四川省内江市白纹伊蚊野外群体电压门控钠离子通道基因突变检测分析

doi:10.11853/j.issn.1003.8280.2023.03.006

摘要/Abstract

摘要： 目的通过对四川省内江市3个白纹伊蚊野外群体电压门控钠离子通道（voltage-gated sodium channel,VGSC）基因检测分析,在分子水平上了解白纹伊蚊对拟除虫菊酯类杀虫剂的抗药性现状。方法 2021年6－7月,在内江市3个县（市、区）白纹伊蚊孳生地使用诱蚊灯法采集野外成蚊,鉴定为白纹伊蚊后,提取基因组DNA,扩增VGSC基因片段,检测VGSC基因抗药性相关位点（1016、1532和1534）的突变类型及其分布频率。结果共收集并成功测序107只白纹伊蚊,检测到1016G突变的有28只,占26.17%,发生1532T突变的有6只,占5.61%,1534位点发生突变（1534C/S）的共100只,占93.46%。对1016、1532、1534位点组合分析发现,仅1个位点为突变型的个体为76个（73.08%）,2个位点为突变型的个体共25个（24.04%）,其中1016位点为野生型、1532和1534位点同时突变的个体为2个（1.92%）,1532位点为野生型、1016和1534位点同时突变的个体为22个（21.15%）,1534位点为野生型、1532和1016位点同时突变的个体共1个（0.96%）。本次现场未发现3个位点同时突变的个体。结论内江市3个白纹伊蚊野外种群存在较高频率的拟除虫菊酯类杀虫剂靶标突变,其中1534位点抗性频率较高,1534S是优势抗性等位基因,提示在化学防治蚊虫时,宜减少拟除虫菊酯类杀虫剂的使用,以延缓抗药性的产生,提高蚊媒控制效果。

关键词: 白纹伊蚊, 突变, 电压门控钠离子通道

Abstract: Objective To investigate the current resistance status of Aedes albopictus to pyrethroid insecticides at the molecular level through the voltage-gated sodium channel (VGSC) genes testing in three field populations of Ae. albopictus in Neijiang, Sichuan province, China.Methods From June to July in 2021, adult mosquitoes were collected by light trapping method in the Ae. albopictus breeding sites of three counties/cities/districts in Neijiang. For mosquitoes identified as Ae. albopictus, the genomic DNA was extracted to amplify VGSC gene fragment using PCR, followed by detection of mutation types and distribution frequencies at VGSC gene resistance-related loci (loci 1016, 1532, and 1534).Results A total of 107 Ae. albopictus mosquitoes were collected and sequenced successfully. Twenty-eight (26.17%) mosquitoes were detected with the 1016G mutation, six (5.61%) with the 1532T mutation, and 100 (93.46%) with the 1534C/S mutations. The combination analysis of the three loci revealed that 76 (73.08%) mosquitoes had mutations at only one locus, and 25 (24.04%) at two loci. There were two (1.92%) mosquitoes with the wild type at 1016 and mutations at both 1532 and 1534, 22 (21.15%) with the wild type at 1532 and mutations at both 1016 and 1534, and one (0.96%) with the wild type at 1534 and mutations at both 1532 and 1016. None had simultaneous mutations at all the three loci.Conclusions The three field Ae. albopictus populations in Neijiang had frequent pyrethroid target-site mutations, with a high frequency of resistance at locus 1534, where 1534S was the dominant resistance allele, suggesting that the use of pyrethroids should be reduced in chemical control to delay resistance and improve mosquito control.

Key words: Aedes albopictus, Mutation, Voltage-gated sodium channel

中图分类号:

R384.1

谭爱, 刘鹃, 王雅伟, 曾佳芮, 刘鹏. 四川省内江市白纹伊蚊野外群体电压门控钠离子通道基因突变检测分析[J]. 中国媒介生物学及控制杂志, 2023, 34(3): 314-318.

TAN Ai, LIU Juan, WANG Ya-wei, ZENG Jia-rui, LIU Peng. Detection of mutations in the voltage-gated sodium channel genes of field Aedes albopictus populations in Neijiang, Sichuan province, China[J]. Chinese Journal of Vector Biology and Control, 2023, 34(3): 314-318.

