基于SSR标记的80份烟草种质指纹图谱的构建及遗传多样性分析
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摘要
利用均匀分布于烟草24个连锁群上的48个SSR标记对80份烟草材料进行分析。结果显示,48个SSR标记共扩增出211个等位基因,平均每个标记4.396个,Shannon′s信息指数I为1.034,多态信息含量值为0.229～0.905,观测杂合度(Ho)、期望杂合度(He)和Nei′s多样性指数(H)平均值分别为0.320、0.572、0.431。聚类结果表明,在遗传距离为0.68时可以将80份烟草种质分为2个类群。5个烟草居群间的遗传一致度在0.643～0.765范围内,遗传距离分布在0.268～0.442。筛选4对核心引物构建了不同烟草种质资源的数字指纹图谱,可将这80份烟草种质资源全部区分开。本研究在分子水平上为筛选优质烟草种质资源、挖掘重要基因以及拓宽烟草育种遗传基础等工作提供科学依据。
80 tobacco germplasm resources were studied using 48 simple sequence repeat(SSR) markers distributeduniformly over 24 linkage groups of tobacco. Results showed that a total of 211 alleles among these germplasm wereidentified at all of 48 SSR markers loci, and an average of 4.396 alleles was observed for each primer pair with averageheterozygosity(Ho) 0.320, average expected heterozygosity(He) 0.572. Shannon′s index was 1.034, Nei′s index was0.431, and the PIC ranged from 0.229 to 0.905. These germplasm were divided into two groups at a genetic similaritycoefficient of 0.68 by UPGMA cluster analysis. The genetic identity among the five tobacco populations ranged from0.643 to 0.765, and the genetic distance ranged from 0.268 to 0.442. Four optimal pairs of core primers were used forthe DNA fingerprint construction. The study can provide reference for further tobacco germplasm resources searchwhich is important for genes discovery and exploitation.
80 tobacco germplasm resources were studied using 48 simple sequence repeat(SSR) markers distributeduniformly over 24 linkage groups of tobacco. Results showed that a total of 211 alleles among these germplasm wereidentified at all of 48 SSR markers loci, and an average of 4.396 alleles was observed for each primer pair with averageheterozygosity(Ho) 0.320, average expected heterozygosity(He) 0.572. Shannon′s index was 1.034, Nei′s index was0.431, and the PIC ranged from 0.229 to 0.905. These germplasm were divided into two groups at a genetic similaritycoefficient of 0.68 by UPGMA cluster analysis. The genetic identity among the five tobacco populations ranged from0.643 to 0.765, and the genetic distance ranged from 0.268 to 0.442. Four optimal pairs of core primers were used forthe DNA fingerprint construction. The study can provide reference for further tobacco germplasm resources searchwhich is important for genes discovery and exploitation.
引文
[1]常爱霞,贾兴华,冯全福,等.我国主要烤烟品种的亲源系谱分析及育种工作建议.中国烟草科学,2013(1):1-6.
[2]王元英,周健.中美主要烟草品种亲源分析与烟草育种.中国烟草学报,1995,2(1):11-22.
[3]Bindler G,Van d H R,Gunduz I,et al. A microsatellite marker basedlinkage map of tobacco. Theoretical&Applied Genetics,2007,114(2):341-349.
[4]刘纪麟.玉米育种学.北京:中国农业出版社,2000:143-175.
[5]郑殿升,杨庆文,刘旭.中国作物种质资源多样性.植物遗传资源学报,2011,12(4):497-500,506.
[6]Ren N,Timko M P. AFLP analysis of genetic polymorphism andevolutionary relationships among cultivated and wild Nicotianaspecies. Genome,2001,44(4):559-571.
[7]Julio E,Verrier J L,Borne F D D. Development of SCAR markerslinked to three disease resistances based on AFLP within Nicotianatabacum L. Theoretical&Applied Genetics,2006,112(2):335-346.
[8]Deng S L,Huang Y L,He H H,et al. Genetic diversity of Aegicerascorniculatum(Myrsinaceae)revealed by amplified fragment lengthpolymorphism(AFLP). Aquatic Botany,2009,90(4):275-281.
[9]Piano,L D,Abet M,Sorrentino C,et al. Genetic variability inNicotiana tabacum and Nicotiana species as revealed by RAPDmarkers:1. Development of the RAPD procedure. Beitr?ge ZurTabakforschung,2014,19(1):1-15.
