苹果EST-SSR引物的开发及部分品种亲缘关系分析Development of apple EST-SSR primers and phylogenetic analysis of some cultivars
宋尚伟;张恒涛;张芳明;王娟;陶永焕;
摘要(Abstract):
【目的】开发苹果EST-SSR引物,评价苹果种质资源的多样性与亲缘关系。【方法】利用NCBI数据库中苹果EST的SSR位点开发了35对EST-SSR引物,建立了苹果EST-SSR体系,并利用筛选出的16对引物对苹果品种资源的亲缘关系进行研究。【结果】118份基因组DNA共扩增出等位基因位点138个,位点数的变化从3到13不等,平均每对引物检测到8.62个位点,总的多态性比率为94.2%,各引物的多态性比例分布为81.8%~100%,相似系数为0.48~1.00。利用UPGMA法构建聚类树状图,在相似性系数0.65处可将118个供试材料分为5大组:其中第1组又可分为2个亚组,包含了绝大部分供试材料;第2、3、4和5组包含的品种数目分别为12个、11个、3个和2个;第5组与其他组亲缘关系较远,相似性系数仅为0.48。【结论】筛选出的16对EST-SSR引物可用于评价苹果种质资源间的遗传多样性。
关键词(KeyWords): 苹果;EST-SSR引物;亲缘关系;聚类分析;相似性系数
基金项目(Foundation): 河南省教育厅科学技术研究重点项目(13A210484)
作者(Author): 宋尚伟;张恒涛;张芳明;王娟;陶永焕;
Email:
DOI: 10.13925/j.cnki.gsxb.2013.04.009
参考文献(References):
- [1]GAO Yuan,LIU Feng-zhi,CAO Yu-fen,WANG Kun.Analysis ofgenetic relationship for Malus germplasm resources by SSR markers[J].Journal of Fruit Science,2007,24(2):129-134.高源,刘凤之,曹玉芬,王昆.苹果属种质资源亲缘关系的SSR分析[J].果树学报,2007,24(2):129-134.
- [2]GIANFRANCESCHI L,SEGLIAS N,TARCHINI R,KOMJANC M,GESSLER C.Simple sequence repeats for the genetic analysis ofApple[J].Theoretical and Applied Genetics,1998,96:1069-1076.
- [3]GALLI Z,HALASZ G,KISS E,HESZKY L,DOBRANSZKI J.Molecular identification of commercial Apple cultivars with mi-crosatellite markers[J].HortScience,2005,40:1974-1977.
- [4]ZHANG Q,LI J,ZHAO Y,KORBAN S S,HAN Y.Evaluation ofgenetic diversity in Chinese wild apple species along with apple cul-tivars using SSR markers[J].Plant Molecular Biology Reporter,2012,30:539-546.
- [5]WANG Li-xin,ZHANG Xiao-jun,SHI Xing-yun,GAO Hua,ZHAOZheng-yang.Establishment of SSR fingerprinting database on majorapple(Malus×domestica)cultivars[J].Journal of Fruit Science,2012,29(6):971-977.王立新,张小军,史星雲,高华,赵政阳.苹果栽培品种SSR指纹图谱的构建[J].果树学报,2012,29(6):971-977.
- [6]SUN Ping,ZONG Yu,LIU Jing,HU Chun-yun,TENG Yuan-wen.Study on genetic diversity of Malus sieboldii in Qingliangfeng Regionbased on SSR markers[J].Journal of Fruit Science,2013,30(1):8-15.孙萍,宗宇,刘晶,胡春云,滕元文.基于SSR标记的清凉峰地区三叶海棠遗传多样性研究[J].果树学报,2013,30(1):8-15.
- [7]RODER M S,KORZUN V,WENDEHAKE K,PLASCHKE J,TIXI-ER M H,LEROY P,GANAL M W.A microsatellite map of wheat[J].Genetics,1998,149:2007-2023.
- [8]KOTA R,VARSHNEY R K,THIEL T,DEHMER KJ,GRANER A.Generation and comparison of EST-derived SSR and SNPs in barley(Hordeum Vulgare L.)[J].Hereditas,2001,135:145-151.
- [9]SCOTT KD,EGGLER P,SEATON G,ROSSETTO M,ABLETT EM,LEE LS,HENRY RJ.Analysis of SSR derived from grape EST[J].Theoretical and Applied Genetics,2000,100:723-726.
- [10]CORDEIRO G M,CASU R,MCINTYRE C L,MANNERS J M,HENRY R J.Microsatellite markers from sugarcane(Saccharumspp)ESTs cross-transferable to Erianthus and Sorghum[J].Plant Sci,2001,160:1115-1123.
