Scripps Institution of Oceanography, Center for Marine Biotechnology and Biomedicine, University of California, San Diego, Mail Code 0204, La Jolla, CA 92093, USA.
Appl Environ Microbiol. 2011 Sep;77(17):5916-25. doi: 10.1128/AEM.00611-11. Epub 2011 Jul 1.
The molecular fingerprinting technique terminal-restriction fragment length polymorphism (T-RFLP) was used in combination with sequence-based approaches to evaluate the geographic distribution of secondary metabolite biosynthetic genes in strains of the marine actinomycete Salinispora arenicola. This study targeted ketosynthase (KS) domains from type I polyketide synthase (PKS) genes and revealed four distinct clusters, the largest of which was comprised of strains from all six global locations sampled. The remaining strains fell into three smaller clusters comprised of strains derived entirely from the Red Sea, the Sea of Cortez, or around the Island of Guam. These results reveal variation in the secondary metabolite gene collectives maintained by strains that are largely clonal at the 16S rRNA level. The location specificities of the three smaller clusters provide evidence that collections of secondary metabolite genes in subpopulations of S. arenicola are endemic to these locations. Cloned KS sequences support the maintenance of distinct sets of biosynthetic genes in the strains associated with each cluster and include four that had not previously been detected in S. arenicola. Two of these new sequences were observed only in strains derived from Guam or the Sea of Cortez. Transcriptional analysis of one of the new KS sequences in conjunction with the production of the polyketide arenicolide A supports a link between this sequence and the associated biosynthetic pathway. From the perspective of natural product discovery, these results suggest that screening populations from distant locations can enhance the discovery of new natural products and provides further support for the use of molecular fingerprinting techniques, such as T-RFLP, to rapidly identify strains that possess distinct sets of biosynthetic genes.
采用末端限制性片段长度多态性(T-RFLP)分子指纹图谱技术与基于序列的方法相结合,评估了海洋放线菌盐生螺旋菌 Salinispora arenicola 中次生代谢物生物合成基因的地理分布。本研究针对 I 型聚酮合酶(PKS)基因中的酮合酶(KS)结构域,并揭示了四个不同的簇,其中最大的簇由来自全球六个采样地点的所有菌株组成。其余的菌株分为三个较小的簇,这些菌株完全来自红海、科尔特斯海或关岛岛。这些结果表明,在很大程度上在 16S rRNA 水平上呈克隆状态的菌株中,次生代谢物基因集合存在变异。三个较小簇的位置特异性为这些位置的 S. arenicola 亚群中的次生代谢物基因集合的地方性提供了证据。克隆的 KS 序列支持与每个簇相关联的菌株中存在不同的生物合成基因集合,其中包括以前在 S. arenicola 中未检测到的四个基因。这四个新序列中的两个仅在源自关岛或科尔特斯海的菌株中观察到。对与一个新的 KS 序列相关联的一个序列的转录分析以及聚酮 arenicolide A 的产生支持该序列与相关生物合成途径之间的联系。从天然产物发现的角度来看,这些结果表明筛选来自遥远地点的种群可以增强对新天然产物的发现,并进一步支持使用分子指纹图谱技术(如 T-RFLP)快速识别具有独特生物合成基因集合的菌株。