基于SSR和SRAP标记的红花玉兰品种遗传关系分析及分子鉴定
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摘要
【目的】为了解析红花玉兰各品种间的遗传多样性和遗传差异等问题,本研究采用SSR和SRAP分子标记方法对红花玉兰不同品种的遗传多样性水平和遗传分化程度进行评估,以建立其分子标记体系。【方法】以35个红花玉兰品种为实验材料,分别进行SSR和SRAP标记扩增,计算出SSR和SRAP分子标记的各遗传参数。利用NTSYS-pc2.1软件计算各品种间的遗传相似系数并用非加权法(UPGMA)进行聚类分析。通过R语言分析筛选出能够完全区分红花玉兰品种的引物对组合。【结果】经筛选共获得18对具有多态性且条带清晰的SSR引物,分别以35个红花玉兰品种基因组DNA为模板,共扩增出DNA条带128条,每对引物扩增条带在2～15之间,平均每对引物扩增7.1条,平均带型数为10.6,平均有效带型数为5.4,平均分辨能力(D)为0.72;不同引物的多态性位点百分比变化范围很大,在0.142 9～1.000 0之间,均值为0.730 2;Shannon’s指数平均为0.304 2,范围是0.086 4～0.433 7;平均期望杂合度为0.248 8,范围是0.059 2～0.282 3。利用筛选获得的11对SRAP引物对35个红花玉兰品种进行鉴定,共扩增156条DNA条带,引物扩增条带数在7～18之间,平均每对引物扩增14.2条,平均带型数为22.7,平均有效带型数为13.2,平均分辨能力(D)为0.93;11对引物的多态性位点百分比平均数为0.815 6,变异范围是0.657 1～1.000 0;平均Shannon’s指数为0.375 9,变异区间在0.287 2～0.455 3;平均期望杂合度为0.240 4,范围在0.180 2～0.311 6之间。【结论】红花玉兰具有较高的遗传多样性,经筛选和优化后的SSR和SRAP引物组合均能将现有红花玉兰品种完全区分开,实现了对红花玉兰品种的简便、快速、准确地鉴定,并为红花玉兰的保护和繁育,以及新品种的选育提供了重要的参考。
[Objective] Genetic diversity and genetic variation of Magnolia wufengensis cultivars were examined by simple sequence repeats(SSR) and sequence-related amplified polymorphism(SRAP)molecular markers for establishing its molecular marker system. [Method] Thirty-five simples of M.wufengensis cultivars were applied to PCR amplification for SSR and SRAP markers. The genetic similarity coefficient of each cultivar was calculated by NTSYS-pc2.1 and cluster analysis was carried out using UPGMA.The locus data of combination of primer pairs that can discriminate all cultivars were screened by R language. [Result] A total of 18 pairs of SSR primers with polymorphism and clear bands were obtained. With the genome DNA of 35 M. wufengensis cultivars as the PCR template, a total of 128 polymorphic bands were generated. The number of polymorphic bands at each polymorphic primer ranged from 2 to 15, with mean band number of 7.1, mean pattern number of 10.6, mean effective pattern number of5.4 and mean D of 0.72.The mean value of the percentage of polymorphic loci was 0.730 2, ranging from0.142 9 ～ 1.000 0. The mean Shannon's index was 0.304 2, ranging from 0.086 4 to 0.433 7. The mean expected heterozygosity among cultivars was 0.248 8, ranging from 0.059 2 to 0.282 3.Eleven pairs of effective SRAP primers were acquired through screening. A total of 156 polymorphic bands were produced with the genome DNA of 35 M.wufengensis cultivars as the template. The number of polymorphic bands at each polymorphic primer ranged from 7 to 18, with mean band number of 14.2, mean pattern number of22.7, mean effective pattern number of 13.2 and mean D of 0.93. The mean value of the percentage of polymorphic loci was 0.815 6, ranging from 0.657 1 ～ 1.000 0. The mean Shannon's index was 0.375 9,ranging from 0.287 2 to 0.455 3. The mean expected heterozygosity among cultivars was 0.240 4, ranging from 0.180 2 to 0.311 6. [Conclusion] M. wufengensis has relatively high genetic diversity. The analysis of molecular variation of both SSR and SRAP marker systems indicates that most genetic diversity is within M.wufengensis cultivars. The locus data of combination of primer pairs can discriminate all cultivars for identifying M. wufengensis cultivars efficiently and accurately.These results provide important references for the protection and breeding of M. wufengensis.
