关键词: 食(药)用菌, 基因组DNA提取

Abstract: Edulis and medicinal fungus were sort of traditional healthy foods of China. Not only Chinese people liked them,but also many people in other countries liked them too. With the increasing of its output and export China, we found more andmore countries asked for the detection of genetically modified ingredients in the fungus. This required the study of transgenicmaterial detection techniques in fungus. The first problem to be resolved was the genomic DNA extraction method and the waysto analyze the quality of the extracted DNA. A high quality extracted genomic DNA was the basis of the following exact resultsof transgenic material detection. So this article set out a study on the genomic DNA extraction method appropriate for most ofthe common edulis and medicinal fungus by analyzing the quality of the extracted genomic DNA.Following 15 species of the common edulis and medicinal fungi(16 samples) were studied here, namely: Pleurotucomucopiae、Volvariella volvacea、Pleurotus sajur-caju、Agrocybe cylindraca maire、Ganoderma lucidum、Russulavinosa Lindbe、Pleurotus eryngii、Lentinus edodes、Agaricus bisporus、Dictyophora indusiata、spore of Ganodermalucidum、Tremella fuciformis Berk、Auricularia auricula、Flammulina velutipes、Pleurotus geesteyanus. Singer andAgaricus blazie. Three commonly used DNA extraction methods were selected, including CTAB method, Food DNA ExtractionKit and Qiagen DNA Extraction Kit. Of them the CTAB method refered for plant introduced in <Short Protocols in MolecularBiology> with a few improvings was adopted. The Food DNA Extraction Kit and Qiagen DNA Extraction Kit refered to theirmethod were provided with the kit. All extracted DNAs were assayed by UV spectrophotometer, agarose electrophoresis andRAPD(Random Amplified Polymorphic DNA) respectively. UV spectrophotometer detection results showed the quality of allextracted genomic DNA was poor as expected. Agarose electrophoresis results showed that all three methods could extract theDNA from the following 7 fungi, Pleurotu comucopiae, Russula vinosa Lindbe, Lentinus edodes, Agaricus bisporus, Dictyophoraindusiata, Auricularia auricula and Agaricus blazie, while CTAB method could extract the DNA of P.geesteyanus.Singer. TheFood DNA Extraction Kit could extract the DNA of V. volvacea and Qiagen DNA Extraction Kit could extract the DNA of T.fuciformis Berk additionally. The RAPD results showed that CTAB method could be used to extract all 15 fungi’ s DNA for PCR,while the other two methods could be used to extract most of the 15 fungi’ s DNA for PCR. According to the above results, wesuggested the choice of the CTAB method for DNA extraction when testing the transgenic ingredients for these edulis and medicinalfungi.All experiment results were confimed by another researcher in our programme.Due to all the values of UV spectrophotometer detection in this study were always abnormal, a comparison with the sameCTAB method on E. coli was made. The results showed that the values of DNA from E. coli were all good. While agaroseelectrophoresis could not be applied to all of the fungi, we had to select RAPD to analyze the DNA for PCR. How to improvethe extraction method to gain high quality DNA from edulis and medicinal fungus if there would be any better method speciallyfor analyzing them would be still needed for further studies.

Key words: Edulis and medicinal fungus, genomic DNA