不同栖息环境的两种啮齿动物Vkorc1基因多态性

doi:10.16829/j.slxb.150692

兽类学报 ›› 2022, Vol. 42 ›› Issue (6): 687-697.DOI: 10.16829/j.slxb.150692

不同栖息环境的两种啮齿动物Vkorc1基因多态性

杨新根¹^,², 王艳龙³, 邹波¹, 常文英¹, 侯玉¹, 赵悠悠⁴, 王庭林¹(), 张健旭⁵^,⁶()

^1.山西农业大学植物保护学院，农业有害生物综合治理山西省重点实验室，太原 030031
^2.山西大学生命科学学院，应用生物学研究所，太原 030006
^3.定襄县林草事务中心，定襄 035400
^4.太原海关技术中心，临汾 043000
^5.中国科学院动物研究所，农业虫害鼠害综合治理研究国家重点实验室，北京 100101
^6.中国科学院大学生物互作卓越创新中心，北京 100049

收稿日期:2022-05-23 接受日期:2022-08-29 出版日期:2022-11-30 发布日期:2022-12-02
通讯作者: 王庭林,张健旭
作者简介:杨新根 (1974- )，男，副研究员，主要从事啮齿动物研究.
基金资助:
科技基础资源调查专项(2019FY100300);农业虫害鼠害综合治理研究国家重点实验室开放课题(ChineseIPM1618);北京自然基金(5192015);中国科学院STS项目(KFJ-STS-ZDTP-2021-002);山西农业大学学术恢复科研专项(2020xshf09)

Vkorc1 polymorphism of two rodent species in different habitats

Xingen YANG¹^,², Yanlong WANG³, Bo ZOU¹, Wenying CHANG¹, Yu HOU¹, Youyou ZHAO⁴, Tinglin WANG¹(), Jianxu ZHANG⁵^,⁶()

^1.College of Plant Protection, Shanxi Agricultural University, Shanxi Key Laboratory of Integrated Pest Management in Agriculture, Taiyuan 030031, China
^2.Institute of Applied Biology, School of Life Science, Shanxi University, Taiyuan 030006, China
^3.Forestry and Grass Affairs Center of Dingxiang County, Dingxiang 035400, China
^4.Technical Center of Taiyuan Customs District, Linfen 043000, China
^5.Institute of Zoology, Chinese Academy of Sciences, The State Key Laboratory of Integrated Management of Pest Insects and Rodents, Beijing 100101, China
^6.CAS Center for Excellence in Biotic Interactions, University of Chinese Academy of Sciences, Beijing 100049, China

Received:2022-05-23 Accepted:2022-08-29 Online:2022-11-30 Published:2022-12-02
Contact: Tinglin WANG,Jianxu ZHANG

摘要/Abstract

摘要：

维生素K环氧化物还原酶复合物亚基 1基因 (Vkorc1) 的变异与啮齿动物对抗凝血灭鼠剂的抗药性密切相关。为掌握Vkorc1基因变异在野栖类和家栖类啮齿动物中的流行情况，从山西省13个县 (市、区) 的农田和14个县 (市、区) 的养殖场采样，检测长尾仓鼠 (Cricetulus longicaudatus) 和黄胸鼠 (Rattus tanezumi) Vkorc1基因编码区的变异位点及携带不同变异位点的个体的分布情况。结果显示，长尾仓鼠在13个采样地均有捕获，整体占野栖类啮齿动物的23.29%；黄胸鼠分布于8个采样地，整体占家栖类啮齿动物的68.63%。在长尾仓鼠样本 (n = 105) 中检测到6个沉默突变位点和5个错义突变位点，其中，沉默突变C438T (His146His) 的变异率最高，为67.62%；共有17只长尾仓鼠样本存在错义突变位点。黄胸鼠样本 (n = 70) 中存在6个沉默突变位点和1个错义突变位点，其中，最常见的沉默突变位点A321C (Ile107Ile) 和T411C (Thr137Thr) 的变异率均达到18.57%；8只黄胸鼠样本存在与其抗药性相关的A416G (Tyr139Cys) 错义突变，其中7只来源于太原市小店区 (XID)，变异率为35.00%。本研究表明在家居和自然环境中均存在Vkorc1基因的变异，并在太原市小店区检测到黄胸鼠抗性种群的存在，需加强对此种群的监测。

关键词: 长尾仓鼠, 黄胸鼠, Vkorc1基因, 抗药性, 沉默突变, 错义突变

Abstract:

