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Preparation of protoplasts from Chlorella protothecoides
Lu, Yongzhong1; Kong, Renqiu2; Hu, Liwei1; Lu, Y.(luyz@yahoo.com)
2012-04-01
Source PublicationWORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY
ISSN0959-3993
Volume28Issue:4Pages:1827-1830
AbstractThis paper describes an enzymatic method for yielding protoplasts from the microalga Chlorella protothecoides. Four kinds of commercially available enzymes were tested. The enzymatic digestion was optimal with 2% cellulase R-10 and 1% snailase prepared in 25 mM Tris buffer (pH 6.0) containing 0.6 M D-mannitol, and the protoplast density could reach the peak after treatment at 30°C for 16 h. Nearly all liberated protoplasts were green in the presence of 0.01% phenosafranin, indicating their high viability. The regeneration rate was about 70% when 0.6 M D-mannitol was used as an osmotic stabilizer in the regeneration medium. This protocol will find useful applications in genetic studies of this algal species.; This paper describes an enzymatic method for yielding protoplasts from the microalga Chlorella protothecoides. Four kinds of commercially available enzymes were tested. The enzymatic digestion was optimal with 2% cellulase R-10 and 1% snailase prepared in 25 mM Tris buffer (pH 6.0) containing 0.6 M D-mannitol, and the protoplast density could reach the peak after treatment at 30A degrees C for 16 h. Nearly all liberated protoplasts were green in the presence of 0.01% phenosafranin, indicating their high viability. The regeneration rate was about 70% when 0.6 M D-mannitol was used as an osmotic stabilizer in the regeneration medium. This protocol will find useful applications in genetic studies of this algal species.
SubtypeArticle
KeywordProtoplast Cell Wall Enzymatic Digestion Chlorella Regeneration
Department(1) Biology Department, Qingdao University of Science and Technology, Qingdao, 266042, China; (2) State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, 430072, China
DOI10.1007/s11274-011-0963-4
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
Indexed BySCI
Language英语
WOS Research AreaBiotechnology & Applied Microbiology
WOS SubjectBiotechnology & Applied Microbiology
WOS IDWOS:000302292500054
WOS KeywordCELL-WALL ; INDUCTION ; ALGAE
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Cited Times:5[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/342005/16686
Collection藻类生物学及应用研究中心_期刊论文
Corresponding AuthorLu, Y.(luyz@yahoo.com)
Affiliation1.Qingdao Univ Sci & Technol, Dept Biol, Qingdao 266042, Peoples R China
2.Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430071, Peoples R China
Recommended Citation
GB/T 7714
Lu, Yongzhong,Kong, Renqiu,Hu, Liwei,et al. Preparation of protoplasts from Chlorella protothecoides[J]. WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY,2012,28(4):1827-1830.
APA Lu, Yongzhong,Kong, Renqiu,Hu, Liwei,&Lu, Y..(2012).Preparation of protoplasts from Chlorella protothecoides.WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY,28(4),1827-1830.
MLA Lu, Yongzhong,et al."Preparation of protoplasts from Chlorella protothecoides".WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY 28.4(2012):1827-1830.
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