Testing two methods to detect voltage-gated sodium channels gene mutations in Tetranychus urticae

doi:10.11686/cyxb20140517

Abstract

Abstract:

Target mutation produces high levels of resistance to pyrethroid insecticides in the spider mite Tetranychus urticae. However, as there are many techniques for detecting target mutations it would be advantageous to identify methods that are both economical and effective. Three mutations (L1022V, A1215D and F1538I) have been identified in the voltage-gated sodium channels (VGSCs) of T. urticae. We have tested the ability of two different methods, PCR product direct sequencing and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), to detect these VGSC gene mutations in two field strains (WW-R and LZ-R) of T. urticae. The results of PCR product direct sequencing showed that the A1215D and F1538I mutations existed in the VGSC genes of both the WW-R and LZ-R strains, but that the F1022V mutation was not detected. The A1215D mutation was located between domains IIS6 and IIIS1 of the VGSCs, while the F1538I mutation was identified in domain IIIS6. The PCR-RFLP method, however, proved able to identify only the A1215D mutation. Our testing of these two methods shows that detecting VGSC mutations in field strains of T. urticae has the potential to develop molecular-based monitoring that can be used to manage the species’ resistance to pyrethroid insecticides.

CLC Number:

S433.7

YANG Shun-yi,YUE Xiu-li,WANG Jin-jun,SHEN Hui-min,GUO Jin-mei,SHEN Yi-fan,ZHOU Xing-long. Testing two methods to detect voltage-gated sodium channels gene mutations in Tetranychus urticae[J]. Acta Prataculturae Sinica, 2014, 23(5): 153-160.

References

Reference: 

[1] Grbic M, Van Leeuwen T, Clark R M, et al. The genome of Tetranychus urticae reveals herbivorous pest adaptations[J]. Nature, 2011, 479: 487-492. 

[2] Zhang T W, Shen H M, Qian X J, et al. Effects of Tetranychus urticae feeding on the chlorophyll content and two kinds of protective enzyme of white clover[J]. Chinese Bulletin of Entomology, 2013, 50(2): 395-400. 

[3] He D H, Zhao X P, Jin Q H, et al. Dispersion of two-spotied spider mite, Tetranychus Urticae Koch, and its selection of host plants on farmland in ningxia[J]. Chinese Journal of Applied & Environment Biology, 2001, 7(5): 447-451. 

[4] Croft B A, Vandebaan H E. Ecological and genetic factors influencing evolution of pesticide resistance in tetranychid and phytoseiid mites[J]. Experimental and Applied Acarology, 1988, 4(3): 277-300. 

[5] Soderlund D M. Pyrethroids, knockdown resistance and sodium channels[J]. Pest Management Science, 2008, 64(6): 610-616. 

[6] Tsagkarakou A, Van Leeuwen T, Khajehali J, et al. Identification of pyrethroid resistance associated mutations in the para sodium channel of the two spotted spider mite Tetranychus urticae (Acari: Tetranychidae)[J]. Insect Molecular Biology, 2009, 18(5): 583-593. 

[7] Kwon D H, Clark J M, Lee S H. Cloning of a sodium channel gene and identification of mutations putatively associated with fenpropathrin resistance in Tetranychus urticae[J]. Pesticide Biochemistry and Physiology, 2010, 97(2): 93-100. 

[8] Feng Y N, Zhao S, Sun W, et al. The sodium channel gene in Tetranychus cinnabarinus (Boisduval): identification and expression analysis of a mutation associated with pyrethroid resistance[J]. Pest Management Science, 2011, 67(8): 904-912. 

[9] Wang R W, Liu Z Q, Dong K, et al. Association of novel mutations in a sodium channel gene with fluvalinate resistance in the mite, Varroa destructor[J]. Journal of Apicultural Research, 2002, 41(12): 17-25. 

[10] Liu Z, Tan J, Huang Z Y, et al. Effect of a fluvalinate resistance associated sodium channel mutation from varroa mites on cockroach sodium channel sensitivity to fluvalinate, a pyrethroid insecticide[J]. Insect Biochemistry and Molecular Biology, 2006, 36(11): 885-889. 

[11] Ingles P J, Adams P M, Knipple D C, et al. Characterization of voltage sensitive sodium channel gene coding sequences from insecticide susceptible and knockdown resistant house fly strains[J]. Insect Biochemistry and Molecular Biology, 1996, 26(4): 319-326. 

[12] Williamson M S, Martinez Torres D, Hick C A, et al. Identification of mutations in the housefly para type sodium channel gene associated with knockdown resistance (kdr) to pyrethroid insecticides[J]. Molecular & General Genetics, 1996, 252(12): 51-60. 

[13] Rinkevich F D, Leichter C A, Lazo T A, et al. Variable fitness costs for pyrethroid resistance alleles in the house fly, Musca domestica, in the absence of insecticide pressure[J]. Pesticide Biochemistry and Physiology, 2013, 105: 161-168. 

