基于COⅠ和Cytb基因的浙江不同抗性水平二化螟种群的遗传结构分析

doi:10.3969/j.issn.1004-1524.2022.11.15

浙江农业学报 ›› 2022, Vol. 34 ›› Issue (11): 2462-2470.DOI: 10.3969/j.issn.1004-1524.2022.11.15

基于COⅠ和Cytb基因的浙江不同抗性水平二化螟种群的遗传结构分析

鲁艳辉(), 郭嘉雯, 田俊策, 薛钊鸿, 郑许松, 吕仲贤()

浙江省农业科学院农产品质量安全危害因子与风险防控国家重点实验室植物保护与微生物研究所,浙江杭州 310021

收稿日期:2021-03-05 出版日期:2022-11-25 发布日期:2022-11-29
通讯作者: 吕仲贤
作者简介:*吕仲贤,E-mail: luzxmh@163.com
鲁艳辉(1978—),女,河北滦县人,博士,副研究员,主要从事昆虫毒理及分子生物学相关研究。E-mail:luyanhui4321@126.com
基金资助:
浙江省重点研发计划(2020C02001)

Population genetic structure of different resistance levels of Chilo suppressalis based on mitochondrial COⅠ and Cytb sequences in Zhejiang, China

LU Yanhui(), GUO Jiawen, TIAN Junce, XUE Zhaohong, ZHENG Xusong, LYU Zhongxian()

State Key Laboratory of Managing Biotic and Chemical Threats to the Quality and Safety of Agro-Products, Institute of Plant Protection and Microbiology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China

Received:2021-03-05 Online:2022-11-25 Published:2022-11-29
Contact: LYU Zhongxian

摘要/Abstract

摘要：

本研究利用线粒体细胞色素氧化酶亚基Ⅰ(COⅠ)和细胞色素b(Cytb)基因的遗传学方法分析了浙江省8个不同抗性水平二化螟地理种群的遗传多样性及种群遗传结构。种群遗传多样性分析表明:PCR 扩增测序分别获得长度为627 bp的COⅠ基因片段和长度为455 bp的Cytb基因片段。355条同源COⅠ序列监测出了68个多态性位点,其中单突变位点22个,简约突变位点46个,共定义了85个单倍型,每个群体的平均单倍型为18.25个,其中瑞安(RA)种群中单倍型最多,为27个单倍型。326条Cytb同源序列监测出了45个多态性位点,其中单突变位点19个,简约突变位点26个,共定义了64个单倍型,每个群体的平均单倍型为14.375个,其中乐清(YQ)种群中单倍型最多,为25个单倍型。此外,各群体中最高的单倍型多样度h分别为0.896 3和0.934 4,反映出二化螟群体较低的遗传多样性水平。种群遗传结构分析表明:二化螟不同地理种群间遗传变异大多数来自于群体内个体间,占80.30%(COⅠ基因)和78.16%(Cytb基因)。较少部分的遗传差异来自于组间,仅占19.43%(COⅠ基因)和21.22%(Cytb基因)。单倍型网络关系图未表现出显著的地理谱系结构,Mantel相关性检测显示,遗传距离与地理距离之间无显著相关性。单倍型邻接树也没有明显分支,未呈现出地域性差异。该研究结果为浙江省不同抗性水平二化螟种群间交流和二化螟的防控提供了基础资料。

关键词: 二化螟, 细胞色素C氧化酶亚基Ⅰ, 细胞色素b, 遗传多样性, 遗传结构, 抗性

Abstract:

In this study, genetic diversity and genetic structure of eight geographical populations with different resistance levels of Chilo suppressalis in Zhejiang Province were analyzed by using genetic methods based on mitochondrial cytochrome oxidase subunit Ⅰ (COⅠ) and cytochrome b (Cytb) genes. The analysis results of population genetic diversity showed that the COⅠ gene fragment with length of 627 bp and the Cytb gene fragment with length of 455 bp were obtained by PCR amplification and sequencing. Among them, 355 homologous COⅠ sequences detected 68 polymorphic sites, including 22 single mutation sites and 46 reduced mutation sites. A total of 85 haplotypes were defined, with an average of 18.25 haplotypes in each population, of which the Ruian (RA) population had the most haplotypes, with 27 haplotypes. Meanwhile, 326 Cytb homologous sequences detected 45 polymorphic sites, including 19 single mutation sites and 26 simple mutation sites. A total of 64 haplotypes were defined. The average haplotype of each population was 14.375, among which the most haplotypes were 25 in Yueqing (YQ) population. The highest haplotype diversity h of the population was 0.896 3 and 0.934 4, respectively. The analysis of population genetic structure showed that the majority of genetic variation existed among different geographical populations of C. suppressalis from individuals within the population, accounting for 80.30% (COⅠ gene) and 78.16% (Cytb gene). Only 19.43% (COⅠ gene) and 21.22% (Cytb gene) of the genetic differences were found among groups. In addition, no clear geographical differentiation was revealed in the haplotype network and on the neighbor-joining tree. Mantel correlation test showed that there was no significant correlation between genetic distance and geographical distance. These results provided basic data for different resistance level population exchange and control of C. suppressalis in Zhejiang Province.

Key words: Chilo suppressalis, mitochondrial cytochrome oxidase subunit Ⅰ, Cytb, genetic diversity, genetic structure, resistance

中图分类号:

S435.112.1

鲁艳辉, 郭嘉雯, 田俊策, 薛钊鸿, 郑许松, 吕仲贤. 基于COⅠ和Cytb基因的浙江不同抗性水平二化螟种群的遗传结构分析[J]. 浙江农业学报, 2022, 34(11): 2462-2470.

LU Yanhui, GUO Jiawen, TIAN Junce, XUE Zhaohong, ZHENG Xusong, LYU Zhongxian. Population genetic structure of different resistance levels of Chilo suppressalis based on mitochondrial COⅠ and Cytb sequences in Zhejiang, China[J]. Acta Agriculturae Zhejiangensis, 2022, 34(11): 2462-2470.

图/表 11

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种群 Popula- tions	采集地点 Collection locations	经纬度 Latitude and longitude	采集时间 Collection time
PH	平湖Pinghu	30°70'N, 121°02'E	2018年3月Mar. 2018
XiaoS	萧山Xiaoshan	30°17'N, 120°27'E	2018年4月Apr. 2018
XS	象山Xiangshan	29°48'N, 121°87'E	2018年4月Apr. 2018
NH	宁海Ninghai	29°28'N, 121°43'E	2018年3月Mar. 2018
YK	永康Yongkang	28°90'N, 120°03'E	2018年4月Apr. 2018
WL	温岭Wenling	28°37'N, 121°37'E	2018年4月Apr. 2018
YQ	乐清Yueqing	28°13'N, 120°95'E	2018年3月Mar. 2018
RA	瑞安Rui’an	27°78'N, 120°63'E	2018年4月Apr. 2018

种群 Popula- tions	采集地点 Collection locations	经纬度 Latitude and longitude	采集时间 Collection time
PH	平湖Pinghu	30°70'N, 121°02'E	2018年3月Mar. 2018
XiaoS	萧山Xiaoshan	30°17'N, 120°27'E	2018年4月Apr. 2018
XS	象山Xiangshan	29°48'N, 121°87'E	2018年4月Apr. 2018
NH	宁海Ninghai	29°28'N, 121°43'E	2018年3月Mar. 2018
YK	永康Yongkang	28°90'N, 120°03'E	2018年4月Apr. 2018
WL	温岭Wenling	28°37'N, 121°37'E	2018年4月Apr. 2018
YQ	乐清Yueqing	28°13'N, 120°95'E	2018年3月Mar. 2018
RA	瑞安Rui’an	27°78'N, 120°63'E	2018年4月Apr. 2018

基因名称 Gene name	引物 Primer sequence(5'-3')	产物长度 Product length/bp
COⅠ	F: TCTATGTGTGATGTTTATTG	700
	R: GATCAGGAATATTTGGGAAC
Cytb	F: TATGTACTACCATGAGGACAAATATC	490
	R: ATTACACCTCCTAATTTATTAGGAAT

