荞麦内转录间隔区(its)的扩增跟序列解析-四川农业大学文档.docVIP

荞麦内转录间隔区（ITS）的扩增及序列分析 生物科学2002级 曾子贤 指导老师 吴 琦 副教授 摘 要：本研究以野生金荞麦、栽培苦荞为材料，采用改进的CTAB法提取荞麦总DNA。设计一对特异性引物，对三份荞麦ITS区序列进行PCR扩增，均获得长约700bp的单一条带。PCR产物测序结果表明，野生金荞麦1号测序片段为699bp，包含ITS区序列为660bp；野生金荞麦2号测序片段为678bp，包含ITS序列为646bp；苦荞测序片段为640bp，包括ITS序列为599bp。将所得到的三份荞麦ITS区序列分别与GenBank中登录的荞麦ITS序列进行比较。结果表明，野生金荞麦1号与已知金荞麦ITS区同源率为90.8%，野生金荞麦2号与已知的金荞麦ITS区序列同源率达到91.6%。供试苦荞与已知苦荞ITS区同源率为99.8%。 关键词：野生金荞麦；苦荞；内转录间隔区（ITS）序列；5.8S rDNA；PCR；序列分析 Amplification and Sequences analysis of Internal Transcribed Spacer of Buckwheat ZENG Zi-xian Biological Science, Grade 2002 Directed by WU Qi (Associate Prof. Ph. D) Abstract：Wildness F. cymosum and cultivated F. tataricum were investigated in this article. By the improving method of CTAB DNA extraction, total DNA from three buckwheat was obtained. A pair of specific primers was designed. The ITS regions of three buckwheat were amplified. The amplified fragments, about 700bp in length, were sequenced directly. The results show that the sequencing fragments are 699bp, 678bp and 640bp in length, including ITS regions of F. cymosum and F. tataricum are 660bp, 646bp and 599bp in length. Comparing F. cymosum and F. tataricum with sequences reported, it suggests that the homology rate of ITS sequence of F. cymosum, NO. 1 is 91.6% and NO. 2 is 90.8% and the homology rate of the ITS sequence of F. tararicum is 99.8%. Keywords: Wildness F. cymosum; F. tataricum; ITS sequences; 5.8S rDNA; Sequence Analysis 荞麦属于蓼科（Polygonaceae）荞麦属（Fagopyrum Mill）植物。荞麦不仅是营养丰富的粮食作物，也是有较好药用价值的药用植物。因此开展荞麦属植物的系统发育和分子进化研究，进一步完善荞麦属植物的分类和新种、野生种的鉴定，对丰富的荞麦遗传资源保护和利用以及荞麦属植物的开发和改良提供系统学资料。 KweonHeo等[1]对荞麦属的野生种进行了分类，将其划分为3大类（表1）。 表1 荞麦属的分类 第1类 F. esculentum F. homotropicum F. esculentum. Ancestralis 第2类 F. cymosum F. tataricum F. tataricum. potanini 第3类 F. callianthum F. capillatum F. gilesii F. gracilipes F. leptopodum F. urophyllum F. lineare F. macrocarpum F .pleioramosum F. rubifolium F. statice 近年来，分子生物学研究技术如RFLP分析、AFLP分析、SSR分析、RAPD分析、DNA序列分析为分子系统学的研究提供了重要的分子标记资料。其中核糖体DNA内转录间隔区（ITS