[Objective] Genetic diversity and genetic variation of Magnolia wufengensis cultivars were examined by simple sequence repeats(SSR) and sequence-related amplified polymorphism(SRAP)molecular markers for establishing its molecular marker system. [Method] Thirty-five simples of M.wufengensis cultivars were applied to PCR amplification for SSR and SRAP markers. The genetic similarity coefficient of each cultivar was calculated by NTSYS-pc2.1 and cluster analysis was carried out using UPGMA.The locus data of combination of primer pairs that can discriminate all cultivars were screened by R language. [Result] A total of 18 pairs of SSR primers with polymorphism and clear bands were obtained. With the genome DNA of 35 M. wufengensis cultivars as the PCR template, a total of 128 polymorphic bands were generated. The number of polymorphic bands at each polymorphic primer ranged from 2 to 15, with mean band number of 7.1, mean pattern number of 10.6, mean effective pattern number of5.4 and mean D of 0.72.The mean value of the percentage of polymorphic loci was 0.730 2, ranging from0.142 9 ～ 1.000 0. The mean Shannon's index was 0.304 2, ranging from 0.086 4 to 0.433 7. The mean expected heterozygosity among cultivars was 0.248 8, ranging from 0.059 2 to 0.282 3.Eleven pairs of effective SRAP primers were acquired through screening. A total of 156 polymorphic bands were produced with the genome DNA of 35 M.wufengensis cultivars as the template. The number of polymorphic bands at each polymorphic primer ranged from 7 to 18, with mean band number of 14.2, mean pattern number of22.7, mean effective pattern number of 13.2 and mean D of 0.93. The mean value of the percentage of polymorphic loci was 0.815 6, ranging from 0.657 1 ～ 1.000 0. The mean Shannon's index was 0.375 9,ranging from 0.287 2 to 0.455 3. The mean expected heterozygosity among cultivars was 0.240 4, ranging from 0.180 2 to 0.311 6. [Conclusion] M. wufengensis has relatively high genetic diversity. The analysis of molecular variation of both SSR and SRAP marker systems indicates that most genetic diversity is within M.wufengensis cultivars. The locus data of combination of primer pairs can discriminate all cultivars for identifying M. wufengensis cultivars efficiently and accurately.These results provide important references for the protection and breeding of M. wufengensis.
引文
[1]马履一,王罗荣,贺随超,等.中国木兰科木兰属一新种[J].植物研究,2006,26(1):4-7.Ma L Y,Wang L R,He S C,et al.A new species of Magnolia(Magnoliaceae)from Hubei China[J].Bulletin of Botanical Research,2006,26(1):4-7.
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[3]Chen L Y,Chen F J,He S C,et al.High genetic diversity and small genetic variation among populations of Magnolia wufengensis(Magnoliaceae),revealed by ISSR and SRAPmarkers[J].Electronic Journal of Biotechnology,2014,17(6):268-274.
[4]Wang L,Xiao A H,Ma L Y,et al.Identification of Magnolia wufengensis(Magnoliaceae)cultivars using phenotypic traits,SSR and SRAP markers:insights into breeding and conservation[J/OL].Genetics and Molecular Research,2017,16(1):gmr16019473[2019-04-05].http://dx.doi.org/10.4238/gmr16019473.
[5]杨杨.红花玉兰幼苗在北京地区生态适应性与越冬技术研究[D].北京:北京林业大学,2015.Yang Y.Studies on the ecological adaptability and overwintering technique of Magnolia wufengensis seedlings in Beijing[D].Beijing:Beijing Forestry University,2015.
[6]Yang Y,Jia Z,Chen F,et al.Comparative analysis of natural cold acclimation and deacclimation of two Magnolia species with different winter hardiness[J].Acta Physiologiae Plantarum,2015,37(7):1-11.
[7]张嘉,刘爱青,张淑玲,等.利用荧光标记SSR绘制中国芍药品种分子身份证[J].北京林业大学学报,2016,38(6):101-109.Zhang J,Liu A Q,Zhang S L,et al.Using the SSR with fluorescent labeling to establish SSR molecular ID code for cultivars of the Chinese herbaceous[J].Journal of Beijing Forestry University,2016,38(6):101-109.
