天麻ITS-1测序及单核苷酸多态性变异位点分析
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摘要
目的:选取天麻3种变型,测定ITS-1序列并进行对比分析,找出变异信息位点,对不同变异类型亲缘关系作出判定。方法:采用改良的CTAB法提取DNA,利用合成的特异引物对ITS区进行扩增,并对目的片段测序分析。结果:ITS-1共计209bp,序列片段上有27个单核苷酸变异位点,ITS-1单核苷酸多态性以碱基转换为主,绿天麻最高达80.8%,乌天麻最低也达70%;黄天麻居中达71.4%。在多态性位点数及多态性频率上,绿天麻均表现为最高分别为26、8.04;黄天麻表现居中为21、9.95;乌天麻表现最低为20、10.45。结论:乌天麻是古老类型,绿天麻是较为进化的类型,黄、绿天麻是由乌天麻进化而来。
Objective:In order to identify the relationship of differences types of variation in three samples of Gastrodia elata,their ITS sequences of rDNA genes were amplified.ITS-1 was sequenced and comparatively analyzed and the variation information sites were identified.Method:Genomic DNA from Gastrodia elata extracted by modified CTAB method were regarded as templates for PCR amplification,cloning,and sequencing by synthetic specific primer the nrDNA ITS sequences.Result:The results showed that there were 27 single nucleotide variable sites in the total of 209 bp.It were confirmed that there was a wealth of genetic diversity in Gastrodia elata.The ITS-1 single nucleotide polymorphisms base conversion in Gastrodia elata,G.elata Bl.f.viridis Mak up to 80.8%,G.elata Bl.f.glauca S.Chow lowest 70%,G.elata Bl.f.flavida S.Chow centered at 71.4%.G.elata Bl.f.viridis Mak was manifested in the top 26,8.04 respectively,G.elata Bl.f.flavida S.Chow centered at 21,9.95,G.elata Bl.f.glauca S.Chow lowest 20,10.45 in Polymorphism of points and polymorphism frequencies.Conclusion:There may be an ancient type.G.elata Bl.f.flavida S.Chow and G.elata Bl.f.viridis Makyellow may be evolved from G.elata Bl.f.glauca S.Chow by 27 single nucleotide mutation sites.
Objective:In order to identify the relationship of differences types of variation in three samples of Gastrodia elata,their ITS sequences of rDNA genes were amplified.ITS-1 was sequenced and comparatively analyzed and the variation information sites were identified.Method:Genomic DNA from Gastrodia elata extracted by modified CTAB method were regarded as templates for PCR amplification,cloning,and sequencing by synthetic specific primer the nrDNA ITS sequences.Result:The results showed that there were 27 single nucleotide variable sites in the total of 209 bp.It were confirmed that there was a wealth of genetic diversity in Gastrodia elata.The ITS-1 single nucleotide polymorphisms base conversion in Gastrodia elata,G.elata Bl.f.viridis Mak up to 80.8%,G.elata Bl.f.glauca S.Chow lowest 70%,G.elata Bl.f.flavida S.Chow centered at 71.4%.G.elata Bl.f.viridis Mak was manifested in the top 26,8.04 respectively,G.elata Bl.f.flavida S.Chow centered at 21,9.95,G.elata Bl.f.glauca S.Chow lowest 20,10.45 in Polymorphism of points and polymorphism frequencies.Conclusion:There may be an ancient type.G.elata Bl.f.flavida S.Chow and G.elata Bl.f.viridis Makyellow may be evolved from G.elata Bl.f.glauca S.Chow by 27 single nucleotide mutation sites.
引文
[1]徐锦堂.中国天麻栽培学[M].北京:北京医科大学出版社,1993:10.
[2]蒋明,陈贝贝,贺蔡明.石豆兰属植物rDNA ITS序列的克隆与分析[J].中草药,2012,43(2):343-347.
[3]李学营,彭建营,白瑞霞,等.基于核rDNA的ITS序列在种子植物系统发育研究中的应用[J].西北植物学报,2005,25(4):829-834.
