文章摘要
王婷,徐燕,谢潮添,纪德华,陈昌生.基于SCAR标记的坛紫菜“闽丰1号”多重PCR鉴定技术的建立[J].水产学报,2013,37(5):688~695
基于SCAR标记的坛紫菜“闽丰1号”多重PCR鉴定技术的建立
Construction of multiplex PCR in variety identification of Porphyra haitanensis “Z-26” based on SCAR markers
投稿时间:2012-06-05  修订日期:2013-03-05
DOI:DOI:10.3724/SP.J.1231.2013.38191
中文关键词: 坛紫菜  随机扩增多态DNA(RAPD)  序列特异扩增区域(SCAR)  多重PCR
英文关键词: Porphyra haitanensis  random amplified polymorphic DNA(RAPD)  sequence characterized amplified region(SCAR)  multiplex PCR
基金项目:国家自然科学基金项目(41176151,41276177);公益性行业(农业)科研专项(200903030);海洋公益性行业科研专项(201105008,201105023);福建省杰出青年基金项目(2010J06016);福建省教育厅新世纪优秀人才项目(JA10186)
作者单位
王婷 集美大学水产学院 
徐燕 集美大学水产学院 
谢潮添 集美大学水产学院 
纪德华 集美大学水产学院 
陈昌生 集美大学水产学院 
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中文摘要:
      为建立坛紫菜“闽丰1号”(Z-26)品系的种质分子鉴定方法,采用300条RAPD引物对6个坛紫菜纯系进行标记扫描,从中筛选出“Z-26”品系的特异性随机扩增多态DNA(RAPD)标记9个,经克隆和测序,其中两个特异性标记成功转化为特异SCAR标记(Z26-600和Z26-360),标记片段大小分别为530 bp和242 bp。并进一步通过4个不同实验验证,确定Z26-600和Z26-360两个标记是坛紫菜“Z-26”品系的特异和稳定性标记。最后为进一步简化种质鉴定程序,建立更为便捷和准确的种质鉴定方法,经过条件优化,以此两个标记为基础建立了坛紫菜“Z-26”品系的多重PCR鉴定方法,该方法可以在一次PCR反应中同时鉴定两个特异性标记。
英文摘要:
      Porphyra haitanensis is one of the most important economic marine crops of China.For any cultivar,the correct identification of species or forma of the cultivated strains is necessary to ensure a well-bred cultivation and good production quality.However,because the gametophytic blade of Porphyra is morphologically simple and marked variations can occur as environmental conditions change,it is very difficult to precisely identify the species or forma of cultivated strains based only on their morphological characteristics.With new advances in molecular biology,molecular markers and DNA fingerprinting techniques have become routine for the identification and classification of many crops,including seaweeds.The strain of “Z-26” of P.haitanensiswas selected by the laboratory of germplasm improvement and the application of P.haitanensis in Jimei University which has the characters of high-temperature tolerance and high yield,and it has been widely cultivated in south China.In order to construct the technology of variety identification for “Z-26”,firstly,300 primers of RAPD were used to scan the specific markers of 6 new strains of P.haitanensis and 9 specific RAPD markers of “Z-26” were selected.After cloning and sequencing,two specific RAPD markers of “Z-26” were transformed into the SCAR markers successfully,the length of the 2 SCAR markers was 540 and 242 bp,respectively.Secondly,after verification by 4 different experiments,we can affirm that the 2 SCAR markers were the specific and stable markers of “Z-26”.At last,based on the 2 SCAR markers,after optimization of experimental conditions,the technology of multiplex PCR which was used to identify the variety of “Z-26” was constructed.The result supplied a simple,fast and reliable technigue for variety identification of “Z-26”.
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