State Key Laboratory of Rice Biology and Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Ministry of Agriculture, Institute of Biotechnology, Zhejiang University, Hangzhou, 310029, China.
Curr Microbiol. 2011 Feb;62(2):650-8. doi: 10.1007/s00284-010-9758-3. Epub 2010 Sep 30.
The purpose of this study was to determine the existence of Burkholderia cepacia complex (Bcc) at species level and the predominant species in the environment of moso bamboo plantations in Hangzhou, China. A total of 423 isolates were recovered from moso bamboo rhizhosphere soil samples of three sites on the selective medium during 2007-2008. Isolates were identified by Bcc-specific PCR assays, followed by recA-restriction fragment length polymorphism assays, species-specific PCR analysis, recA gene sequencing, multilocus sequence typing (MLST) scheme, and BOX-PCR fingerprinting for genomic diversity. Out of 423 isolates, 278 isolates were assigned to the following Bcc species, eight B. stabilis, 26 B. anthina, 193 B. pyrrocinia, and 51 B. arboris, which indicated B. pyrrocinia as the most dominant species followed by B. arboris. Moreover, false positives were observed in certain isolates of B. arboris while performing species-specific PCR test. Furthermore, the results of recA gene sequence similarity and MLST data demonstrated that nine isolates formed a single discrete cluster but were PCR negative to species-specific primers representing novel species may exist within the Bcc. In addition, BOX-PCR fingerprinting for all the Bcc isolates also showed the strain diversity. It is the first report of the existence of B. arboris and predominance of B. pyrrocinia in the moso bamboo environment.
本研究旨在确定中国杭州毛竹种植环境中伯克霍尔德氏菌复合群(Bcc)在种水平上的存在情况和主要物种。2007 年至 2008 年期间,在选择性培养基上从三个地点的毛竹根际土壤样本中总共回收了 423 株分离物。通过 Bcc 特异性 PCR 检测、recA 限制酶切片段长度多态性检测、种特异性 PCR 分析、recA 基因测序、多位点序列分型(MLST)方案和基因组多样性 BOX-PCR 指纹图谱对分离物进行鉴定。在 423 株分离物中,278 株被分配到以下 Bcc 物种,8 株 B. stabilis、26 株 B. anthina、193 株 B. pyrrocinia 和 51 株 B. arboris,表明 B. pyrrocinia 是最主要的物种,其次是 B. arboris。此外,在进行种特异性 PCR 检测时,某些 B. arboris 分离物出现了假阳性。此外,recA 基因序列相似性和 MLST 数据的结果表明,有九个分离物形成了一个单一的离散聚类,但对代表新种的种特异性引物呈 PCR 阴性,可能存在 Bcc 内的新种。此外,所有 Bcc 分离物的 BOX-PCR 指纹图谱也显示了菌株多样性。这是首次报道 B. arboris 的存在和 B. pyrrocinia 在毛竹环境中的优势。