江树勋1 邵碧英1 陈文炳1 * 李寿崧1 王泽生2 朱晓南1 廖剑华2
（1 福建出入境检验检疫局技术中心，福州 350001
2 福建省轻工业研究所双孢蘑菇菌种研究推广站,福州 350005）
摘要：本文研究了15种主要食（药）用菌共16个样品的3种总DNA提取方法之比较，它们分别是CTAB法、食品DNA提取试剂盒法和QIAGEN试剂盒法。提取结果分别经过核酸蛋白分析仪、电泳以及RAPD检测。紫外检测结果表明3种方法提取到的总DNA质量均不理想，电泳结果表明3种方法均能提取出小平菇、红菇、香菇、双孢蘑菇、竹荪、黑木耳、姬松茸等7种食（药）用菌总DNA，RAPD检测结果表明CTAB法适用于所有15种食（药）用菌提取用于PCR的总DNA。根据以上结果，我们认为可以选用CTAB法作为在检测这些食（药）用菌的转基因成分时的总DNA提取方法。
Abstract：Edulis and medicinal fungus was a kind of traditional healthy food of China, not only Chinese people liked it, but also many people in other country liked it too. With its output and export increasing in China, we found more and more countries asked for the detection of genetically modified ingredients in fungus, this improved the study of transgenic material detection technique in fungus. The first problem to be resolved was the DNA extraction method and how to analyze the quality of the extracted DNA. A high quality extracted genomic DNA was the basis of the following exact result of transgenic material detection. So this article set out to study the DNA extraction method appropriate for most of the common edulis and medicinal fungus by analyzing the quality of the extracted DNA.
Following 15 kinds of the common edulis and medicinal fungi(16 samples) were studied here , Pleurotu comucopiae、Volvariella volvacea、Pleurotus sajur-caju、Agrocybe cylindraca maire、Ganoderma lucidum、Russula vinosa Lindbe、Pleurotus eryngii、Lentinus edodes、Agaricus bisporus、Dictyophora indusiata、spore of Ganoderma lucidum、Tremella fuciformis Berk、Auricularia auricula、Flammulina velutipes、Pleurotus geesteyanus.Singer、Agaricus blazie. Three common used DNA extraction method were selected , including CTAB method, Food DNA Extraction Kit and Qiagen DNA Extraction Kit ,of which the CTAB method refered to the CTAB method for plant introduced in <Short Protocols in Molecular Biology> except a few improvings , Food DNA Extraction Kit and Qiagen DNA Extraction Kit refered to their method provided with the kit . All extracted DNA were detected by UV spectrophotometer, agarose electrophoresis and RAPD(Random Amplified Polymorphic DNA) respectively .UV spectrophotometer detection results showed the quality of all extracted genomic DNA was poor as expected , Agarose electrophoresis results showed that all three methods could extract the DNA from below 7 fungi, Pleurotu comucopiae, Russula vinosa Lindbe, Lentinus edodes, Agaricus bisporus, Dictyophora indusiata, Auricularia auricula and Agaricus blazie, while CTAB method could extract the DNA of P.geesteyanus.Singer, Food DNA Extraction Kit could extract the DNA of V. volvacea and Qiagen DNA Extraction Kit could extract the DNA of T.fuciformis Berk additionally. RAPD results showed that CTAB method could be used to extract all 15 fungi’ s DNA for PCR, while other two methods could be used to extract most of the 15 fungi’ s DNA for PCR. According to above results, we suggested to select the CTAB method for DNA extraction when testing the transgenic ingredients for these edulis and medicinal fungi.All experiment results were confimed by another researcher in our programme.