[8]Jing Z B,Wang X P,Cheng J M.Analysis of genetic diversity among Chinese wild Vitis species revealed with SSR and SRAPmarkers[J/OL].Genetics and Molecular Research,2013,12(2):1962-1973[2019-04-05].http://dx.doi.org/10.4238/2013.June.13.5.
[9]吴钰滢,周璇,徐庭亮,等.现代月季品种‘赞歌’和粉团蔷薇杂交后代鉴定与评价[J].北京林业大学学报,2019,41(3):124-133.Wu Y Y,Zhou X,Xu T L,et al.Identification and evaluation of F1 hybrids between Rosa‘Sanka’×R.multiflora var.cathayensis[J].Journal of Beijing Forestry University,2019,41(3):124-133.
[10]Xu M,Li Z,Wang J,et al.RNA sequencing and SSR marker development for genetic diversity research in Woonyoungia septentrionalis,(Magnoliaceae)[J].Conservation Genetics Resources,2017,10(4):1-6.
[11]Gy?rgy Z,Vouillamoz J F,H?hn M.Microsatellite markers reveal common east Alpine-Carpathian genepool for the arctic-alpine Rhodiola rosea(Crassulaceae)[J].Plant Systematics and Evolution,2016,302(6):721-730.
[12]周鹏,林玮,朱芹,等.基于SRAP分子标记的刨花润楠遗传多样性分析[J].北京林业大学学报,2016,38(9):16-24.Zhou P,Lin W,Zhu Q,et al.Genetic diversity of Machilus pauhoi assessed by SRAP markers[J].Journal of Beijing Forestry University,2016,38(9):16-24.
[13]Francis C Y,Rong C Y,Boyle T.Popgene microsoft windowsbased freeware for population genetic analysis[J].University of Alberta,1999:1-31.
[14]Rohlf F J.NTSYSpc numerical taxonomy and multivariate analysis system version 2.0 user guide[Z].New York:Applied Biostatistics Incorporated,1998.
[15]邱帅,沈柏春,李婷婷,等.基于随机森林算法和SRAP分子标记的桂花品种鉴定方法[J].林业科学,2018,54(1):32-45.Qiu S,Shen B C,Li T T,et al.A method of Osmanthus fragrans cultivars identification based on random forest algorithm and SRAP molecular markers[J].Scientia Silvae Sinicae,2018,54(1):32-45.
[16]Zhao X F,Sun W B,Yang J B,et al.Isolation and characterization of 12 microsatellite loci for Michelia coriacea(Magnoliaceae),a critically endangered endemic to Southeast Yunnan,China[J].Conservation Genetics,2009,10(5):1583-1585.
[17]Sun Y,Liu Y F,Wang J,et al.Ten polymorphic microsatellite markers in Michelia maudiae(Magnoliaceae)[J/OL].American Journal of Botany,2010,97(12):e157-e158[2019-04-12].https://doi.org/10.3732/ajb.1000332.
[18]林燕芳.利用微卫星分子标记检测单性木兰的种群遗传结构与基因流[D].桂林:广西师范大学,2012.Lin Y F.The population genetic structure and gene flow of Kmeria septentrionalis assessed by microsatellite markers[D].Guilin:Guangxi Normal University,2012.
[19]熊敏,王静,张志荣,等.濒危植物华木莲核基因组微卫星引物开发研究[J].植物分类与资源学报,2011,33(5):535-539.Xiong M,Wang J,Zhang Z R,et al.The development of nuclear microsatellite markers for an endangered plant,Sinoman glietiaglauca(Magnoliaceae)[J].Plant Diversity,2011,33(5):535-539.
[20]IsagiY,Kanazahi T,Suzuki W,et al.Polymorphic microsatellite DNA markers for Magnolia obovata Thunb.and their utility in related species[J].Molecular Ecology,1999,8(4):698-700.
[21]Solmaz I,Ka?ar Y A,Sar?N,et al.Genetic diversity within Turkish watermelon(Citrullus lanatus Thunb.)Matsumura&Nakai)accessions revealed by SSR and SRAP markers[J].Turkish Journal of Agriculture and Forestry,2016,40(3):407-419.
[22]Yuan X,Mingyue T U,Yali H E,et al.Analysis of genetic diversity in 73 Kentucky bluegrass materials by SSR and SRAPmarkers[J].Notulae Botanicae Horti Agrobotanici Cluj-Napoca,2018,46(2):327-335.
