家蚕地方品种遗传多样性及其分子系统学研究

doi:10.3724/SP.J.1003.2013.11035

生物多样性 ›› 2013, Vol. 21 ›› Issue (3): 315-325. DOI: 10.3724/SP.J.1003.2013.11035

家蚕地方品种遗传多样性及其分子系统学研究

杜周和^1,², 刘俊凤^1,^*(), 刘彬斌¹, 左艳春¹, 吴建梅¹, 陈义安¹, 张剑飞¹, 鲁成^2,^*()

1 四川省农科院蚕业研究所, 四川南充 637000
2 西南大学蚕学与系统生物学研究所, 重庆 400716

收稿日期:2013-02-04 接受日期:2013-04-26 出版日期:2013-05-20 发布日期:2013-06-05
通讯作者: 刘俊凤,鲁成
基金资助:
国家重点基础研究发展计划(2005CB121000)、四川省应用基础(2013JY0113)和省农科院优秀论文基金(2013LWJJ-008)

Genetic diversity and molecular genealogy of local silkworm varieties

Zhouhe Du^1,², Junfeng Liu^1,^*, Binbin Liu¹, Yanchun Zuo¹, Jianmei Wu¹, Yi-an Chen¹, Jianfei Zhang¹, Cheng Lu^2,^*()

1 Institute of Sericulture, Sichuan Academy of Agricultural Sciences, Nanchong, Sichuan 637000
2 Institute of Sericulture and Systems Biology, Southwest University, Chongqing 400716

Received:2013-02-04 Accepted:2013-04-26 Online:2013-05-20 Published:2013-06-05
Contact: Liu Junfeng,Lu Cheng

摘要/Abstract

摘要：

为检测家蚕(Bombyx mori)地方品种间的遗传差异, 分析其遗传多样性和系统分化模式, 为家蚕的品种鉴定和杂交育种亲本选择提供分子依据, 本文对72个家蚕地方品种的淀粉酶基因(Bmamy2)片段进行测序, 采用DNA序列分析技术探究了家蚕地方品种在核苷酸序列水平上的遗传差异及其系统分化特征。结果显示, 家蚕地方品种在Bmamy2基因变异丰富, 碱基突变率为5.6-8.2%, 单倍型多样度为0.8294, 核苷酸多样度为0.0236±0.00122, 表明Bmamy2是一个具有丰富序列多样性的标记基因, 能够较好地区分家蚕地方品种间的遗传多样性。系统发育分析、分子方差分析和Network分析等多种分析方法均检测到家蚕地方品种不按地理系统或生态类型聚类, 各聚类种群均由来自不同地理系统或生态类型的品种构成, 而来自同一地理系统或生态类型的品种则分别归属于不同的聚类种群。推测在家蚕的进化过程中并没有形成按地理系统或生态类型归类的分化格局, 家蚕可能具有多种化性混合驯化起源的遗传背景和系统分化特征。

关键词: 家蚕, 种质资源, 遗传多样性, 分子系统学

Abstract

In order to explore the genetic diversity and systematic differentiation pattern among silkworm varieties, aiming to guide hybridization breeding, we sequenced a total of 72 Bmamy2gene fragments from local silkworm varieties. The analysis of nucleotide sequence diversity and systematic differentiation indicated that there was rich genovariation in the sequencing region of Bmamy2gene, and the base mutation rate is 5.6- 8.2%, the haplotype diversity is 0.8294, and the nucleotide diversity is 0.0236±0.00122, suggesting Bmamy2being a better marking gene with rich nucleotide sequence diversity, based on which the genetic diversity among different local silkworm varieties can be identified. The same heredity population structure is proclaimed by several analysis methods that every clade consisting of varieties from different geosystems and ecological types, while the varieties from the same geosystem and ecotype belong to different clades in the phylogeny. There is no population structure pattern that different varieties claded together according to geosystem or ecotype. It can be speculated that the silkworm origins from mixture of kinds of several voltinism mulberry silkworm, Bombyx mandarina, while the domestication events took place in several regions, from which the domesticated mulberry silkworms are all devoting to the domesticated silkworm population of today.