基因名称 Gene name	引物 Primer sequence(5'-3')	产物长度 Product length/bp
COⅠ	F: TCTATGTGTGATGTTTATTG	700
	R: GATCAGGAATATTTGGGAAC
Cytb	F: TATGTACTACCATGAGGACAAATATC	490
	R: ATTACACCTCCTAATTTATTAGGAAT

药剂 Insecticides	种群 Populations	斜率±标准误 Slope±SE	卡方(自由度) χ² (df)	致死中浓度(95%置信限) LC₅₀ (95% FL)/(mg·L^-1)	抗性倍数 Resistance ratio
氯虫苯甲酰胺	PH	2.73±0.28	0.97(3)	80.58(67.16~98.32)	57.84
Chlorantraniliprole	XiaoS	2.11±0.36	0.87(3)	205.258(188.45~230.98)	147.35
	XS	2.87±0.41	3.93(3)	136.93 (118.09~162.22)	98.30
	NH	2.45±0.43	4.89(3)	178.403(155.42~210.04)	128.07
	YK	2.21±0.23	2.72(3)	196.68(171.15~215.16)	141.19
	WL	2.44±0.28	3.88(3)	301.96(256.53~347.25)	216.77
	YQ	2.59±0.26	5.67(3)	194.94(119.89~246.76)	139.94
	RA	1.75±0.21	4.07(3)	94.47(62.64~137.58)	67.82
阿维菌素	PH	2.51±0.39	2.60 (3)	17.84(15.37~21.98)	19.16
Abamectin	XiaoS	1.97±0.25	1.96(3)	13.92(11.36~17.42)	14.95
	XS	2.14±0.27	1.38(3)	9.11(7.32~12.04)	9.79
	NH	2.45±0.29	2.87(3)	13.53(11.70~16.39)	14.53
	YK	2.24±0.34	3.51(3)	12.88(9.52~18.06)	13.83
	WL	2.49±0.24	2.65(3)	8.57(6.38~10.83)	9.21
	YQ	2.61±0.36	1.94(3)	15.66(12.82~18.28)	16.82
	RA	2.28±0.25	1.83(3)	8.90(6.47~9.64)	9.56
甲氧虫酰肼	PH	2.11±0.28	2.32(3)	81.77(68.41~99.04)	112.17
Methoxyfenozide	XiaoS	1.74±0.21	2.15(3)	99.59(78.46~125.21)	136.61
	XS	2.35±0.25	1.89(3)	67.84(56.93~80.98)	93.06
	NH	2.74±0.27	1.54(3)	41.88(34.18~49.97)	57.45
	YK	2.23±0.24	3.09(3)	79.79(67.28~93.05)	109.45
	WL	2.43±0.21	2.92(3)	36.91(29.45~42.27)	50.47
	YQ	2.67±0.28	3.60(3)	73.13(58.94~91.63)	100.32
	RA	1.71±0.22	1.82(3)	88.76(69.63~112.35)	121.76

基于COⅠ和Cytb基因的浙江不同抗性水平二化螟种群的遗传结构分析

Population genetic structure of different resistance levels of Chilo suppressalis based on mitochondrial COⅠ and Cytb sequences in Zhejiang, China

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基因 Genes	种群 Populations	样本数量 N	单倍型数 H	多态位点数 s	单倍型多样度 h	平均核苷酸差异数 k	核苷酸多样性指数 π
COⅠ	PH	34	15	24	0.887 7	5.841 4	0.009 3
	XiaoS	46	13	28	0.861 8	5.963 3	0.009 5
	XS	34	20	28	0.930 5	5.260 3	0.008 5
	NH	49	9	15	0.511 1	1.821 4	0.002 9
	YK	49	15	20	0.682 0	3.649 7	0.005 9
	WL	46	19	22	0.858 9	4.637 7	0.007 4
	YQ	49	26	26	0.897 1	4.850 3	0.007 7
	RA	48	27	24	0.896 3	4.681 7	0.007 5
Cytb	PH	36	15	19	0.888 9	4.819 1	0.010 7
	XiaoS	40	9	16	0.669 2	3.152 6	0.006 9
	XS	32	16	19	0.907 3	4.300 4	0.009 5
	NH	49	6	12	0.266 2	1.821 4	0.003 4
	YK	49	18	17	0.651 4	3.331 6	0.007 3
	WL	52	17	21	0.865 0	4.070 1	0.009 0
	YQ	48	25	25	0.934 4	4.103 7	0.009 0
	RA	20	9	10	0.794 7	3.200 0	0.007 0