To explore the polymorphism of vitamin K epoxide reductase complex subunit 1 gene (Vkorc1), which is closely related to the resistance to anticoagulant rodenticides in the long-tailed hamster (Cricetulus longicaudatus)and the Asian house rat (Rattus tanezumi),the mutation sites of Vkorc1 and their distribution were detected. In present study, samples were collected using the trapping methods from farms at 14 sites and agricultural fields at 13 sites. The exons of Vkorc1 were amplified and sequenced. The sequences were aligned using MEGA X to visually detect the mutation sites in exons and their distribution. The results showed that C. longicaudatus samples werecaptured in all the 13 sampling sites and accounted for 23.29% of wild rodents. R.tanezumi were distributed in 8 sampling sites, accounting for 68.63% of commensal rodents. Six silent mutation sites and 5 missense mutation sites in Vkorc1 were detected in C. longicaudatus samples (n = 105). Among them, silent mutation C438T (His146His) had the highest mutation rate of 67.62%. A total of 17 C. longicaudatus samples existed missense mutation sites. We found 6 silent mutation sites and 1 missense mutation site in R. tanezumi samples (n = 70). The mutation rates of the most common silent mutation sites A321C (Ile107Ile) and T411C (Thr137Thr) was up 18.57%. The missense mutation A416G (Tyr139Cys) associated with drug resistance was found in 8 R. tanezumi samples, of which 7 were captured from Xiaodian, Taiyuan, with a mutation rate of 35.00%. In conclusion, Vkorc1 mutations were detected both in anthropogenic habitats and natural ecosystems. We should strengthen the monitoring of R. tanezumi in Xiaodian, Taiyuan, since the formation of the resistant population.

Key words: Cricetulus longicaudatus, Rattus tanezumi, Vkorc1, Resistance, Silent mutation, Missense mutation

中图分类号:

Q346

杨新根, 王艳龙, 邹波, 常文英, 侯玉, 赵悠悠, 王庭林, 张健旭. 不同栖息环境的两种啮齿动物Vkorc1基因多态性[J]. 兽类学报, 2022, 42(6): 687-697.

Xingen YANG, Yanlong WANG, Bo ZOU, Wenying CHANG, Yu HOU, Youyou ZHAO, Tinglin WANG, Jianxu ZHANG. Vkorc1 polymorphism of two rodent species in different habitats[J]. ACTA THERIOLOGICA SINICA, 2022, 42(6): 687-697.

图/表 7

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样本采集地 Sampling sites	采样生境 Sampling habitats	地理位置 Geographic coordinates	海拔 Elevation （m）
运城市永济市 Yongji City， Yuncheng City (YOJ)	A	110.43°E， 34.86°N	392
晋城市陵川县 Lingchuan County， Jincheng City (LIC)	A，B	113.27°E， 35.74°N	1 080
临汾市洪洞县 Hongtong County， Linfen City （HOT）	C	111.59°E， 36.25°N	450
临汾市隰县 Xi County， Linfen City （XIX）	A，D	110.85°E， 36.50°N	1 020
临汾市永和县 Yonghe County， Linfen City （YOH）	D	110.63°E， 36.76°N	890
长治市沁县 Qin County， Changzhi City （QNX）	A，B	112.69°E， 36.74°N	1 140
晋中市左权县 Zuoquan County， Jinzhong City （ZOQ）	A，C	113.31°E， 37.05°N	1 080
晋中市祁县 Qi County， Jinzhong City （QIX）	A	112.31°E， 37.35°N	770
吕梁市中阳县 Zhongyang County， Lvliang City （ZHY）	B	111.17°E， 37.36°N	1 450
吕梁市离石区 Lishi District， Lvliang City （LIS）	A，B	111.98°E， 37.52°N	1 230
太原市小店区 Xiaodian District， Taiyuan City （XID）	C	113.27°E， 37.80°N	780
太原市娄烦县 Loufan County， Taiyuan City （LOF）	A，C	111.86°E， 38.05°N	1 320
太原市阳曲县 Yangqu County， Taiyuan City （YAQ）	B	112.67°E， 38.29°N	800
阳泉市盂县 Yu County， Yangquan City （YUX）	C，E	113.35°E， 37.71°N	1 250
忻州市静乐县 Jingle County， Xinzhou City （JNL）	B	111.95°E， 38.39°N	1 260
忻州市五台县 Wutai County， Xinzhou City （WUT）	C，E	113.20°E， 38.64°N	1 060
朔州市朔城区 Shuocheng District， Shuozhou City （SOC）	C	112.30°E， 39.11°N	1 140
大同市浑源县 Hunyuan County， Datong City （HNY）	C，E	113.96°E， 39.85°N	1 550

