基于SRAP分子标记的华顶杜鹃遗传多样性

doi:10.3969/j.issn.1004-1524.20221828

浙江农业学报 ›› 2024, Vol. 36 ›› Issue (1): 127-133.DOI: 10.3969/j.issn.1004-1524.20221828

基于SRAP分子标记的华顶杜鹃遗传多样性

吴倩¹(), 汤紫依¹, 田盛野¹, 何海叶¹, 潘伟伟¹, 王军峰², 鲍洪华³, 张慧娟¹, 蒋明¹^,^*()

1.台州学院生命科学学院,浙江台州 318000
2.华东药用植物园科研管理中心,浙江丽水 323000
3.台州市生态环境保护局路桥分局,浙江台州 318050

收稿日期:2022-12-26 出版日期:2024-01-25 发布日期:2024-02-18
作者简介:吴倩(2000—),女,湖南衡阳人,本科生,研究方向为植物分子生物学。E-mail:1625197596@qq.com。
通讯作者: * 蒋明,E-mail:jiangming1973@139.com
基金资助:
台州市大学生科技创新项目(2021年度);国家级大学生创新创业训练计划(202110350015)

Genetic diversity of Rhododendron huadingense based on SARP molecular marker

WU Qian¹(), TANG Ziyi¹, TIAN Shengye¹, HE Haiye¹, PAN Weiwei¹, WANG Junfeng², BAO Honghua³, ZHANG Huijuan¹, JIANG Ming¹^,^*()

1. College of Life Sciences, Taizhou University, Taizhou 318000, Zhejiang, China
2. Scientific Research Management Center, East China Medicinal Botanical Garden, Lishui 323000, Zhejiang, China
3. Luqiao Branch of Taizhou Municipal Ecology and Environment Bureau, Taizhou 318050, Zhejiang, China

Received:2022-12-26 Online:2024-01-25 Published:2024-02-18

摘要/Abstract

摘要：

华顶杜鹃是浙江特有的珍稀濒危野生植物,分布区域十分狭窄,植株数量稀少,该植物亟待保护。以不同居群华顶杜鹃的叶片为材料,采用相关序列扩增多态性(sequence-related amplified polymorphism,SRAP)分子标记技术进行遗传多样性研究。结果表明,从140个组合中筛选出8对扩增多态性高、条带清晰、重复性好的SRAP引物,利用这些引物对50份华顶杜鹃基因组DNA进行PCR扩增,共得到263个条带,其中多态性位点261个,多态性比率(PPB)为99.24%;在物种水平上,观测等位基因数(N_a)为1.992 4,有效等位基因数(N_e)为1.340 1,Nei’s基因多样性指数(H)为0.221 5,Shannon's信息指数(I)为0.357 9,表明华顶杜鹃在物种水平上有较高的遗传多样性;在居群水平上,5个自然群体的遗传分化系数(G_st)为0.459 6,基因流(N_m)为0.587 8,表明居群间存在一定的基因分化和基因交流,而大部分遗传变异出现在居群内;UPGMA聚类结果表明,5个居群的样品可分为5组,与主成分分析结果一致。研究结果为后续开展该珍稀物种的保护和利用奠定了基础。

关键词: 华顶杜鹃, 遗传多样性, 亲缘关系

Abstract:

Rhododendron huadingense is a rare and endangered wild plant species endemic to Zhejiang Province, China, with narrow distribution and very few individuals, thus this species needs to be protected. In this study, leaves of R. huadingense from different populations were collected, and SRAP (sequence related amplified polymorphism) molecular marker technology was applied to study its genetic diversity. Fifty genomic DNA samples were used for PCR amplification. The results indicated 8 pairs of primers with high levels of polymorphism, clear bands, and high reliability were screened from 140 primer pairs. Totally, 263 bands were detected, including 261 polymorphic sites, and the percentage of the polymorphic band reached 99.24%. On species level, the number of observed alleles (N_a), number of effective alleles (N_e), Nei’s gene diversity (H), and Shannon’s information index (I) were 1.992 4, 1.340 1, 0.221 5, and 0.357 9, respectively, revealing its high genetic diversity on species level. On population level, the coefficient of genetic differentiation (G_st) and gene flow (N_m) were 0.459 6 and 0.587 8, respectively, indicating genetic differentiation and gene flow have occurred among populations, and most of the variation existed within populations. UPGMA results showed samples from 5 populations could be divided into 5 groups, which were consistent with principal component analysis results. Studies on genetic diversity of R. huadingense provided insights on protection and utilization of this rare plant species.

Key words: Rhododendron huadingense, genetic diversity, relationship

中图分类号:

S685.21

吴倩, 汤紫依, 田盛野, 何海叶, 潘伟伟, 王军峰, 鲍洪华, 张慧娟, 蒋明. 基于SRAP分子标记的华顶杜鹃遗传多样性[J]. 浙江农业学报, 2024, 36(1): 127-133.

WU Qian, TANG Ziyi, TIAN Shengye, HE Haiye, PAN Weiwei, WANG Junfeng, BAO Honghua, ZHANG Huijuan, JIANG Ming. Genetic diversity of Rhododendron huadingense based on SARP molecular marker[J]. Acta Agriculturae Zhejiangensis, 2024, 36(1): 127-133.

