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KMID : 0043320120350040701
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Archives of Pharmacal Research
2012 Volume.35 No. 4 p.701 ~ p.708
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Authentication of an endangered herb Changium smyrnioides from different producing areas based on rDNA ITS sequences and allele-specific PCR
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Sun Xiaoqin
Wei Yanglian
Qin Minjian
Guo Qiaosheng
Guo Jianlin
Zhou Yifeng
Hang Yueyu
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Abstract
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The rDNA ITS region of 18 samples of Changium smyrnioides from 7 areas and of 2 samples of Chuanminshen violaceum were sequenced and analyzed. The amplified ITS region of the samples, including a partial sequence of ITS1 and complete sequences of 5.8S and ITS2, had a total length of 555 bp. After complete alignment, there were 49 variable sites, of which 45 were informative, when gaps were treated as missing data. Samples of C. smyrnioides from different locations could be identified exactly based on the variable sites. The maximum parsimony (MP) and neighbor joining (NJ) tree constructed from the ITS sequences based on Kumar’s two-parameter model showed that the genetic distances of the C. smyrnioides samples from different locations were not always related to their geographical distances. A specific primer set for Allele-specific PCR authentication of C. violaceum from Jurong of Jiangsu was designed based on the SNP in the ITS sequence alignment. C. violaceum from the major genuine producing area in Jurong of Jiangsu could be identified exactly and quickly by Allele-specific PCR.
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KEYWORD
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Endangered medicinal plant, Changium smyrnioides, ITS, Allele-specific PCR, Genuine producing area
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