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Chloroplasts Isolation from Chlamydomonas reinhardtii under Nitrogen Stress
Yang, Miao1,2,3; Jiang, Jun-Peng1,2; Xie, Xi4; Chu, Ya-Dong1; Fan, Yan1,2; Cao, Xu-Peng1; Xue, Song1; Chi, Zhan-You3
关键词Chlamydomonas Chloroplast Isolation Triacylglycerol Nitrogen Stress Fatty Acid Biomarkers
刊名FRONTIERS IN PLANT SCIENCE
2017-08-29
DOI10.3389/fpls.2017.01503
8
收录类别SCI
文章类型Article
WOS标题词Science & Technology ; Life Sciences & Biomedicine
类目[WOS]Plant Sciences
研究领域[WOS]Plant Sciences
关键词[WOS]INTACT CHLOROPLASTS ; INORGANIC CARBON ; HEAT-STRESS ; LOW CO2 ; CELLS ; BIOSYNTHESIS ; LIPIDS ; PHOTOSYNTHESIS ; FRACTIONATION ; ACCUMULATION
英文摘要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 mu g 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.
语种英语
WOS记录号WOS:000408660800001
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文献类型期刊论文
条目标识符http://cas-ir.dicp.ac.cn/handle/321008/149874
专题中国科学院大连化学物理研究所
作者单位1.Chinese Acad Sci, Dalian Inst Chem Phys, Marine Bioengn Grp, Dalian, Peoples R China
2.Univ Chinese Acad Sci, Beijing, Peoples R China
3.Dalian Univ Technol, Sch Life Sci & Biotechnol, Dalian, Peoples R China
4.Liaoning Ocean & Fisheries Sci Res Inst, Dalian, Peoples R China
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GB/T 7714
Yang, Miao,Jiang, Jun-Peng,Xie, Xi,et al. Chloroplasts Isolation from Chlamydomonas reinhardtii under Nitrogen Stress[J]. FRONTIERS IN PLANT SCIENCE,2017,8.
APA Yang, Miao.,Jiang, Jun-Peng.,Xie, Xi.,Chu, Ya-Dong.,Fan, Yan.,...&Chi, Zhan-You.(2017).Chloroplasts Isolation from Chlamydomonas reinhardtii under Nitrogen Stress.FRONTIERS IN PLANT SCIENCE,8.
MLA Yang, Miao,et al."Chloroplasts Isolation from Chlamydomonas reinhardtii under Nitrogen Stress".FRONTIERS IN PLANT SCIENCE 8(2017).
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