[10]Bai D,Reeleder R,Brandie J E. Identification of two RAPD markerstightly linked with the Nicotiana debneyi gene for resistance to blackroot rot of tobacco. Theoretical&Applied Genetics,1995,91(8):1184-1189.
[11]祁建民,梁景霞,陈美霞,等.应用ISSR与SRAP分析烟草种质资源遗传多样性及遗传演化关系.作物学报,2012,38(8):1425-1434.
[12]Denduang,Boripant J,Setaphan S,et al. Determination of localtobacco cultivars using ISSR molecular marker. Chiang Mai Journalof Science,2009,37(2):293-303.
[13]Bindler G,Plieske J,Bakaher N,et al. A high density geneticmap of tobacco(Nicotiana tabacum L.)obtained from large scalemicrosatellite marker development. Theoretical&Applied Genetics,2011,123(2):219-230.
[14]Moon H S,Nicholson J S,Lewis R S. Use of transferable Nicotianatabacum L. microsatellite markers for investigating genetic diversityin the genus Nicotiana. Genome,2008,51(8):547-559.
[15]陈杰,杨静,陈建军,等.烤烟种质资源形态学标记及SSR标记的多样性研究.华南农业大学学报,2013(4):450-457.
[16]丁浩,郑唐春,梁德洋,等.一种简易高效提取多种植物纯净DNA的方法.植物研究,2015,35(3):457-461.
[17]童治军.烟草微卫星标记的开发与利用.杭州:浙江大学,2012.
[18]NY/T 1433-2007.水稻品种鉴定-DNA指纹方法.北京:中国标准出版社,2007.
[19]NY/T 1432-2007.玉米品种鉴定-DNA指纹方法.北京:中国标准出版社,2007.
[20]宋海斌,崔喜波,马鸿艳,等.基于SSR标记的甜瓜品种(系)DNA指纹图谱库的构建.中国农业科学,2012,45(13):2676-2689.
[21]刘国祥,邹昆晏,任民,等. 33份晒烟种质资源SSR标记的指纹图谱构建.中国烟草学报,2017,23(5):57-64.
[22]陈婷婷,李显富,贾春雷,等.云烟87突变体新品系的特征特性及其DNA指纹图谱鉴定.浙江大学学报(农业与生命科学版),2016,42(2):179-189.
[23]李显富,陈婷婷,安雪琴,等.烤烟K326突变体新品系‘片片黄’的性状表现及其DNA指纹图谱构建.西南农业学报,2015,28(3):1300-1305.
[24]黄莉莎,曹恒春,王毅,等.烟草主栽品种的SSR指纹图谱的构建及其应用.山东农业大学学报(自然科学版),2013,44(4):509-515.
[25]徐军,刘艳华,任民,等.普通烟草种质资源的SSR标记与指纹图谱分析.中国烟草科学,2011,32(2):62-65.
[26]陈世军,张明泽,姚玉仙,等.基于SSR标记的黔南茶树种质资源DNA指纹图谱构建.植物遗传资源学报,2017,18(1):106-111.
[27]肖炳光,卢江平,卢秀萍,等.烤烟品种的RAPD分析.中国烟草学报,2000,6(2):10-15.
[28]杨柳,汪斌,童治军,等. 25份普通烟草种质资源遗传多样性的SSR标记分析.福建农林大学学报(自然科学版),2013,42(2):171-175.
[29]杨本超,肖炳光,陈学军,等.基于ISSR标记的烤烟种质遗传多样性研究.遗传,2005,27(5):753-758.
[2]王元英,周健.中美主要烟草品种亲源分析与烟草育种.中国烟草学报,1995,2(1):11-22.
[3]Bindler G,Van d H R,Gunduz I,et al. A microsatellite marker basedlinkage map of tobacco. Theoretical&Applied Genetics,2007,114(2):341-349.
[4]刘纪麟.玉米育种学.北京:中国农业出版社,2000:143-175.
[5]郑殿升,杨庆文,刘旭.中国作物种质资源多样性.植物遗传资源学报,2011,12(4):497-500,506.
[6]Ren N,Timko M P. AFLP analysis of genetic polymorphism andevolutionary relationships among cultivated and wild Nicotianaspecies. Genome,2001,44(4):559-571.
