基于SSR标记的冬瓜种质资源遗传多样性分析
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摘要
为了加强冬瓜种质资源的保护和评价,明确资源间的亲缘关系,本研究利用SSR标记在分子水平上对111份冬瓜种质资源进行遗传多样性分析。结果表明,19对SSR引物共扩增出182条条带,其中多态性条带169条,多态率92.86%;通过POPGENE 1.31软件计算每对引物的平均有效等位基因数(Ne)为1.393 2,平均Nei's遗传多样性指数(He)为0.260 1,平均Shannon's信息指数(I)为0.418 7。采用Jaccard相似系数进行种质间的聚类分析,在遗传距离为0.70的位置可将全部种质分为6个类群。通过聚类结果可知,种质材料未按照地理来源划分,但相同果型、籽型的种质资源间亲缘关系较近。本研究基于SSR标记分析冬瓜种质资源遗传多样性并进行亲缘关系的聚类,将为冬瓜种质的保存和利用提供理论依据。
In order to strengthen the protection and evaluation of wax gourd germplasm resources, and define the kinship between resources, this study analyzed the genetic diversity of 111 wax gourd germplasm resources at the molecular level by SSR markers. The results showed that 182 bands were amplified by 19 pairs of markers, 169 of which were polymorphic, with a polymorphism rate of 92.86%. When calculated by POPGENE 1.31 software, the average effective allele number(Ne) of each pair primer was 1.393 2. The average Nei's genetic diversity index(He)was 0.260 1, and the average shannon's information index(I) was 0.418 7. The Jaccard similarity coefficient was used for cluster analysis of these germplasm, and all the germplasm were divided into 6 groups at the genetic distance of 0.70. According to the clustering results, the germplasm materials were not classified according to geographical origin, however the germplasm resources of the same fruit type and seed type had a close genetic relationship. This study analyzed the genetic diversity of wax gourd germplasm resources and clustered genetic relationships based on SSR markers, which would provide a theoretical basis for the preservation and utilization of wax gourd germplasm.
In order to strengthen the protection and evaluation of wax gourd germplasm resources, and define the kinship between resources, this study analyzed the genetic diversity of 111 wax gourd germplasm resources at the molecular level by SSR markers. The results showed that 182 bands were amplified by 19 pairs of markers, 169 of which were polymorphic, with a polymorphism rate of 92.86%. When calculated by POPGENE 1.31 software, the average effective allele number(Ne) of each pair primer was 1.393 2. The average Nei's genetic diversity index(He)was 0.260 1, and the average shannon's information index(I) was 0.418 7. The Jaccard similarity coefficient was used for cluster analysis of these germplasm, and all the germplasm were divided into 6 groups at the genetic distance of 0.70. According to the clustering results, the germplasm materials were not classified according to geographical origin, however the germplasm resources of the same fruit type and seed type had a close genetic relationship. This study analyzed the genetic diversity of wax gourd germplasm resources and clustered genetic relationships based on SSR markers, which would provide a theoretical basis for the preservation and utilization of wax gourd germplasm.
引文
An Z.Y.,Xie D.S.,Peng Q.W.,and He X.M.,2011,Comparison of gemomic DNA extraction methods in Chieh-qua(Benincasa hispida),Zhongguo Guacai(China Cucurbits and Vegetables),24(5):1-4(安重莹,谢大森,彭庆务,何晓明,2011节瓜基因组DNA提取方法比较研究,中国瓜菜,24(5)1-4)