- [11]EUJAYL I,SORRELLS M E,BAUM M,WOLTERS P,POWELL W.Isolation of EST-derived microsatellite markers for genotyping the Aand B genomes of wheat[J].Theoretical and Applied Genetics,2002,104(2-3):399-407.
- [12]THIEL T,MICHALEK W,VARSHNEY RK,GRANER A.Exploit-ing EST databases for the development and characterization of gene-derived SSR-markers in barley(Hordeum vulgare L.)[J].Theoreticaland Applied Genetics,2003,106:411-412.
- [13]SONG Q J,MAREK L F,SHOEMAKER RC,LARK KG,CON-CIBIDO VC,DELANNAY X.A new integrated genetic linkage mapof the soybean[J].Theoretical and Applied Genetics,2004,109:122-128.
- [14]KANTETY R V,LA ROTA M L,MATTHEWS D E,SORRELLS M E.Data mining for simple sequence repeats in expressed sequence tagsfrom barley,maize,rice,sorghum and wheat[J].Plant Molecular Biol-ogy,2002,48:501-510.
- [15]YAO Li-hua,CAI Dan-ying,ZHENG Xiao-yan,TENG Yuan-wen.Developing EST-SSR Markers from ESTs of Malus Mill.[C]//Progressof Horticulture Shanghai:Shanghai Jiao Tong University Press,2008:160-162.姚利华,蔡丹英,郑小艳,滕元文.从苹果属EST数据库中开发EST-SSR标记[C]//园艺学进展,上海:上海交通大学出版社,2008:160-162.
- [16]GASIC K,HAN Y P,KERTBUNDIT S,SHULAEVV,IEZZONI AF,Ed STOVER W,BELL R L,WISNIEWSKI M E,KORBAN S S.Characteristics and transferability of new apple EST-derived SSRsto other Rosaceae species[J].Molecular Breeding,2009,23(3):397-411.
- [17]CHEN Kun-song,LI Fang,XU Chang-jie,ZHANG Shang-long,FU Cheng-xin.An efficient macro-method of genomic DNA isolationfrom actinidia Chinensis leaves[J].Hereditas,2004,26(4):529-531.陈昆松,李方,徐昌杰,张上隆,傅承新.改良CTAB法用于多年生植物组织基因组DNA的大量提取[J].遗传,2004,26(4):529-531.
- [18]BASSAM B J,CAETANO-ANOLLE G,GRESSHOFF P M.Fastand sensitive silver staining of DNA in polyacrylamide gels[J].AnalBiochem,1991,196:80-83.
- [19]AYERS N M,MCCLUNG A M,LARKIN P D.Microsatellites and asingle nucleotide polymorphism diferentiate apparent amylose class-es in an extended pedigree of US rice germplasm[J].Theoretical andApplied Genetics,1997,94:773-781.
- [20]BAO J S,CORKE H,SUN M.Mierosatellites in starch synthesizinggenes in relation to starch Physiochemical Properties in waxy rice(Oryza statiiva L.)[J].Theoretical and Applied Genetics,2002,105(6-7):898-905.
- [21]HOKANSON S C,LAMBCY,W F,SZEWC-MCFADDEN A K.Microsatellite(SSR)variation in a collection of Malus(apple)speciesand hybrids[J].Euphytica,2001,118:281-294.
- [22]THIEL T,MICHALEK W,VARSHNEY R K.Exploiting EST databas-es for the development and characterization of gene-derived SSR-markers in barley(Hordeum vulgare L.)[J].Theoretical and AppliedGenetics,2003,106:411-422.
- [23]CUI Huai-yu,GAO Mei-xiu,HE Shui-tao.Dwarf dense plantingcultivation in apple varieties[M].Zhengzhou:Henan Science andTechnology Press,1995:21-24.崔怀玉,高梅秀,何水涛.苹果优良品种矮化密植栽培[M].郑州:河南科学技术出版社,1995:21-24.
- [24]WANG Jing-yan,JIA Ding-xian,MI Wen-guang.New cultivar andcultivation key points of apple[M].Shaanxi:Shaanxi Science and Tech-nology Press,1992:65-67.汪景彦,贾定贤,米文广.苹果新品种及其栽培要点[M].陕西:陕西科学技术出版社,1992:65-67.
- [25]GUO Zhao-nian.Dwarf dense planting cultivation in apple varieties[M].Beijing:Jindun Press,2006:25.郭兆年.苹果新品种及矮化密植技术[M].北京:金盾出版社,2006:25.
- [26]SHU Huai-rui.Pomology[M].Beijing:Agricultural Press of China,1999:119-121.束怀瑞.苹果学[M].北京:中国农业出版社,1999:119-121.