黏虫谷胱甘肽S-转移酶基因对两种杀虫剂的响应
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摘要
谷胱甘肽S-转移酶是昆虫体内重要解毒酶,对昆虫抗药性具有重要作用。通过转录组测序获得黏虫谷胱甘肽S-转移酶cDNA序列,命名为MsGSTe1(GenBank登录号:MH700949)。cDNA全长1 082 bp,包含一个651 bp开放阅读框,编码216个氨基酸,氨基酸等电点为4.75,分子质量为24.679 ku,具有一个GST N端结构域和一个GST C端结构域。经不同剂量氯虫苯甲酰胺处理不同时间后,MsGSTe1基因表达量均显著下调。处理24和48 h,谷胱甘肽S-转移酶活性显著提高。不同剂量高效氯氟氰菊酯处理不同时间后,MsGSTe1表达量和GST酶活性存在显著差异。处理24 h,各剂量处理MsGSTe1基因表达量均显著上调,且酶活性提高。处理48 h,与对照相比,各剂量处理酶活性显著提高,但基因表达量无显著差异,表明MsGSTe1参与黏虫对高效氯氟氰菊酯的解毒代谢。
Glutathione S-transferase was an important detoxifying enzyme in insects. It played an important role in the development of insect resistance. In this study, a novel glutathione S-transferases cDNA sequence named MsGSTe1(GenBank accession No. MH700949) was obtained from Mythimna separata by transcriptome sequencing technology. The full-length c DNA sequence was 1 082 bp, the ORF length of MsGSTe1 was 651 bp and encoded a 216 amino acids sequence, the molecular mass was about 24.679 ku, and the isoelectric point was about 4.75. The amino acids sequence included a GST N-terminal domain and a GST C-terminal domain. After treatment with different doses of chlorantraniliprole for different times, the expression level of MsGSTe1 gene was significantly downregulated. However, glutathione S-transferase activity was significantly increased after 24 and 48 h treatment. After treatment with different doses of lambda-cyhalothrin for different times, the expression level of MsGSTe1 and enzyme activities of GST were significantly different. At 24 h of treatment, the expression levels of MsGSTe1 were significantly up-regulated and the enzyme activity was increased.At 48 h of treatment, the enzyme activity of each treatment was significantly increased compared with that of the control, but there was no significant difference in expression level of MsGSTe1 mRNA. It indicated that MsGSTe1 might play an important role in the detoxification of lambda-cyhalothrin.
Glutathione S-transferase was an important detoxifying enzyme in insects. It played an important role in the development of insect resistance. In this study, a novel glutathione S-transferases cDNA sequence named MsGSTe1(GenBank accession No. MH700949) was obtained from Mythimna separata by transcriptome sequencing technology. The full-length c DNA sequence was 1 082 bp, the ORF length of MsGSTe1 was 651 bp and encoded a 216 amino acids sequence, the molecular mass was about 24.679 ku, and the isoelectric point was about 4.75. The amino acids sequence included a GST N-terminal domain and a GST C-terminal domain. After treatment with different doses of chlorantraniliprole for different times, the expression level of MsGSTe1 gene was significantly downregulated. However, glutathione S-transferase activity was significantly increased after 24 and 48 h treatment. After treatment with different doses of lambda-cyhalothrin for different times, the expression level of MsGSTe1 and enzyme activities of GST were significantly different. At 24 h of treatment, the expression levels of MsGSTe1 were significantly up-regulated and the enzyme activity was increased.At 48 h of treatment, the enzyme activity of each treatment was significantly increased compared with that of the control, but there was no significant difference in expression level of MsGSTe1 mRNA. It indicated that MsGSTe1 might play an important role in the detoxification of lambda-cyhalothrin.
引文
[1]马俊宁,代鲁鲁,张然然,等.华山松大小蠹谷胱甘肽S-转移酶基因DaGSTe1的克隆与表达[J].西北农林科技大学学报:自然科学版, 2015, 43(8):116-122.
