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Molecular Cloning of Rice and Arabidopsis Ceramidase and Identification of Mutants

Author: LiJian
Tutor: YaoZuo
School: Sun Yat-sen University
Course: Biotechnology
Keywords: Arabidopsis Sphingolipid protoplasts ceramide ceramidase
CLC: S511
Type: Master's thesis
Year: 2011
Downloads: 6
Quote: 0
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Abstract


Sphingolipid is not only the important component of eukaryotic membranes, but also as an important signal molecule to regulate cell differentiation and death. However, the current understanding about the plant sphingolipid metabolism is very limited. Ceramide is the core metabolite of the sphingolipid pathway which plays a decisive role in mediating the level of sphingolipid in plants. Ceramidases can hydrolyze ceramide into fatty acids and sphingosines.Rice is an important crop. The research about the genes related sphingolipid metabolism has important theoretical and practical value. In this thesis, we first cloned the full length of OsCDase, and then used two methods of Agrobacterium-mediated rice transformation to obtain OsCDase over-expression and RNAi transgenic rice, which can be used as good materials for follow-up study about the gene function.Meanwhile, we purchased three single Arabidopsis ceramidase mutants from ABRC. After identification of those single mutants, we crossed mutants to obtain double homologous mutants, and also got the triple homologous mutants called Atcdase. In addition, we isolated the protoplasts from three single Arabidopsis ceramidase mutants and wild-type plants, then treated with C2-ceramide to detect the cell viability. We found that C2-ceramide could promote the cell death in AtCDase-2 mutants, while the other two single mutants have no significant difference when compared with wild-type, indicating AtCDase-2 is the key ceramidase in Arabidopsis.

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CLC: > Agricultural Sciences > Crop > Cereal crops > Rice
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