中国5个地方牦牛品种遗传多样性的微卫星分析

doi:10.3724/SP.J.1003.2008.07292

生物多样性 ›› 2008, Vol. 16 ›› Issue (2): 156-165. DOI: 10.3724/SP.J.1003.2008.07292

中国5个地方牦牛品种遗传多样性的微卫星分析

廖信军¹^,³, 常洪³, 张桂香²^,^*(), 王冬蕾³, 宋卫涛⁴, 韩旭², 张自富⁵

1 井冈山大学生命科学学院, 江西吉安 343009
2 全国畜牧兽医总站畜禽牧草种质资源保存利用中心, 北京 100094
3 扬州大学动物科学与技术学院, 扬州 225009
4 中国农业科学院家禽研究所, 扬州 225003
5 中国农业大学农学与生物技术学院, 北京 100094

收稿日期:2007-09-17 接受日期:2007-12-24 出版日期:2008-03-20 发布日期:2008-02-20
通讯作者: 张桂香
作者简介:*E-mail:gxzhang27@hotmail.com
基金资助:
农业部中国地方牛种遗传距离测定项目(农财发[2002]9号)

Genetic diversity of five native Chinese yak breeds based on microsatellite DNA markers

Xinjun Liao¹^,³, Hong Chang³, Guixiang Zhang²^,^*(), Donglei Wang³, Weitao Song⁴, Xu Han², Zifu Zhang⁵

1 College of Life Sciences, Jinggangshan University, Ji’an, Jiangxi 343009
2 Center of Preservation and Utilization of Genetic Resources of Domestic Animals and Forage, National Animal Husbandry and Veterinary Service, Beijing 100094
3 College of Animal Science and Technology, Yangzhou University, Yangzhou 225009
4 Institute of Poultry Science, Chinese Academy of Agricultural Sciences, Yangzhou 225003
5 College of Agriculture and Biotechnology, China Agricultural University, Beijing 100094

Received:2007-09-17 Accepted:2007-12-24 Online:2008-03-20 Published:2008-02-20
Contact: Guixiang Zhang

摘要/Abstract

摘要：

本文以大额牛(Bos frontalis)为外群, 应用16对微卫星DNA标记结合荧光-多重PCR技术, 评估了5个中国地方牦牛(Bos grunniens) (帕里牦牛、斯布牦牛、西藏高山牦牛、麦洼牦牛和九龙牦牛)品种内遗传变异和品种间遗传关系。6个群体的16个微卫星座位上共检测到159个等位基因, 其中有33个等位基因为5个牦牛品种所特有。6个群体的有效等位基因数(N_e)在2.2043-3.2754之间, 平均杂合度(H)在0.4858-0.6153之间, 平均多态信息含量(PIC)在0.4230-0.5711之间。5个牦牛品种的微卫星座位有丰富的遗传多样性; 而大额牛的遗传多样性相对较贫乏。5个牦牛群体间的遗传分化系数(Gst)为0.0527, 表明牦牛亚群体间遗传分化水平很低。采用邻近结合法构建聚类图和模糊聚类分析表明, 5个牦牛品种分为两大类, 其中斯布牦牛、西藏高山牦牛、帕里牦牛和麦洼牦牛为一大类, 九龙牦牛为一类。研究结果将为中国地方牦牛品种的保护和利用提供重要的理论依据。

关键词: 牦牛, 遗传分化, 资源保护, 遗传关系

Abstract

Genetic variations within and relationships between five indigenous Chinese yak (Bos grunniens) breeds (Pali, Sibu, Tibetan High Mountainous yak, Maiwa and Jiulong) were investigated using 16 microsatellite markers with the fluorescence-multiplex PCR technique. The gayal (Bos frontalis) was used as outgroup. For the six populations studied, number of effective alleles (N_e) ranged from 2.2043 to 3.2754, mean heterozygosity (H) ranged from 0.4858 to 0.6153, and mean polymorphism information content (PIC) ranged from 0.4230 to 0.5711. The five indigenous Chinese yak breeds had high genetic diversity at the microsatellite DNA level, while the gayal showed less genetic variation. The genetic differentiation among the populations was very low, with a Gst value of 0.0527. In neighbor-joining and fuzzy clustering trees, the five yak breeds grouped into two clades, one comprising Jiulong yaks and the other including Pali, Sibu, Tibetan High Mountainous yak and Maiwa yaks. Our results may provide a scientific basis for future conservation and exploitation of native Chinese yak breeds.

Key words: yak, genetic differentiation, resource conservation, genetic relationship

廖信军, 常洪, 张桂香, 王冬蕾, 宋卫涛, 韩旭, 张自富 (2008) 中国5个地方牦牛品种遗传多样性的微卫星分析. 生物多样性, 16, 156-165. DOI: 10.3724/SP.J.1003.2008.07292.