[2] Booth J, Boyland E, Sims P. An enzyme from rat liver catalysing conjugations with glutathione[J]. Biochemical Journal, 1961, 79(3):516-524.
[3] Jowsey I R, Flanagan J U, Hayes R D. Glutathione transferases[J].Arabidopsis Book, 2005, 8(45):51-88.
[4] Liu S, Zhang Y X, Wang W L, et al. Identification and characterisation of seventeen glutathione S-transferase genes from the cabbage white butterfly Pieris rapae[J]. Pesticide Biochemistry and Physiology, 2017, 143(11):102-110.
[5] Ding Y, Ortelli F, Rossiter L C, et al. The Anopheles gambiae glutathione transferase supergene family:Annotation, phylogeny and expression profiles[J]. Bmc Genomics, 2003, 4(1):35-51.
[6] Friedman R. Genomic organization of the glutathione S-transferase family in insects[J]. Molecular Phylogenetics and Evolution,2011, 61(3):924-932.
[7]李帅,苏丽,李伯辽,等.梨小食心虫谷胱甘肽S-转移酶GmolGST6的基因克隆、原核表达和酶学特征[J].昆虫学报,2018, 61(4):398-409.
[8]左伟东,余泉友,李斌,等.家蚕谷胱甘肽转移酶基因的克隆、序列分析及其表达模式[J].农业生物技术学报, 2007, 15(4):595-601.
[9] Enayati A A, Ranson H, Hemingway J. Insect glutathione transferases and insecticide resistance[J]. Insect Molecular Biology,2005, 14(1):3-8.
[10] Ivarsson Y, Mackey A J, Edalat M, et al. Identification of residues in glutathione transferase capable of driving functional diversification in evolution. A novel approach to protein redesign[J]. Journal of Biological Chemistry, 2003, 278(10):8733-8738
[11] Brugger K E, Cole P G, Newman I C, et al. Selectivity of chlorantraniliprole to parasitoid wasps[J]. Pest Management Science,2010, 66(10):1075-1081.
[12]唐振华.新型二酰胺类杀虫剂对鱼尼丁受体作用的分子机理[J].昆虫学报, 2008, 51(6):646-651.
[13]沈登荣,何超,宋文菲,等.高效氯氟氰菊酯亚致死浓度对番石榴实蝇代谢酶的影响[J].西南师范大学学报:自然科学版,2015(8):58-61.
[14]吕敏,刘惠霞,吴文君.谷胱甘肽S-转移酶与昆虫抗药性的关系[J].应用昆虫学报, 2003, 40(3):204-207.
[15] Lumjuan N, Stevenson B J, Prapanthadara L A, et al. The Aedes aegypti glutathione transferase family[J]. Insect Biochemistry and Molecular Biology, 2007, 37(10):1026-1035.
[16] You Y, Xie M, Ren N, et al. Characterization and expression profiling of glutathione S-transferases in the diamondback moth,Plutella xylostella(L.)[J]. Bmc Genomics, 2015, 16(1):1-13.
[17] Zhou W W, Liang Q M, Xu Y, et al. Genomic insights into the glutathione S-transferase gene family of two rice planthoppers,Nilaparvata lugens and Sogatella furcifera(Horváth)(Hemiptera:Delphacidae)[J]. PloS One, 2013, 8(2):e56604.
[18]金燕璐,张邦贤,林华峰.二化螟谷胱甘肽S-转移酶基因的鉴定与表达模式分析[J].应用昆虫学报, 2018, 55(3):349-359.
[19] Han J B, Li G Q, Wan P J, et al. Identification of glutathione Stransferase genes in Leptinotarsa decemlineata and their expression patterns under stress of three insecticides[J]. Pesticide Biochemistry and Physiology, 2016, 133(10):26-34.
[20] Chen X, Zhang Y L. Identification and characterization of multiple glutathione S-transferase genes from the diamondback moth,Plutella xylostella[J]. Pest Management Science, 2015, 71(4):592-600.
