Centre of Marine Microbiology and Genetics, Australian Institute of Marine Science, Townsville, Queensland, Australia.
ISME J. 2011 Mar;5(3):559-64. doi: 10.1038/ismej.2010.143. Epub 2010 Sep 2.
Factors that facilitate the onset of black band disease (BBD) of corals remain elusive, though anoxic conditions under the complex microbial mat and production of sulfide are implicated in necrosis of underlying coral tissues. This study investigated the diversity and quantitative shifts of sulfate-reducing bacterial (SRB) populations during the onset of BBD using real-time PCR (RT-PCR) and cloning approaches targeting the dissimilatory (bi)sulfite reductase (dsrA) gene. A quantitative-PCR (qPCR) assay targeting the 16S rRNA gene also provided an estimate of total bacteria, and allowed the relative percentage of SRB within the lesions to be determined. Three Montipora sp. coral colonies identified with lesions previously termed cyanobacterial patches (CPs) (comprising microbial communities unlike those of BBD lesions), were tagged and followed through time as CP developed into BBD. The dsrA-targeted qPCR detected few copies of the gene in the CP samples (<65 per ng DNA), though copy numbers increased in BBD lesions (>2500 per ng DNA). SRB in CP samples were less than 1% of the bacterial population, though represented up to 7.5% of the BBD population. Clone libraries also demonstrated a shift in the dominant dsrA sequences as lesions shifted from CP into BBD. Results from this study confirm that SRB increase during the onset of BBD, likely increasing sulfide concentrations at the base of the microbial mat and facilitating the pathogenesis of BBD.
尽管缺氧条件下复杂微生物席下产生的硫化物与珊瑚组织坏死有关,但促进珊瑚黑带病(BBD)发生的因素仍难以捉摸。本研究使用实时 PCR(RT-PCR)和针对异化(双)亚硫酸盐还原酶(dsrA)基因的克隆方法,调查了 BBD 发生过程中硫酸盐还原菌(SRB)种群的多样性和数量变化。针对 16S rRNA 基因的定量-PCR(qPCR)检测法还提供了总细菌的估计值,并允许确定病变内 SRB 的相对百分比。三个 Montipora sp.珊瑚群落,先前被标记为蓝藻斑块(CPs)(包含与 BBD 病变不同的微生物群落)的病变,随着 CP 发展为 BBD,进行了标记并随时间进行了跟踪。dsrA 靶向 qPCR 在 CP 样本中检测到该基因的拷贝数很少(<65 个拷贝/ng DNA),尽管在 BBD 病变中拷贝数增加(>2500 个拷贝/ng DNA)。CP 样本中的 SRB 不到细菌总数的 1%,但代表了高达 7.5%的 BBD 种群。克隆文库还表明,随着病变从 CP 转变为 BBD,dsrA 序列的主导序列也发生了转变。本研究的结果证实,SRB 在 BBD 发生过程中增加,可能会增加微生物席底部的硫化物浓度,并促进 BBD 的发病机制。