Xinjun Liao, Hong Chang, Guixiang Zhang, Donglei Wang, Weitao Song, Xu Han, Zifu Zhang (2008) Genetic diversity of five native Chinese yak breeds based on microsatellite DNA markers. Biodiversity Science, 16, 156-165. DOI: 10.3724/SP.J.1003.2008.07292.

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链接本文: https://www.biodiversity-science.net/CN/10.3724/SP.J.1003.2008.07292

https://www.biodiversity-science.net/CN/Y2008/V16/I2/156

图/表 10

参考文献 26

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代号 Code	品种 Breeds	数量 Sample size	采集地点 Collection location
PL	帕里牦牛 Pali yak	62	西藏自治区帕里镇 Pali Town, Tibet
SB	斯布牦牛 Sibu yak	57	西藏自治区斯布乡斯布牦牛场 Sibu Yak Farm of Sibu Town, Tibet
GS	西藏高山牦牛 Tibetan High Mountainous yak	61	西藏自治区那曲地区那曲县布巴镇 Buba Town, Naqu County, Tibet
JL	九龙牦牛 Jiulong yak	60	四川九龙县 Jiulong County, Sichuan
MW	麦洼牦牛 Maiwa yak	59	四川阿坝州畜牧局牦牛选育基地 Yak Breeding Center of Aba State Livestock Bureau, Sichuan
GY	大额牛 Gayal	20	云南泸水县老窝乡 Laowo Town, Lushui County, Yunnan

代号 Code	品种 Breeds	数量 Sample size	采集地点 Collection location
PL	帕里牦牛 Pali yak	62	西藏自治区帕里镇 Pali Town, Tibet
SB	斯布牦牛 Sibu yak	57	西藏自治区斯布乡斯布牦牛场 Sibu Yak Farm of Sibu Town, Tibet
GS	西藏高山牦牛 Tibetan High Mountainous yak	61	西藏自治区那曲地区那曲县布巴镇 Buba Town, Naqu County, Tibet
JL	九龙牦牛 Jiulong yak	60	四川九龙县 Jiulong County, Sichuan
MW	麦洼牦牛 Maiwa yak	59	四川阿坝州畜牧局牦牛选育基地 Yak Breeding Center of Aba State Livestock Bureau, Sichuan
GY	大额牛 Gayal	20	云南泸水县老窝乡 Laowo Town, Lushui County, Yunnan

座位 Locus	引物序列 (5’-3’) Primer sequence	退火温度 Annealing temp.(℃)	片段大小 Allele Size (bp)	观察等位基因数 Observed allele number	荧光标记 Fluorescence	所在染色体 Chromosome location
SPS115	F: AAAGTGACACAACAGCTTCTCCAG R: AACGAGTGTCCTAGTTTGGCTGTG	55-60	234-258	11	Vic	15
TGLA122	F: CCCTCCTCCAGGTAAATCAGC R: AATCACATGGCAAATAAGTACATAC	55-58	136-184	12	Ned	21
TGLA53	F: GCTTTCAGAAATAGTTTGCATTCA R: ATCTTCACATGATATTACAGCAGA	55	143-191	13	Pet	16
TGLA126	F: CTAATTTAGAATGAGAGAGGCTTCT R: TTGGTCTCTATTCTCTGAATATTCC	55-58	115-131	7	Vic	20
TGLA227	F: CGAATTCCAAATCTGTTAATTTGCT R: ACAGACAGAAACTCAATGAAAGCA	55-56	75-105	10	Pet	18
HEL13	F: TAAGGACTTGAGATAAGGAG R: CCATCTACCTCCATCTTAAC	51.6	165-191	11	Pet	11
INRA005	F: CAATCTGCATGAAGTATAAATAT R: CTTCAGGCATACCCTACACC	55	120-160	10	Ned	12
BM1824	F: GAGCAAGGTGTTTTTCCAATC R: CATTCTCCAACTGCTTCCTTG	57	180-200	7	6fam	1
HEL5	F: GCAGGATCACTTGTTAGGGA R: AGACGTTAGTGTACATTAAC	54	149-165	7	Vic	21
BM2113	F: GCTGCCTTCTACCAAATACCC R: CTTAGACAACAGGGGTTTGG	55-60	116-146	13	6fam	2
TGLA57	F: GCTTCCAAAACTTTACAATATGTAT R: GCTTTTTAATCCTCAGCTTGCTG	55	77-99	9	6fam	1
ILSTS013	F: ACACAAAATCAGATCAGTGG R: CTTGATCCTTATAGAACTGG	58	121-135	11	Vic	9
ILSTS050	F: AAATCAGACACCCAGTTTCC R: GTTTTTCTACACGAGTTGGC	55	161-183	13	Pet	21
AGLA293	F: GAAACTCAACCCAAGACAACTCAAG R: ATGACTTTATTCTCCACCTAGCAGA	55	210-240	11	6fam	5
ILSTS008	F: GAATCATGGATTTTCTGGGG R: TAGCAGTGAGTGAGGTTGGC	58	175-187	7	6fam	14
TGLA73	F: GAGAATCACCTAGAGAGAGGCA R: CTTTCTCTTTAAATTCTATATGGT	55	111-143	14	6fam	9

