新疆野樱桃李EST-SSR引物开发及通用性评价
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摘要
野生樱桃李是中国濒临灭绝的树种之一。本研究从分子水平对野樱桃李进行研究,以期为制定保护策略提供理论依据。从NCBI数据库中下载樱桃李EST序列,用SSR Hunter 1.3查找SSR位点,用Primer 6.0设计了30对引物并验证引物的通用性。结果表明:有20对引物能扩增出有效条带,扩增率为66.67%,其中9对引物能扩增出多态性条带,多态性比率为45%。用20对有效引物在欧洲李和天山樱桃中进行扩增,均表现良好的通用性,且同属植物间的通用性明显高于同科植物间,说明引物通用性的高低与基因组的差异有关,亲缘关系越近引物通用性越高。
Wild type Prunus cerasifera is one of the endangered species in China. In this research, wild type P.cerasifera was studied from the molecular level, so as to provide a theoretical basis for making protection strategies. The EST sequence of P. cerasifera was downloaded from NCBI database, the SSR loci was found with SSR Hunter 1.3, 30 pairs of primers were designed with Primer 6.0 and their universality were validated.The results showed that 20 pairs of primers could amplify effective bands, and the amplification rate was 66.67%, 9 pairs of which could amplify polymorphic bands, and the polymorphic rate was 45%. Besides, all the 20 pairs of effective primers amplified in P. domestica and Cerasus tianschanica had good universality, and the universality among the same generic plants was significantly higher than that among the same family plants,which indicated that the universality level of primer depended on the differences of genome, and the closer the genetic relationship was, the higher universality the primer had.
Wild type Prunus cerasifera is one of the endangered species in China. In this research, wild type P.cerasifera was studied from the molecular level, so as to provide a theoretical basis for making protection strategies. The EST sequence of P. cerasifera was downloaded from NCBI database, the SSR loci was found with SSR Hunter 1.3, 30 pairs of primers were designed with Primer 6.0 and their universality were validated.The results showed that 20 pairs of primers could amplify effective bands, and the amplification rate was 66.67%, 9 pairs of which could amplify polymorphic bands, and the polymorphic rate was 45%. Besides, all the 20 pairs of effective primers amplified in P. domestica and Cerasus tianschanica had good universality, and the universality among the same generic plants was significantly higher than that among the same family plants,which indicated that the universality level of primer depended on the differences of genome, and the closer the genetic relationship was, the higher universality the primer had.
引文
[1]廖康.新疆野生果树资源研究[M].乌鲁木齐:新疆人民出版社,2013.
[2]兰士波.樱桃李的研究进展及开发利用前景[J].中国林副特产,2008(5):89-91.
[3]周龙,廖康,王磊,等.低温胁迫对新疆野生樱桃李电解质渗出率和丙二醛含量的影响[J].新疆农业大学学报,2006,29(1):47-50.
[4]林培钧,林德佩,王磊,等.新疆果树的野生近缘植物[J].八一农学院学报,1984(4):25-32.
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[6]张士康,肖正春,张卫明,等.中国野生樱桃李的生态学研究[J].中国野生植物资源,2004,23(2):1-3.
[7]刘影,张相锋,赵玉,等.新疆濒危野生樱桃李的种群结构与动态[J].生态学杂志,2013,32(7):1762-1769
[8]周龙,胡建芳,许正,等.野生樱桃李天然群体表型多样性研究[J].新疆农业大学学报,2011(3):222-225.
[9]周龙,胡建芳,许正,等.野生樱桃李天然群体果实形态多样性分析[J].吉林农业大学学报,2011,33(6):637-642,653.
[10]周龙,胡建芳,卢崇烈,等.野生樱桃李天然群体的遗传多样性分析[J].新疆农业大学学报,2013,36(3):213-217.
[11]林元震,郭海,刘纯鑫,等.EST-SSR标记在木本植物中的开发和应用[J].植物生理学通讯,2009,45(12):1221-1225.
[12]上官凌飞,李晓颖,宁宁,等.杏EST-SSR标记的开发[J].园艺学报,2011,38(1):43-54.
[13]廖娇,黄春辉,辜青青,等.猕猴桃EST-SSR引物筛选及通用性分析[J].果树学报,2011,28(6):1111-1116.
[14]罗纯.柿EST-SSR标记在柿属植物中的通用性研究[J].湖北农业科学,2014,53(22):5434-5437.
[15]王娟.葡萄EST-SSR引物的开发及部分种质聚类分析[J].华北农学报,2014,29(2):121-126.
[16]王西成,姜淑苓,上官凌飞,等.梨EST-SSR标记的开发及其在梨品种遗传多样性分析中的应用评价[J].中国农业科学,2010,43(24):5079-5087.
[17]Aranzanam J,Garcia-mas J,Carbo J,et al.Development and variability analysis of microsatellite markers in peach[J].Plant Breed,2002,121:87-92.
[18]Jung S,Abbott A,Jesudural C,et al.Frequency,type,distribution and annotation of simple sequence repeats in Rosaceae ESTs[J].Functional&Integrative Genomics,2005,5(3):136-143.
[19]Poncet V,Rondeau M,Tranchant C,et al.SSR mining in coffee tree EST databases:potential use of EST-SSRs as markers for the Coffea genus[J].Molecular Genetics and Genomics,2006,276(5):436-449.