Guo L.F.,Zou M.H.,Zeng H.,Du L.Q.,and Lu C.Z.,2007,High quality DNA extraction from mature leaves of macadamia by fied CTAB protocols,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),5(6):187-190(郭凌飞,邹明宏,曾辉,杜丽清,陆超忠,2007,利用改良CTAB法提取澳洲坚果成熟叶片高质量DNA,分子植物育种,5(6):187-190)
Jiang B.,Liu W.R.,Xie D.S.,He X.M.,and Peng Q.W.,2013,Phylogenetic relationships of wax gourd(Benincasa hispida)germplasms based on ISSR markers,Redai Zuowu Xuebao(Chinese Journal of Tropical Crops),34(4):616-620(江彪,刘文睿,谢大森,何晓明,彭庆务,2013,冬瓜种质资源亲缘关系的ISSR分析,热带作物学报,34(4):616-620)
Jiang B.,Xie D.S.,Liu W.R.,Peng Q.W.,and He X.M.,2013,De novo assembly and characterization of the transcriptome,and development of SSR markers in wax gourd(Benicasa hispida),PLoS One,8(8):e71054
Meng X.D.,Wei Y.Y.,Ma H.,Zhang W.H.,and Li J.R.,1996,I-dentification of Chinese wax gourd and Chieh-qua cultivars using RAPD markers,Shanghai Nongye Xuebao(Acta A-griculturae Shanghai),12(4):45-49(孟祥栋,魏佑营,马红,张卫华,李纪蓉,1996,RAPD技术在冬瓜和节瓜品种鉴定中的应用,上海农业学报,12(4):45-49)
Pandey S.,Kumar S.,Mishra U.,Rai A.,Singh M.,and Rai M.2008,Genetic diversity in Indian ash gourd(Benincasa hispida)accessions as revealed by quantitative traits and RAPDmarkers,Sci.Hortic.,118(1):80-86
Qiao Y.C.,Lin J.Y.,Xie W.P.,Xie L.F.,and Li L.F.,2014,Genetic relationship of Chieh-qua and wax-gourd on the morphological phenotypes and SRAP markers,Zhiwu Yichuan Ziyuan Xuebao(Journal of Plant Genetic Resources),15(5)1150-1155(乔燕春,林锦英,谢伟平,谢丽芳,李莲芳,2014节瓜、冬瓜重要性状多样性及亲缘关系的SRAP分析,植物遗传资源学报,15(5):1150-1155)
Song S.W.,Li Z.,Liu H.C.,Sun G.W.,and Chen R.Y.,2010RAPD analysis of genetic diversity of wax gourd and Chiehqua germplasm,Zhongguo Shucai(China Vegetables),1(22):47-53(宋世威,李珍,刘厚诚,孙光闻,陈日远,2010冬瓜和节瓜种质资源遗传多样性的RAPD分析,中国蔬菜,1(22):47-53)
Verma V.K.,Behera T.K.,Munshi A.D.,Parida S.K.,and Mohapatra T.,2007,Genetic diversity of ash gourd(Benincasa hispida,(Thunb.)Cogn.)inbred lines based on RAPDand ISSR markers and their hybrid performance,Sci.Hortic.,113(3):231-237
Wang P.,Liu Z.G.,Huang C.Q.,Chen Y.,and Tan H.P.,2015,Genetic diversity of 36 Chieh-qua inbred lines based on IS-SR,Zhiwu Yichuan Ziyuan Xuebao(Journal of Plant Genetic Resources),16(5):1123-1127(王鹏,刘政国,黄成秋,陈勇,谭海平,2015,节瓜自交系遗传多样性的ISSR分析,植物遗传资源学报,16(5):1123-1127)
Xie D.S.,He X.M.,Peng Q.W.,and Zhao Q.,2010,Studies on DNA extraction methods form winter gourd,Zhongguo Shucai(China Vegetables),1(2):38-41(谢大森,何晓明,彭庆务,赵芹,2010,冬瓜DNA提取方法的研究,中国蔬菜,1(2):38-41)
Zhang J.J.,Liu S.G.,Yu M.Q.,and Ao Q.Y.,2009,Research on wax gourd agronomic characters and genetic diversity,Sichuan Daxue Xuebao(Journal of Sichuan University(Natural Science Edition)),46(6):1855-1861(张建军,刘世贵,余懋群,敖清艳,2009,100份中国冬瓜种质资源农艺性状与遗传多样性研究,四川大学学报(自然科学版),46(6):1855-1861)
Zhu D.D.,Liu Z.G.,Bao Z.Y.,Jiao X.X.,and Wang X.Y.,2016,Analysis of ISSR markers linked to the fruit color trait gene in Chieh-qua,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),35(10):2781-2787(朱冬冬,刘政国,鲍遵宇,焦贤贤,王先裕,2016,节瓜果皮颜色相关基因的ISSR标记分析,基因组学与应用生物学,35(10):2781-2787)