[4]郑伟,陈龙清.基于ITS序列探讨珍珠菜属过路黄组的系统关系[J].广西植物,2012,32(2):163-166.
[5]徐红,李晓波,丁小余,等.中药黄草石斛rDNA ITS序列分析[J].药学学报,2001,36(10):777-783.
[6]谢渊,张小蕾,李毅,等.天麻rDNAITS序列测定及特点的初步分析[J].中药材,2009,32(12):1814-1815.
[7]吴波,刘勇.柑橘类植物GPAT基因片段克隆和SNP分析[J].江西农业大学学报,2010,32(1):51-56.
[8]严明理,刘显军,官春云,等.芥菜型油菜TT1基因的克隆和SNP分析[J].作物学报,2010,36(10):1634-1641.
[9]赵雪,谢华,马荣才.植物功能基因组研究中出现的新型分子标记[J].中国生物工程杂志,2007,27(8):104-110.
[10]王德信.天麻ITS序列分析及变异类型鉴定[J].生物技术,2010,20(6):33-35.
[11]王德信.随机扩增多态DNA技术在天麻遗传多样性分析中的应用[J].中国农学通报,2010,26(4):19-24.
[12]于颢,闫旭,郭卫东,等.水稻micro RNA单核苷酸多态性[J].中国水稻科学,2011,25(5):467-474.
[13]赵辉,李启寨,李俊,等.相邻碱基组分与产生SNP的转换或颠换在植物基因组中的研究[J].中国科学C辑:生命科学,2006,36(1):1-8.
[14]许亮,谷丽艳,赵丹玉,等.基于ITS序列的植物类中药材鉴定研究及展望[J].中华中医药学刊,2010,28(4):737-738.
[15]赵欢,吴卫,郑有良,等.核糖体DNA ITS序列分析在药用植物研究中的应用[J].时珍国医国药,2009,20(4):959-962.
[2]蒋明,陈贝贝,贺蔡明.石豆兰属植物rDNA ITS序列的克隆与分析[J].中草药,2012,43(2):343-347.
[3]李学营,彭建营,白瑞霞,等.基于核rDNA的ITS序列在种子植物系统发育研究中的应用[J].西北植物学报,2005,25(4):829-834.
[4]郑伟,陈龙清.基于ITS序列探讨珍珠菜属过路黄组的系统关系[J].广西植物,2012,32(2):163-166.
[5]徐红,李晓波,丁小余,等.中药黄草石斛rDNA ITS序列分析[J].药学学报,2001,36(10):777-783.
[6]谢渊,张小蕾,李毅,等.天麻rDNAITS序列测定及特点的初步分析[J].中药材,2009,32(12):1814-1815.
[7]吴波,刘勇.柑橘类植物GPAT基因片段克隆和SNP分析[J].江西农业大学学报,2010,32(1):51-56.
[8]严明理,刘显军,官春云,等.芥菜型油菜TT1基因的克隆和SNP分析[J].作物学报,2010,36(10):1634-1641.
[9]赵雪,谢华,马荣才.植物功能基因组研究中出现的新型分子标记[J].中国生物工程杂志,2007,27(8):104-110.
[10]王德信.天麻ITS序列分析及变异类型鉴定[J].生物技术,2010,20(6):33-35.
[11]王德信.随机扩增多态DNA技术在天麻遗传多样性分析中的应用[J].中国农学通报,2010,26(4):19-24.
[12]于颢,闫旭,郭卫东,等.水稻micro RNA单核苷酸多态性[J].中国水稻科学,2011,25(5):467-474.
[13]赵辉,李启寨,李俊,等.相邻碱基组分与产生SNP的转换或颠换在植物基因组中的研究[J].中国科学C辑:生命科学,2006,36(1):1-8.
[14]许亮,谷丽艳,赵丹玉,等.基于ITS序列的植物类中药材鉴定研究及展望[J].中华中医药学刊,2010,28(4):737-738.
[15]赵欢,吴卫,郑有良,等.核糖体DNA ITS序列分析在药用植物研究中的应用[J].时珍国医国药,2009,20(4):959-962.