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Transcriptome Analysis of Pyrethroid-Resistant Chrysodeixis includens (Lepidoptera: Noctuidae) Reveals Overexpression of Metabolic Detoxification Genes
Journal of Economic Entomology ( IF 2.2 ) Pub Date : 2020-11-02 , DOI: 10.1093/jee/toaa233
Clerison R Perini 1 , Christine A Tabuloc 2 , Joanna C Chiu 2 , Frank G Zalom 2 , Regis F Stacke 1 , Oderlei Bernardi 1 , David R Nelson 3 , Jerson C Guedes 1
Affiliation  

Chrysodeixis includens (Walker, [1858]) is one of the most important defoliator of soybean in Brazil because of its extensive geographical distribution and high tolerance to insecticides compared with other species of caterpillars. Because of this, we conducted bioassays to evaluate the efficacy of pyrethroid λ-cyhalothrin on a C. includens resistant strain (MS) and a susceptible (LAB) laboratory strain. High throughput RNA sequencing (RNA-seq) of larval head and body tissues were performed to identify potential molecular mechanisms underlying pyrethroid resistance. Insecticide bioassays showed that MS larvae exhibit 28.9-fold resistance to pyrethroid λ-cyhalothrin relative to LAB larvae. RNA-seq identified evidence of metabolic resistance in the head and body tissues: 15 cytochrome P450 transcripts of Cyp6, Cyp9, Cyp4, Cyp304, Cyp307, Cyp337, Cyp321 families, 7 glutathione-S-transferase (Gst) genes, 7 α-esterase genes from intracellular and secreted catalytic classes, and 8 UDP-glucuronosyltransferase (Ugt) were overexpressed in MS as compared with LAB larvae. We also identified overexpression of GPCR genes (CiGPCR64-like and CiGPCRMth2) in the head tissue. To validate RNA-seq results, we performed RT-qPCR to assay selected metabolic genes and confirmed their expression profiles. Specifically, CiCYP9a101v1, CiCYP6ae149, CiCYP6ae106v2, CiGSTe13, CiCOE47, and CiUGT33F21 exhibited significant overexpression in resistant MS larvae. In summary, our findings detailed potential mechanisms of metabolic detoxification underlying pyrethroid resistance in C. includens.

中文翻译:

拟除虫菊酯抗性 Chrysodeixis includens(鳞翅目:夜蛾科)的转录组分析揭示代谢解毒基因的过度表达

Chrysodeixis includens (Walker, [1858]) 是巴西最重要的大豆脱叶剂之一,因为与其他种类的毛虫相比,它具有广泛的地理分布和对杀虫剂的高耐受性。因此,我们进行了生物测定以评估拟除虫菊酯 λ-三氯氟氰菊酯对 C. includens 抗性菌株 (MS) 和易感 (LAB) 实验室菌株的功效。对幼虫头部和身体组织进行高通量 RNA 测序 (RNA-seq),以确定潜在的拟除虫菊酯抗性的分子机制。杀虫剂生物测定表明,MS 幼虫对拟除虫菊酯 λ-三氯氟氰菊酯的抗性是 LAB 幼虫的 28.9 倍。RNA-seq 鉴定了头部和身体组织中代谢抗性的证据:Cyp6、Cyp9、Cyp4、Cyp304、Cyp307、Cyp337 的 15 个细胞色素 P450 转录物,与 LAB 幼虫相比,Cyp321 家族、7 个谷胱甘肽-S-转移酶 (Gst) 基因、7 个来自细胞内和分泌催化类的 α-酯酶基因和 8 个 UDP-葡萄糖醛酸转移酶 (Ugt) 在 MS 中过表达。我们还确定了头部组织中 GPCR 基因(CiGPCR64 样和 CiGPCRMth2)的过度表达。为了验证 RNA-seq 结果,我们进行了 RT-qPCR 来检测选定的代谢基因并确认它们的表达谱。具体而言,CiCYP9a101v1、CiCYP6ae149、CiCYP6ae106v2、CiGSTe13、CiCOE47 和 CiUGT33F21 在抗性 MS 幼虫中表现出显着的过度表达。总而言之,我们的研究结果详细说明了潜在的潜在代谢解毒机制,导致 C. includens 中拟除虫菊酯抗性。和 8 UDP-葡萄糖醛酸转移酶 (Ugt) 与 LAB 幼虫相比在 MS 中过表达。我们还确定了头部组织中 GPCR 基因(CiGPCR64 样和 CiGPCRMth2)的过度表达。为了验证 RNA-seq 结果,我们进行了 RT-qPCR 来检测选定的代谢基因并确认它们的表达谱。具体而言,CiCYP9a101v1、CiCYP6ae149、CiCYP6ae106v2、CiGSTe13、CiCOE47 和 CiUGT33F21 在抗性 MS 幼虫中表现出显着的过度表达。总而言之,我们的研究结果详细说明了潜在的潜在代谢解毒机制,导致 C. includens 中拟除虫菊酯抗性。和 8 UDP-葡萄糖醛酸转移酶 (Ugt) 与 LAB 幼虫相比在 MS 中过表达。我们还确定了头部组织中 GPCR 基因(CiGPCR64 样和 CiGPCRMth2)的过度表达。为了验证 RNA-seq 结果,我们进行了 RT-qPCR 来检测选定的代谢基因并确认它们的表达谱。具体而言,CiCYP9a101v1、CiCYP6ae149、CiCYP6ae106v2、CiGSTe13、CiCOE47 和 CiUGT33F21 在抗性 MS 幼虫中表现出显着的过度表达。总而言之,我们的研究结果详细说明了潜在的潜在代谢解毒机制,导致 C. includens 中拟除虫菊酯抗性。我们进行了 RT-qPCR 来检测选定的代谢基因并确认它们的表达谱。具体而言,CiCYP9a101v1、CiCYP6ae149、CiCYP6ae106v2、CiGSTe13、CiCOE47 和 CiUGT33F21 在抗性 MS 幼虫中表现出显着的过度表达。总而言之,我们的研究结果详细说明了潜在的潜在代谢解毒机制,导致 C. includens 中拟除虫菊酯抗性。我们进行了 RT-qPCR 来检测选定的代谢基因并确认它们的表达谱。具体而言,CiCYP9a101v1、CiCYP6ae149、CiCYP6ae106v2、CiGSTe13、CiCOE47 和 CiUGT33F21 在抗性 MS 幼虫中表现出显着的过表达。总而言之,我们的研究结果详细说明了潜在的潜在代谢解毒机制,导致 C. includens 中拟除虫菊酯抗性。
更新日期:2020-11-02
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