Guo L.F.,Zou M.H.,Zeng H.,Du L.Q.,and Lu C.Z.,2007,High quality DNA extraction from mature leaves of macadamia by fied CTAB protocols,Fenzi Zhiwu Yuzhong(Molecular Plant Breeding),5(6):187-190(郭凌飞,邹明宏,曾辉,杜丽清,陆超忠,2007,利用改良CTAB法提取澳洲坚果成熟叶片高质量DNA,分子植物育种,5(6):187-190)
Jiang B.,Liu W.R.,Xie D.S.,He X.M.,and Peng Q.W.,2013,Phylogenetic relationships of wax gourd(Benincasa hispida)germplasms based on ISSR markers,Redai Zuowu Xuebao(Chinese Journal of Tropical Crops),34(4):616-620(江彪,刘文睿,谢大森,何晓明,彭庆务,2013,冬瓜种质资源亲缘关系的ISSR分析,热带作物学报,34(4):616-620)
Jiang B.,Xie D.S.,Liu W.R.,Peng Q.W.,and He X.M.,2013,De novo assembly and characterization of the transcriptome,and development of SSR markers in wax gourd(Benicasa hispida),PLoS One,8(8):e71054
Meng X.D.,Wei Y.Y.,Ma H.,Zhang W.H.,and Li J.R.,1996,I-dentification of Chinese wax gourd and Chieh-qua cultivars using RAPD markers,Shanghai Nongye Xuebao(Acta A-griculturae Shanghai),12(4):45-49(孟祥栋,魏佑营,马红,张卫华,李纪蓉,1996,RAPD技术在冬瓜和节瓜品种鉴定中的应用,上海农业学报,12(4):45-49)
Pandey S.,Kumar S.,Mishra U.,Rai A.,Singh M.,and Rai M.2008,Genetic diversity in Indian ash gourd(Benincasa hispida)accessions as revealed by quantitative traits and RAPDmarkers,Sci.Hortic.,118(1):80-86
Qiao Y.C.,Lin J.Y.,Xie W.P.,Xie L.F.,and Li L.F.,2014,Genetic relationship of Chieh-qua and wax-gourd on the morphological phenotypes and SRAP markers,Zhiwu Yichuan Ziyuan Xuebao(Journal of Plant Genetic Resources),15(5)1150-1155(乔燕春,林锦英,谢伟平,谢丽芳,李莲芳,2014节瓜、冬瓜重要性状多样性及亲缘关系的SRAP分析,植物遗传资源学报,15(5):1150-1155)
Song S.W.,Li Z.,Liu H.C.,Sun G.W.,and Chen R.Y.,2010RAPD analysis of genetic diversity of wax gourd and Chiehqua germplasm,Zhongguo Shucai(China Vegetables),1(22):47-53(宋世威,李珍,刘厚诚,孙光闻,陈日远,2010冬瓜和节瓜种质资源遗传多样性的RAPD分析,中国蔬菜,1(22):47-53)
Verma V.K.,Behera T.K.,Munshi A.D.,Parida S.K.,and Mohapatra T.,2007,Genetic diversity of ash gourd(Benincasa hispida,(Thunb.)Cogn.)inbred lines based on RAPDand ISSR markers and their hybrid performance,Sci.Hortic.,113(3):231-237
Wang P.,Liu Z.G.,Huang C.Q.,Chen Y.,and Tan H.P.,2015,Genetic diversity of 36 Chieh-qua inbred lines based on IS-SR,Zhiwu Yichuan Ziyuan Xuebao(Journal of Plant Genetic Resources),16(5):1123-1127(王鹏,刘政国,黄成秋,陈勇,谭海平,2015,节瓜自交系遗传多样性的ISSR分析,植物遗传资源学报,16(5):1123-1127)
Xie D.S.,He X.M.,Peng Q.W.,and Zhao Q.,2010,Studies on DNA extraction methods form winter gourd,Zhongguo Shucai(China Vegetables),1(2):38-41(谢大森,何晓明,彭庆务,赵芹,2010,冬瓜DNA提取方法的研究,中国蔬菜,1(2):38-41)
Zhang J.J.,Liu S.G.,Yu M.Q.,and Ao Q.Y.,2009,Research on wax gourd agronomic characters and genetic diversity,Sichuan Daxue Xuebao(Journal of Sichuan University(Natural Science Edition)),46(6):1855-1861(张建军,刘世贵,余懋群,敖清艳,2009,100份中国冬瓜种质资源农艺性状与遗传多样性研究,四川大学学报(自然科学版),46(6):1855-1861)
Zhu D.D.,Liu Z.G.,Bao Z.Y.,Jiao X.X.,and Wang X.Y.,2016,Analysis of ISSR markers linked to the fruit color trait gene in Chieh-qua,Jiyinzuxue Yu Yingyong Shengwuxue(Genomics and Applied Biology),35(10):2781-2787(朱冬冬,刘政国,鲍遵宇,焦贤贤,王先裕,2016,节瓜果皮颜色相关基因的ISSR标记分析,基因组学与应用生物学,35(10):2781-2787)
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