title: Bacteria abundance and diversity of different life stages of Plutella xylostella (Lepidoptera: Plutellidae), revealed by bacteria culture-dependent and PCR-DGGE methods.
abstract: Microbial abundance and diversity of different life stages (fourth instar larvae, pupae and adults) of the diamondback moth, Plutella xylostella L., collected from field and reared in laboratory, were investigated using bacteria culture-dependent method and PCR-DGGE analysis based on the sequence of bacteria 16S rRNA V3 region gene. A large quantity of bacteria was found in all life stages of P. xylostella. Field population had higher quantity of bacteria than laboratory population, and larval gut had higher quantity than pupae and adults. Culturable bacteria differed in different life stages of P. xylostella. Twenty-five different bacterial strains were identified in total, among them 20 strains were presented in larval gut, only 8 strains in pupae and 14 strains in adults were detected. Firmicutes bacteria, Bacillus sp., were the most dominant species in every life stage. 15 distinct bands were obtained from DGGE electrophoresis gel. The sequences blasted in GenBank database showed these bacteria belonged to six different genera. Phylogenetic analysis showed the sequences of the bacteria belonged to the Actinobacteri, Proteobacteria and Firmicutes. Serratia sp. in Proteobacteria was the most abundant species in larval gut. In pupae, unculturable bacteria were the most dominant species, and unculturable bacteria and Serratia sp. were the most dominant species in adults. Our study suggested that a combination of molecular and traditional culturing methods can be effectively used to analyze and to determine the diversity of gut microflora. These known bacteria may play important roles in development of P. xylostella.
authors.affiliation: State Key Laboratory of Crop Stress Biology for the Arid Areas, and Key Laboratory of Northwest Loess Plateau Crop Pest Management of Ministry of Agriculture, Northwest A&F University, Yangling, Shaanxi, China.
authors.forename: Xiao-Li
authors.initials: XL
authors.lastname: Lin
authors.affiliation: State Key Laboratory of Crop Stress Biology for the Arid Areas, and Key Laboratory of Northwest Loess Plateau Crop Pest Management of Ministry of Agriculture, Northwest A&F University, Yangling, Shaanxi, China.
authors.forename: Qin-Jian
authors.initials: QJ
authors.lastname: Pan
authors.affiliation: State Key Laboratory of Crop Stress Biology for the Arid Areas, and Key Laboratory of Northwest Loess Plateau Crop Pest Management of Ministry of Agriculture, Northwest A&F University, Yangling, Shaanxi, China.
authors.forename: Hong-Gang
authors.initials: HG
authors.lastname: Tian
authors.affiliation: Department of Entomology and Department of Molecular Biology & Genetics, Cornell University, Ithaca, NY, 14853, USA.
authors.forename: Angela E
authors.initials: AE
authors.lastname: Douglas
authors.affiliation: State Key Laboratory of Crop Stress Biology for the Arid Areas, and Key Laboratory of Northwest Loess Plateau Crop Pest Management of Ministry of Agriculture, Northwest A&F University, Yangling, Shaanxi, China.
authors.forename: Tong-Xian
authors.initials: TX
authors.lastname: Liu
mesh_terms: D000818:Animals; D001419:Bacteria / Q000235:genetics / Q000302:isolation & purification*; D015169:Colony Count, Microbial; D004269:DNA, Bacterial / Q000032:analysis; D041981:Gastrointestinal Tract / Q000382:microbiology; D007814:Larva / Q000382:microbiology; D009036:Moths / Q000254:growth & development / Q000382:microbiology*; D010802:Phylogeny; D011679:Pupa / Q000382:microbiology; D012336:RNA, Ribosomal, 16S / Q000032:analysis
chemical_list: D004269:DNA, Bacterial; D012336:RNA, Ribosomal, 16S
keywords: 16S rRNA; diamondback moth; gut flora; insect bacteria; microbial diversity