座位 Locus	引物序列 (5’-3’) Primer sequence	退火温度 Annealing temp.(℃)	片段大小 Allele Size (bp)	观察等位基因数 Observed allele number	荧光标记 Fluorescence	所在染色体 Chromosome location
SPS115	F: AAAGTGACACAACAGCTTCTCCAG R: AACGAGTGTCCTAGTTTGGCTGTG	55-60	234-258	11	Vic	15
TGLA122	F: CCCTCCTCCAGGTAAATCAGC R: AATCACATGGCAAATAAGTACATAC	55-58	136-184	12	Ned	21
TGLA53	F: GCTTTCAGAAATAGTTTGCATTCA R: ATCTTCACATGATATTACAGCAGA	55	143-191	13	Pet	16
TGLA126	F: CTAATTTAGAATGAGAGAGGCTTCT R: TTGGTCTCTATTCTCTGAATATTCC	55-58	115-131	7	Vic	20
TGLA227	F: CGAATTCCAAATCTGTTAATTTGCT R: ACAGACAGAAACTCAATGAAAGCA	55-56	75-105	10	Pet	18
HEL13	F: TAAGGACTTGAGATAAGGAG R: CCATCTACCTCCATCTTAAC	51.6	165-191	11	Pet	11
INRA005	F: CAATCTGCATGAAGTATAAATAT R: CTTCAGGCATACCCTACACC	55	120-160	10	Ned	12
BM1824	F: GAGCAAGGTGTTTTTCCAATC R: CATTCTCCAACTGCTTCCTTG	57	180-200	7	6fam	1
HEL5	F: GCAGGATCACTTGTTAGGGA R: AGACGTTAGTGTACATTAAC	54	149-165	7	Vic	21
BM2113	F: GCTGCCTTCTACCAAATACCC R: CTTAGACAACAGGGGTTTGG	55-60	116-146	13	6fam	2
TGLA57	F: GCTTCCAAAACTTTACAATATGTAT R: GCTTTTTAATCCTCAGCTTGCTG	55	77-99	9	6fam	1
ILSTS013	F: ACACAAAATCAGATCAGTGG R: CTTGATCCTTATAGAACTGG	58	121-135	11	Vic	9
ILSTS050	F: AAATCAGACACCCAGTTTCC R: GTTTTTCTACACGAGTTGGC	55	161-183	13	Pet	21
AGLA293	F: GAAACTCAACCCAAGACAACTCAAG R: ATGACTTTATTCTCCACCTAGCAGA	55	210-240	11	6fam	5
ILSTS008	F: GAATCATGGATTTTCTGGGG R: TAGCAGTGAGTGAGGTTGGC	58	175-187	7	6fam	14
TGLA73	F: GAGAATCACCTAGAGAGAGGCA R: CTTTCTCTTTAAATTCTATATGGT	55	111-143	14	6fam	9

座位 Locus	帕里牦牛 Pali yak		斯布牦牛 Sibu yak	西藏高山牦牛 Tibetan High Mountainous yak	九龙牦牛 Jiulong yak	麦洼牦牛 Maiwa yak	大额牛 Gayal
BM1824				192 (0.009)
BM2113						116 (0.009)	120 (0.632)
						138 (0.009) .	132 (0.261)
HEL5	149 (0.010)			141 (0.010)	155 (0.021)
INRA005			136 (0.026)				132 (0.500)
			148 (0.009)
TGLA126					127(0.009)
TGLA53				161 (0.009)		179 (0.009)
				175 (0.009)
HEL13	83 (0.009)					165 (0.009)	191 (0.375)
	193 (0.096)
SPS115	230 (0.018)						256 (0.103)
TGLA122	138 (0.018)		142 (0.008)		166 (0.009)
					168 (0.017)
TGLA227		87 (0.009)		99 (0.010)	101 (0.020)	97 (0.009)	81 (0.125)
AGLA293	202 (0.009)			238 (0.009)	220 (0.017)		234 (0.059)
				242 (0.019)
ILSTS013	105 (0.009)						133(0.231)
	109 (0.009)
ILSTS050							147 (0.184) 149 (0.105) 151 (0.316)
							153 (0.316) 155 (0.079)
TGLA57		31 (0.017)				71 (0.008)	97 (0.017)
TGLA73					193 (0.017)