Key words: Bombyx mori, germplasm resources, genetic diversity, molecule genealogy

杜周和, 刘俊凤, 刘彬斌, 左艳春, 吴建梅, 陈义安, 张剑飞, 鲁成 (2013) 家蚕地方品种遗传多样性及其分子系统学研究. 生物多样性, 21, 315-325. DOI: 10.3724/SP.J.1003.2013.11035.

Zhouhe Du,Junfeng Liu,Binbin Liu,Yanchun Zuo,Jianmei Wu,Yi-an Chen,Jianfei Zhang,Cheng Lu (2013) Genetic diversity and molecular genealogy of local silkworm varieties. Biodiversity Science, 21, 315-325. DOI: 10.3724/SP.J.1003.2013.11035.

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链接本文: https://www.biodiversity-science.net/CN/10.3724/SP.J.1003.2013.11035

https://www.biodiversity-science.net/CN/Y2013/V21/I3/315

图/表 9

参考文献 38

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序号 No.	品种 Variety	代号 Code	生态类型 Ecotype	序号 No.	品种 Variety	代号 Code	生态类型 Ecotype
1	小石丸 Xiaoshiwan	XSW	CV1	37	萨尼斯 Sanisi	SNS	EV1
2	二毛 Ermao	EM	CV1	38	罗尼7 Luoni7	LN7	EV1
3	土三虎斑 Tusanhuban	TSHB	CV1	39	法408 Fa408	F408	EV1
4	中土 Zhongtu	ZT	CV1	40	欧18 Ou18	O18	EV1
5	达县土种 Daxiantuzhong	DXTZ	CV1	41	意16 Yi16	Y16	EV1
6	土一肉黄圆 Tuyirouhuangyuan	TYRHY	CV1	42	乌D WuD	WD	EV2
7	土八 Tuba	Tba	CV1	43	乌G WuG	WG	EV2
8	鲁1 Lu1	L1	CV1	44	乌B WuB	WB	EV2
9	Re黑 Rehei	RH	CV1	45	乌2 Wu2	W2	EV2
10	鲁10 Lu10	L10	CV1	46	日110 Ri110	R110	T
11	土种01 Tuzhong01	TZ01	CV1	47	大造 Dazao	DZ	T
12	诸桂 Zhugui	ZG	CV1	48	高花 Gaohua	GH	T
13	温州种 Wenzhouzhong	WZZ	CV1	49	第五白卵 Diwubailuan	DWBL	T
14	芙蓉灰卵 Furonghuiluan	FRHL	CV1	50	演金黄白 Yanjinhuangbai	YJHB	T
15	灰色卵 Huiseluan	HSL	CV1	51	15-100	15-100	T
16	盱眙种 Xuyizhong	XYZ	CV1	52	07-310	07-310	T
17	余杭24 Yuhang24	YH24	CV1	53	高白 Gaobai	GB	T
18	沛县种 Peixianzhong	PXZ	CV1	54	琼山海南 Qiongshanhainan	QSHN	T
19	邯郸种 Handanzhong	HDZ	CV1	55	655	655	T
20	中4010 Zhong4010	Z4010	CV2	56	防四 Fangsi	FS	T
21	泰 Tai	T	CV2	57	印度种 Yinduzhong	YDZ	T
22	玉溪2号 Yuxi2hao	YX	CV2	58	加秋 Jiaqiu	JQ	T
23	平 Ping	P	CV2	59	白夏B BaixiaB	BXB	T
24	C108	C108	CV2	60	天龙青白 Tianlongqingbai	TLQB	JV1
25	余杭2 Yuhang2	YH2	CV2	61	赤熟 Chishu	Cshu	JV1
26	土白 Tubai	Tbai	CV2	62	日9 Ri9	R9	JV1
27	松花形吴 Songhuaxingwu	SHXW	CV2	63	姬蚕 Jican	JC	JV1
28	保黄 Baohuang	BH	CVd	64	黑子 Heizi	HZ	JV1
29	MCH101	MCH101	CVd	65	日10 Ri10	R10	JV1
30	BT924	BT924	CVd	66	绵蚕 Miancan	MC	JV1
31	BH863	BH863	CVd	67	日限2 Rixian2	RX2	JV2
表1 (续) Table 1 (continued)
序号 No.	品种 Variety	代号 Code	生态类型 Ecotype	序号 No.	品种 Variety	代号 Code	生态类型 Ecotype
32	琼十 Qiongshi	QS	CVd	68	赢纹形 Yingwenxing	YWX	JV2
33	四海 Sihai	SH	CVd	69	J115	J115	JV2
34	兰十 Lanshi	LanS	CVd	70	830	830	JV2
35	罗萨 Luosa	LuoS	EV1	71	春四 Chunsi	CS	JV2
36	阿利可斯 Alikesi	ALKS	EV1	72	T8	T8	JV2