样本采集地 Sampling sites	采样生境 Sampling habitats	地理位置 Geographic coordinates	海拔 Elevation （m）
运城市永济市 Yongji City， Yuncheng City (YOJ)	A	110.43°E， 34.86°N	392
晋城市陵川县 Lingchuan County， Jincheng City (LIC)	A，B	113.27°E， 35.74°N	1 080
临汾市洪洞县 Hongtong County， Linfen City （HOT）	C	111.59°E， 36.25°N	450
临汾市隰县 Xi County， Linfen City （XIX）	A，D	110.85°E， 36.50°N	1 020
临汾市永和县 Yonghe County， Linfen City （YOH）	D	110.63°E， 36.76°N	890
长治市沁县 Qin County， Changzhi City （QNX）	A，B	112.69°E， 36.74°N	1 140
晋中市左权县 Zuoquan County， Jinzhong City （ZOQ）	A，C	113.31°E， 37.05°N	1 080
晋中市祁县 Qi County， Jinzhong City （QIX）	A	112.31°E， 37.35°N	770
吕梁市中阳县 Zhongyang County， Lvliang City （ZHY）	B	111.17°E， 37.36°N	1 450
吕梁市离石区 Lishi District， Lvliang City （LIS）	A，B	111.98°E， 37.52°N	1 230
太原市小店区 Xiaodian District， Taiyuan City （XID）	C	113.27°E， 37.80°N	780
太原市娄烦县 Loufan County， Taiyuan City （LOF）	A，C	111.86°E， 38.05°N	1 320
太原市阳曲县 Yangqu County， Taiyuan City （YAQ）	B	112.67°E， 38.29°N	800
阳泉市盂县 Yu County， Yangquan City （YUX）	C，E	113.35°E， 37.71°N	1 250
忻州市静乐县 Jingle County， Xinzhou City （JNL）	B	111.95°E， 38.39°N	1 260
忻州市五台县 Wutai County， Xinzhou City （WUT）	C，E	113.20°E， 38.64°N	1 060
朔州市朔城区 Shuocheng District， Shuozhou City （SOC）	C	112.30°E， 39.11°N	1 140
大同市浑源县 Hunyuan County， Datong City （HNY）	C，E	113.96°E， 39.85°N	1 550

样本 Samples	引物名称 Primer names	引物序列（5′‒3′） Primer sequences （5′‒3′）	PCR产物大小 Size of PCR products （bp）	目标外显子 Target exons
长尾仓鼠 Cricetulus longicaudatus	C.l.-Vk1/F/e1-2	AGAATGGCAGCCGAGGCGGGTCAACCTTC	约1 300	1，2
	C.l.-Vk1/R/e1-2	CTTCCTGAACTAACTAGGATGCTGGTGTC	约1 300	1，2
	C.l.-Vk1/F/e3	GAATATAGACTTTAGCAGGAGTACTTACT	约500	3
	C.l.-Vk1/R/e3	GTGTGGTTATGGAGGGGGACTGGAATGAAG	约500	3
黄胸鼠 Ruttus tanezumi	R.t.-Vk1/F/e1-2	GTTTACCAGCCCAGCATTCCTAGCTGTCA	约 1 400	1，2
	R.t.-Vk1/R/e1-2	TTTGAGGGCCCTTCAAGCCTCTGGCTACC	约 1 400	1，2
	R.t.-Vk1/F/e3	ACTCTGGGTCCCAAGAATTATTGAACAG	约 400	3
	R.t.-Vk1/R/e3	GGCAAAGCAAGTCATGTCAGCCTGGCAT	约 400	3

样本 Samples	引物名称 Primer names	引物序列（5′‒3′） Primer sequences （5′‒3′）	PCR产物大小 Size of PCR products （bp）	目标外显子 Target exons
长尾仓鼠 Cricetulus longicaudatus	C.l.-Vk1/F/e1-2	AGAATGGCAGCCGAGGCGGGTCAACCTTC	约1 300	1，2
	C.l.-Vk1/R/e1-2	CTTCCTGAACTAACTAGGATGCTGGTGTC	约1 300	1，2
	C.l.-Vk1/F/e3	GAATATAGACTTTAGCAGGAGTACTTACT	约500	3
	C.l.-Vk1/R/e3	GTGTGGTTATGGAGGGGGACTGGAATGAAG	约500	3
黄胸鼠 Ruttus tanezumi	R.t.-Vk1/F/e1-2	GTTTACCAGCCCAGCATTCCTAGCTGTCA	约 1 400	1，2
	R.t.-Vk1/R/e1-2	TTTGAGGGCCCTTCAAGCCTCTGGCTACC	约 1 400	1，2
	R.t.-Vk1/F/e3	ACTCTGGGTCCCAAGAATTATTGAACAG	约 400	3
	R.t.-Vk1/R/e3	GGCAAAGCAAGTCATGTCAGCCTGGCAT	约 400	3