中国5个地方牦牛品种遗传多样性的微卫星分析

Genetic diversity of five native Chinese yak breeds based on microsatellite DNA markers

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座位 Locus	帕里牦牛 Pali yak	斯布牦牛 Sibu yak	西藏高山牦牛 Tibetan High Mountainous yak	九龙牦牛 Jiulong yak	麦洼牦牛 Maiwa yak	大额牛 Gayal
BM1824					180 (0.562)	186 (0.925)
BM2113	128 (0.772)		128 (0.583)		128 (0.612)	120 (0.632)
HEL5	137 (0.696)	137 (0.962)	137 (0.781)	137 (0.894)	137 (0.726)
INRA005	134 (0.658)				134 (0.543)	132 (0.500), 134 (0.500)
TGLA126					117 (0.586)	119 (0.700)
TGLA53				153 (0.500)		159 (0.500), 165 (0.500)
HEL13			183 (0.644)	183 (0.628)	183 (0.603)	197 (0.625)
SPS115	240 (0.465)					250 (0.667)
TGLA122					152 (0.388)
TGLA227	75 (0.632)	75 (0.573)	73 (0.620)	73 (0.561)	73 (0.602)	75 (0.850)
AGLA293	208 (0.860)	208 (0.890)	208 (0.833)	208 (0.627)	208 (0.737)	226 (0.529)
ILSTS008	181 (0.553)			181 (0.686)	181 (0.647)	171 (0.895)
ILSTS013			121 (0.593)	121 (0.585)		117 (0.410)
ILSTS050				169 (0.696)		151 (0.316), 153 (0.316)
TGLA57	73 (0.658)	73 (0.746)	73 (0.741)	73 (0.636)	73 (0.746)	81 (0.450)
TGLA73	115 (0.246)					125 (0.600)

座位 Locus	5个牦牛群体共有等位基因(bp) Common alleles in five yak populations	6个品种共有等位基因(bp) Common alleles in six breeds	5个牦牛群体特有等位基因数 No. of unique allele in five yak populations	大额牛特有等位基因数 No. of unique allele in Gayal
BM1824	178, 180, 186, 190	186	1	0
BM2113	118, 124, 128, 134, 144	124, 128	2	2
HEL5	137, 139		3	0
INRA005	134, 152	134	2	1
TGLA126	109, 115, 117, 119	119	1	0
TGLA53	153, 157, 159, 165, 169	165	3	0
HEL13	183, 189, 197, 199	197	3	1
SPS115	232, 238, 240, 242, 244, 246		1	1
TGLA122	140, 146, 148, 150, 152, 164		4	0
TGLA227	73, 75	75	4	1
AGLA293	208, 214, 226	226	4	1
ILSTS008	179, 181		0	0
ILSTS013	119, 121, 123, 125, 127	119	2	1
ILSTS050	167, 169, 171		0	5
TGLA57	73, 77, 81, 83	77, 81	2	1
TGLA73	115, 117, 119, 123, 125, 127, 129, 193	115, 117, 123, 125	1	0

品种 Breeds	帕里牦牛 PL	斯布牦牛 SB	西藏高山牦牛 GS	九龙牦牛 JL	麦洼牦牛 MW	大额牛 GY
帕里牦牛 PL		0.0861	0.0707	0.1266	0.0944	0.7136
斯布牦牛 SB	0.0793		0.0715	0.1071	0.0615	0.7052
西藏高山牦牛 GS	0.0827	0.0649		0.0970	0.0723	0.6955
九龙牦牛 JL	0.1632	0.1659	0.1168		0.0987	0.6515
麦洼牦牛 MW	0.0838	0.0738	0.0937	0.1239		0.6836
大额牛 GY	1.1508	1.4356	1.6049	1.2457	1.4628

品种 Breeds	帕里牦牛 PL	斯布牦牛 SB	西藏高山牦牛 GS	九龙牦牛 JL	麦洼牦牛 MW	大额牛 GY
帕里牦牛 PL	1.0000	0.9530	0.9530	0.9340	0.9530	0.3450
斯布牦牛 SB	0.9530	1.0000	0.9600	0.9340	0.9560	0.3450
西藏高山牦牛 GS	0.9510	0.9600	1.0000	0.9340	0.9560	0.3450
九龙牦牛 JL	0.9110	0.9100	0.9340	1.0000	0.9340	0.3450
麦洼牦牛 MW	0.9510	0.9560	0.9460	0.9310	1.0000	0.3450
大额牛 GY	0.2420	0.2500	0.1650	0.3450	0.2370	1.0000

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