序号 No.	品种 Variety	代号 Code	生态类型 Ecotype	序号 No.	品种 Variety	代号 Code	生态类型 Ecotype
1	小石丸 Xiaoshiwan	XSW	CV1	37	萨尼斯 Sanisi	SNS	EV1
2	二毛 Ermao	EM	CV1	38	罗尼7 Luoni7	LN7	EV1
3	土三虎斑 Tusanhuban	TSHB	CV1	39	法408 Fa408	F408	EV1
4	中土 Zhongtu	ZT	CV1	40	欧18 Ou18	O18	EV1
5	达县土种 Daxiantuzhong	DXTZ	CV1	41	意16 Yi16	Y16	EV1
6	土一肉黄圆 Tuyirouhuangyuan	TYRHY	CV1	42	乌D WuD	WD	EV2
7	土八 Tuba	Tba	CV1	43	乌G WuG	WG	EV2
8	鲁1 Lu1	L1	CV1	44	乌B WuB	WB	EV2
9	Re黑 Rehei	RH	CV1	45	乌2 Wu2	W2	EV2
10	鲁10 Lu10	L10	CV1	46	日110 Ri110	R110	T
11	土种01 Tuzhong01	TZ01	CV1	47	大造 Dazao	DZ	T
12	诸桂 Zhugui	ZG	CV1	48	高花 Gaohua	GH	T
13	温州种 Wenzhouzhong	WZZ	CV1	49	第五白卵 Diwubailuan	DWBL	T
14	芙蓉灰卵 Furonghuiluan	FRHL	CV1	50	演金黄白 Yanjinhuangbai	YJHB	T
15	灰色卵 Huiseluan	HSL	CV1	51	15-100	15-100	T
16	盱眙种 Xuyizhong	XYZ	CV1	52	07-310	07-310	T
17	余杭24 Yuhang24	YH24	CV1	53	高白 Gaobai	GB	T
18	沛县种 Peixianzhong	PXZ	CV1	54	琼山海南 Qiongshanhainan	QSHN	T
19	邯郸种 Handanzhong	HDZ	CV1	55	655	655	T
20	中4010 Zhong4010	Z4010	CV2	56	防四 Fangsi	FS	T
21	泰 Tai	T	CV2	57	印度种 Yinduzhong	YDZ	T
22	玉溪2号 Yuxi2hao	YX	CV2	58	加秋 Jiaqiu	JQ	T
23	平 Ping	P	CV2	59	白夏B BaixiaB	BXB	T
24	C108	C108	CV2	60	天龙青白 Tianlongqingbai	TLQB	JV1
25	余杭2 Yuhang2	YH2	CV2	61	赤熟 Chishu	Cshu	JV1
26	土白 Tubai	Tbai	CV2	62	日9 Ri9	R9	JV1
27	松花形吴 Songhuaxingwu	SHXW	CV2	63	姬蚕 Jican	JC	JV1
28	保黄 Baohuang	BH	CVd	64	黑子 Heizi	HZ	JV1
29	MCH101	MCH101	CVd	65	日10 Ri10	R10	JV1
30	BT924	BT924	CVd	66	绵蚕 Miancan	MC	JV1
31	BH863	BH863	CVd	67	日限2 Rixian2	RX2	JV2
表1 (续) Table 1 (continued)
序号 No.	品种 Variety	代号 Code	生态类型 Ecotype	序号 No.	品种 Variety	代号 Code	生态类型 Ecotype
32	琼十 Qiongshi	QS	CVd	68	赢纹形 Yingwenxing	YWX	JV2
33	四海 Sihai	SH	CVd	69	J115	J115	JV2
34	兰十 Lanshi	LanS	CVd	70	830	830	JV2
35	罗萨 Luosa	LuoS	EV1	71	春四 Chunsi	CS	JV2
36	阿利可斯 Alikesi	ALKS	EV1	72	T8	T8	JV2

