太岁DNA提取方法差异及对测知细菌结构的影响

Differences in Taisui DNA extraction methods and the effects on the result of bacterial structure

以太岁取样的加水热溶解为特点设计4种提取方法,将提取效果好的2个DNA样品用Illumina MiSeq高通量测序平台测序。结果表明:滤膜法(SI3)和研磨法均可获得显示电泳条带的DNA,但前者能将太岁材料彻底破碎和滤除多糖;滤膜法样品测得优质序列46676条,聚类OTU 1095个,比研磨法多154个,只包含1个序列相对丰度大于1%的共有OTU,特有1个序列相对丰度41.14%的OTU,是研磨法的237倍,但滤膜法样品香农指数反比研磨法低1.37%,并导致两者菌群结构在各分类水平上有着明显差别。太岁DNA提取方法深刻影响细菌结构测定结果,滤膜法DNA菌群信息更真实,据其测知的优势结构为假单胞菌属(Pseudomonas)和不动杆菌属(Acinetobacter)。

The aim of this study was to compare the differences of Taisui bacterial DNA extracted by different methods and the resulting changes in bacterial community structure.Four DNA extraction methods were based on grinding and thermal dissolution.Meanwhile,2 DNA samples of the obvious target band were selected for Illumina MiSeq high-throughput sequencing.Both the liquid nitrogen grinding kit method(Grinding method)and the hot-dissolved membrane Filtering CTAB method(Filtering method,SI3)could extract DNA with the target band,however,the SI3 can completely break Taisui tissue and remove polysaccharide.Sample SI3 obtained a total of 46676 valid reads and 1095 OTUs,154 more than that of Sample 9.In addition,there was only one OTU in both samples for which the abundance was greater than 1%.There was an OTU in S13,the abundance was 41.14%,237 times of that in 9.The shannon index of S13 was 1.37%less than 9.The result of bacterial structures of the two samples was different at each classification level.The difference in Taisui DNA extraction methods had a profound influence on the bacterial structure.The filtering method was better than the grinding method.The bacterial information of the former was more comprehensive.The dominant bacteria were Pseudomonas and Acinetobacter.