毛茛叶报春的遗传多样性及遗传结构

doi:10.3724/SP.J.1003.2013.09098

生物多样性 ›› 2013, Vol. 21 ›› Issue (5): 601-609. DOI: 10.3724/SP.J.1003.2013.09098

毛茛叶报春的遗传多样性及遗传结构

王德元¹, 彭婕¹, 陈雅静¹, 吕国胜¹, 张小平^1,², 邵剑文^1,^3,^*()

1 安徽师范大学生命科学学院, 安徽芜湖 241000
2 生物环境与生态安全安徽省高校重点实验室, 安徽芜湖 241000
3 安徽省重要生物资源保护与利用研究重点实验室, 安徽芜湖 241000

收稿日期:2013-04-18 接受日期:2013-07-01 出版日期:2013-09-20 发布日期:2013-10-08
通讯作者: 邵剑文
基金资助:
国家自然科学基金面上项目(31170317);安徽高校省级科学研究重点项目(KJ2011A132)

Genetic diversity and genetic structure of the rare and endangered species, Primula ranunculoides

Deyun Wang¹, Jie Peng¹, Yajing Chen¹, Guosheng Lü¹, Xiaoping Zhang^1,², Jianwen Shao^1,^3,^*()

1 College of Life Science, Anhui Normal University, Wuhu, Anhui 241000
2 Key Laboratory of Biotic Environment and Ecological Safety in Anhui Province, Anhui Normal University, Wuhu, Anhui 241000
3 Key Laboratory of Conservation and Employment of Biological Resources of Anhui, Wuhu, Anhui 241000

Received:2013-04-18 Accepted:2013-07-01 Online:2013-09-20 Published:2013-10-08
Contact: Shao Jianwen

摘要/Abstract

摘要：

毛茛叶报春(Primula ranunculoides)是我国特有的珍稀濒危花卉, 具较好的园艺开发潜力, 其野生种群已十分稀少。本文利用9对微卫星引物对7个自然种群共222个个体的遗传多样性和遗传结构进行了研究。结果表明, 与近缘种相比, 毛茛叶报春具较低的遗传多样性, 种群间分化明显。种群的平均观察杂合度为0.286, 期望杂合度为0.330, 种群遗传多样性与植株密度有显著的正相关性, 而与种群大小和面积无显著相关性。种群间的平均基因流N_m = 0.730。AMOVA分析表明毛茛叶报春有48.08%的变异存在于种群间, 51.92%出现在种群内。毛茛叶报春种下的遗传结构与地形关系密切, 山脉间的两湖平原(江汉平原和洞庭湖平原)及武宁和修水低洼农耕区等异质生境是其主要的基因流屏障。建议将毛茛叶报春划分为九岭山、幕阜山、七姊妹山和银炉4个单元进行遗传多样性管理。

关键词: Primula ranunculoides, 遗传多样性, 遗传结构, 微卫星, 保护建议

Abstract

The rare and endangered species Primula ranunculoides is endemic to China and is of reasonably high potential horticultural value. The genetic diversity and structure of the species was examined by surveying variation at nine microsatellite loci across 222 individuals sampled from seven natural populations. The results indicated that there was relatively low genetic diversity present within populations (H_e = 0.330, H_o = 0.286) and high genetic differentiation among populations. Levels of population genetic diversity and effective population size were both correlated with plant density, rather than population census size or population area. Gene flow between populations was low (N_m = 0.730) and an AMOVA analysis showed that 48.08% of the total genetic diversity was attributable to among populations and the rest (51.92%) to variation within populations. Bayesian assignment and principal coordinate analyses supported clustering of the seven populations into four groups, which were correlated with topographical features. This suggested that Lianghu Plain (Dongtinghu Plain and Jianghan Plain) and the farming area of Xiushui and Wuning counties were major barriers to gene flow, causing high divergence between different mountain populations. Based on the genetic structure of P. ranunculoides, four management units for conservation purposes are proposed, i.e. the Jiulingshan, Mufushan, Qizimeishan and Yinlu units.

Key words: Primula ranunculoides, genetic diversity, genetic structure, microsatellites, conservation proposal

王德元, 彭婕, 陈雅静, 吕国胜, 张小平, 邵剑文 (2013) 毛茛叶报春的遗传多样性及遗传结构. 生物多样性, 21, 601-609. DOI: 10.3724/SP.J.1003.2013.09098.

Deyun Wang,Jie Peng,Yajing Chen,Guosheng Lü,Xiaoping Zhang,Jianwen Shao (2013) Genetic diversity and genetic structure of the rare and endangered species, Primula ranunculoides. Biodiversity Science, 21, 601-609. DOI: 10.3724/SP.J.1003.2013.09098.