[7]Julio E,Verrier J L,Borne F D D. Development of SCAR markerslinked to three disease resistances based on AFLP within Nicotianatabacum L. Theoretical&Applied Genetics,2006,112(2):335-346.
[8]Deng S L,Huang Y L,He H H,et al. Genetic diversity of Aegicerascorniculatum(Myrsinaceae)revealed by amplified fragment lengthpolymorphism(AFLP). Aquatic Botany,2009,90(4):275-281.
[9]Piano,L D,Abet M,Sorrentino C,et al. Genetic variability inNicotiana tabacum and Nicotiana species as revealed by RAPDmarkers:1. Development of the RAPD procedure. Beitr?ge ZurTabakforschung,2014,19(1):1-15.
[10]Bai D,Reeleder R,Brandie J E. Identification of two RAPD markerstightly linked with the Nicotiana debneyi gene for resistance to blackroot rot of tobacco. Theoretical&Applied Genetics,1995,91(8):1184-1189.
[11]祁建民,梁景霞,陈美霞,等.应用ISSR与SRAP分析烟草种质资源遗传多样性及遗传演化关系.作物学报,2012,38(8):1425-1434.
[12]Denduang,Boripant J,Setaphan S,et al. Determination of localtobacco cultivars using ISSR molecular marker. Chiang Mai Journalof Science,2009,37(2):293-303.
[13]Bindler G,Plieske J,Bakaher N,et al. A high density geneticmap of tobacco(Nicotiana tabacum L.)obtained from large scalemicrosatellite marker development. Theoretical&Applied Genetics,2011,123(2):219-230.
[14]Moon H S,Nicholson J S,Lewis R S. Use of transferable Nicotianatabacum L. microsatellite markers for investigating genetic diversityin the genus Nicotiana. Genome,2008,51(8):547-559.
[15]陈杰,杨静,陈建军,等.烤烟种质资源形态学标记及SSR标记的多样性研究.华南农业大学学报,2013(4):450-457.
[16]丁浩,郑唐春,梁德洋,等.一种简易高效提取多种植物纯净DNA的方法.植物研究,2015,35(3):457-461.
[17]童治军.烟草微卫星标记的开发与利用.杭州:浙江大学,2012.
[18]NY/T 1433-2007.水稻品种鉴定-DNA指纹方法.北京:中国标准出版社,2007.
[19]NY/T 1432-2007.玉米品种鉴定-DNA指纹方法.北京:中国标准出版社,2007.
[20]宋海斌,崔喜波,马鸿艳,等.基于SSR标记的甜瓜品种(系)DNA指纹图谱库的构建.中国农业科学,2012,45(13):2676-2689.
[21]刘国祥,邹昆晏,任民,等. 33份晒烟种质资源SSR标记的指纹图谱构建.中国烟草学报,2017,23(5):57-64.
[22]陈婷婷,李显富,贾春雷,等.云烟87突变体新品系的特征特性及其DNA指纹图谱鉴定.浙江大学学报(农业与生命科学版),2016,42(2):179-189.
[23]李显富,陈婷婷,安雪琴,等.烤烟K326突变体新品系‘片片黄’的性状表现及其DNA指纹图谱构建.西南农业学报,2015,28(3):1300-1305.
[24]黄莉莎,曹恒春,王毅,等.烟草主栽品种的SSR指纹图谱的构建及其应用.山东农业大学学报(自然科学版),2013,44(4):509-515.
[25]徐军,刘艳华,任民,等.普通烟草种质资源的SSR标记与指纹图谱分析.中国烟草科学,2011,32(2):62-65.
[26]陈世军,张明泽,姚玉仙,等.基于SSR标记的黔南茶树种质资源DNA指纹图谱构建.植物遗传资源学报,2017,18(1):106-111.
[27]肖炳光,卢江平,卢秀萍,等.烤烟品种的RAPD分析.中国烟草学报,2000,6(2):10-15.
[28]杨柳,汪斌,童治军,等. 25份普通烟草种质资源遗传多样性的SSR标记分析.福建农林大学学报(自然科学版),2013,42(2):171-175.
[29]杨本超,肖炳光,陈学军,等.基于ISSR标记的烤烟种质遗传多样性研究.遗传,2005,27(5):753-758.