[23]Archak S,Gaikwad A B,Gautam D,et al.Comparative assessment of DNA fingerprinting techniques(RAPD,ISSR and AFLP)for genetic analysis of cashew(Anacardium occidentale L.)accessions of India[J].Genome,2003,46(3):362-369.
[24]毛秀红,郑勇奇,孙百友,等.基于SSR的刺槐无性系遗传多样性分析和指纹图谱构建[J].林业科学,2017,53(10):83-92.Mao X H,Zheng Y Q,Sun B Y,et al.Genetic diversity and fingerprints of Robinia pseudoacacia clones based on SSRmarkers[J].Scientia Silvae Sinicae,2017,53(10):83-92.
[25]邹自征.苎麻分子标记方法比较和遗传图谱的初步构建[D].北京:中国农业科学院,2012.Zou Z Z.Comparison among different methods of molecularmarkers and preliminary construction of molecular genetic map in ramie(Boehmeria nivea(L.)Gaud.)[D].Beijing:Chinese Academy of Agricultural Sciences Dissertation,2012.
[26]Chen J Q,Ci X Q,Li Q M,et al.Genetic diversity of Litseas zemaois,an endangered species endemic to China,detected by inter-simple sequence repeat(ISSR)[J].Biodivers Science,2006,14(5):410-420.
[27]Yu H H,Yan Z L,Liu R N,et al.Genetic diversity and relationship of endangered plant Magnolia officinalis(Magnoliacee)assessed with ISSR polymorphisms[J].Biochemical Systematics and Ecology,2011,39(2):71-78.
[28]杨梅,张敏,师守国,等.武当木兰种群遗传结构的ISSR分析[J].林业科学,2014,50(1):76-81.Yang M,Zhang M,Shi S G,et al.Analysis of genetic structure of Magnolia sprengeri populations based on ISSR markers[J].Scientia Silvae Sinicae,2014,50(1):76-81.
[29]Nybom H.Comparison of different nuclear DNA markers for estimating intraspecific genetic diversity in plants[J].Molecular Ecology,2010,13(5):1143-1155.
[30]Tang S Q,Bin X Y,Zhou J Y,et al.Assessment of genetic diversity in cultivars and wild accessions of Luohanguo(Siraitia grosvenorii),a species with edible and medicinal sweet fruits endemic to southern China,using RAPD and AFLP markers[J].Genetic Resources and Crop Evolution,2007,54(5):1053-1061.
[31]He J,Chen L,Huang B,et al.Population structure and genetic diversity distribution in wild and cultivated populations of the traditional Chinese medicinal plant Magnolia officinalis subsp.biloba[J].Genetica,2009,135(2):233-243.
[32]Zhang X M,Wen J,Motley T J,et al.Genetic variation and conservation assessment of Chinese populations of Magnolia cathcartii(Magnoliaceae),a rare evergreen tree from the SouthCentral China hotspot in the eastern Himalayas[J].Journal of Plant Research,2010,123(3):321-331.
[33]Zhang Y X,Zhang X R,Hua W,et al.Analysis of genetic diversity among indigenous landraces from sesame(Sesamum indicum L.)core collection in China as revealed by SRAP and SSR markers[J].Genes and Genomics,2010,32(3):207-215.
[2]贺随超,马履一,陈发菊.红花玉兰种质资源遗传多样性初探[J].西北植物学报,2007,27(12):2421-2428.He S C,Ma L Y,Chen F J.Genetic diversity of Magnolia wufengensis based on AFLP[J].Acta Botanica BorealiOccidentalia Sinica,2007,27(12):2421-2428.
[3]Chen L Y,Chen F J,He S C,et al.High genetic diversity and small genetic variation among populations of Magnolia wufengensis(Magnoliaceae),revealed by ISSR and SRAPmarkers[J].Electronic Journal of Biotechnology,2014,17(6):268-274.
[4]Wang L,Xiao A H,Ma L Y,et al.Identification of Magnolia wufengensis(Magnoliaceae)cultivars using phenotypic traits,SSR and SRAP markers:insights into breeding and conservation[J/OL].Genetics and Molecular Research,2017,16(1):gmr16019473[2019-04-05].http://dx.doi.org/10.4238/gmr16019473.