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链接本文: https://www.biodiversity-science.net/CN/10.3724/SP.J.1003.2013.09098

https://www.biodiversity-science.net/CN/Y2013/V21/I5/601

图/表 8

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种群 Population	地点 Locality	生境 Habitat	地理位置 Geographical location	海拔 Altitude (m)	面积 Area (m²)	花朵类型 Flower type	种群大小 Population size
杨家坪A YJPA	江西省修水县杨家坪 Yangjiaping, Xiushui County, Jiangxi	林下溪边或路边 Brook-side or roadside under forests	28°47'45"N, 114°43'56"E	300-375	720	两型花 Distyle	~2,000
杨家坪B YJPB	江西省修水县杨家坪 Yangjiaping, Xiushui County, Jiangxi	林下溪边或路边 Brook-side or roadside, under forest	28°47'04"N, 114°44'13"E	350-360	1,060	两型花 Distyle	~6,000
毛竹山 MZS	江西省修水县毛竹山 Maozhushan, Xiushui County, Jiangxi	林下溪边或路边 Brook-side or roadside, under forest	28°50'31"N, 114°52'00"E	390-410	1,650	两型花 Distyle	~1,500
三界A SJA	湖北省通山县三界 Sanjie, Tongshan County, Hubei	林下溪边或路边 Brook-side or roadside, under forest	29°24'16"N, 114°27'15"E	255-285	545	两型花 Distyle	~1,000
三界B SJB	湖北省通山县三界 Sanjie, Tongshan County, Hubei	林下溪边或路边 Brook-side or roadside, under forest	29°22'57"N, 114°27'08"E	265-280	385	两型花 Distyle	~700
七姊妹山 JPT	湖北省宣恩县七姊妹山 Qizimei Mount, Xuanen County, Hubei	林下溪边 Brook-side, under forest	30°02'09"N, 109°43'57"E	1,310-1,365	1,200	同型花 Monostyle	~600
银炉 YL	江西省武宁县银炉 Yinlu, Wuning County, Jiangxi	溪边石壁 Rock near brook-side	28°59'31''N, 114°49'39''E	420	50	两型花 Distyle	~200

种群 Population	地点 Locality	生境 Habitat	地理位置 Geographical location	海拔 Altitude (m)	面积 Area (m²)	花朵类型 Flower type	种群大小 Population size
杨家坪A YJPA	江西省修水县杨家坪 Yangjiaping, Xiushui County, Jiangxi	林下溪边或路边 Brook-side or roadside under forests	28°47'45"N, 114°43'56"E	300-375	720	两型花 Distyle	~2,000
杨家坪B YJPB	江西省修水县杨家坪 Yangjiaping, Xiushui County, Jiangxi	林下溪边或路边 Brook-side or roadside, under forest	28°47'04"N, 114°44'13"E	350-360	1,060	两型花 Distyle	~6,000
毛竹山 MZS	江西省修水县毛竹山 Maozhushan, Xiushui County, Jiangxi	林下溪边或路边 Brook-side or roadside, under forest	28°50'31"N, 114°52'00"E	390-410	1,650	两型花 Distyle	~1,500
三界A SJA	湖北省通山县三界 Sanjie, Tongshan County, Hubei	林下溪边或路边 Brook-side or roadside, under forest	29°24'16"N, 114°27'15"E	255-285	545	两型花 Distyle	~1,000
三界B SJB	湖北省通山县三界 Sanjie, Tongshan County, Hubei	林下溪边或路边 Brook-side or roadside, under forest	29°22'57"N, 114°27'08"E	265-280	385	两型花 Distyle	~700
七姊妹山 JPT	湖北省宣恩县七姊妹山 Qizimei Mount, Xuanen County, Hubei	林下溪边 Brook-side, under forest	30°02'09"N, 109°43'57"E	1,310-1,365	1,200	同型花 Monostyle	~600
银炉 YL	江西省武宁县银炉 Yinlu, Wuning County, Jiangxi	溪边石壁 Rock near brook-side	28°59'31''N, 114°49'39''E	420	50	两型花 Distyle	~200

