三种一般钩藤ITS序列的RFLP分析

目的以核糖体转录间隔区(rDNA ITS)序列为分子标记,对采集自3个不同地区的一般钩藤Uncaria rhynchophylla(Miq.)Jack.进行RFLP遗传分析,研究不同地理区域一般钩藤的rDNA ITS序列是否具有生物地理差异。方法通过改良CTAB法提取一般钩藤总DNA,利用通用引物对其rDNA ITS序列进行PCR扩增,并对连接转化后扩增得到的rDNAITS序列进行限制性片段长度多态性(Restriction Fragment Length Polymorphism,RFLP)分析。结果获得采集自广东韶关阳山、广东梅州大埔、广西植物园的3种一般钩藤两种限制性内切酶(Msp I&HaeⅢ)酶切图谱。结论通过基于rDNAITS序列进行的RFLP分析,表明这3个地区的一般钩藤rDNA ITS序列无明显差异,不同区域的一般钩藤其遗传本质不具有地理差异。

ITS Sequence Analysis of Uncaria rhynchophylla (Miq.) Jack.from Three Different Regions

Objective In order to illustrate the geographical differences of Uncaria rhynchophylla(Miq.) Jack.,genetic analysis was carried out for three different regions of Uncaria rhynchophylla(Miq.) Jack.by using the rDNA ITS sequence as molecular marker. Methods Total DNA was extracted with modified CTAB method and thereby rDNA ITS regions were amplified with universally conserved primer.The rDNA ITS amplicon was characterized by RFLP(Restriction Fragment Length Polymorphism,RFLP)analyses. Results The Msp I-and Hae III-based RFLP patterns of rDNA ITS from three kinds of Uncaria rhynchophylla(Miq.) Jack.were obtained,which collected from Yangshan Shaoguan,Guangdong and Dabu Meizhou,Guangdong and Guangxi Botanical Garden. Conclusion Based on molecular biological methods of rDNA ITS region RFLP analysis,Uncaria rhynchophylla(Miq.) Jack from the three regions have no significant geographical differences.