The relationship between pigmentations of pupae and insecticide resistance of adults of Bactrocera dorsalis (Hendel)
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Abstract
The oriental fruit fly Bactrocera dorsalis (Hendel) is a worldwide pest leading to crop production. The management of B. dorsalis is becoming more and more difficult because of the emerging insecticide resistance. Searching for new and effective methods in managing this pest has become an urgent task. Here, we explored the relationship between pigmentation and insecticide resistance through conducting studies on 5 strains of B. dorsalis (CK, CZ, CB, LC and LX) with various traits of pigmentation and insecticide resistance. Strain CK was a wild strain with brown pupae coloration while strain CZ was a beta-cypermethrin-resistant strain with pupae coloration was brown. Strain CB was a pigmentation mutation strain with white pupae coloration. Strain LC was the female of pigmentation sex genetic strain whose pupae coloration was white. Strain LX was the male of pigmentation sex genetic strain whose pupae coloration was brown. Efficacy tests were employed to evaluate the resistance performance of the 5 trains, transcriptome sequencing and gene quantifications were used to explore the potential genes in both pigmentation and detoxification pathway. The brief of results were as follow:
Results of efficacy tests of the 5 strains with beta-cypermethrin, abamectin and trichlorfon respectively showed that resistances to these three insecticide of strains LX and LC were very low. The corrective mortalities of strains LX and LC in 128 ug/ml of abamectin were 49.48±3.01%, 59.06±1.89% respectively while that of strain CK was 11.80%±1.37. The corrective mortality of strain CZ were lowest in all concentrations level of all test reagent. The resistance of strain CK was similar to CB. Transcriptomic analysis showed that 24 genes and 165 genes were associated with pigmentation and detoxification. Gene expression pattern of strain CK was similar to strain CB while these two strains were different to strains LC and LX, and the gene expression pattern of strain CZ was more similar to strains LC and LX. Results of gene quantifications in B. dorsalis pupae showed that the relative expressions of yellow gene MK529913, DDC gene MK515141, laccase2 gene MK515143 and PO gene MK515140 in brown pupae (CK, CZ, LX) were significantly higher than those in white pupae (CB, LC) while the relative expressions of AANAT gene MK515144 in white pupae were significantly higher than those in yellow pupae. In summary, analyses of gene quantifications and efficacy tests indicated that high expressions of P450 gene, AANAT gene might contribute to resistances of adults of B. dorsalis to beta-cypermethrin and a bamectin; high expression of yellow gene might contribute to resistances of adults which emergenced from brown pupae to beta-cypermethrin and abamectin; high expressions of yellow genes, laccase2 gene, PO gene and DDC gene might contribute to the resistance of B. dorsalis to trichlorfon.
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2019. The relationship between pigmentations of pupae and insecticide resistance of adults of Bactrocera dorsalis (Hendel) PeerJ Preprints 7:e27597v1 https://doi.org/10.7287/peerj.preprints.27597v1Author comment
This is a preprint submission to PeerJ Preprints.
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Supplemental Information
Figure 2 Respondings of B. dorsalis adults to beta-cypermethrin, abamectin and trichlorfon
(A) Corrected mortalities of 5 strains in beta-cypermethrin. (B) Corrected mortalities of 5 strains in abamectin. (C) Corrected mortalities of 5 strains in trichlorfon. Notes: X-axis standed for strains of B. dorsalis. Y-axis standed for corrected mortalities of genes. Length of column standed for the average of corrected mortalitie. Line on columns standed for standard error. The difference of letters on columns standed for significant by DMRT (a=0.05) analysis of variance.
Figure 3 The relationship of 5 strains in biological process, cell component and molecular function
(A) Numbers of biological process associated with detoxifications and pigmentations, (B) Numbers of cell component associated with detoxifications and pigmentations, (C) Numbers of molecular function associated with detoxifications and pigmentations. Note: “Det” and “Pig” in the figure were abbreviations of Detoxification and Pigmentation respectively.
Figure 4 Relationships of gene expressions of 5 strains
(A) All genes of 5 strains, (B) 165 genes associated with detoxification enzymes of 5 strains, (C) 24 genes associated with pigmentations of 5 strains, (D) 116 genes up-regulated or down-regulated in 10 differential comparison groups of 5 strains. Notes: The row name was the family name of gene. The number in the expansion was the accession number of gene in GenBank. Genes associated with pigmentations and marked with accession numbers were subjected to real-time PCR amplifications.
