[Fingerprints identification of Gynostemma pentaphyllum by RAPD and cloning and analysis of its specific DNA fragment].
OBJECTIVE
To identify the resources of Gynostemma pentaphyllum and its spurious breed plant Cayratia japonica at level of DNA.


METHODS
Two random primers ( WGS001, WGS004) screened were applied to do random amplification with genomic DNA extracted from Gynostemma pentaphyllum and Cayratia japonica which were collected from different habitats. After amplificated with WGS004, one characteristic fragment about 500 bp which was common to all Gynostemma pentaphyllum samples studied but not to Cayratia japonica was cloned and sequenced. Then these sequences obtained were analyzed for identity and compared by Blastn program in GenBank.


RESULTS
There were obvious different bands amplified by above two primers in their fingerprints of genomic DNA. On the basis of these different bands of DNA fingerprints, they could distinguish Gynostemma pentaphyllum and Cayratia japonica obviously. Sequence alignment of seven cloned bands showed that their identities ranged from 45.7% - 94.5%. There was no similar genome sequences searched in GenBank. This indicated that these seven DNA fragments had not been reported before and they should be new sequences.


CONCLUSIONS
RAPD technique can be used for the accurate identification of Gynostemma pentaphyllum and its counterfeit goods Cayratia japonica. Besides, these specific DNA sequences for Gynostemmna pentaphyllum in this study are useful for the further research on identification of species and assisted selection breeding in Gynostemma pentaphyllum.
Jiang
Jun-fu
JF
Department of Biochemistry and Molecular Biology, Guangxi Medical University, Nanning 530021, China. jiangjunfu2002@163.com
Li
Xiong-ying
XY
Wu
Yao-sheng
YS
Luo
Yu
Y
Zhao
Rui-qiang
RQ
Lan
Xiu-wan
XW
D003001:Cloning, Molecular; D017931:DNA Primers; D018744:DNA, Plant / Q000235:genetics*; D004340:Drug Contamination; D005819:Genetic Markers; D029774:Gynostemma / Q000145:classification / Q000235:genetics*; D010946:Plants, Medicinal / Q000235:genetics*; D019105:Random Amplified Polymorphic DNA Technique; D017422:Sequence Analysis, DNA; D027844:Vitaceae / Q000145:classification / Q000235:genetics*
D017931:DNA Primers; D018744:DNA, Plant; D005819:Genetic Markers