样本采集地 Sampling sites	采样生境 Sampling habitats	布夹（笼）数 Number of clips （cages）	捕获数 Number of captures	捕获率 Capture rate （%）	啮齿动物占比 Proportions of rodents （%）
样本采集地 Sampling sites	采样生境 Sampling habitats	布夹（笼）数 Number of clips （cages）	捕获数 Number of captures	捕获率 Capture rate （%）	黄胸鼠 Ruttus tanezumi	长尾仓鼠 Cricetulus longicaudatus	其他啮齿动物 Others
YOJ	A	200	10	5.00	50.00		50.00
LIC	B	450	41	9.11		12.20	87.80
	A	180	13	7.22	76.92		23.08
HOT	C	200	9	4.50	77.78		22.22
XIX	D	440	65	14.77		13.85	86.15
	A	240	3	1.25	0.00		100
YOH	D	400	38	9.50		21.05	78.95
QNX	B	450	43	9.56		25.58	74.42
	A	210	10	4.76	90.00		10.00
ZOQ	B	425	41	9.65		21.95	78.05
	C	160	2	1.25	0.00		100
QIX	A	210	8	3.81	75.00		25.00
ZHY	B	450	34	7.56		32.35	67.65
LIS	B	540	37	6.85		21.62	78.38
	A	330	10	3.03	70.00		30.00
XID	C	750	23	3.07	86.96		13.04
LOF	B	440	34	7.73		32.35	67.65
	C	200	1	0.50	0.00		100
YAQ	B	550	42	7.64		26.19	73.81
YUX	E	400	29	7.25		34.48	65.52
	C	160	1	0.63	0.00		100
JNL	B	440	42	9.55		19.04	80.96
WUT	E	400	36	9.00		30.56	69.44
	C	460	9	1.96	66.67		33.33
SOC	C	270	2	0.74	0.00		100
HNY	E	420	29	6.90		24.14	75.86
	C	270	1	0.37	0.00		100
合计 Total		9 665	613

不同栖息环境的两种啮齿动物Vkorc1基因多态性

Vkorc1 polymorphism of two rodent species in different habitats

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Metrics

样本 Samples	样本数 Number of samples	变异类型 Mutation types	核苷酸变异 Nucleotide mutations	氨基酸变异 Amino acid mutations	外显子 Exons	携带变异位点个体数 Number of samples with mutations*	变异率 Mutation rate （%）
长尾仓鼠 Cricetulus longicaudatus	105	沉默突变 Silent mutation	G78A（C）	Ala26Ala	1	16 （LIC1， QNX5， ZOQ1， ZHY2，LOF5， YUX1， JNL1）	15.24
			G135A	Val45Val	1	9 （LIC3， XIX1， ZOQ1， ZHY1，YAQ1， HNY2）	8.57
			C159A	Arg53Arg	1	11 （ZHY2， LIS2， YAQ4， YUX3）	10.48
			C222T	Ser74Ser	2	10 （LIC5， XIX1， ZOQ1， ZHY1，HNY2）	9.52
			C342T	Phe114Phe	3	30 （LIC1， QNX5， ZOQ3， ZHY1， LIS1， LOF4， YUX4， JNL5， WUT6）	28.57
			C438T	His146His	3	71 （LIC3， XIX8， YOH5， QNX2， ZOQ5， ZHY8， LIS7， LOF4， YAQ8， YUX4， JNL5， WUT6， HNY6）	67.62
		错义突变 Missense mutation	C8T	Thr3Ile	1	9 （LIC3， ZOQ1， ZHY1， LOF1，YAQ1， HNY2）	8.57
			G106A	Asp36Asn	1	3 （LIC1， ZOQ1， LOF1）	2.86
			A203G	His68Arg	2	10 （LIC5， XIX1， ZOQ1，ZHY1，HNY2）	9.52
			A203T	His68Leu	2	2 （XIX2）	1.90
			G346A	Gly116Ser	3	1 （YUX1）	0.95
黄胸鼠 Ruttus tanezumi	70	沉默突变 Silent mutation	A36G	Arg12Arg	1	11 （HOT1， LIS5， XID3， WUT2）	15.71
			G123A	Ala41Ala	1	4 （QIX2， XID2）	5.71
			T204C	His68His	2	8 （HOT1， LIS5， XID1， WUT1）	11.43
			A246T	Ile82Ile	2	3 （QIX1， XID2）	4.29
			A321C	Ile107Ile	3	13 （HOT1， QIX2， LIS5， XID3， WUT2）	18.57
			T411C	Thr137Thr	3	13 （HOT1， QIX2， LIS5，XID3，WUT2）	18.57
		错义突变 Missense mutation	A416G	Tyr139Cys	3	8 （QIX1， XID7）	11.43

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