Chloroplasts Isolation from Chlamydomonas reinhardtii under Nitrogen Stress.
Chloroplasts Isolation from Chlamydomonas reinhardtii under Nitrogen Stress.
复制标题
氮胁迫下莱茵衣藻叶绿体的分离
DOI：
10.3389/fpls.2017.01503
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发表时间：
2017
影响因子：
5.6
通讯作者：
Chi ZY
中科院分区：
文献类型：
--
作者：
Yang M;Jiang JP;Xie X;Chu YD;Fan Y;Cao XP;Xue S;Chi ZY
在一些暴露于压力的微藻中，通过叶绿体和内质网途径大量产生三酰甘油，尽管任一途径的相对贡献仍然难以捉摸。这些途径的表征需要分离的细胞器。在这项研究中，一个有效的和可重复的方法，包括均质分批培养的氮剥夺藻细胞的光生物反应器，温和的细胞破碎使用一个简单的定制的破碎机与机械剪切力，优化差速离心和Percoll密度梯度离心，从衣藻叶绿体受到氮胁迫。使用这种方法，最大的限制应力持续时间为4小时，并强调细胞表现出19%和32%的细胞内叶绿素和氮含量，分别下降。从含10 mg叶绿素的胁迫细胞中成功地分离出含48 - 300 μ g叶绿素的叶绿体。这些压力叶绿体出现完整的，作为监测的超微结构观察和一种新的质量控制方法，涉及脂肪酸生物标志物。这种方法可以为微藻的亚细胞生化研究提供足够数量的完整胁迫叶绿体。
Triacylglycerols are produced in abundance through chloroplast and endoplasmic reticulum pathways in some microalgae exposed to stress, though the relative contribution of either pathway remains elusive. Characterization of these pathways requires isolation of the organelles. In this study, an efficient and reproducible approach, including homogenous batch cultures of nitrogen-deprived algal cells in photobioreactors, gentle cell disruption using a simple custom-made disruptor with mechanical shear force, optimized differential centrifugation and Percoll density gradient centrifugation, was developed to isolate chloroplasts from Chlamydomonas reinhardtii subjected to nitrogen stress. Using this approach, the maximum limited stress duration was 4 h and the stressed cells exhibited 19 and 32% decreases in intracellular chlorophyll and nitrogen content, respectively. Chloroplasts with 48 – 300 μg chlorophyll were successfully isolated from stressed cells containing 10 mg chlorophyll. These stressed chloroplasts appeared intact, as monitored by ultrastructure observation and a novel quality control method involving the fatty acid biomarkers. This approach can provide sufficient quantities of intact stressed chloroplasts for subcellular biochemical studies in microalgae.
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影响因子：
3.7
作者：
BOLLIVAR, DW;BEALE, SI
通讯作者：
BEALE, SI
影响因子：
6.9
作者：
Baginsky, S;Gruissem, W
通讯作者：
Gruissem, W
DOI：
10.1073/pnas.78.4.2310
发表时间：
1981-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
BELKNAP, WR;TOGASAKI, RK
通讯作者：
TOGASAKI, RK
影响因子：
7.4
作者：
KLEIN, U;CHEN, CG;PLATTALOIA, KA
通讯作者：
PLATTALOIA, KA
影响因子：
7.3
作者：
Legeret, B.;Schulz-Raffelt, M.;Li-Beisson, Y.
通讯作者：
Li-Beisson, Y.