基于SSR标记的白化和黄化茶树品种遗传多样性分析及指纹图谱构建
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摘要
利用62对SSR引物对16个白化、黄化茶树品种资源的遗传多样性进行了分析,初步明确了不同白化、黄化品种的遗传结构,以及SSR标记在白化、黄化品种鉴定上的适用性,为此类茶树品种资源的鉴定评价提供了理论依据。经过对引物筛选和扩增条带的分析,结果显示:具有多态性的55对引物中,共检测出169个等位基因,每对引物检测出的等位基因数为2~5个,平均为3.07个;多态信息含量(PIC)和Shannon信息指数(I)的平均值分别是0.40和0.79;169个等位基因出现频率在3.12%~96.88%之间;16个参试品种的遗传距离在0.086~0.532,品种间遗传结构差异明显;当D≈0.18时,可将16个品种划分为3类,其中大部分亲缘关系相近或地理位置一致的品种聚为一类。此外,笔者根据不同引物的等位基因带型构建了16个白化和黄化茶树品种的分子指纹图谱,并筛选出3对核心引物(TM156、TM508、MSG0380)用于不同白化、黄化茶树品种的鉴定。
To differentiate and identify different albino tea cultivars, sixty two SSR primers were used to analyze the genetic diversity of 16 tea cultivars that exhibit the chlorina phenotype. The result showed that a total of 169 alleles were amplified by 55 SSR primers with good polymorphism, and the number of alleles per primer ranged from 2 to 5, with an average of 3.07. The average value of polymorphism information content(PIC) and Shannon's information index(I) was 0.40 and 0.79 respectively, which indicate these albino and yellow-leaf tea cultivars having a moderate level of diversity. And the occurrence frequency of 169 alleles ranged from 3.12% to 96.88%. It suggested the difference of genetic structure among tested varieties is obvious. The Nei′s genetic distance(D) of sixteen tested cultivars ranged from 0.086 to 0.532. These cultivars could be divided into three categories when D was 0.18, and the cultivars with close relatives or similar geographical origin were clustered into one group. Lastly, three primers(TM156, TM508, MSG0380) with easy identification, good stability and high polymorphism, were finally chosen to establish the fingerprint. The 16 albino and yellow-leaf tea cultivars could be effectively distinguished by the primers. This study provided an important basis for the identification of albino tea cultivars, and the evaluation and utilization of these germplasm resources.
To differentiate and identify different albino tea cultivars, sixty two SSR primers were used to analyze the genetic diversity of 16 tea cultivars that exhibit the chlorina phenotype. The result showed that a total of 169 alleles were amplified by 55 SSR primers with good polymorphism, and the number of alleles per primer ranged from 2 to 5, with an average of 3.07. The average value of polymorphism information content(PIC) and Shannon's information index(I) was 0.40 and 0.79 respectively, which indicate these albino and yellow-leaf tea cultivars having a moderate level of diversity. And the occurrence frequency of 169 alleles ranged from 3.12% to 96.88%. It suggested the difference of genetic structure among tested varieties is obvious. The Nei′s genetic distance(D) of sixteen tested cultivars ranged from 0.086 to 0.532. These cultivars could be divided into three categories when D was 0.18, and the cultivars with close relatives or similar geographical origin were clustered into one group. Lastly, three primers(TM156, TM508, MSG0380) with easy identification, good stability and high polymorphism, were finally chosen to establish the fingerprint. The 16 albino and yellow-leaf tea cultivars could be effectively distinguished by the primers. This study provided an important basis for the identification of albino tea cultivars, and the evaluation and utilization of these germplasm resources.
引文
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[4]Feng L,Gao MJ,Hou RY,et al.Determination of quality constituents in the young leaves of albino tea culti vars[J].Food Chemistry,2014,155(11):98-104.
[5]卢翠,沈程文.茶树白化变异研究进展[J].茶叶科学,2016,36(5):445-451.
[6]马春雷,姚明哲,王新超,等.茶树叶绿素合成相关基因克隆及在白叶1号不同白化阶段的表达分析[J].作物学报,2015,41(2):240-250.
[7]梁明山,曾宇,周翔,等.遗传标记及其在作物品种鉴定中的作用[J].植物学报,2001,18(3):257-265.
[8]马建强,姚明哲,陈亮.茶树种质资源研究进展[J].茶叶科学,2015,35(1):11-16.