位点 Locus	序列 Repeat motif	引物 Primer (5'-3')	退火温度(℃) Annealing temperature	荧光标记 Fluorescent label	等位基因数 No. of alleles
Pm1	(TC)₁₀(CT)₃	F:ATCTTTGAGGTCCTTTTA	50	FAM	7
		R: ATCGCCCAATGGAGTGAA
Pm2	(AG)₁₄	F: CGCCTACAGTGTTTGGGA	55	FAM	2
		R: CTATCTCACCTGCGTTCT
Pm7	(AG)₃GG(AG)₈	F: TTGTTCACCGACGCATAC	54	HEX	18
		R: TTACACGCACCAAATCAT
Pm9	(TTTC)₂(TC)₆	F: AGACTCACGAGGAATACG	50	HEX	6
		R: AGAAAAGGAGGAGACAAA
Pm12	(TC)₁₀(CT)₃	F: TAAAACTCCTGGAGGGGTAC	51	TAMRA	10
		R: ATCGCCCAATGGAGTGAA
Pm13	(GATAGG)₂GAT(AG)₇	F: GAGGACAGGCACCACAGA	54	TAMRA	6
		R: TCCCCAACTTCATGCTCTT
Pm14	(AG)₃(GA)₁₂	F: TCGCCCAATGGAGTGAAC	50	TAMRA	9
		R: TCTTTGAGGTCCTTTTAT
Pm16	(GAGGGA)₃(GA)₃	F: AACCACTCGTCGTCCTAA	51	FAM	5
		R: CGATAGATTGCCTTACCC
Pm17	(GA)₉	F: TAAATCAAGGTAGCAACT	51	HEX	9
		R: TACCTACCATTACTCCC

位点 Locus	序列 Repeat motif	引物 Primer (5'-3')	退火温度(℃) Annealing temperature	荧光标记 Fluorescent label	等位基因数 No. of alleles
Pm1	(TC)₁₀(CT)₃	F:ATCTTTGAGGTCCTTTTA	50	FAM	7
		R: ATCGCCCAATGGAGTGAA
Pm2	(AG)₁₄	F: CGCCTACAGTGTTTGGGA	55	FAM	2
		R: CTATCTCACCTGCGTTCT
Pm7	(AG)₃GG(AG)₈	F: TTGTTCACCGACGCATAC	54	HEX	18
		R: TTACACGCACCAAATCAT
Pm9	(TTTC)₂(TC)₆	F: AGACTCACGAGGAATACG	50	HEX	6
		R: AGAAAAGGAGGAGACAAA
Pm12	(TC)₁₀(CT)₃	F: TAAAACTCCTGGAGGGGTAC	51	TAMRA	10
		R: ATCGCCCAATGGAGTGAA
Pm13	(GATAGG)₂GAT(AG)₇	F: GAGGACAGGCACCACAGA	54	TAMRA	6
		R: TCCCCAACTTCATGCTCTT
Pm14	(AG)₃(GA)₁₂	F: TCGCCCAATGGAGTGAAC	50	TAMRA	9
		R: TCTTTGAGGTCCTTTTAT
Pm16	(GAGGGA)₃(GA)₃	F: AACCACTCGTCGTCCTAA	51	FAM	5
		R: CGATAGATTGCCTTACCC
Pm17	(GA)₉	F: TAAATCAAGGTAGCAACT	51	HEX	9
		R: TACCTACCATTACTCCC

种群 Population	取样数 Sample size	有效种群大小 Effective population size	期望杂合度(H_e) Expected heterozygosity	观察杂合度(H_o) Observed heterozygosity	稀有等位基因 Rare alleles	近交系数(F_is) Inbreeding coefficient
YJPA	32	166	0.430	0.424	2	0.014 ^ns
YJPB	31	231	0.412	0.399	4	0.032^**
MZS	30	105	0.297	0.155	2	0.484^***
SJA	33	169	0.348	0.330	4	0.055^***
SJB	31	149	0.281	0.230	1	0.184^***
JPT	33	127	0.203	0.132	2	0.350^***
YL	32	153	0.340	0.328	2	0.037^**
平均 Mean	31.7	157.1	0.330	0.286	2.4

毛茛叶报春的遗传多样性及遗传结构

Genetic diversity and genetic structure of the rare and endangered species, Primula ranunculoides

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	YJPA	YJPB	MZS	SJA	SJB	JPT	YL
YJPA	-	0.091	0.208	0.420	0.528	0.875	0.702
YJPB	0.096	-	0.167	0.311	0.363	0.793	0.588
MZS	0.256	0.222	-	0.336	0.378	0.827	0.742
SJA	0.301	0.262	0.399	-	0.030	0.921	0.473
SJB	0.385	0.332	0.476	0.047	-	0.929	0.467
JPT	0.510	0.526	0.645	0.569	0.599	-	0.318
YL	0.445	0.424	0.536	0.392	0.428	0.367	-

变异来源 Source	自由度 d.f.	方差和 Sum of squares	变异组分 Variance components	占总变异比例% Percentage of variation	显著性检验 Significance tests
种群间 Among populations	6	225.635	0.6596	48.08	P<0.001
种群内 Within populations	215	310.851	0.7124	51.92	P<0.001
总和 Total	222	536.486	1.3720	100%

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