摘要
为揭示高温对鹿角杯形珊瑚(Pocillopora damicornis)氨同化作用的影响,阐明造礁石珊瑚的热适应机制,本研究鉴定并克隆了1个鹿角杯形珊瑚氨转运体基因PdRhp-1,其开放阅读框(ORF)全长1 410 bp,编码一条由469 aa组成的多肽链。序列分析结果表明,PdRhp-1编码一种疏水性跨膜蛋白,含有12个跨膜结构域,属于Rhesus型氨转运体,其氨基酸序列与人(Homo sapiens)的氨转运体RhCG的一致性为44.14%。为解析PdRhp-1的生物学功能,将其重组表达载体转染至HEK293T细胞,并在培养基中添加氯化铵,发现表达PdRhp-1的细胞总氨摄取率显著高于对照组,说明PdRhp-1具有氨转运功能。同时,分析了高温处理鹿角杯形珊瑚的转录组数据,发现高温显著抑制了PdRhp-1及部分氨同化相关基因的表达。以上结果确认,鹿角杯形珊瑚PdRhp-1编码一个Rhesus型氨转运体,高温可能通过抑制该蛋白介导的氨转运过程影响珊瑚与虫黄藻的共生稳态。
To reveal the effect of high temperature on the ammonia assimilation of Pocillopora damicornis and elucidate the thermal adaptation mechanism of scleractinian coral,we identified and cloned an ammonia transporter gene PdRhp-1 from P.damicornis.The open reading frame(ORF)of PdRhp-1 is 1410 bp,encoding a polypeptide chain composed of 469 amino acid residues.Sequence analysis showed that PdRhp-1 encodes a hydrophobic transmembrane protein with 12 transmembrane domains and belongs to the Rhesus-type ammonia transporter.Its amino acid sequence shares 44.14%identity with Homo sapiens ammonia transporter RhCG.In order to analyze the biological function of PdRhp-1,its recombinant expression vector was transfected into HEK293T cells,and ammonium chloride was added to the culture medium.It was found that the total ammonia uptake rate in the cells expressing PdRhp-1 was significantly higher than that in the control group,indicating that PdRhp-1 has ammonia transport function.At the same time,the transcriptome data of high temperature treated P.damicornis were analyzed,and it was found that high temperature significantly inhibited the expression of PdRhp-1 and some genes related to ammonia assimilation.The above results demonstrate that PdRhp-1 encodes a Rhesus-type ammonia transporter,and high temperature may affect the symbiotic homeostasis of corals and zooxanthellae by inhibiting the ammonia transport process mediated by this protein.
作者
曾赛男
秦郅暄
王艺
刘兆群
周智
Zeng Sainan;Qin Zhixuan;Wang Yi;Liu Zhaoqun;Zhou Zhi(School of Marine Biology and Fisheries,Hainan University,Haikou 570228,China;School of Marine Science and Engineering,Hainan University,Haikou 570228,China)
出处
《海洋学报》
北大核心
2025年第11期84-94,共11页
基金
海南省重点研发计划项目(ZDYF2022SHFZ324)。
关键词
鹿角杯形珊瑚
氨转运体
氨同化作用
高温
Pocillopora damicornis
ammonia transporter
ammonia assimilation
high temperature