[9]黄丹娟,马建强,陈亮.茶树DNA分子指纹图谱研究进展[J].茶叶科学,2015,35(6):513-519.
[10]陈新民,何中虎,史建荣,等.利用SSR标记进行优质东小麦品种(系)的遗传多样性研究[J].作物学报,2003,29(1):13-19.
[11]程保山,徐海风,顾正中,等.淮安地区主栽小麦品种指纹图谱的构建及遗传多样性分析[J].浙江农业学报,2011,23(1):20-24.
[12]谭君,丁仲芳,孙仕贤,等.西南常用玉米自交系SSR指纹图谱构建[J].西南农业学报,2003,16(2):1-6.
[13]李丽华,魏昕,潘光堂.20个骨干玉自交系的SSR指纹图谱构建[J].西南农业学报,2007,20(6):1172-1175.
[14]肖小余,王玉平,张建勇,等.四川省主要杂交稻亲本的SSR多态性分析和指纹图谱的构建与应用[J].中国水稻科学,2006,20(1):1-7.
[15]陆徐忠,从夕汉,刘海珍,等.杂交水稻亲本分子身份证及SSR指纹数据库的建立[J].核农学报,2012,26(6):853-861.
[16]藕丹,樊军锋,周永学,等.美洲黑杨×青杨派杂种无性系SSR指纹图谱的构建与遗传差异性分析[J].西北林学院学报,2017(2):112-116.
[17]艾呈祥,张力思,魏海蓉,等.甜樱桃品种SSR指纹图谱数据库的建立[J].中国农学通报,2007,23(5):55-58.
[18]王星星,周琦,陶圆圆,等.48个果用银杏品种SSR指纹图谱构建与遗传多样性分析[J].分子植物育种,2017,15(5):1963-1970.
[19]李莉,宋书锋,刘全胜,等.利用SSR分子标记建立杂交水稻指纹图谱[J].长江大学学报(自然科学版):农学卷,2008,5(4):67-70.
[20]王立新,张小军,史星雲,等.苹果栽培品种SSR指纹图谱的构建[J].果树学报,2012(6):971-977.
[21]乔小燕,乔婷婷,周艳华,等.基于EST-SSR的广东御广西茶树资源遗传结构和遗传分化比较分析[J].中国农业科学,2011,44(16):3297-3311.
[22]黄丹娟,马建强,陈亮.茶树PVP申请品种的SSR分子标记鉴定和系谱关系分析[J].茶叶科学,2016,36(1):68-76.
[23]Dellaporta S L,Wood J,Hicks J B.A plant DNA mini preparation:version II[J].Plant Molecular Biology Reporter,1983(1):19-21.
[24]Charters Y M,Wilkinson M J.The use of self-pollinated progenies as‘in-groups’for the genetic characterization of cocoa germplasm[J].Theoretical and Applied Genetics,2000,100(1):160-166.
[25]Yeh F C,Yang R C,Boyle T B,et al.POPGENE,the user-friendly shareware for population genetic analysis[R].Edmonton:Molecular Biology and Biotechnology Centre,University of Alberta,1997:112-118.
[26]Liu K,Muse S V.Power Marker an integrated analysis environment for genetic marker analysis[J].Bioinformatics,2005,21(9):2128-2129.
[27]Takezaki H,Nei M.Genetic distances and reconstruction of phylogenetic trees from microsatellite DNA[J].Genetics,1996,144(1):389-399.
[28]杨阳,刘振,赵洋,等.湖南省主要茶树品种分子指纹图谱的构建[J].茶叶科学,2010,30(5):367-373.
[29]张兰,李鑫,魏吉鹏,等.白叶一号的低温抗逆特性分析[J].茶叶科学,2017,37(1):71-77.
[30]王开荣,李明,梁月荣,等.茶树新品种黄金芽选育研究[J].中国茶叶,2008,30(4):21-23.
[31]夏功敏,唐茜,张淑娟,等.黄金芽等五个引进特色茶树品种春梢生化品质特性初探[J].西南农业学报,2014,27(3):978-983.
[32]黄茶育种协作组.特异优质茶树新品种“中黄1号”选育研究[J].中国茶叶,2014,36(9):8-9.
[33]虞富莲.中黄1号、中黄2号的特异性、一致性和稳定性[J].中国茶叶,2016,38(3):14-16.
