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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
KeywordChlamydomonas Chloroplast Isolation Triacylglycerol Nitrogen Stress Fatty Acid Biomarkers
Source PublicationFRONTIERS IN PLANT SCIENCE
2017-08-29
DOI10.3389/fpls.2017.01503
Volume8
Indexed BySCI
SubtypeArticle
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
WOS SubjectPlant Sciences
WOS Research AreaPlant Sciences
WOS KeywordINTACT CHLOROPLASTS ; INORGANIC CARBON ; HEAT-STRESS ; LOW CO2 ; CELLS ; BIOSYNTHESIS ; LIPIDS ; PHOTOSYNTHESIS ; FRACTIONATION ; ACCUMULATION
AbstractTriacylglycerols 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.
Language英语
WOS IDWOS:000408660800001
Citation statistics
Cited Times:3[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/149974
Collection中国科学院大连化学物理研究所
Affiliation1.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
Recommended Citation
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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