	序列长度 Sequence length (bp)	碱基插入/缺失位点数 Insert/delete (%)	变异位点数 Variable site (%)	单个突变位点 Singleton variable site (%)	简约信息位点 Parsimony variable site (%)
外显子 Exon	493	2(0.4)*	10(2.0)	2(0.4)	8(1.6)
内含子 Intron	670	139(20.7)	55(8.2)	7(1.0)	48(7.2)
全序列 Entire	1,163	141(12.1)	65(5.6)	9(0.8)	56(4.8)

	序列长度 Sequence length (bp)	碱基插入/缺失位点数 Insert/delete (%)	变异位点数 Variable site (%)	单个突变位点 Singleton variable site (%)	简约信息位点 Parsimony variable site (%)
外显子 Exon	493	2(0.4)*	10(2.0)	2(0.4)	8(1.6)
内含子 Intron	670	139(20.7)	55(8.2)	7(1.0)	48(7.2)
全序列 Entire	1,163	141(12.1)	65(5.6)	9(0.8)	56(4.8)

	CV2	EV1	T	CV1M3	JV2	CV1M4	JV1	CVd
CV2
EV1	0.025
T	0.027	0.024
CV1M3	0.026	0.015	0.023
JV2	0.026	0.012	0.024	0.016
CV1M4	0.026	0.013	0.024	0.017	0.014
JV1	0.027	0.025	0.024	0.025	0.026	0.025
CVd	0.025	0.025	0.024	0.025	0.026	0.025	0.024
EV2	0.025	0.044	0.029	0.039	0.044	0.042	0.026	0.026

家蚕地方品种遗传多样性及其分子系统学研究

Genetic diversity and molecular genealogy of local silkworm varieties

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聚类支 Clade	地理生态类型 Ecotype	品种数(个) No. of varieties	占各支百分比 %
A	中系一化 CV1	15	36.59
	中系二化 CV2	4	9.76
	中系多化 CVd	3	7.32
	日系一化 JV1	3	7.32
	日系二化 JV2	5	12.20
	欧系一化 EV1	6	14.63
	热带系统 T	5	12.20
	小计 Sum	41
B	中系一化 CV1	6	33.33
	中系多化 CVd	2	11.11
	日系一化 JV1	2	11.11
	日系二化 JV2	1	5.56
	欧系一化 EV1	1	5.46
	热带系统 T	6	33.33
	小计 Sum	18
C	中系二化 CV2	4	30.77
	中系多化 CVd	2	15.38
	日系一化 JV1	2	14.38
	欧系二化 EV2	4	30.77
	热带系统 T	1	7.69
	小计 Sum	13
合计	Total	72

变异来源 Source of variance	自由度 df	方差分量 Variance component	变异百分比 (%)	P-value
地理生态类型组群间 Among geo-voltings	7	6.93028	19.86	> 0.05
地理生态类型组群内 Among population within geo-voltings	1	1.23267	3.53	> 0.05
种群内 Within populations	63	26.73978	76.61	< 0.01

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