摘要： 本研究测得油桐叶片和种仁总糖含量分别为75.20,32.80 mg/g,总酚含量分别为15.75,0.57 mg/g,在此基础上对TRIzol法、CTAB-LiCl法进行改良,分别提取出高质量的种仁、叶片总RNA,并与异硫氰酸胍法及NOGEN试剂盒法进行比较,结果表明:改良TRIzol法提取的油桐种仁总RNA OD260/OD280稳定在1.9～2.0,28S rRNA和18S rRNA条带清晰完整,RNA产率较高,改良CTAB-LiCl法得到的种仁总RNA有部分降解,异硫氰酸胍法则对小分子盐类去除不够充分,而试剂盒提取出的油桐种仁总RNA有少许降解且有DNA杂质污染。上述4种方法中只有改良CTAB-LiCl法能成功提取出质量较高的油桐叶片总RNA。因此,改良TRIzol法和改良CTAB-LiCl法可分别作为提取油桐种仁和叶片总RNA的首选方法。

关键词: 油桐, 总RNA提取, 改良TRIzol法, 改良CTAB-LiCl法, 异硫氰酸胍法

Abstract: The content of total sugar and total phenols were determined to be 75.20 and 32.80 mg/g,15.75 and 0.57 mg/g respectively in leaves and kernels of tung tree.On that basis,high-quality total RNA of leaves and kernels were respectively extracted by modified TRIzol method and modified CTAB-LiCl method,and the effects of the above methods,Guanidinium idothiocyanate method and NOGEN RNAprep pure kit method were compared.The results showed that not only the OD260/OD280 value of 1.9-2.0 of the kernel of RNA isolated by modified TRIzol method stabilized,but also the bands of 28S rRNA and 18S rRNA were clear and complete,and higher yields was even more important.The total RNA of kernels isolated by modified CTAB-LiCl method were degradation in some degrees,and small molecule in the RNA extracted by Guanidinium idothiocyanate method could not be removed fully.Moreover the total RNA of kernels with DNA and degradation in some degrees could be obtained via NOGEN RNAprep pure kit method.In above four methods,only modified CTAB-LiCl method could extract the higher quality total RNA of leaves of tung tree.For these reasons,modified TRIzol method and modified CTAB-LiCl method could be regarded as preferred method for the RNA isolated respectively from leaves and kernels of tung tree.

Key words: Tung tree, Total RNA isolation Modified, TRIzol method Modified, CTAB-LiCl method, Guanidinium idothiocyanate method

中图分类号: 

• Q75