入侵植物一年蓬基因组DNA提取方法优化  

Optimization of DNA extraction methods from invasive plant Erigeron annuus

作　　者：王仕文 杨子琼 魏淑雅 丁彤 顾婉如 陈雨蓓 屈利利 童跃伟 WANG Shiwen;YANG Ziqiong;WEI Shuya;DING Tong;GU Wanru;CHEN Yubei;QU Lili;TONG Yuewei(Research Center of Aquatic Organism Conservation and Water Ecosystem Restoration/School of Life Science,Anqing Normal University,Anqing 246133,China)

机构地区：[1]安庆师范大学生命科学学院/水生生物保护与水生态修复安徽省高等学校工程技术研究中心,安徽安庆246133

出　　处：《安徽农学通报》2024年第13期66-70,共5页Anhui Agricultural Science Bulletin

基　　金：安徽省自然科学基金面上项目“入侵植物一年蓬的谱系分化格局与适应性进化研究”(1908085MC58);安徽省高等学校省级质量工程项目“《植物学》野外实习虚拟仿真实验教学”(2021xnfzxm062);安庆师范大学校级质量工程项目(2022aqnujyxm45)。

摘　　要：一年蓬为菊科飞蓬属植物,是分布较广、危害较重的一种外来入侵杂草。为优选该植物基因组DNA的提取方法,本研究采取四因素三水平正交试验设计,对一年蓬基因组DNA的十六烷基三甲基溴化铵(CTAB)提取方法进行优化,并与常规的植物基因组试剂盒法进行对比。结果表明,一年蓬基因组DNA较优的提取方法为改良CTAB法,即一年蓬的干燥叶片经CTAB-free去除多糖,并加入β-巯基乙醇防止酚氧化,用氯仿∶异戊醇(24∶1)抽提3次。改良CTAB法提取的一年蓬基因组DNA质量较高,微量分光光度计检测的A_(260)/A_(280)数值接近1.8~2.0,且凝胶电泳DNA条带清晰,无明显拖带。该方法为该植物的遗传多样性和入侵机制等研究提供参考。Erigeron annuus,which belongs to the genus Suaeda of Asteraceae family,is one of the most widely distributed and seriously harmful alien invasive weeds.To select a better method for extracting genomic DNA of E.annuus,the CTAB method for extracting genomic DNA of E.annuus was optimized by orthogonal experiment with four factors and three levels,and compared with the common plant kit method.The results showed that the modified CTAB method was used to extract genomic DNA of E.annuus.This involved treating the dried leaves of E.annuus with CTABfree to remove polysaccharides,addingβ-mercaptoethanol to prevent phenol oxidation,and extracting with chloroform:isopentanol(24∶1)three times.The quality of the DNA extracted by the optimized scheme was good,with A_(260)/A_(280) values measured by a microspectrophotometer close to 1.8 to 2.0,and the gel electrophoresis image band was clear without obvious drag.This method laid a foundation for studies on the genetic diversity and invasion mechanisms of E.annuus.

关 键 词：菊科 一年蓬 DNA提取方法 改良CTAB法 正交试验 

分 类 号：Q949.783.5[生物学—植物学] Q75