参考文献

[1] 赵春春. 我国白纹伊蚊抗药性及kdr基因分布研究[D]. 北京:中国疾病预防控制中心,2019.Zhao CC. Study on insectcides resistance and kdr gene distribution of Aedes albopictus in China[D]. Beijing:Chinese Center for Disease Control and Prevention,2019. (in Chinese)
[2] 王明强,徐勇,刘鹃,等. 内江市建成区白纹伊蚊幼虫对5种杀虫剂的抗药性调查[J]. 中华卫生杀虫药械,2018,24(1):99-100. DOI:10.19821/j.1671-2781.2018.01.032.Wang MQ,Xu Y,Liu J,et al. Resistance of Aedes albopictus larvae to five insecticides in build-up area district of Neijiang city[J]. Chin J Hyg Insect Equip,2018,24(1):99-100. DOI:10.19821/j.1671-2781.2018.01.032.(in Chinese)
[3] 姜陆,邹玉梅,高玉峰. 蚊虫抗药性研究主要进展[J]. 口岸卫生控制,2019,24(6):59-62. DOI:10.3969/j.issn.1008-5777. 2019.06.018.Jiang L,Zou YM,Gao YF. Major advances in mosquito resistance research[J]. Port Health Control,2019,24(6):59-62. DOI:10.3969/j.issn.1008-5777.2019.06.018.(in Chinese)
[4] 邱星辉. 白纹伊蚊抗药性分子机制研究进展[J]. 寄生虫与医学昆虫学报,2019,26(3):194-198. DOI:10.3969/j.issn.1005-0507.2019.03.009.Qiu XH. Current knowledge about the molecular mechanisms underlying insecticide resistance in Aedes albopictus[J]. Acta Parasitol Med Entomol Sin,2019,26(3):194-198. DOI:10.3969/j.issn.1005-0507.2019.03.009.(in Chinese)
[5] 王冠熙,李雅姝,李月月,等. 江苏省白纹伊蚊对溴氰菊酯抗性及击倒抗性突变分析[J]. 中国寄生虫学与寄生虫病杂志,2022,40(4):468-474. DOI:10.12140/j.issn.1000-7423.2022. 04.008.Wang GX,Li YS,Li YY,et al. Resistance to deltamethrin and knockdown resistance mutation in Aedes albopictus from Jiangsu province[J]. Chin J Parasitol Parasit Dis,2022,40(4):468-474. DOI:10.12140/j.issn.1000-7423.2022.04.008.(in Chinese)
[6] 曹一鸥,吕强,张佳珂,等. 2019年四川省首例登革热本地病例调查与疫区处置情况分析[J]. 预防医学情报杂志,2020,36(11):1499-1503.Cao YO,Lyu Q,Zhang JK,et al. Investigation and disposal of the first local case of dengue fever in Sichuan province[J]. J Prev Med Inf,2020,36(11):1499-1503. (in Chinese)
[7] 姬淑红,张杰,陆崇华,等. 上海市杨浦区白纹伊蚊幼虫对5种常用杀虫剂的抗药性调查[J]. 中华卫生杀虫药械,2017,23(4):335-336. DOI:10.19821/j.1671-2781.2017.04.012.Ji SH,Zhang J,Lu CH,et al. Resistance investigation of Aedes albopictus to five kinds of common used insecticides in Yangpu district of Shanghai[J]. Chin J Hyg Insect Equip,2017,23(4):335-336. DOI:10.19821/j.1671-2781.2017.04.012.(in Chinese)
[8] 古灵满,郭建宇,杨雪琴,等. 2020年中山市区白纹伊蚊抗药性调查[J]. 现代预防医学,2021,48(17):3097-3100.Gu LM,Guo JY,Yang XQ,et al. Investigation on insecticide resistance of Aedes albopictus in Zhongshan city,2020[J]. Mod Prev Med,2021,48(17):3097-3100. (in Chinese)
[9] 张要锋,梅树林,齐振文. 2020年焦作市某县白纹伊蚊对常用卫生杀虫剂抗药性调查[J]. 河南预防医学杂志,2022,33(4):291-293. DOI:10.13515/j.cnki.hnjpm.1006-8414.2022.04.014.Zhang YF,Mei SL,Qi ZW. Investigation of Aedes albopictus resistance to sanitary insecticides in Jiaozuo,2020[J]. Henan J Prev Med,2022,33(4):291-293. DOI:10.13515/j.cnki.hnjpm.1006-8414.2022.04.014.(in Chinese)
