Hao Juan, Jiao Kaili, Yu Chenliang, Guo Hong, Zhu Yujia, Yang Xiao, Zhang Siyang, Zhang Lei, Feng Shangguo, Song Yaobin, Dong Ming, Wang Huizhong, Shen Chenjia
College of Life and Environmental Sciences, Hangzhou Normal University, Hangzhou, 310036, China.
Zhejiang Provincial Key Laboratory for Genetic Improvement and Quality Control of Medicinal Plants, Hangzhou Normal University, Hangzhou, 310036, China.
Biochem Genet. 2018 Jun;56(3):255-266. doi: 10.1007/s10528-018-9842-0. Epub 2018 Jan 31.
Taxus media is an important species in the family Taxaceae with high medicinal and commercial value. Overexploitation and illegal trade have led T. media to a severe threat of extinction. In addition, T. media and other Taxus species have similar morphological traits and are easily misidentified, particularly during the seedling stage. The purpose of this study is to develop a species-specific marker for T. media. Through a screening of 36 start codon targeted (SCoT) polymorphism primers, among 15 individuals of 4 Taxus species (T. media, T. chinensis, T. cuspidate and T. fuana), a clear species-specific DNA fragment (amplified by primer SCoT3) for T. media was identified. After isolation and sequencing, a DNA sequence with 530 bp was obtained. Based on this DNA fragment, a primer pair for the sequence-characterized amplified region marker was designed and named MHSF/MHSR. PCR analysis with primer pair MHSF/MHSR revealed a clear amplified band for all individuals of T. media but not for T. chinensis, T. cuspidate and T. fuana. Therefore, this marker can be used as a quick, efficient and reliable tool to identify T. media among other related Taxus species. The results of this study will lay an important foundation for the protection and management of T. media as a natural resource.
曼地亚红豆杉是红豆杉科的一个重要物种,具有很高的药用和商业价值。过度开发和非法贸易已使曼地亚红豆杉面临严重的灭绝威胁。此外,曼地亚红豆杉与其他红豆杉物种具有相似的形态特征,容易被误认,尤其是在幼苗期。本研究的目的是开发一种曼地亚红豆杉的物种特异性标记。通过对36个起始密码子靶向(SCoT)多态性引物进行筛选,在4种红豆杉(曼地亚红豆杉、中国红豆杉、东北红豆杉和南方红豆杉)的15个个体中,鉴定出了一条清晰的曼地亚红豆杉物种特异性DNA片段(由引物SCoT3扩增)。经过分离和测序,获得了一条530 bp的DNA序列。基于该DNA片段,设计了一对用于序列特征扩增区域标记的引物,并命名为MHSF/MHSR。用引物对MHSF/MHSR进行PCR分析,结果显示曼地亚红豆杉的所有个体均出现一条清晰的扩增带,而中国红豆杉、东北红豆杉和南方红豆杉则没有。因此,该标记可作为一种快速、高效、可靠的工具,用于在其他相关红豆杉物种中鉴定曼地亚红豆杉。本研究结果将为曼地亚红豆杉作为一种自然资源的保护和管理奠定重要基础。