Figure 5 Expresions of 9 genes in pupae of 6 strains of B. dorsalis
(A) Expresion of gene MK529913(yellow). (B) Expresion of gene MK529914(yellow). (C) Expresion of gene MK529912(yellow). (D) Expresion of gene MK529915(yellow). (E) Expresion of gene MK515143(laccase2). (F) Expresion of gene MK515144(AANAT). (G) Expresion of gene MK515140(PO). (H) Expresion of gene MK515141(DDC). (I) Expresion of gene MK515142(P450). Notes: X-axis standed for strains of B. dorsalis. Y-axis standed for relative expression of genes. Length of column standed for the average of relative expression of gene. Line on column standed for standard error. The difference of letters on columns standed for significant by DMRT (a=0.05) analysis of variance.
Figure 6 Expresions of 9 genes in adults of 6 strains of B. dorsalis
(A) Expresion of gene MK529913(yellow). (B) Expresion of gene MK529914(yellow). (C) Expresion of gene MK529912(yellow). (D) Expresion of gene MK529915(yellow). (E) Expresion of gene MK515143(laccase2). (F) Expresion of gene MK515144(AANAT). (G) Expresion of gene MK515140(PO). (H) Expresion of gene MK515141(DDC). (I) Expresion of gene MK515142(P450). Notes: X-axis standed for strains of B. dorsalis. Y-axis standed for relative expression of genes. Length of column standed for the average of relative expression of gene. Line on column standed for standard error. The difference of letters on columns standed for significant by DMRT(a=0.05) analysis of variance.
Supplemental file 1 The protein sequences and alignments of 127 P450 genes
Supplemental file 2 The protein sequences and alignments of 33 GST genes
Supplemental file 3 The protein sequences and alignments of 1 AcE gene
Supplemental file 4 The protein sequences and alignments of 4 CarE genes
Supplemental file 5 The protein sequences and alignments of 10 yellow genes
Supplemental file 6 The protein sequences and alignments of 1 AANAT gene
Supplemental file 7 The protein sequences and alignments of 2 laccase2 genes
Supplemental file 8 The protein sequences and alignments of 1 ebony gene
Supplemental file 9 The protein sequences and alignments of 1 tan gene
Supplemental file 10 The protein sequences and alignments of 2 PO genes
Supplemental file 11 The protein sequences and alignments of 2 ADC genes
Supplemental file 12 The protein sequences and alignments of 2 DDC genes
Supplemental file 13 The protein sequences and alignments of 1 PAH gene
Supplemental file 14 The protein sequences and alignments of 1 TH gene
Additional Information
Competing Interests
Gu-Qian Wang is employed by Jiangxi Yang's Fruit CO., Ltd.
Author Contributions
Yu-Peng Chen conceived and designed the experiments, performed the experiments, analyzed the data, contributed reagents/materials/analysis tools, prepared figures and/or tables, authored or reviewed drafts of the paper, approved the final draft.
Gu-Qian Wang conceived and designed the experiments, performed the experiments, contributed reagents/materials/analysis tools.
Ting Yu conceived and designed the experiments, performed the experiments, contributed reagents/materials/analysis tools, authored or reviewed drafts of the paper.
Xiao-Yan Peng performed the experiments, contributed reagents/materials/analysis tools.
Tao Li performed the experiments, contributed reagents/materials/analysis tools.
Zhou Gao performed the experiments, contributed reagents/materials/analysis tools.
Xue Bai performed the experiments, contributed reagents/materials/analysis tools.
Chun-Yan Zheng performed the experiments, contributed reagents/materials/analysis tools.
Guang-Wen Liang conceived and designed the experiments, authored or reviewed drafts of the paper, approved the final draft.
Yong-Yue Lu conceived and designed the experiments, authored or reviewed drafts of the paper, approved the final draft.
Data Deposition
The following information was supplied regarding data availability:
Supplemental files 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 and 14 were used for identification of genes in different gene families. Protein sequences and alignments of P450 genes are shown in supplemental file 1. Protein sequences and alignments of GST genes were showed in supplemental file 2. Protein sequences and alignments of AcE genes were showed in supplemental file 3. Protein sequences and alignments of CarE genes were showed in supplemental file 4. Protein sequences and alignments of yellow genes, AANAT genes, laccase2 genes, ebony genes, tan genes, PO genes, ADC genes, DDC genes, PAH genes and TH genes were showed in supplemental files 5, 6, 7, 8, 9, 10, 11, 12, 13 and 14 respectively.
Funding
The authors received no funding for this work.
