基于SRAP分子标记技术研究浙产栀子遗传多样性
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摘要
为探究浙产栀子种质遗传多样性,进一步开展该物种保护及品种选育,利用SRAP分子标记技术,从多态性检测、遗传多样性水平及聚类分析揭示栀子种质遗传多样性。结果表明:从88对SRAP引物中筛选6对引物用于浙江居群、华中居群、西南居群的21份栀子的SRAP-PCR,共获得98条扩增条带,其中多态性条带87条,在物种水平上,多态性比率(PPB)为88.78%,平均有效等位基因数(Ne)为1.515 2,Nei's基因多样性指数(H)为0.308 9,Shannon's指数(I)为0.465 1;在居群水平上,PPB为53.06%～77.55%,H为0.212 0～0.258 1,I为0.309 4～0.390 4;3个居群的基因分化系数(Gst)为0.235 6,即76.44%的遗传变异在种群内进行,基因流(Nm)为1.621 8,证明居群间存在一定的基因流动(Nm>1);UPGMA聚类结果表明,21份样品分成4组,华中和西南居群各聚成一支,浙江居群被分成二支。本研究表明,栀子具有较高的遗传多样性水平,居群间存在一定的基因交流,可为栀子新品种选育工作提供参考。
In order to study the genetic diversity of Gardenia jasminoides and provide the information for the protection and new variety breeding, SRAP was used to assess the polymorphisms, genetic diversity, and cluster analysis of G. jasminoides. The results showed that 87(88.78%) of the 98 bands were polymorphic by 6 pairs of SRAP primers in 21 G. jasminoides materials from 3 populations. At the species level, the effective number of allele(Ne) was 1.515 2, Nei's genetic diversity(H) and Shannon's information index(I) were 0.308 9 and 0.465 1, respectively. At the population level, the PPB ranged from 53.06% to 77.55%, H and I were 0.212 0～0.258 1 and 0.309 4～0.390 4, respectively. The coefficient of gene differentiation(Gst) for population was 0.235 6, which indicated that the within-population component accounted for 76.44%. The average mean of gene flow was 1.621 8(Nm>1), which suggested that there was a certain gene flow among populations. The UPGMA clustering results indicated that 21 samples were divided into 4 groups, including Central China and Southwest China populations were clustered into the same clade and the Zhejiang population was divided into two groups. The research showed that the gene diversity of G. jasminoides populations was at a high level, and the genetic differentiation that occurred mainly within populations, which provided a reference for breeding new variety of G. jasminoides.
In order to study the genetic diversity of Gardenia jasminoides and provide the information for the protection and new variety breeding, SRAP was used to assess the polymorphisms, genetic diversity, and cluster analysis of G. jasminoides. The results showed that 87(88.78%) of the 98 bands were polymorphic by 6 pairs of SRAP primers in 21 G. jasminoides materials from 3 populations. At the species level, the effective number of allele(Ne) was 1.515 2, Nei's genetic diversity(H) and Shannon's information index(I) were 0.308 9 and 0.465 1, respectively. At the population level, the PPB ranged from 53.06% to 77.55%, H and I were 0.212 0～0.258 1 and 0.309 4～0.390 4, respectively. The coefficient of gene differentiation(Gst) for population was 0.235 6, which indicated that the within-population component accounted for 76.44%. The average mean of gene flow was 1.621 8(Nm>1), which suggested that there was a certain gene flow among populations. The UPGMA clustering results indicated that 21 samples were divided into 4 groups, including Central China and Southwest China populations were clustered into the same clade and the Zhejiang population was divided into two groups. The research showed that the gene diversity of G. jasminoides populations was at a high level, and the genetic differentiation that occurred mainly within populations, which provided a reference for breeding new variety of G. jasminoides.
引文
[1] 国家药典委员会. 中国药典: 一部[M]. 北京: 中国医药科技出版社, 2015: 248.