[20]邓传良,高俊,曹莹,等.菠菜性别相关EST-SSR标记的开发及应用[J].基因组学与应用生物学,2012(5):467-472.
[21]李美芹,潘叶羽,钱萍仙,等.杜鹃花EST-SSR标记的开发及遗传多样性分析[J].植物生理学报,2016(3):356-364.
[22]林范学,程水明,李安政,等.香菇EST-SSR引物筛选[J].农业生物技术学报,2007,15(2):358-359.
[23]贾新平.鸟巢蕨EST-SSR标记的开发与应用[J].华北农学报,2015,30(6):134-139.
[24]孙清明,马文朝,马帅鹏,等.荔枝EST资源的SSR信息分析及ESTSSR标记开发[J].中国农业科学,2011,44(19):4037-4049.
[25]崔海荣,刘金义,佟兆国,等.砂梨EST-SSR引物开发及其应用[J].西北植物学报,2010,30(8):1551-1556.
[26]忻雅,崔海瑞,张明龙,等.白菜EST-SSR标记的通用性[J].细胞生物学杂志,2006,28:248-252.
[27]李红英,李康琴,胥猛,等.北美鹅掌楸EST-SSR跨属间通用性[J].东北林业大学学报,2011(2):28-30,42.
[2]兰士波.樱桃李的研究进展及开发利用前景[J].中国林副特产,2008(5):89-91.
[3]周龙,廖康,王磊,等.低温胁迫对新疆野生樱桃李电解质渗出率和丙二醛含量的影响[J].新疆农业大学学报,2006,29(1):47-50.
[4]林培钧,林德佩,王磊,等.新疆果树的野生近缘植物[J].八一农学院学报,1984(4):25-32.
[5]朱保志,雷新英.野生樱桃李的适生性[J].新疆林业,2001(3):22-22.
[6]张士康,肖正春,张卫明,等.中国野生樱桃李的生态学研究[J].中国野生植物资源,2004,23(2):1-3.
[7]刘影,张相锋,赵玉,等.新疆濒危野生樱桃李的种群结构与动态[J].生态学杂志,2013,32(7):1762-1769
[8]周龙,胡建芳,许正,等.野生樱桃李天然群体表型多样性研究[J].新疆农业大学学报,2011(3):222-225.
[9]周龙,胡建芳,许正,等.野生樱桃李天然群体果实形态多样性分析[J].吉林农业大学学报,2011,33(6):637-642,653.
[10]周龙,胡建芳,卢崇烈,等.野生樱桃李天然群体的遗传多样性分析[J].新疆农业大学学报,2013,36(3):213-217.
[11]林元震,郭海,刘纯鑫,等.EST-SSR标记在木本植物中的开发和应用[J].植物生理学通讯,2009,45(12):1221-1225.
[12]上官凌飞,李晓颖,宁宁,等.杏EST-SSR标记的开发[J].园艺学报,2011,38(1):43-54.
[13]廖娇,黄春辉,辜青青,等.猕猴桃EST-SSR引物筛选及通用性分析[J].果树学报,2011,28(6):1111-1116.
[14]罗纯.柿EST-SSR标记在柿属植物中的通用性研究[J].湖北农业科学,2014,53(22):5434-5437.
[15]王娟.葡萄EST-SSR引物的开发及部分种质聚类分析[J].华北农学报,2014,29(2):121-126.
[16]王西成,姜淑苓,上官凌飞,等.梨EST-SSR标记的开发及其在梨品种遗传多样性分析中的应用评价[J].中国农业科学,2010,43(24):5079-5087.
[17]Aranzanam J,Garcia-mas J,Carbo J,et al.Development and variability analysis of microsatellite markers in peach[J].Plant Breed,2002,121:87-92.
[18]Jung S,Abbott A,Jesudural C,et al.Frequency,type,distribution and annotation of simple sequence repeats in Rosaceae ESTs[J].Functional&Integrative Genomics,2005,5(3):136-143.
[19]Poncet V,Rondeau M,Tranchant C,et al.SSR mining in coffee tree EST databases:potential use of EST-SSRs as markers for the Coffea genus[J].Molecular Genetics and Genomics,2006,276(5):436-449.
[20]邓传良,高俊,曹莹,等.菠菜性别相关EST-SSR标记的开发及应用[J].基因组学与应用生物学,2012(5):467-472.
[21]李美芹,潘叶羽,钱萍仙,等.杜鹃花EST-SSR标记的开发及遗传多样性分析[J].植物生理学报,2016(3):356-364.
[22]林范学,程水明,李安政,等.香菇EST-SSR引物筛选[J].农业生物技术学报,2007,15(2):358-359.
[23]贾新平.鸟巢蕨EST-SSR标记的开发与应用[J].华北农学报,2015,30(6):134-139.
[24]孙清明,马文朝,马帅鹏,等.荔枝EST资源的SSR信息分析及ESTSSR标记开发[J].中国农业科学,2011,44(19):4037-4049.
[25]崔海荣,刘金义,佟兆国,等.砂梨EST-SSR引物开发及其应用[J].西北植物学报,2010,30(8):1551-1556.
[26]忻雅,崔海瑞,张明龙,等.白菜EST-SSR标记的通用性[J].细胞生物学杂志,2006,28:248-252.
[27]李红英,李康琴,胥猛,等.北美鹅掌楸EST-SSR跨属间通用性[J].东北林业大学学报,2011(2):28-30,42.