引用本文:	刘雪莹,王广策,张宝玉,宫相忠.条斑紫菜丙酮酸羧化酶基因表达对不同碳源的响应[J].海洋科学,2022,46(12):138-147.

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条斑紫菜丙酮酸羧化酶基因表达对不同碳源的响应
刘雪莹^1,2,3,4, 王广策^2,3,4, 张宝玉^2,3,4, 宫相忠¹
1.中国海洋大学海洋生命学院, 山东青岛 266003;2.中国科学院海洋研究所实验海洋生物学重点实验室, 山东青岛 266071;3.青岛海洋科学与技术试点国家实验室海洋生物学与生物技术功能实验室, 山东青岛 266237;4.中国科学院海洋大科学研究中心, 山东青岛 266071

摘要:

丙酮酸羧化酶(pyruvate carboxylase, PYC)在非光合生物中催化丙酮酸羧化生成草酰乙酸(oxaloacetate, OAA), 作为糖异生的第一步, 在动物维持代谢稳态中具有重要作用。研究发现, PYC在光合生物中也发挥重要作用。为了探究PYC在潮间带大型海藻中的作用, 我们从条斑紫菜叶状体中扩增获得PYC基因全长序列(命名为PyPYC), 并分析了其序列特征。通过对系统进化树分析表明PyPYC与来自细菌的PYC具有较近亲缘关系。针对条斑紫菜叶状体生长环境所面临的碳源变化, 设置了不同类型和不同浓度的无机碳培养条件, 采用实时荧光定量(RT-qPCR)检测了该基因对这些无机碳源的响应。结果表明, 高浓度的CO₂能显著上调PyPYC基因的表达, 而高浓度的HCO₃^?对其影响较小。由此, 我们初步认为, 当紫菜叶状体暴露在空气中时, PYC在其无机碳利用中发挥一定作用。

关键词: 丙酮酸羧化酶条斑紫菜无机碳利用

DOI：10.11759/hykx20220422001

分类号:Q71

基金项目:国家自然科学基金(41876163); 中国科学院海洋大科学研究中心重点部署项目(COMS2019Q02); 山东省“泰山学者”工程专项经费资助项目(tspd20210316); 财政部和农业农村部: 国家现代化农业产业技术体系(CARS-50)

Expression of the pyruvate carboxylase gene in Pyropia yezoensis in response to different inorganic carbon sources

LIU Xue-ying^1,2,3,4, ZHANG Bao-yu^2,3,4, WANG Guang-ce^2,3,4, GONG Xiang-zhong¹

1.College of Marine Life Sciences, Ocean University of China, Qingdao 266003, China;2.Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, Qingdao 266071, China;3.Laboratory for Marine Biology and Biotechnology, Pilot National Laboratory for Marine Science and Technology (Qingdao), Qingdao 266237, China;4.Center for Ocean Mega-Science, Chinese Academy of Sciences, Qingdao 266071, China

Abstract:

Pyruvate carboxylase (PYC) catalyzes the carboxylation of pyruvate to oxaloacetate in non-photosynthetic organisms. PYC plays an important role in maintaining metabolic homeostasis in animals as the first step in gluconeogenesis. PYC also functions in photosynthetic organisms, including microalgae and plants, but its function in macroalgae inhabiting the intertidal zone is unknown. In this study, the complete sequence of the PYC gene (namely PyPYC) was obtained from the thallus cDNA of Pyropia yezoensis, and the sequence characteristics were analyzed. The phylogenetic tree based on amino acid sequences indicated that PyPYC was closely related to PYC from prokaryotes. Different types and concentrations of inorganic carbon sources were employed during the cultivation of the thallus based on the changes in carbon sources that wild thallus of P. yezoensis encounter in the natural intertidal zone. The relative expression level of PyPYC in thallus cultivated under these inorganic carbon conditions was evaluated by quantitative real-time polymerase chain reaction. The results demonstrated that the transcription of PyPYC was significantly induced by a high concentration of CO₂ compared with the normal inorganic carbon concentration. The change in transcription was not as obvious in response to a high concentration of HCO₃^–. Therefore, we propose that PYC probably plays a particular role in inorganic carbon utilization when P. yezoensis thallus is exposed to the atmosphere.

Key words: Pyruvate carboxylase Pyropia yezoensis inorganic carbon utilization