[10] 陈翰明. 我国白纹伊蚊群体遗传结构和击倒抗性突变检测研究[D]. 上海:海军军医大学,2019.Chen HM. Detection of genetic structure and knockdown resistance mutations in Aedes albopictus (Diptera:Culicidae) populations from China[D]. Shanghai:Naval Medical University,2019. (in Chinese)
[11] 朱彩英,赵春春,伦辛畅,等. 云南省景洪市2018-2019年白纹伊蚊击倒抗性基因型分布研究[J]. 中国媒介生物学及控制杂志,2020,31(1):7-11. DOI:10.11853/j.issn.1003.8280. 2020.01.002.Zhu CY,Zhao CC,Lun XC,et al. Distribution of knockdown resistance genotypes in Aedes albopictus in Jinghong,Yunnan province,China,2018-2019[J]. Chin J Vector Biol Control,2020,31(1):7-11. DOI:10.11853/j.issn.1003.8280.2020. 01.002.(in Chinese)
[12] 周小洁,赵宇晗,刘念,等. 北京市两个白纹伊蚊种群kdr抗性突变检测分析[J]. 中华卫生杀虫药械,2021,27(4):304-307. DOI:10.19821/j.1671-2781.2021.04.003.Zhou XJ,Zhao YH,Liu N,et al. Detection and analysis of kdr resistance mutations in two populations of Aedes albopictus in Beijing[J]. Chin J Hyg Insect Equip,2021,27(4):304-307. DOI:10.19821/j.1671-2781.2021.04.003.(in Chinese)
[13] 陈翰明,高景鹏,姜进勇,等. 我国白纹伊蚊现场群体击倒抗性基因I1532和F1534突变检测及I1532T突变等位基因报告[J]. 中国媒介生物学及控制杂志,2018,29(2):120-125. DOI:10.11853/j.issn.1003.8280.2018.02.002.Chen HM,Gao JP,Jiang JY,et al. Detection of the I1532 and F1534 kdr mutations and a novel mutant allele I1532T in VGSC gene in the field populations of Aedes albopictus from China[J]. Chin J Vector Biol Control,2018,29(2):120-125. DOI:10.11853/j.issn.1003.8280.2018.02.002.(in Chinese)
[14] 刘鲁宏. 济宁市白纹伊蚊kdr基因突变检测[J]. 中华卫生杀虫药械,2022,28(3):267-269. DOI:10.19821/j.1671-2781.2022. 03.020.Liu LH. Detection of knockdown resistance gene mutation in Aedes albopictus of Jining city[J]. Chin J Hyg Insect Equip,2022,28(3):267-269. DOI:10.19821/j.1671-2781.2022.03.020.(in Chinese)
[15] Chen HY,Li KL,Wang XH,et al. First identification of kdr allele F1534S in VGSC gene and its association with resistance to pyrethroid insecticides in Aedes albopictus populations from Haikou city,Hainan island,China[J]. Infect Dis Poverty,2016,5:31.DOI:10.1186/s40249-016-0125-x.
[16] Guo YJ,Zhou JN,Zhao YJ,et al. CRISPR/Cas9-mediated F1534S substitution in the voltage-gated sodium channel reveals its necessity and sufficiency for deltamethrin resistance in Aedes albopictus[J] J Pest Sci,2022(96):1173-1186. DOI:10.1007/s10340-022-01557-6.
[17] Zhu CY,Zhao CC,Wang YG,et al. Establishment of an innovative and sustainable PCR technique for 1534 locus mutation of the knockdown resistance (kdr) gene in the dengue vector Aedes albopictus[J]. Parasit Vectors,2019,12(1):603. DOI:10.1186/s13071-019-3829-5.
[18] 师灿南. 景洪市登革热媒介伊蚊对常用杀虫剂的抗药性及机制初步研究[D]. 北京:中国疾病预防控制中心,2017.Shi CN. Insecticides resistance and its underlying mechanisms for dengue vectors Aedes albopictus and Ae. aegypti in Jinghong city,Yunnan province[D]. Beijing:Chinese Center for Disease Control and Prevention,2017. (in Chinese)