Division of Bio Science, Dongguk University, Kyungju, Kyongbuk 780-714, Korea.
J Microbiol Biotechnol. 2011 Apr;21(4):333-40.
Herbicide-tolerant Zoysia grass has been previously developed through Agrobacterium-mediated transformation. We investigated the effects of genetically modified (GM) Zoysia grass and the associated herbicide application on bacterial community structure by using culture-independent approaches. To assess the possible horizontal gene transfer (HGT) of transgenic DNA to soil microorganisms, total soil DNAs were amplified by PCR with two primer sets for the bar and hpt genes, which were introduced into the GM Zoysia grass by a callus-type transformation. The transgenic genes were not detected from the total genomic DNAs extracted from 1.5 g of each rhizosphere soils of GM and non-GM Zoysia grasses. The structures and diversities of the bacterial communities in rhizosphere soils of GM and non-GM Zoysia grasses were investigated by constructing 16S rDNA clone libraries. Classifier, provided in the RDP II, assigned 100 clones in the 16S rRNA gene sequences library into 11 bacterial phyla. The most abundant phyla in both clone libraries were Acidobacteria and Proteobacteria. The bacterial diversity of the GM clone library was lower than that of the non- GM library. The former contained four phyla, whereas the latter had seven phyla. Phylogenetic trees were constructed to confirm these results. Phylogenetic analyses of the two clone libraries revealed considerable difference from each other. The significance of difference between clone libraries was examined with LIBSHUFF statistics. LIBSHUFF analysis revealed that the two clone libraries differed significantly (P〈0.025), suggesting alterations in the composition of the microbial community associated with GM Zoysia grass.
耐除草剂百喜草先前已经通过根癌农杆菌介导的转化方法得到了培育。我们采用非培养方法研究了遗传修饰（GM）百喜草和相关除草剂的应用对细菌群落结构的影响。为了评估转基因 DNA 向土壤微生物发生水平基因转移（HGT）的可能性，我们使用两对引物通过 PCR 对总土壤 DNA 进行了扩增，这两对引物分别针对导入 GM 百喜草的 bar 和 hpt 基因。从 GM 和非 GM 百喜草根际土壤中提取的总基因组 DNA 中未检测到转基因基因。通过构建 16S rDNA 克隆文库，研究了 GM 和非 GM 百喜草根际土壤中细菌群落的结构和多样性。在 RDP II 中提供的分类器将 16S rRNA 基因序列文库中的 100 个克隆分配到 11 个细菌门中。这两个克隆文库中最丰富的门都是酸杆菌门和变形菌门。GM 克隆文库的细菌多样性低于非 GM 文库。前者包含四个门，而后者有七个门。构建系统发育树来证实这些结果。两个克隆文库的系统发育分析彼此之间存在很大差异。使用 LIBSHUFF 统计数据检查了克隆文库之间差异的显著性。LIBSHUFF 分析表明，两个克隆文库存在显著差异（P〈0.025），这表明与 GM 百喜草相关的微生物群落组成发生了变化。