[5]杨杨.红花玉兰幼苗在北京地区生态适应性与越冬技术研究[D].北京:北京林业大学,2015.Yang Y.Studies on the ecological adaptability and overwintering technique of Magnolia wufengensis seedlings in Beijing[D].Beijing:Beijing Forestry University,2015.
[6]Yang Y,Jia Z,Chen F,et al.Comparative analysis of natural cold acclimation and deacclimation of two Magnolia species with different winter hardiness[J].Acta Physiologiae Plantarum,2015,37(7):1-11.
[7]张嘉,刘爱青,张淑玲,等.利用荧光标记SSR绘制中国芍药品种分子身份证[J].北京林业大学学报,2016,38(6):101-109.Zhang J,Liu A Q,Zhang S L,et al.Using the SSR with fluorescent labeling to establish SSR molecular ID code for cultivars of the Chinese herbaceous[J].Journal of Beijing Forestry University,2016,38(6):101-109.
[8]Jing Z B,Wang X P,Cheng J M.Analysis of genetic diversity among Chinese wild Vitis species revealed with SSR and SRAPmarkers[J/OL].Genetics and Molecular Research,2013,12(2):1962-1973[2019-04-05].http://dx.doi.org/10.4238/2013.June.13.5.
[9]吴钰滢,周璇,徐庭亮,等.现代月季品种‘赞歌’和粉团蔷薇杂交后代鉴定与评价[J].北京林业大学学报,2019,41(3):124-133.Wu Y Y,Zhou X,Xu T L,et al.Identification and evaluation of F1 hybrids between Rosa‘Sanka’×R.multiflora var.cathayensis[J].Journal of Beijing Forestry University,2019,41(3):124-133.
[10]Xu M,Li Z,Wang J,et al.RNA sequencing and SSR marker development for genetic diversity research in Woonyoungia septentrionalis,(Magnoliaceae)[J].Conservation Genetics Resources,2017,10(4):1-6.
[11]Gy?rgy Z,Vouillamoz J F,H?hn M.Microsatellite markers reveal common east Alpine-Carpathian genepool for the arctic-alpine Rhodiola rosea(Crassulaceae)[J].Plant Systematics and Evolution,2016,302(6):721-730.
[12]周鹏,林玮,朱芹,等.基于SRAP分子标记的刨花润楠遗传多样性分析[J].北京林业大学学报,2016,38(9):16-24.Zhou P,Lin W,Zhu Q,et al.Genetic diversity of Machilus pauhoi assessed by SRAP markers[J].Journal of Beijing Forestry University,2016,38(9):16-24.
[13]Francis C Y,Rong C Y,Boyle T.Popgene microsoft windowsbased freeware for population genetic analysis[J].University of Alberta,1999:1-31.
[14]Rohlf F J.NTSYSpc numerical taxonomy and multivariate analysis system version 2.0 user guide[Z].New York:Applied Biostatistics Incorporated,1998.
[15]邱帅,沈柏春,李婷婷,等.基于随机森林算法和SRAP分子标记的桂花品种鉴定方法[J].林业科学,2018,54(1):32-45.Qiu S,Shen B C,Li T T,et al.A method of Osmanthus fragrans cultivars identification based on random forest algorithm and SRAP molecular markers[J].Scientia Silvae Sinicae,2018,54(1):32-45.
[16]Zhao X F,Sun W B,Yang J B,et al.Isolation and characterization of 12 microsatellite loci for Michelia coriacea(Magnoliaceae),a critically endangered endemic to Southeast Yunnan,China[J].Conservation Genetics,2009,10(5):1583-1585.
[17]Sun Y,Liu Y F,Wang J,et al.Ten polymorphic microsatellite markers in Michelia maudiae(Magnoliaceae)[J/OL].American Journal of Botany,2010,97(12):e157-e158[2019-04-12].https://doi.org/10.3732/ajb.1000332.
[18]林燕芳.利用微卫星分子标记检测单性木兰的种群遗传结构与基因流[D].桂林:广西师范大学,2012.Lin Y F.The population genetic structure and gene flow of Kmeria septentrionalis assessed by microsatellite markers[D].Guilin:Guangxi Normal University,2012.