[14] Cao X M, Zhao T Y. Molecule-based techniques for the detection of insecticde resistance in pests[J]. Acta Parasitologica et Medica Entomologica Sinica, 2006, 13(1): 57-63. 

[15] Gao M, Shen R P, Zhang P, et al. Detection of beta-cypermethrin resistance in Shandong populations of Spodoptera exigua by competitive PCR amplification of specific allele[J]. Acta Phytophylacica Sinica, 2013, 40(4): 363-368. 

[16] Zhao C J, Li N, Deng X M. The establishment of method for identifying SNP genotype by CRS-PCR[J]. Hereditas(Beijing), 2003, 25(3): 327-329. 

[17] Liao X Y, Zhang Y F, Gu X F. Technique of PCR-ACRS for the detection of CYP21 gene mutations[J]. Chinese Journal of Medical Genetics, 2003, 20(5): 449-451. 

[18] Van Leeuwen T, Vontas J, Tsagkarakou A, et al. Acaricide resistance mechanisms in the two spotted spider mite Tetranychus urticae and other important Acari: A review[J]. Insect Biochemistry and Molecular Biology, 2010, 40(8): 563-572. 

[19] He H, Chen A C, Davey R B, et al. Identification of a point mutation in the para type sodium channel gene from a pyrethroid-resistant cattle tick[J]. Biochemical and Biophysical Research Communications, 1999, 261(3): 558-561. 

[20] Tan J G, Liu Z Q, Wang R W, et al. Identification of amino acid residues in the insect sodium channel critical for pyrethroid binding[J]. Molecular Pharmacology, 2005, 67(2): 513-522. 

[21] O’Reilly A O, Khambay B P, Williamson M S, et al. Modelling insecticide binding sites in the voltage gated sodium channel[J]. Biochemical Journal, 2006, 396(2): 255-263. 

[22] Davies T G E, O'Reilly A O, Field L M, et al. Knockdown resistance to DDT and pyrethroids: from target-site mutations to molecular modelling[J].Pest Management Science, 2008, 64(11): 1126-1130. 

[23] Gao X J. Study on resistance mechanisms and enzymology resistance monitoring of Tetranychus urticae koch to fenpropathrin[D]. Lanzhou: Gansu Agricultural University, 2012. 

[24] Van Leeuwen T, Dermauw W, Grbic M, et al. Spider mite control and resistance management: does a genome help?[J] Pest Management Science, 2013, 69(2): 156-159. 

[25] Wybouw N, Balabanidou V, Ballhorn D J, et al. A horizontally transferred cyanase gene in the spider mite Tetranychus urticae is involved in cyanate metabolism and is differentially expressed upon host plant change[J]. Insect Biochemistry and Molecular Biology, 2012, 42(12): 881-889. 

[26] Dermauw W, Wybouw N, Rombauts S, et al. A link between host plant adaptation and pesticide resistance in the polyphagous spider mite Tetranychus urticae[J]. Proceedings of the National Academy of Sciences of the United States of America, 2013, 110(2): 113-122. 

[27] Whittall R A, Scartezini M, Li K, et al. Development of a high-resolution melting method for mutation detection in familial hyperch olesterolaemia patients[J]. Annals of Clinical Biochemistry, 2010, 47: 44-55. 

[28] Xiong L, Liang C F, Chen C, et al. Application of qPCR-HRM curve analysis and common-PCR with DNA sequencing in detection of EGFR mutations in glioma tissues[J]. New Medicine, 2012, 43(5): 327-330. 

参考文献：

[1] Grbic M, Van Leeuwen T, Clark R M, et al. The genome of Tetranychus urticae reveals herbivorous pest adaptations[J]. Nature, 2011, 479: 487-492. 

[2] 张廷伟, 沈慧敏, 钱秀娟, 等. 二斑叶螨刺吸胁迫对白三叶叶绿素含量和两种保护酶的影响[J]. 应用昆虫学报, 2013, 50(2): 395-400. 

[3] 贺达汉, 赵晓萍, 靳巧红, 等. 宁夏地区二斑叶螨的寄主植物选择及其季节转移[J]. 环境与应用生物学报, 2001, 7(5): 447-451. 

[4] Croft B A, Vandebaan H E. Ecological and genetic-factors influencing evolution of pesticide resistance in tetranychid and phytoseiid mites[J]. Experimental and Applied Acarology, 1988, 4(3): 277-300. 

[5] Soderlund D M. Pyrethroids, knockdown resistance and sodium channels[J]. Pest Management Science, 2008, 64(6): 610-616. 

[6] Tsagkarakou A, Van Leeuwen T, Khajehali J, et al. Identification of pyrethroid resistance associated mutations in the para sodium channel of the two-spotted spider mite Tetranychus urticae (Acari: Tetranychidae)[J]. Insect Molecular Biology, 2009, 18(5): 583-593. 

[7] Kwon D H, Clark J M, Lee S H. Cloning of a sodium channel gene and identification of mutations putatively associated with fenpropathrin resistance in Tetranychus urticae[J]. Pesticide Biochemistry and Physiology, 2010, 97(2): 93-100. 