[2] 韩建萍, 陈士林, 张文生, 等. 栀子遗传多样性及遗传分化的RAPD分析[J]. 中国药学杂志, 2007, 42(23): 1774-1778. HAN J P, CHEN S L, ZHANG W S, et al. Study on genetic diversity and differentiation of Gardenia jasminoides Ellis using RAPD markers [J]. Chinese Pharmaceutical Journal, 2007, 42(23): 1774-1778. (in Chinese with English abstract)
[3] 邓邵勇, 朱培林, 温强, 等. 基于EST-SSR分子标记的栀子野生群体多样性研究[J]. 中草药, 2018, 49(2): 431-438.DENG S Y, ZHU P L, WEN Q, et al. Genetic diversity of natural Gardenia jasminoides population based on EST-SSR analysis [J]. Chinese Traditional and Herbal Drugs, 2018, 49(2): 431-438. (in Chinese with English abstract)
[4] 罗跃龙, 周日宝, 贺又舜, 等. 湖南省栀子种植的概况与分析[J]. 湖南中医药导报, 2004, 10(3): 54-56.LUO Y L, ZHOU R B, HE Y S, et al. The condition and analysis about implantation of Cape jasmine fruit in Hunan Province[J]. Hunan Guiding Journal of Traditional Chinese Medicine and Pharmacology, 2004, 10(3): 54-56. (in Chinese with English abstract)
[5] 曹岚, 梁芳, 戴泽霞. 江西新干产栀子种质资源研究初报[J]. 时珍国医国药, 2008, 19(4): 794-795.CAO L, LIANG F, DAI Z X. Primary reports of research on the germplasm resources of Gardenia jasminoides in Jiangxi Xingan[J].Lishizhen Medicine and Materia Medica Research, 2008, 19(4): 794-795. (in Chinese with English abstract)
[6] 周昌华, 张兴翠, 罗健, 等. 山栀子品种资源的综合评价[J]. 中国中药杂志, 1998, 23(3): 141-143.ZHOU C H, ZHANG X C, LUO J, et al. Comprehensive evaluation of Gardenia jasminoides variety resources[J]. China Journal of Chinese Materia Medica, 1998, 23(3): 141-143. (in Chinese)
[7] 雷栗, 王益, 赵阿曼, 等. 栀子道地药材遗传关系的ISSR证据[J]. 中草药, 2009, 40(1): 116-120.LEI L, WANG Y, ZHAO A M, et al. Genetic relationship of Gardenia jasminoides among plantations revealed by ISSR[J]. Chinese Traditional and Herbal Drugs, 2009, 40(1): 116-120. (in Chinese with English abstract)
[8] 何峰, 唐灿, 王子雯, 等. 巴中栀子遗传多样性的ISSR分析[J]. 中国医院药学杂志, 2014, 34(10): 782-786.HE F, TANG C, WANG Z W, et al. ISSR analysis on the genetic diversity of Fructus gardeniae from Bazhong producing area[J]. Chinese Hospital of Pharmacy Journal, 2014, 34(10): 782-786. (in Chinese with English abstract)
[9] 朱欣, 蒋晓颖, 金魏佳, 等. 基于SRAP分子标记技术研究浙江产细叶石仙桃的遗传多样[J]. 中草药, 2018, 49(13): 3116-3121.ZHU X, JIANG X Y, JIN W J, et al. Genetic diversity analysis of Pholidota cantonensis in Zhejiang based on SRAP molecular marker technique[J]. Chinese Traditional and Herbal Drugs, 2018, 49(13): 3116-3121. (in Chinese with English abstract)
[10] 张安世, 司清亮, 刘莹, 等. 珍稀濒危植物太行菊遗传多样性的SRAP分析[J]. 分子植物育种, 2014, 12(3): 517-524.ZHANG A S, SI Q L, LIU Y, et al. Genetic diversity of the rare and endangered plant Opisthopapus taihangensis(Ling) Shih detected by SRAP analysis[J]. Molecular Plant Breeding, 2014, 12(3): 517-524. (in Chinese with English abstract)
