiotic-like insecticides. The present study aimed to determine the diversity of gut microflora in DBM, quantify esterase activity and elucidate their possible role in degradation of indoxacarb. We screened 11 geographic populations of DBM in India and analyzed them for bacterial diversity. The culturable gut bacterial flora underwent molecular characterization with 16S rRNA. We obtained 25 bacterial isolates from larvae (n=13) and adults (n=12) of DBM. In larval gut isolates, gammaproteobacteria was the most abundant (76%), followed by bacilli (15.4%). Molecular characterization placed adult gut bacterial strains into three major classes based on abundance: gammaproteobacteria (66%), bacilli (16.7%) and flavobacteria (16.7%). Esterase activity from 19 gut bacterial isolates ranged from 0.072 to 2.32μmol/min/mg protein. Esterase bands were observed in 15 bacterial strains and the banding pattern differed in Bacillus cereus - KC985225 and Pantoea agglomerans - KC985229. The bands were characterized as carboxylesterase with profenofos used as an inhibitor. Minimal media study showed that B. cereus degraded indoxacarb up to 20%, so it could use indoxacarb for metabolism and growth. Furthermore, esterase activity was greater with minimal media than control media: 1.87 versus 0.26μmol/min/mg protein. Apart from the insect esterases, bacterial carboxylesterase may aid in the degradation of insecticides in DBM.
authors.affiliation: ICAR-National Bureau of Agricultural Insect Resources, Hebbal, Bangalore, Karnataka, India.
authors.forename: Shanivarsanthe Leelesh
authors.initials: SL
authors.lastname: Ramya
authors.affiliation: ICAR-National Bureau of Agricultural Insect Resources, Hebbal, Bangalore, Karnataka, India. Electronic address: tvenkat12@gmail.com.
authors.forename: Thiruvengadam
authors.initials: T
authors.lastname: Venkatesan
authors.affiliation: ICAR-National Bureau of Agricultural Insect Resources, Hebbal, Bangalore, Karnataka, India.
authors.forename: Kottilingam
authors.initials: K
authors.lastname: Srinivasa Murthy
authors.affiliation: ICAR-National Bureau of Agricultural Insect Resources, Hebbal, Bangalore, Karnataka, India.
authors.forename: Sushil Kumar
authors.initials: SK
authors.lastname: Jalali
authors.affiliation: ICAR-National Bureau of Agricultural Insect Resources, Hebbal, Bangalore, Karnataka, India.
authors.forename: Abraham
authors.initials: A
authors.lastname: Verghese
mesh_terms: D000818:Animals; D001419:Bacteria / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000302:isolation & purification; D043182:Carboxylesterase / Q000235:genetics / Q000378:metabolism*; D004950:Esterases / Q000235:genetics / Q000378:metabolism*; D005260:Female; D000069196:Gastrointestinal Microbiome; D041981:Gastrointestinal Tract / Q000382:microbiology; D007194:India; D007306:Insecticides / Q000378:metabolism*; D008297:Male; D009036:Moths / Q000382:microbiology*; D010078:Oxazines / Q000378:metabolism*; D010802:Phylogeny
chemical_list: D007306:Insecticides; D010078:Oxazines; C401104:indoxacarb; D004950:Esterases; D043182:Carboxylesterase
keywords: 16S rRNA; Bacteria; Carboxylesterase; Indoxacarb; Phylogeny; Plutella xylostella