基因 Genes	种群 Populations	NH	PH	RA	WL	XiaoS	XS	YK	YQ
COⅠ	NH		0.009	0.009	0.008	0.008	0.008	0.005	0.009
	PH	0.354 39		0.010	0.009	0.011	0.010	0.008	0.009
	RA	0.451 48	0.112 08		0.008	0.011	0.008	0.008	0.008
	WL	0.334 68	0.089 83	0.015 68		0.010	0.008	0.007	0.008
	XiaoS	0.195 27	0.126 09	0.249 14	0.165 40		0.011	0.008	0.011
	XS	0.333 75	0.080 46	0.009 94	0.010 91	0.160 04		0.008	0.008
	YK	0.201 08	0.104 97	0.182 93	0.076 34	0.074 51	0.085 20		0.008
	YQ	0.407 02	0.086 50	0.011 48	0.007 13	0.217 52	-0.002 90	0.143 95
Cytb	NH		0.009	0.012	0.01	0.006	0.011	0.006	0.011
	PH	0.207 71		0.011	0.011	0.010	0.011	0.009	0.011
	RA	0.586 93	0.152 12		0.008	0.012	0.008	0.011	0.008
	WL	0.360 78	0.073 16	0.037 60		0.011	0.009	0.010	0.009
	XiaoS	0.075 21	0.131 28	0.439 96	0.276 78		0.012	0.008	0.012
	XS	0.418 74	0.065 35	0.005 43	0.004 03	0.320 59		0.010	0.009
	YK	0.062 62	0.042 35	0.309 33	0.153 98	0.065 49	0.181 29		0.010
	YQ	0.414 33	0.071 82	0.000 76	0.001 64	0.317 15	-0.015 99	0.193 50

基因 Genes	分组 Grouping	变异来源 Variation source	自由度 DF	方差总和 Variance sum	方差组分 Variance components	变异百分比 Variation percentage/%	F_st	P
COⅠ	(NH)&(PH)&(XiaoS)&	组间Between groups	4	144.863	0.550 85 Va	19.43	0.194 40	0.010 8
	(YK)&(RA,WL,XS,YQ)	组内群体间	3	7.828	0.007 58 Vb	0.27	0.003 30	0.338 2
		Intra population and inter group
		群体内个体间	347	789.757	2.275 96 Vc	80.30	0.197 00	0.000 1
		Between individuals within the population
		总和Total	354	942.448	2.834 39
Cytb	(NH)&(PH)&(XiaoS)&	组间Between groups	4	142.151	0.565 62 Va	21.22	0.212 20	0.014 06
	(YK)&(RA,WL,XS,YQ)	组内群体间	3	8.069	0.016 57 Vb	0.62	0.007 90	0.308 22
		Intra population and inter group
		群体内个体间	318	662.626	2.083 73 Vc	78.16	0.218 38	0.000 01
		Between individuals within the population
		总和Total	325	812.845	2.665 92