[19]熊敏,王静,张志荣,等.濒危植物华木莲核基因组微卫星引物开发研究[J].植物分类与资源学报,2011,33(5):535-539.Xiong M,Wang J,Zhang Z R,et al.The development of nuclear microsatellite markers for an endangered plant,Sinoman glietiaglauca(Magnoliaceae)[J].Plant Diversity,2011,33(5):535-539.
[20]IsagiY,Kanazahi T,Suzuki W,et al.Polymorphic microsatellite DNA markers for Magnolia obovata Thunb.and their utility in related species[J].Molecular Ecology,1999,8(4):698-700.
[21]Solmaz I,Ka?ar Y A,Sar?N,et al.Genetic diversity within Turkish watermelon(Citrullus lanatus Thunb.)Matsumura&Nakai)accessions revealed by SSR and SRAP markers[J].Turkish Journal of Agriculture and Forestry,2016,40(3):407-419.
[22]Yuan X,Mingyue T U,Yali H E,et al.Analysis of genetic diversity in 73 Kentucky bluegrass materials by SSR and SRAPmarkers[J].Notulae Botanicae Horti Agrobotanici Cluj-Napoca,2018,46(2):327-335.
[23]Archak S,Gaikwad A B,Gautam D,et al.Comparative assessment of DNA fingerprinting techniques(RAPD,ISSR and AFLP)for genetic analysis of cashew(Anacardium occidentale L.)accessions of India[J].Genome,2003,46(3):362-369.
[24]毛秀红,郑勇奇,孙百友,等.基于SSR的刺槐无性系遗传多样性分析和指纹图谱构建[J].林业科学,2017,53(10):83-92.Mao X H,Zheng Y Q,Sun B Y,et al.Genetic diversity and fingerprints of Robinia pseudoacacia clones based on SSRmarkers[J].Scientia Silvae Sinicae,2017,53(10):83-92.
[25]邹自征.苎麻分子标记方法比较和遗传图谱的初步构建[D].北京:中国农业科学院,2012.Zou Z Z.Comparison among different methods of molecularmarkers and preliminary construction of molecular genetic map in ramie(Boehmeria nivea(L.)Gaud.)[D].Beijing:Chinese Academy of Agricultural Sciences Dissertation,2012.
[26]Chen J Q,Ci X Q,Li Q M,et al.Genetic diversity of Litseas zemaois,an endangered species endemic to China,detected by inter-simple sequence repeat(ISSR)[J].Biodivers Science,2006,14(5):410-420.
[27]Yu H H,Yan Z L,Liu R N,et al.Genetic diversity and relationship of endangered plant Magnolia officinalis(Magnoliacee)assessed with ISSR polymorphisms[J].Biochemical Systematics and Ecology,2011,39(2):71-78.
[28]杨梅,张敏,师守国,等.武当木兰种群遗传结构的ISSR分析[J].林业科学,2014,50(1):76-81.Yang M,Zhang M,Shi S G,et al.Analysis of genetic structure of Magnolia sprengeri populations based on ISSR markers[J].Scientia Silvae Sinicae,2014,50(1):76-81.
[29]Nybom H.Comparison of different nuclear DNA markers for estimating intraspecific genetic diversity in plants[J].Molecular Ecology,2010,13(5):1143-1155.
[30]Tang S Q,Bin X Y,Zhou J Y,et al.Assessment of genetic diversity in cultivars and wild accessions of Luohanguo(Siraitia grosvenorii),a species with edible and medicinal sweet fruits endemic to southern China,using RAPD and AFLP markers[J].Genetic Resources and Crop Evolution,2007,54(5):1053-1061.
[31]He J,Chen L,Huang B,et al.Population structure and genetic diversity distribution in wild and cultivated populations of the traditional Chinese medicinal plant Magnolia officinalis subsp.biloba[J].Genetica,2009,135(2):233-243.
[32]Zhang X M,Wen J,Motley T J,et al.Genetic variation and conservation assessment of Chinese populations of Magnolia cathcartii(Magnoliaceae),a rare evergreen tree from the SouthCentral China hotspot in the eastern Himalayas[J].Journal of Plant Research,2010,123(3):321-331.
[33]Zhang Y X,Zhang X R,Hua W,et al.Analysis of genetic diversity among indigenous landraces from sesame(Sesamum indicum L.)core collection in China as revealed by SRAP and SSR markers[J].Genes and Genomics,2010,32(3):207-215.
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