不同来源莲rDNA ITS的PCR扩增、克隆及序列分析PCR,clone and sequence analysis of rDNA-ITS of Nelumbo nucifera from different geographical origins in China
林珊;郑伟文;吴锦忠;周莉娟;宋亚娜;
LIN Shan1,2,ZHENG Wei-wen2,WU Jin-zhong1,ZHOU Li-juan2,SONG Ya-na2(1.Department of Pharmacy,Fujian University of Traditional Chinese Medicine,Fuzhou 350003,China;2.Biotechnology Institute,Fujian Academy of Agricultural Sciences,Fuzhou 350003,China)
摘要(Abstract):
目的:通过分析不同来源莲的ITS序列,为莲的鉴别提供DNA分子标记。方法:利用特异性引物进行PCR扩增和克隆、测序技术对莲的rDNA ITS区间碱基序列进行测定,比较其差异。结果:得到参试的11个莲栽培品种的rDNA的ITS及5.8S rDNA全序列,18S和26S rDNA部分序列,共约750 bp。显示了不同产地、不同栽培品种莲的rDNA ITS的差异。结论:本研究为利用ITS区序列差异,鉴别不同来源的莲提供了依据,此法可用于莲种间及真伪品鉴别。
Objective:To provide DNA molecular marker for identification of Nelumbo nucifera by exploring the differences of nrDNA-ITS sequence of N.nucifera originated from different habitats.Method: To compare nrDNA-ITS base sequence using specific PCR-ITS. Result:The completed sequence of ITS and 5.8 S rDNA,and the partial sequences of 18S rDNA and 26S rDNA,totally 750 bp,from N.nucifera were obtained.The differences among N.nucifera from different habitats and from different cultivars were found.Conclusion: The method can be used to identify N.nucifera among different species and to distinguish their fakes.It provided the basis for identifying N.nucifera from different geographical regions by comparison of their ITS sequences.
关键词(KeyWords):
分子鉴定;莲;ITS序列;序列分析
molecular identification;Nelumbo nucifera; ITS region;sequence analysis
基金项目(Foundation): 福建省科技重大项目(2002Y006);; 福建省科技项目(2004Y007,2001Z026)
作者(Authors):
林珊;郑伟文;吴锦忠;周莉娟;宋亚娜;
LIN Shan1,2,ZHENG Wei-wen2,WU Jin-zhong1,ZHOU Li-juan2,SONG Ya-na2(1.Department of Pharmacy,Fujian University of Traditional Chinese Medicine,Fuzhou 350003,China;2.Biotechnology Institute,Fujian Academy of Agricultural Sciences,Fuzhou 350003,China)
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- 林珊
- 郑伟文
- 吴锦忠
- 周莉娟
- 宋亚娜
LIN Shan1- 2
- ZHENG Wei-wen2
- WU Jin-zhong1
- ZHOU Li-juan2
- SONG Ya-na2(1.Department of Pharmacy
- Fujian University of Traditional Chinese Medicine
- Fuzhou 350003
- China
- 2.Biotechnology Institute
- Fujian Academy of Agricultural Sciences
- Fuzhou 350003
- China)
- 林珊
- 郑伟文
- 吴锦忠
- 周莉娟
- 宋亚娜
LIN Shan1- 2
- ZHENG Wei-wen2
- WU Jin-zhong1
- ZHOU Li-juan2
- SONG Ya-na2(1.Department of Pharmacy
- Fujian University of Traditional Chinese Medicine
- Fuzhou 350003
- China
- 2.Biotechnology Institute
- Fujian Academy of Agricultural Sciences
- Fuzhou 350003
- China)