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响应珊瑚损伤,AOS-LOXa表达上调且类花生酸合成增加。

Up-regulated expression of AOS-LOXa and increased eicosanoid synthesis in response to coral wounding.

作者信息

Lõhelaid Helike, Teder Tarvi, Tõldsepp Kadri, Ekins Merrick, Samel Nigulas

机构信息

Department of Chemistry, Tallinn University of Technology, Tallinn, Estonia.

Sessile Marine Invertebrates, Queensland Museum, Brisbane, Queensland, Australia.

出版信息

PLoS One. 2014 Feb 14;9(2):e89215. doi: 10.1371/journal.pone.0089215. eCollection 2014.

Abstract

In octocorals, a catalase-like allene oxide synthase (AOS) and an 8R-lipoxygenase (LOX) gene are fused together encoding for a single AOS-LOX fusion protein. Although the AOS-LOX pathway is central to the arachidonate metabolism in corals, its biological function in coral homeostasis is unclear. Using an acute incision wound model in the soft coral Capnella imbricata, we here test whether LOX pathway, similar to its role in plants, can contribute to the coral damage response and regeneration. Analysis of metabolites formed from exogenous arachidonate before and after fixed time intervals following wounding indicated a significant increase in AOS-LOX activity in response to mechanical injury. Two AOS-LOX isoforms, AOS-LOXa and AOS-LOXb, were cloned and expressed in bacterial expression system as active fusion proteins. Transcription levels of corresponding genes were measured in normal and stressed coral by qPCR. After wounding, AOS-LOXa was markedly up-regulated in both, the tissue adjacent to the incision and distal parts of a coral colony (with the maximum reached at 1 h and 6 h post wounding, respectively), while AOS-LOXb was stable. According to mRNA expression analysis, combined with detection of eicosanoid product formation for the first time, the AOS-LOX was identified as an early stress response gene which is induced by mechanical injury in coral.

摘要

在八放珊瑚中,一种过氧化氢酶样丙二烯氧化物合酶(AOS)和一个8R-脂氧合酶(LOX)基因融合在一起,编码一种单一的AOS-LOX融合蛋白。尽管AOS-LOX途径是珊瑚中花生四烯酸代谢的核心,但它在珊瑚体内平衡中的生物学功能尚不清楚。我们利用软珊瑚交织拟杯珊瑚的急性切口伤口模型,来测试LOX途径是否与其在植物中的作用类似,有助于珊瑚的损伤反应和再生。对受伤后固定时间间隔前后由外源性花生四烯酸形成的代谢物进行分析,结果表明机械损伤会使AOS-LOX活性显著增加。克隆了两种AOS-LOX同工型,即AOS-LOXa和AOS-LOXb,并在细菌表达系统中作为活性融合蛋白进行表达。通过qPCR测量正常和应激珊瑚中相应基因的转录水平。受伤后,AOS-LOXa在切口附近的组织和珊瑚群体的远端部分均显著上调(分别在受伤后1小时和6小时达到最大值),而AOS-LOXb则保持稳定。根据mRNA表达分析,并首次结合类二十烷酸产物形成的检测,AOS-LOX被鉴定为一种早期应激反应基因,它在珊瑚中由机械损伤诱导产生。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/abcd/3925239/b11aee0a17fc/pone.0089215.g001.jpg

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