Project description:We used our newly ultra deep sequence data and bioinformatics to re-annotate P. xylostella genome for high confidence miRNAs with the correct 5p and 3p arm features. Furthermore, the whole genome was screened to identify potential miRNA binding sites using three target-predicting algorithms. Totally, 203 mature miRNAs were annotated, including 33 novel miRNAs. Two geographical populations of Diamondback moth larvae from Queensland (Gatton) and South Australia (Waite) were collected and reared on the cabbage plant at the University of Queensland in Australia. Total RNA was extracted from fifteen 3rd instar larval samples using Triazol® following the manufacturerâs protocol (Life Technologies). The small RNA libraries were generated from both populations with three biological replicates using the Illumina Truseq small RNA preparation kit at the Australian Genome Research Facility (AGRF-Melbourne, Australia). The purified cDNA libraries were sequenced on Illumina HiSeq and raw sequencing reads (50 nt) were obtained using Illuminaâs Sequencing Control Studio software.
Project description:Amplicon sequencing analysis using Illumina MiSeq for monitoring a cyantraniliprole resistance-related mutation in the diamondback moth populations
Project description:In insects, male accessory gland proteins (ACPs) are important reproductive proteins secreted by male accessory glands (MAGs) of the internal male reproductive system. During mating, ACPs were transferred along with sperms inside female bodies and have a significant impact on the physiology of female reproduction. Under sexual selection pressures, the ACPs exhibit remarkably rapid and divergent evolution and varies from species to species. The diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae), is a major insect pest of cruciferous vegetables worldwide. The reproductive physiology on post-mating state of this species is still largely unknown, which is important for management of this pest. In this study, the ACPs transferred into females during mating were identified by using a tandem mass tags quantitative proteomic analysis. The MAGs were compared before and after mating immediately. In total, we identified 123 putative secreted ACPs, including most important physiological: regulators of proteolysis, transporters and protein export machinery, signal transduction and immunity. Comparing P. xylostella with other four insect ACPs, trypsins were the only ACPs detected in all insect species. This was the first time to identify and analyze ACPs in P. xylostella. Our results have provided an important list of putative secreted ACPs, and have set the stage for further explore functions of these putative proteins in P. xylostella reproduction.
Project description:Amplicon sequencing analysis using next generation sequencer for the genomic diagnosis of pesticide resistance in the diamondback moth populations
Project description:In insects, male accessory gland proteins (ACPs) are important reproductive proteins secreted by male accessory glands (MAGs) of the internal male reproductive system. During mating, ACPs were transferred along with sperms inside female bodies and have a significant impact on the physiology of female reproduction. Under sexual selection pressures, the ACPs exhibit remarkably rapid and divergent evolution and varies from species to species. The diamondback moth, Plutella xylostella (L.) (Lepidoptera: Plutellidae), is a major insect pest of cruciferous vegetables worldwide. The reproductive physiology on post-mating state of this species is still largely unknown, which is important for management of this pest. In this study, the ACPs transferred into females during mating were identified by using a tandem mass tags quantitative proteomic analysis. The MAGs were compared before and after mating immediately. The proteomes of copulatory bursas (CB) in mated females shortly after mating were also analyzed by shotgun LC-MS/MS technique. In total, we identified 123 putative secreted ACPs, including most important physiological: regulators of proteolysis, transporters and protein export machinery, signal transduction and immunity. Comparing P. xylostella with other four insect ACPs, trypsins were the only ACPs detected in all insect species. We also identified some new insect ACPs, including protein with chitin binding Peritrophin-A domain, PMP-22/EMP/MP20/Claudin tight junction domain-containing protein, netrin-1, type II inositol 1,4,5-trisphosphate 5-phosphatase, two spaetzles, allatostatin-CC and cuticular protein. This was the first time to identify and analyze ACPs in P. xylostella. Our results have provided an important list of putative secreted ACPs, and have set the stage for further explore functions of these putative proteins in P. xylostella reproduction.
Project description:miRNAs play important roles in various biological processes through post-transcriptional regulation of gene expression. We previously identified 203 mature miRNAs in Diamondback moth, Plutella xylostella. This species has developed extremely high levels of resistance to chlorantraniliprole and other class of insecticides in the field. In this study, we examined the miRNA profile of P. xylostella in response to chlorantraniliprole exposure. The smRNA-seq data analyses showed that insecticide treatment caused significant changes in the abundance of some miRNAs. Increasing exposure time (6h to 24h) and insecticide concentration (0.01 to 0.1 ppm) induced more dysregulated miRNAs in DBM larvae.
Project description:UV-B radiation regulates numerous morphogenic, biochemical and physiological responses in plants, and can stimulate some responses typically associated with other abiotic and biotic stimuli, including invertebrate herbivory. Removal of UV-B from the growing environment of various plant species has been found to increase their susceptibility to consumption by invertebrate pests, however, to date, little research has been conducted to investigate the effects of UV-B on crop susceptibility to field pests. Here, we report findings from a multi-omic and genetic-based study investigating the mechanisms of UV-B-stimulated resistance of the crop, Brassica napus (oilseed rape), to herbivory from an economically important lepidopteran specialist of the Brassicaceae, Plutella xylostella (diamondback moth). The UV-B photoreceptor, UV RESISTANCE LOCUS 8 (UVR8), was not found to mediate resistance to this pest. RNA-Seq and untargeted metabolomics identified components of the sinapate/lignin biosynthetic pathway that were similarly regulated by UV-B and herbivory. Arabidopsis mutants in genes encoding two enzymes in the sinapate/lignin biosynthetic pathway, CAFFEATE O-METHYLTRANSFERASE 1 (COMT1) and ELICITOR-ACTIVATED GENE 3-2 (ELI3-2), retained UV-B-mediated resistance to P. xylostella herbivory. However, overexpression of B. napus COMT1 in Arabidopsis further reduced plant susceptibility to P. xylostella herbivory in a UV-B-dependent manner. These findings demonstrate that overexpression of a component of the sinapate/lignin biosynthetic pathway in a member of the Brassicaceae can enhance UV-B-stimulated resistance to herbivory from P. xylostella.
Project description:Purpose: To investigate the molecular mechanism underlying the function of DMNT function on P. xylostella larvar, we performed a whole genome transcriptome sequencing (RNA-seq) analysis