[34]王开荣,梁月荣,李明,等.白化和紫花茶种质资源开发进展[J].中国茶叶加工,2015(3):5-8.
[35]王开荣,梁月荣,张龙杰,等.白化茶种质资源的分类及特性[J].中国茶叶,2008,30(8):9-11.
[36]王开荣,梁月荣,李明,等.白化茶骨干亲本及其家系种质性状分析[J].茶叶,2015,41(3):130-132.
[37]刘新龙,马丽,陈学宽,等.云南甘蔗自育品种DNA指纹身份证构建[J].作物学报,2010,36(2):202-210.
[38]王让剑,杨军,孔祥瑞,等.福建15个茶树品种SSR遗传差异分析与指纹图谱建立[J].福建农业学报,2014,29(10):970-975.
[39]Ujihara T,Ohta R,Hayashi N,et al.Identification of Japanese and Chinese green tea cultivars by using simple sequence repeat markers to encourage proper labeling[J].Bioscience Biotechnology and Biochemistry,2009,73(1):15-20.
[40]张成才,刘园,姜燕华,等.SSR标记鉴定浙江省主要无性系茶树品种的研究[J].植物遗传资源学报,2014,15(5):926-931.
[41]章志芳,马建强.基于SSR标记的茶树新品种遗传多样性分析及指纹图谱构建[J].湖南农业科学,2012(19):1-4.
[42]郭燕,刘声传,曹雨,等.基于SSR标记贵州古茶树资源的遗传多样性分析及指纹图谱构建[J].西南农业学报,2016,29(3):491-497.
[2]Li CF,Yao MZ,Ma CL,et al.Differential metabolic profiles during the albescent stages of'Anji Baicha'(Camellia sinensis)[J].PLo S One,2015,10(10):e0139996.
[3]Ma CL,Chen L,Wang XC,et al.Differential expression analysis of different albescent stages of‘Anji Baicha’(Camellia sinensis(L.)O.Kuntze)using c DNA microarray[J].Scientia Horticulturae,2012,148(4):246-254.
[4]Feng L,Gao MJ,Hou RY,et al.Determination of quality constituents in the young leaves of albino tea culti vars[J].Food Chemistry,2014,155(11):98-104.
[5]卢翠,沈程文.茶树白化变异研究进展[J].茶叶科学,2016,36(5):445-451.
[6]马春雷,姚明哲,王新超,等.茶树叶绿素合成相关基因克隆及在白叶1号不同白化阶段的表达分析[J].作物学报,2015,41(2):240-250.
[7]梁明山,曾宇,周翔,等.遗传标记及其在作物品种鉴定中的作用[J].植物学报,2001,18(3):257-265.
[8]马建强,姚明哲,陈亮.茶树种质资源研究进展[J].茶叶科学,2015,35(1):11-16.
[9]黄丹娟,马建强,陈亮.茶树DNA分子指纹图谱研究进展[J].茶叶科学,2015,35(6):513-519.
[10]陈新民,何中虎,史建荣,等.利用SSR标记进行优质东小麦品种(系)的遗传多样性研究[J].作物学报,2003,29(1):13-19.
[11]程保山,徐海风,顾正中,等.淮安地区主栽小麦品种指纹图谱的构建及遗传多样性分析[J].浙江农业学报,2011,23(1):20-24.
[12]谭君,丁仲芳,孙仕贤,等.西南常用玉米自交系SSR指纹图谱构建[J].西南农业学报,2003,16(2):1-6.
[13]李丽华,魏昕,潘光堂.20个骨干玉自交系的SSR指纹图谱构建[J].西南农业学报,2007,20(6):1172-1175.
[14]肖小余,王玉平,张建勇,等.四川省主要杂交稻亲本的SSR多态性分析和指纹图谱的构建与应用[J].中国水稻科学,2006,20(1):1-7.
[15]陆徐忠,从夕汉,刘海珍,等.杂交水稻亲本分子身份证及SSR指纹数据库的建立[J].核农学报,2012,26(6):853-861.
[16]藕丹,樊军锋,周永学,等.美洲黑杨×青杨派杂种无性系SSR指纹图谱的构建与遗传差异性分析[J].西北林学院学报,2017(2):112-116.
[17]艾呈祥,张力思,魏海蓉,等.甜樱桃品种SSR指纹图谱数据库的建立[J].中国农学通报,2007,23(5):55-58.
