AFLP法分析染料对黄孢原毛平革菌的损伤效应

Evaluation of the Damaging Effect of Dyestuff on Phanerochaete chrysosporium with Amplified Fragment Length Polymorphism (AFLP) Analysis

利用AFLP对DNA碱基序列分辨率高的特性,研究纯培养条件下染料对黄孢原毛平革菌(P.chrysosporium)菌株DNA的损伤效应.在对黄孢原毛平革菌(P.chrysosporium)AFLP最佳引物筛选的基础上,通过分析最佳引物扩增结果中DNA相似性、染料浓度与DNA相似性关系以及构建UPGMA相似性树状图,评价染料对P.chrysosporiumDNA损伤效应.结果表明,AFLP法可较好反映染料对DNA损伤效应,在优选的3种引物组合中,E-AAC/M-CAA引物组合AFLP扩增效果最好,共获得35条DNA扩增条带,其中有12条具有特异性,多态性比率34.3%;在进行DNA相似性分析和UPGMA法聚类分析后发现,AFLP技术较好地表达了DNA相似性与染料浓度的正相关关系,并且显示P.chrysosporium对染料损伤效应敏感,染料浓度不宜高于50 mg/L,当染料浓度达到300 mg/L即可对P.chrysosporium产生较大损伤作用,使之基本丧失原有的处理功能.

The AFLP analysis was carried out in order to evaluate the DNA damaging effects of dyestuff of different concentrations on Pharterochaete chrysosporium. The DNA similarity and UPGMA cluster analysis were conducted to determine the relationships between the concentrations of dyestuff and DNA damaging effects. The results showed that the AFLP analysis could reflect the polymorphisms of DNA well, and the primer pairs E-AAC/M-CAA could give more amplified DNA fragments than others primers. From the DNA maps of best primer pairs, 35 clear and stable DNA bands were observed, including 12 polymorphous bands which were accounted for 34.3 % diversity. It was indicated from the DNA similarity and UPGMA cluster analysis that DNA polymorphisms correlated with dyestuff＇s concentrations well. If the concentration of dyestuff was not higher than 50 mg/L, the damaging effect on DNA was finite. If the concentrations of dyestuff were up to 300 mg/L, the DNA of Phanerochaete chrysosporium would have been injured greatly, and would have been bankrupted in its intrinsic function.