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[2]	杨春, 孟泽洪, 李帅, 梁思慧, 乔大河, 陈正武. 十二个茶树品种对茶棍蓟马、茶小绿叶蝉抗性表现及抗性成分初步鉴定[J]. 浙江农业学报, 2022, 34(8): 1713-1724.
[3]	陆景伟, 陈国康, 周娜, 魏捷, 胡燕, 郑阳, 陶伟林. 六个丝瓜品种对南方根结线虫的抗性[J]. 浙江农业学报, 2022, 34(5): 959-965.
[4]	刘苡含, 牟青山, 陈珊宇, 阮关海, 胡晋, 关亚静. 基于SSR-HRM技术的向日葵DNA指纹图谱构建[J]. 浙江农业学报, 2022, 34(4): 678-686.
[5]	裴芸, 徐秀红, 陆锦彪, 陈阿敏, 张万萍. 151份贵州地方樱桃番茄资源的遗传多样性分析[J]. 浙江农业学报, 2022, 34(2): 310-316.
[6]	曹联飞, 施金虎, 徐雅岚, 苏晓玲, 胡福良, 郑火青. 基于线粒体DNA tRNA^leu-COⅡ序列的浙江省东方蜜蜂遗传多样性研究[J]. 浙江农业学报, 2022, 34(11): 2395-2403.
[7]	汪宝根, 董君暘, 汪颖, 李素娟, 王尖, 鲁忠富, 吴晓花, 李国景, 吴新义. 浙江省地方菜豆种质资源鉴定与遗传多样性分析[J]. 浙江农业学报, 2022, 34(11): 2416-2427.
[8]	王郅琪, 孙建, 梁俊超, 赵云燕, 颜廷献, 颜小文, 危文亮, 乐美旺. 基于分子标记的江西省芝麻地方种质遗传多样性分析[J]. 浙江农业学报, 2021, 33(9): 1565-1580.
[9]	马杰, 屈雯, 陈春艳, 王磊, 马俊, 刘针杉, 马维, 周平, 何远宽, 孙勃. 基于转录组序列的叶用芥菜奶奶青菜EST-SSR标记开发与遗传多样性分析[J]. 浙江农业学报, 2021, 33(9): 1640-1649.
[10]	刘士力, 卞玉玲, 贾永义, 迟美丽, 李飞, 郑建波, 程顺, 顾志敏. 基于线粒体CO Ⅰ 基因序列的红螯螯虾养殖群体遗传结构分析[J]. 浙江农业学报, 2021, 33(8): 1385-1392.
[11]	黄宣, 金林灿, 叶朝辉, 姜洁锋, 施贤波. 浙江近年育成粳稻新品种(系)部分抗病虫基因的分子检测与育种应用[J]. 浙江农业学报, 2021, 33(7): 1159-1169.
[12]	张静珍, 王连军, 雷剑, 柴沙沙, 杨新笋, 张文英. 基于cpSSR标记的山药种质资源DNA指纹图谱构建及遗传多样性分析[J]. 浙江农业学报, 2021, 33(7): 1222-1233.
[13]	安红卫, 宋勤飞, 牛素贞. 贵州茶树种质资源遗传多样性、群体结构和遗传分化研究[J]. 浙江农业学报, 2021, 33(7): 1234-1243.
[14]	杨梅, 胡小兰, 申涛, 谭康, 刘代铃, 邱红波. 玉米第8染色体单片段代换系的构建与灰斑病抗性材料筛选[J]. 浙江农业学报, 2021, 33(3): 383-389.
[15]	王贤, 刘放, 魏小红, 朱晓林, 王宝强. 不同种质番茄材料抗番茄黄化曲叶病毒病特性研究[J]. 浙江农业学报, 2021, 33(11): 2085-2097.

药剂 Insecticides		单倍型多样度 h		核苷酸多样性指数 π
药剂 Insecticides		COⅠ	Cytb	COⅠ	Cytb
氯虫苯甲酰胺Chlorantraniliprole	R	-0.184	0.098	-0.198	-0.136
	P	0.663	0.818	0.638	0.748
阿维菌素Abamectin	R	-0.146	0.089	0.095	0.106
	P	0.730	0.835	0.823	0.803
甲氧虫酰肼Methoxyfenozide	R	0.431	0.270	0.604	0.207
	P	0.286	0.518	0.113	0.623

药剂 Insecticides		单倍型多样度 h		核苷酸多样性指数 π
药剂 Insecticides		COⅠ	Cytb	COⅠ	Cytb
氯虫苯甲酰胺Chlorantraniliprole	R	-0.184	0.098	-0.198	-0.136
	P	0.663	0.818	0.638	0.748
阿维菌素Abamectin	R	-0.146	0.089	0.095	0.106
	P	0.730	0.835	0.823	0.803
甲氧虫酰肼Methoxyfenozide	R	0.431	0.270	0.604	0.207
	P	0.286	0.518	0.113	0.623