Gilbert Jack A, Thomas Simon, Cooley Natalie A, Kulakova Anna, Field Dawn, Booth Tim, McGrath John W, Quinn John P, Joint Ian
Plymouth Marine Laboratory, Prospect Place, The Hoe, Plymouth PL1 3DH, UK.
Environ Microbiol. 2009 Jan;11(1):111-25. doi: 10.1111/j.1462-2920.2008.01745.x. Epub 2008 Sep 8.
Phosphonates are organic compounds that contain a C-P bond and are a poorly characterized component of the marine phosphorus cycle. They may represent a potential source of bioavailable phosphorus, particularly in oligotrophic conditions. This study has investigated the distribution of the phnA gene which encodes phosphonoacetate hydrolase, the enzyme that mineralizes phosphonoacetate. Using newly designed degenerate primers targeting the phnA gene we analysed the potential for phosphonoacetate utilization in DNA and cDNA libraries constructed from a phytoplankton bloom in the Western English Channel during July 2006. Total RNA was isolated and reverse transcribed and phosphonoacetate hydrolase (phnA) transcripts were PCR amplified from the cDNA with the degenerate primers, cloned and sequenced. Phylogenetic analysis demonstrated considerable diversity with 14 sequence types yielding five unique phnA protein groups. We also identified 28 phnA homologues in a 454-pyrosequencing metagenomic and metatranscriptomic study from a coastal marine mesocosm, indicating that > 3% of marine bacteria in this study contained phnA. phnA homologues were also present in a metagenomic fosmid library from this experiment. Finally, cultures of four isolates of potential coral pathogens belonging to the Vibrionaceae contained the phnA gene. In the laboratory, these isolates were able to grow with phosphonoacetate as sole P and C source. The fact that the capacity to utilize phosphonoacetate was evident in each of the three coastal environments suggests the potential for widespread utilization of this bioavailable P source.
膦酸盐是含有碳 - 磷键的有机化合物,是海洋磷循环中特征描述较少的一个组成部分。它们可能是生物可利用磷的潜在来源,特别是在贫营养条件下。本研究调查了编码膦酰乙酸水解酶(该酶可使膦酰乙酸矿化)的phnA基因的分布情况。我们使用新设计的针对phnA基因的简并引物,分析了2006年7月在英吉利海峡西部浮游植物大量繁殖期间构建的DNA和cDNA文库中利用膦酰乙酸的潜力。分离出总RNA并进行逆转录,然后用简并引物从cDNA中PCR扩增膦酰乙酸水解酶(phnA)转录本,进行克隆和测序。系统发育分析表明存在相当大的多样性,14种序列类型产生了5个独特的phnA蛋白组。我们还在一项来自沿海海洋中宇宙的454焦磷酸测序宏基因组和宏转录组研究中鉴定出28个phnA同源物,表明该研究中超过3%的海洋细菌含有phnA。phnA同源物也存在于该实验的宏基因组fosmid文库中。最后,属于弧菌科的四种潜在珊瑚病原体分离株的培养物中含有phnA基因。在实验室中,这些分离株能够以膦酰乙酸作为唯一的磷和碳源生长。在这三个沿海环境中的每一个中都明显存在利用膦酰乙酸的能力,这一事实表明这种生物可利用磷源具有广泛利用的潜力。