图/表 8

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居群代号 Population code	样本数 Sample size	海拔 Altitude/m	经度 Longitude	纬度 Latitude
TGJ	10	936~949	121°0'72″~121°0'90″	28°45'124″~28°45'45″
DCJ	10	705~817	121°2'110″~121°2'381″	28°48'11″~28'48'51″
CZ	10	717~781	121°0'103″~121°0'948″	28°45'290″~28°45'376″
KCS	10	706~787	120°56'192″~120°56'410″	28°48'032″~28°48'956″
TTS	10	903~918	121°05'257″~121°05'283″	29°15'106″~29°15'179″

居群代号 Population code	样本数 Sample size	海拔 Altitude/m	经度 Longitude	纬度 Latitude
TGJ	10	936~949	121°0'72″~121°0'90″	28°45'124″~28°45'45″
DCJ	10	705~817	121°2'110″~121°2'381″	28°48'11″~28'48'51″
CZ	10	717~781	121°0'103″~121°0'948″	28°45'290″~28°45'376″
KCS	10	706~787	120°56'192″~120°56'410″	28°48'032″~28°48'956″
TTS	10	903~918	121°05'257″~121°05'283″	29°15'106″~29°15'179″

编号 Code	引物组合 Primer combinations	上游引物 Forward primer (5'→3')	下游引物 Reverse primer (5'→3')
1	ME 2/em 6	ME 2:TGAGTCCAAACCGGAGC	em 6:GACTGCGTACGAATTGCA
2	ME 3/em 6	ME 3:TGAGTCCAAACCGGAAT	em 6:GACTGCGTACGAATTGCA
3	ME 3/em 14	ME 3:TGAGTCCAAACCGGAAT	em 14:GACTGCGTACGAATTCAG
4	ME 4/em 11	ME 4:TGAGTCCAAACCGGACC	em 11:GACTGCGTACGAATTTCC
5	ME 4/em 14	ME 4:TGAGTCCAAACCGGACC	em 14:GACTGCGTACGAATTCAG
6	ME 4/em 17	ME 4:TGAGTCCAAACCGGACC	em 17:GACTGCGTACGAATTCCA
7	ME 5/em 4	ME 5:TGAGTCCAAACCGGAAG	em 4:GACTGCGTACGAATTTGA
8	ME 8/em 1	ME 8:TGAGTCCAAACCGGTGT	em 1:GACTGCGTACGAATTAAT

编号 Code	引物组合 Primer combinations	上游引物 Forward primer (5'→3')	下游引物 Reverse primer (5'→3')
1	ME 2/em 6	ME 2:TGAGTCCAAACCGGAGC	em 6:GACTGCGTACGAATTGCA
2	ME 3/em 6	ME 3:TGAGTCCAAACCGGAAT	em 6:GACTGCGTACGAATTGCA
3	ME 3/em 14	ME 3:TGAGTCCAAACCGGAAT	em 14:GACTGCGTACGAATTCAG
4	ME 4/em 11	ME 4:TGAGTCCAAACCGGACC	em 11:GACTGCGTACGAATTTCC
5	ME 4/em 14	ME 4:TGAGTCCAAACCGGACC	em 14:GACTGCGTACGAATTCAG
6	ME 4/em 17	ME 4:TGAGTCCAAACCGGACC	em 17:GACTGCGTACGAATTCCA
7	ME 5/em 4	ME 5:TGAGTCCAAACCGGAAG	em 4:GACTGCGTACGAATTTGA
8	ME 8/em 1	ME 8:TGAGTCCAAACCGGTGT	em 1:GACTGCGTACGAATTAAT

居群 Population	居群个体数 Individuals in a population	多态位点数 Number of polymorphic bands	多态位点百分率 Percentage of polymorphic bands%	观测等位基因数 Observed number of alleles	有效等位基因数 Effective number of alleles	Nei’s基因多样性指数 Nei’s gene diversity index	Shannon's信息指数 Shannon’s information index
TGJ	10	93	35.36	1.353 6	1.201 7	0.117 8	0.177 7
DCJ	10	102	38.78	1.387 8	1.220 5	0.130 0	0.196 2
CZ	10	95	36.12	1.36 12	1.199 8	0.118 6	0.179 8
KCS	10	108	41.06	1.410 6	1.235 1	0.138 1	0.207 9
TTS	10	88	33.46	1.334 6	1.148 6	0.093 7	0.147 0
平均Average	10	99.5	36.96	1.369 6	1.201 1	0.119 9	0.181 7
物种水平	50	261	99.24	1.992 4	1.340 1	0.221 5	0.357 9
Species level

基于SRAP分子标记的华顶杜鹃遗传多样性

Genetic diversity of Rhododendron huadingense based on SARP molecular marker

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居群 Population	TGJ	DCJ	CZ	KCS	TTS
TGJ		0.869 6	0.844 1	0.839 4	0.874 7
DCJ	0.139 8		0.889 9	0.846 8	0.889 2
CZ	0.169 5	0.116 7		0.804 7	0.858 2
KCS	0.175 0	0.166 3	0.217 3		0.837 9
TTS	0.133 9	0.117 5	0.152 9	0.176 9

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