Identification,Phylogenetic Relationship Analysis of Lycium Based on rbcL-a and ITS Sequence and the Discovery of ITS Pseudogene

doi:10.13560/j.cnki.biotech.bull.1985.2017.05.018

Abstract

Abstract: Identification and phylogenetic relationship analysis of 14 species or cultivars of Lycium and 1 adulterant of Nitraria tangutorum were studied using rbcL-a and ITS as molecular markers. Total DNA was extracted by CTAB method. With universal primers,rbcL-a and ITS were amplified,cloned,sequenced and analyzed via PCR. T cloning technique was used to deal with the unsuccessful sequencing of ITS sequence of some samples. rbcL-a and ITS sequences with universal primers were amplified successfully in all samples. The lengths of rbcL-a in Lycium and N. tangutorum were 643 bp,and the sequences of rbcL-a were the same in all samples of Lycium,but existed 45 bp variation sites（7%）compared with N. tangutorum. The lengths of ITS in 14 species or cultivars of Lycium ranged from 513 - 692 bp. The UPGMA phylogenetic tree was constructed based on ITS sequence to distinguish samples of Lycium and N. tangutorum well. ITS sequences of L. ruthenicum and L. barbarum were clustered into two groups,respectively. L. yunnanense,L. chinense and L. barbarum var.auranticarpum were classed as the same branch of L. barbarum. The marker of rbcL-a can effectively identify Lycium from its adulterant. The molecular marker of ITS is applicable in variety identification and genetic relationship analysis of Lycium. This study revealed the polymorphism of ITS in L. barbarum L. varieties and individuals for the first time,and the presence of ITS pseudogenes in L. barbarum L.,which offers some new views for systematic evolution research of Lycium using ITS sequence.

Key words: Lycium, identification, rbcL-a, ITS, pseudogene

CHEN Jin-jin, ZHAO Ming-xia, JIANG Shu, CAO Li-li, ZHAO Qing-sheng, ZHAO Bing, WANG Xiao-dong. Identification,Phylogenetic Relationship Analysis of Lycium Based on rbcL-a and ITS Sequence and the Discovery of ITS Pseudogene[J]. Biotechnology Bulletin, 2017, 33(5): 123-130.

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