[8] Feng Y N, Zhao S, Sun W, et al. The sodium channel gene in Tetranychus cinnabarinus (Boisduval): identification and expression analysis of a mutation associated with pyrethroid resistance[J]. Pest Management Science, 2011, 67(8): 904-912. 

[9] Wang R W, Liu Z Q, Dong K, et al. Association of novel mutations in a sodium channel gene with fluvalinate resistance in the mite, Varroa destructor[J]. Journal of Apicultural Research, 2002, 41(1-2): 17-25. 

[10] Liu Z, Tan J, Huang Z Y, et al. Effect of a fluvalinate-resistance-associated sodium channel mutation from varroa mites on cockroach sodium channel sensitivity to fluvalinate, a pyrethroid insecticide[J]. Insect Biochemistry and Molecular Biology, 2006, 36(11): 885-889. 

[11] Ingles P J, Adams P M, Knipple D C, et al. Characterization of voltage-sensitive sodium channel gene coding sequences from insecticide-susceptible and knockdown-resistant house fly strains[J]. Insect Biochemistry and Molecular Biology, 1996, 26(4): 319-326. 

[12] Williamson M S, Martinez-Torres D, Hick C A, et al. Identification of mutations in the housefly para-type sodium channel gene associated with knockdown resistance (kdr) to pyrethroid insecticides[J]. Molecular & General Genetics, 1996, 252(1-2): 51-60. 

[13] Rinkevich F D, Leichter C A , Lazo T A, et al. Variable fitness costs for pyrethroid resistance alleles in the house fly, Musca domestica, in the absence of insecticide pressure[J]. Pesticide Biochemistry and Physiology, 2013, 105: 161-168. 

[14] 曹晓梅, 赵彤言. 害虫抗药性分子检测技术发展现状[J]. 寄生虫与医学昆虫学报, 2006, 13(1): 57-63. 

[15] 高明, 申瑞平, 张鹏, 等. 竞争性特异等位基因PCR检测山东甜菜夜蛾对高效氯氰菊酯的抗性[J]. 植物保护学报, 2013, 40(4): 363-368. 

[16] 赵春江, 李宁, 邓学梅. 应用创造酶切位点法检测单碱基突变[J]. 遗传, 2003, 25(3): 327-329. 

[17] 廖相云, 张雅芬, 顾学范. 扩增引进限制性酶切位点技术对两种CYP21基因突变的检测[J]. 中华医学遗传学杂志, 2003, 20(5): 449-451. 

[18] Van Leeuwen T, Vontas J, Tsagkarakou A, et al. Acaricide resistance mechanisms in the two-spotted spider mite Tetranychus urticae and other important Acari: A review[J]. Insect Biochemistry and Molecular Biology, 2010, 40(8): 563-572. 

[19] He H, Chen A C, Davey R B, et al. Identification of a point mutation in the para-type sodium channel gene from a pyrethroid-resistant cattle tick[J]. Biochemical and Biophysical Research Communications, 1999, 261(3): 558-561. 

[20] Tan J G, Liu Z Q, Wang R W, et al. Identification of amino acid residues in the insect sodium channel critical for pyrethroid binding[J]. Molecular Pharmacology, 2005, 67(2): 513-522. 

[21] O’Reilly A O, Khambay B P, Williamson M S, et al. Modelling insecticide-binding sites in the voltage-gated sodium channel[J]. Biochemical Journal, 2006, 396(2): 255-263. 

[22] Davies T G E, O’Reilly A O, Field L M, et al. Knockdown resistance to DDT and pyrethroids: from target-site mutations to molecular modelling[J]. Pest Management Science, 2008, 64(11): 1126-1130. 

[23] 高新菊. 二斑叶螨对甲氰菊酯的抗性分子机理研究[D]. 兰州: 甘肃农业大学, 2012. 

[24] Van Leeuwen T, Dermauw W, Grbic M, et al. Spider mite control and resistance management: does a genome help?[J] Pest Management Science, 2013, 69(2): 156-159. 

[25] Wybouw N, Balabanidou V, Ballhorn D J, et al. A horizontally transferred cyanase gene in the spider mite Tetranychus urticae is involved in cyanate metabolism and is differentially expressed upon host plant change[J]. Insect Biochemistry and Molecular Biology, 2012, 42(12): 881-889. 

[26] Dermauw W, Wybouw N, Rombauts S, et al. A link between host plant adaptation and pesticide resistance in the polyphagous spider mite Tetranychus urticae[J]. Proceedings of the National Academy of Sciences of the United States of America, 2013, 110(2): 113-122. 

[27] Whittall R A, Scartezini M, Li K, et al. Development of a high-resolution melting method for mutation detection in familial hyperch -olesterolaemia patients[J]. Annals of Clinical Biochemistry, 2010, 47: 44-55. 

[28] 熊亮, 梁朝峰, 陈川, 等. qPCR-HRM曲线分析技术与普通PCR加直接测序法检测胶质瘤EGFR突变比较[J]. 新医学, 2012, 43(5): 327-330.