دورية أكاديمية
Development of a forward genetic screen to isolate oil mutants in the green microalga Chlamydomonas reinhardtii
| العنوان: | Development of a forward genetic screen to isolate oil mutants in the green microalga Chlamydomonas reinhardtii |
|---|---|
| المؤلفون: | Cagnon, Caroline, Mirabella, Boris, Nguyen, Hoa Mai, Beyly-Adriano, Audrey, Bouvet, Séverine, Cuiné, Stéphan, Beisson, Fred, Peltier, G., Li-Beisson, Yonghua |
| المساهمون: | Environnement, Bioénergie, Microalgues et Plantes (EBMP), Institut de Biosciences et Biotechnologies d'Aix-Marseille (ex-IBEB) (BIAM), Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Aix Marseille Université (AMU)-Centre National de la Recherche Scientifique (CNRS)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), ANR-12-BIME-0001,DIESALG,Production de biodiesel par microalgues(2012) |
| المصدر: | ISSN: 1754-6834 ; Biotechnology for Biofuels ; https://hal.science/hal-03378490Test ; Biotechnology for Biofuels, 2013, 6 (1), ⟨10.1186/1754-6834-6-178⟩. |
| بيانات النشر: | HAL CCSD BioMed Central |
| سنة النشر: | 2013 |
| المجموعة: | HAL-CEA (Commissariat à l'énergie atomique et aux énergies alternatives) |
| مصطلحات موضوعية: | Chlamydomonas mutants, Flow cytometry, Genetic screen, Lipid remobilization, Microalgal oil, Nile red, [SDV]Life Sciences [q-bio] |
| الوصف: | International audience ; Oils produced by microalgae are precursors to biodiesel. To achieve a profitable production of biodiesel from microalgae, identification of factors governing oil synthesis and turnover is desirable. The green microalga Chlamydomonas reinhardtii is amenable to genetic analyses and has recently emerged as a model to study oil metabolism. However, a detailed method to isolate various types of oil mutants that is adapted to Chlamydomonas has not been reported.ResultsWe describe here a forward genetic approach to isolate mutants altered in oil synthesis and turnover from C. reinhardtii. It consists of a three-step screening procedure: a primary screen by flow cytometry of Nile red stained transformants grown in 96-deep-well plates under three sequential conditions (presence of nitrogen, then absence of nitrogen, followed by oil remobilization); a confirmation step using Nile red stained biological triplicates; and a validation step consisting of the quantification by thin layer chromatography of oil content of selected strains. Thirty-one mutants were isolated by screening 1,800 transformants generated by random insertional mutagenesis (1.7%). Five showed increased oil accumulation under the nitrogen-replete condition and 13 had altered oil content under nitrogen-depletion. All mutants were affected in oil remobilization.ConclusionThis study demonstrates that various types of oil mutants can be isolated in Chlamydomonas based on the method set-up here, including mutants accumulating oil under optimal biomass growth. The strategy conceived and the protocol set-up should be applicable to other microalgal species such as Nannochloropsis and Chlorella, thus serving as a useful tool in Chlamydomonas oil research and algal biotechnology. |
| نوع الوثيقة: | article in journal/newspaper |
| اللغة: | English |
| العلاقة: | hal-03378490; https://hal.science/hal-03378490Test; https://hal.science/hal-03378490/documentTest; https://hal.science/hal-03378490/file/Cagnon2013.pdfTest |
| DOI: | 10.1186/1754-6834-6-178 |
| الإتاحة: | https://doi.org/10.1186/1754-6834-6-178Test https://hal.science/hal-03378490Test https://hal.science/hal-03378490/documentTest https://hal.science/hal-03378490/file/Cagnon2013.pdfTest |
| حقوق: | http://creativecommons.org/licenses/byTest/ ; info:eu-repo/semantics/OpenAccess |
| رقم الانضمام: | edsbas.95016B45 |
| قاعدة البيانات: | BASE |
| DOI: | 10.1186/1754-6834-6-178 |
|---|