基于ITS条形码序列枸杞属植物鉴定.docVIP

基于ITS条形码序列枸杞属植物鉴定 　　摘要：采用改进CTAB法提取枸杞（Lycium Chinense Mill.）叶片DNA，利用合成的特异引物对其DNA中nrDNA ITS区进行扩增，利用ITS条形码序列，对枸杞属种质资源进行鉴定，分析其亲缘关系。结果表明，测序得到了17份枸杞属近缘种的ITS条形码序列，整个ITS序列长度变异范围为603～632 bp，平均为624 bp，整个转录间隔区（ITS1+ITS2）对位排列后总长度为480 bp，有194个变异位点，占40%；保守位点288个，占60%。聚类分析结果表明，17份种质资源可分为5个大类群。基于ITS条形码序列分析在鉴定枸杞属种质遗传多样性及其亲缘关系具有一定的优越性。 　　关键词：枸杞属（Lycium L.）；ITS序列；DNA测序；鉴定 　　中图分类号：S567.1+9 文献标识码：A 文章编号：0439-8114（2016）22-5966-03 　　DOI：10.14088/j.cnki.issn0439-8114.2016.22.059 　　Identification of Lycium L. Germplasm Resources Based on nrDNA ITS Sequence 　　SHI Zhi-gang，WAN Ru，LI Yan-long，WANG Ya-jun 　　（Ningxia Academy of Agriculture and Forestry Sciences， Yinchuan 750002，China） 　　Abstract：Adopting improved CTAB to acquire DNA of leaf of Lycium Chinense Mill.，and making use of composite special primers to propagate and clone nrDNA ITS area of DNA，and then sequencing analysis of the target segment. To identify genetic diversity of Lycium L. germplasm resources by the nrDNA ITS sequence. The results showed that the nrDNA ITS sequence of 17 Lycium L. was gained for the first time；the variation length of the whole ITS sequence was 603 to 632 bp，and the average was 624 bp； the total length of whole transcribe partition area（ITS1 and ITS2） sequence alignmentwas 480 bp，and there were 194 variable sites accounting for 40.4%， and 286 conserved sitesaccounting for 59.6%. Based on the nrDNA ITS sequence analysis of 17 Lycium L. germplasm resources，it was obvious that they could be divided into 5 groups. The nrDNA ITS sequence analysis is a advantageous way to study genetic diversity among Lycium L. germplasm resources. 　　Key words：Lycium L.； Internal transcribed spacer； measure the sequence of DNA； identification 　　枸杞（Lycium Chinense Mill.）?儆谇芽疲?Solanaceae）族（Solaneae Reichb.）枸杞属（Lycium L.），枸杞属植物在全球的分布约有80种，多分布于南、北美洲，以美国亚利桑那州和阿根廷形成两个分布中心；欧亚大陆约10余种。中国多数种类分布于西北与华北，中国分布的枸杞属有7种3变种[1]，宁夏农林科学院建成世界惟一的枸杞种质资源圃，保存了国内外分布的7种3变种2 000余份、近2万余株枸杞种质资源。收集保存的枸杞属种质资源种类繁多，传统的单纯依靠形态学的方法难以做出精确鉴别。由于ITS序列进化快，能更精确地对