[18]王星星,周琦,陶圆圆,等.48个果用银杏品种SSR指纹图谱构建与遗传多样性分析[J].分子植物育种,2017,15(5):1963-1970.
[19]李莉,宋书锋,刘全胜,等.利用SSR分子标记建立杂交水稻指纹图谱[J].长江大学学报(自然科学版):农学卷,2008,5(4):67-70.
[20]王立新,张小军,史星雲,等.苹果栽培品种SSR指纹图谱的构建[J].果树学报,2012(6):971-977.
[21]乔小燕,乔婷婷,周艳华,等.基于EST-SSR的广东御广西茶树资源遗传结构和遗传分化比较分析[J].中国农业科学,2011,44(16):3297-3311.
[22]黄丹娟,马建强,陈亮.茶树PVP申请品种的SSR分子标记鉴定和系谱关系分析[J].茶叶科学,2016,36(1):68-76.
[23]Dellaporta S L,Wood J,Hicks J B.A plant DNA mini preparation:version II[J].Plant Molecular Biology Reporter,1983(1):19-21.
[24]Charters Y M,Wilkinson M J.The use of self-pollinated progenies as‘in-groups’for the genetic characterization of cocoa germplasm[J].Theoretical and Applied Genetics,2000,100(1):160-166.
[25]Yeh F C,Yang R C,Boyle T B,et al.POPGENE,the user-friendly shareware for population genetic analysis[R].Edmonton:Molecular Biology and Biotechnology Centre,University of Alberta,1997:112-118.
[26]Liu K,Muse S V.Power Marker an integrated analysis environment for genetic marker analysis[J].Bioinformatics,2005,21(9):2128-2129.
[27]Takezaki H,Nei M.Genetic distances and reconstruction of phylogenetic trees from microsatellite DNA[J].Genetics,1996,144(1):389-399.
[28]杨阳,刘振,赵洋,等.湖南省主要茶树品种分子指纹图谱的构建[J].茶叶科学,2010,30(5):367-373.
[29]张兰,李鑫,魏吉鹏,等.白叶一号的低温抗逆特性分析[J].茶叶科学,2017,37(1):71-77.
[30]王开荣,李明,梁月荣,等.茶树新品种黄金芽选育研究[J].中国茶叶,2008,30(4):21-23.
[31]夏功敏,唐茜,张淑娟,等.黄金芽等五个引进特色茶树品种春梢生化品质特性初探[J].西南农业学报,2014,27(3):978-983.
[32]黄茶育种协作组.特异优质茶树新品种“中黄1号”选育研究[J].中国茶叶,2014,36(9):8-9.
[33]虞富莲.中黄1号、中黄2号的特异性、一致性和稳定性[J].中国茶叶,2016,38(3):14-16.
[34]王开荣,梁月荣,李明,等.白化和紫花茶种质资源开发进展[J].中国茶叶加工,2015(3):5-8.
[35]王开荣,梁月荣,张龙杰,等.白化茶种质资源的分类及特性[J].中国茶叶,2008,30(8):9-11.
[36]王开荣,梁月荣,李明,等.白化茶骨干亲本及其家系种质性状分析[J].茶叶,2015,41(3):130-132.
[37]刘新龙,马丽,陈学宽,等.云南甘蔗自育品种DNA指纹身份证构建[J].作物学报,2010,36(2):202-210.
[38]王让剑,杨军,孔祥瑞,等.福建15个茶树品种SSR遗传差异分析与指纹图谱建立[J].福建农业学报,2014,29(10):970-975.
[39]Ujihara T,Ohta R,Hayashi N,et al.Identification of Japanese and Chinese green tea cultivars by using simple sequence repeat markers to encourage proper labeling[J].Bioscience Biotechnology and Biochemistry,2009,73(1):15-20.
[40]张成才,刘园,姜燕华,等.SSR标记鉴定浙江省主要无性系茶树品种的研究[J].植物遗传资源学报,2014,15(5):926-931.
[41]章志芳,马建强.基于SSR标记的茶树新品种遗传多样性分析及指纹图谱构建[J].湖南农业科学,2012(19):1-4.
[42]郭燕,刘声传,曹雨,等.基于SSR标记贵州古茶树资源的遗传多样性分析及指纹图谱构建[J].西南农业学报,2016,29(3):491-497.