[2] Booth J, Boyland E, Sims P. An enzyme from rat liver catalysing conjugations with glutathione[J]. Biochemical Journal, 1961, 79(3):516-524.
[3] Jowsey I R, Flanagan J U, Hayes R D. Glutathione transferases[J].Arabidopsis Book, 2005, 8(45):51-88.
[4] Liu S, Zhang Y X, Wang W L, et al. Identification and characterisation of seventeen glutathione S-transferase genes from the cabbage white butterfly Pieris rapae[J]. Pesticide Biochemistry and Physiology, 2017, 143(11):102-110.
[5] Ding Y, Ortelli F, Rossiter L C, et al. The Anopheles gambiae glutathione transferase supergene family:Annotation, phylogeny and expression profiles[J]. Bmc Genomics, 2003, 4(1):35-51.
[6] Friedman R. Genomic organization of the glutathione S-transferase family in insects[J]. Molecular Phylogenetics and Evolution,2011, 61(3):924-932.
[7]李帅,苏丽,李伯辽,等.梨小食心虫谷胱甘肽S-转移酶GmolGST6的基因克隆、原核表达和酶学特征[J].昆虫学报,2018, 61(4):398-409.
[8]左伟东,余泉友,李斌,等.家蚕谷胱甘肽转移酶基因的克隆、序列分析及其表达模式[J].农业生物技术学报, 2007, 15(4):595-601.
[9] Enayati A A, Ranson H, Hemingway J. Insect glutathione transferases and insecticide resistance[J]. Insect Molecular Biology,2005, 14(1):3-8.
[10] Ivarsson Y, Mackey A J, Edalat M, et al. Identification of residues in glutathione transferase capable of driving functional diversification in evolution. A novel approach to protein redesign[J]. Journal of Biological Chemistry, 2003, 278(10):8733-8738
[11] Brugger K E, Cole P G, Newman I C, et al. Selectivity of chlorantraniliprole to parasitoid wasps[J]. Pest Management Science,2010, 66(10):1075-1081.
[12]唐振华.新型二酰胺类杀虫剂对鱼尼丁受体作用的分子机理[J].昆虫学报, 2008, 51(6):646-651.
[13]沈登荣,何超,宋文菲,等.高效氯氟氰菊酯亚致死浓度对番石榴实蝇代谢酶的影响[J].西南师范大学学报:自然科学版,2015(8):58-61.
[14]吕敏,刘惠霞,吴文君.谷胱甘肽S-转移酶与昆虫抗药性的关系[J].应用昆虫学报, 2003, 40(3):204-207.
[15] Lumjuan N, Stevenson B J, Prapanthadara L A, et al. The Aedes aegypti glutathione transferase family[J]. Insect Biochemistry and Molecular Biology, 2007, 37(10):1026-1035.
[16] You Y, Xie M, Ren N, et al. Characterization and expression profiling of glutathione S-transferases in the diamondback moth,Plutella xylostella(L.)[J]. Bmc Genomics, 2015, 16(1):1-13.
[17] Zhou W W, Liang Q M, Xu Y, et al. Genomic insights into the glutathione S-transferase gene family of two rice planthoppers,Nilaparvata lugens and Sogatella furcifera(Horváth)(Hemiptera:Delphacidae)[J]. PloS One, 2013, 8(2):e56604.
[18]金燕璐,张邦贤,林华峰.二化螟谷胱甘肽S-转移酶基因的鉴定与表达模式分析[J].应用昆虫学报, 2018, 55(3):349-359.
[19] Han J B, Li G Q, Wan P J, et al. Identification of glutathione Stransferase genes in Leptinotarsa decemlineata and their expression patterns under stress of three insecticides[J]. Pesticide Biochemistry and Physiology, 2016, 133(10):26-34.
[20] Chen X, Zhang Y L. Identification and characterization of multiple glutathione S-transferase genes from the diamondback moth,Plutella xylostella[J]. Pest Management Science, 2015, 71(4):592-600.