Identification of genes for chlorophyll biosynthesis and its roles in stay green phenotype and cell death

叶绿素生物合成基因的鉴定及其在保持绿色表型和细胞死亡中的作用

• 批准号: 15370015
• 负责人: TANAKA Ayumi
• 金额: $ 9.66万
• 依托单位: HOKKAIDO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15370015/
• 关键词: Chlorophyll; Photosynthesis; Metabolism; Chloroplast; 光エネルギー; 発育

Recently, significant progress has also been made toward elucidating the linkages between chlorophyll metabolism and other cellular processes including leaf senescence, programmed cell death, and plastid signaling. The purpose of the present research is to complete chlorophyll metabolic pathway and to identify all the genes involved in the pathway. Chlorophyll metabolism has been extensively studied with various organisms and almost all of the chlorophyll biosynthetic genes have been identified in higher plants. However, only the gene for 3, 8-divinyl protochlorophyllide a 8-vinyl reductase (DVR), which is indispensable for the photosynthetic organisms to produce monovinylchlorophyll or dinvinyl chlorophyll, has not been identified yet. In this study, we isolated an Arabidopsis mutant that accumulated divinyl chlorophyll instead of monovinyl chlorophyll by ethyl-methane sulfonate mutagenesis. Map-based cloning ofthis mutant resulted in the identification of a gene (At5g18660) that shows sequence similarity with isoflavon reductase genes. The mutant phenotype was complemented by the transformation with the wild-type gene. A recombinant protein encoded by At5g18660 was expressed in E.coli and found to catalyze the conversion of divinyl chlorophyllide to monovinyl chlorophyllide ; thereby indicating that the gene encodes a functional DVR. DVR is encoded by a single-copy gene in the Arabidopsis genome. With the identification of DVR, finally all genes required for chlorophyll biosynthesis have been identified in higher plants. Analysis of the complete genome of Arabidopsis showed that it has 15 enzymes encoded by 27 genes for chlorophyll biosynthesis from 3 glutamyl-tRNAglu to chlorophyll b.

最近，也取得了重大进展，阐明叶绿素代谢和其他细胞过程，包括叶片衰老，程序性细胞死亡，质体信号之间的联系。本研究的目的是完善叶绿素代谢途径，并对参与该途径的所有基因进行鉴定。叶绿素代谢已被广泛研究与各种生物和几乎所有的叶绿素生物合成基因已被确定在高等植物。然而，只有3，8-二乙烯基原叶绿素a 8-乙烯基还原酶（DVR）的基因尚未被鉴定，DVR是光合生物产生单乙烯基叶绿素或二乙烯基叶绿素所必需的。在这项研究中，我们分离出一个拟南芥突变体，积累二乙烯基叶绿素，而不是单乙烯基叶绿素的乙基甲烷磺酸诱变。基于图位克隆的突变体导致在鉴定的基因（At 5g 18660），显示序列相似性与葡萄糖醛酸还原酶基因。突变体表型通过用野生型基因转化来补充。在大肠杆菌中表达了由At 5g 18660编码的重组蛋白，发现其催化二乙烯基叶绿素转化为单乙烯基叶绿素;从而表明该基因编码功能性DVR。DVR由拟南芥基因组中的单拷贝基因编码。随着DVR的鉴定，最终在高等植物中鉴定了所有叶绿素生物合成所需的基因。对拟南芥全基因组的分析表明，拟南芥有15个酶，由27个基因编码，从3个谷氨酰-tRNAglu到叶绿素B。

项目成果

The N-terminal domain of chlorophyllide a oxygenase confer protein instability in response to chlorophyll b accumulation.

叶绿素 a 加氧酶的 N 末端结构域赋予蛋白质响应叶绿素 b 积累的不稳定性。

• 发表时间: 2005
• 期刊: The Plant Cell. 17(5)
• 作者: Yamasato;A.;Nagata;N.;Tanaka;R.;Tanaka;A.
• 通讯作者: A.

Unique constitution of photosystem I with a novel subunit in the cyanobacterium Gloeobacter violaceus PCC 7421

• DOI: 10.1016/j.febslet.2004.11.013
• 发表时间: 2004-12-17
• 期刊: FEBS LETTERS
• 影响因子: 3.5
• 作者: Inoue, H;Tsuchiya, T;Mimuro, M
• 通讯作者: Mimuro, M

The in vivo and in vitro reconstitution of pigment-protein complexes, and its implication in acquiring a new system.

色素-蛋白质复合物的体内和体外重建及其对获得新系统的意义。

• 发表时间: 2004
• 期刊: Photosynth.Res. 81
• 作者: Mimuro;M.;A.Tanaka
• 通讯作者: A.Tanaka

Tanaka, R: "The Arabidopsis-accelerated cell death gene ACDI is involved in oxygenation of pheophorbide a : Inhibition of the pheophorbide a oxygenase activity does not lead to the "stay-green" phenotype in Arabidopsis"Plant Cell Physiol.. 44. 1266-1274 (

Tanaka, R：“拟南芥加速细胞死亡基因 ACDI 参与脱镁叶绿酸 a 的氧化：抑制脱镁叶绿酸 a 氧化酶活性不会导致拟南芥中的“保持绿色”表型”植物细胞生理学.. 44. 1266

Ultrafast Excitation Relaxation Dynamics of Lutein in Solution and in the Light-Harvestion Complexes II Isolated from Arabidopsis thaliana.

溶液中和从拟南芥中分离的光捕获复合物 II 中叶黄素的超快激发弛豫动力学。

• 发表时间: 2005
• 期刊: The Journal of Physical Chemistry B 109(25)
• 作者: Akimoto;S.;Yokono;M.;Ohmae;M.;Yamazaki;I.;Tanaka;A.;Higuchi;M.;Tsuchiya;T.;Miyashita;H.;Mimuro;M.
• 通讯作者: M.