[2] 韩建萍, 陈士林, 张文生, 等. 栀子遗传多样性及遗传分化的RAPD分析[J]. 中国药学杂志, 2007, 42(23): 1774-1778. HAN J P, CHEN S L, ZHANG W S, et al. Study on genetic diversity and differentiation of Gardenia jasminoides Ellis using RAPD markers [J]. Chinese Pharmaceutical Journal, 2007, 42(23): 1774-1778. (in Chinese with English abstract)
[3] 邓邵勇, 朱培林, 温强, 等. 基于EST-SSR分子标记的栀子野生群体多样性研究[J]. 中草药, 2018, 49(2): 431-438.DENG S Y, ZHU P L, WEN Q, et al. Genetic diversity of natural Gardenia jasminoides population based on EST-SSR analysis [J]. Chinese Traditional and Herbal Drugs, 2018, 49(2): 431-438. (in Chinese with English abstract)
[4] 罗跃龙, 周日宝, 贺又舜, 等. 湖南省栀子种植的概况与分析[J]. 湖南中医药导报, 2004, 10(3): 54-56.LUO Y L, ZHOU R B, HE Y S, et al. The condition and analysis about implantation of Cape jasmine fruit in Hunan Province[J]. Hunan Guiding Journal of Traditional Chinese Medicine and Pharmacology, 2004, 10(3): 54-56. (in Chinese with English abstract)
[5] 曹岚, 梁芳, 戴泽霞. 江西新干产栀子种质资源研究初报[J]. 时珍国医国药, 2008, 19(4): 794-795.CAO L, LIANG F, DAI Z X. Primary reports of research on the germplasm resources of Gardenia jasminoides in Jiangxi Xingan[J].Lishizhen Medicine and Materia Medica Research, 2008, 19(4): 794-795. (in Chinese with English abstract)
[6] 周昌华, 张兴翠, 罗健, 等. 山栀子品种资源的综合评价[J]. 中国中药杂志, 1998, 23(3): 141-143.ZHOU C H, ZHANG X C, LUO J, et al. Comprehensive evaluation of Gardenia jasminoides variety resources[J]. China Journal of Chinese Materia Medica, 1998, 23(3): 141-143. (in Chinese)
[7] 雷栗, 王益, 赵阿曼, 等. 栀子道地药材遗传关系的ISSR证据[J]. 中草药, 2009, 40(1): 116-120.LEI L, WANG Y, ZHAO A M, et al. Genetic relationship of Gardenia jasminoides among plantations revealed by ISSR[J]. Chinese Traditional and Herbal Drugs, 2009, 40(1): 116-120. (in Chinese with English abstract)
[8] 何峰, 唐灿, 王子雯, 等. 巴中栀子遗传多样性的ISSR分析[J]. 中国医院药学杂志, 2014, 34(10): 782-786.HE F, TANG C, WANG Z W, et al. ISSR analysis on the genetic diversity of Fructus gardeniae from Bazhong producing area[J]. Chinese Hospital of Pharmacy Journal, 2014, 34(10): 782-786. (in Chinese with English abstract)
[9] 朱欣, 蒋晓颖, 金魏佳, 等. 基于SRAP分子标记技术研究浙江产细叶石仙桃的遗传多样[J]. 中草药, 2018, 49(13): 3116-3121.ZHU X, JIANG X Y, JIN W J, et al. Genetic diversity analysis of Pholidota cantonensis in Zhejiang based on SRAP molecular marker technique[J]. Chinese Traditional and Herbal Drugs, 2018, 49(13): 3116-3121. (in Chinese with English abstract)
[10] 张安世, 司清亮, 刘莹, 等. 珍稀濒危植物太行菊遗传多样性的SRAP分析[J]. 分子植物育种, 2014, 12(3): 517-524.ZHANG A S, SI Q L, LIU Y, et al. Genetic diversity of the rare and endangered plant Opisthopapus taihangensis(Ling) Shih detected by SRAP analysis[J]. Molecular Plant Breeding, 2014, 12(3): 517-524. (in Chinese with English abstract)