Isolation of Chlamydomonas reinhardtii mutants with altered mitochondrial respiration by chlorophyll fluorescence measurement
Isolation of Chlamydomonas reinhardtii mutants with altered mitochondrial respiration by chlorophyll fluorescence measurement
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DOI：
10.1016/j.jbiotec.2015.05.009
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发表时间：
2015-12-10
影响因子：
4.1
通讯作者：
Cardol, Pierre
中科院分区：
文献类型：
--
作者：
Massoz, Simon;Larosa, Veronique;Cardol, Pierre
单细胞绿色衣藻是研究能量代谢的模式生物。迄今为止，大多数线粒体功能缺陷突变体的特征在于它们在异养条件下生长能力降低的基础上被分离出来。线粒体的缺陷通常通过调节光合作用活性而得到部分补偿，尤其是通过向状态2的转变。在这项工作中，我们探索了机会，选择呼吸受损和/或改变在黑暗中的代谢通过测量最大光合效率叶绿素荧光分析（F-V/F-M）。在从野生型或从状态转换缺陷的突变株（stt 7株）产生的约2900个潮霉素抗性插入突变体中，发现22个在异养条件下生长缓慢，其中8个还显示出较低的F-V/F-M值。鉴定了几个被破坏的编码序列，包括编码呼吸链复合物I的三个不同亚基（NU 09、NUOA 9、NUOP 4）或异柠檬酸裂合酶（ICL 1）的基因。总之，在野生型或stt 7遗传背景中获得的呼吸突变体的比较表明，Fv/Fm值可用于分离在暗代谢中严重受损的突变体。(C)2015 Elsevier B. V.版权所有。
The unicellular green alga Chlamydomonas reinhardtii is a model organism for studying energetic metabolism. Most mitochondrial respiratory-deficient mutants characterized to date have been isolated on the basis of their reduced ability to grow in heterotrophic conditions. Mitochondrial deficiencies are usually partly compensated by adjustment of photosynthetic activity and more particularly by transition to state 2. In this work, we explored the opportunity to select mutants impaired in respiration and/or altered in dark metabolism by measuring maximum photosynthetic efficiency by chlorophyll fluorescence analyses (F-V/F-M). Out of about 2900 hygromycin-resistant insertional mutants generated from wild type or from a mutant strain deficient in state transitions (stt7 strain), 22 were found to grow slowly in heterotrophic conditions and 8 of them also showed a lower F-V/F-M value. Several disrupted coding sequences were identified, including genes coding for three different subunits of respiratory-chain complex I (NU09, NUOA9, NUOP4) or for isocitrate lyase (ICL1). Overall, the comparison of respiratory mutants obtained in wild-type or stt7 genetic backgrounds indicated that the Fv/Fm value can be used to isolate mutants severely impaired in dark metabolism. (C) 2015 Elsevier B.V. All rights reserved.