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Pistacia Chinensis Transcriptome Sequencing Analysis And Expression Analysis Of PcWRI1 And PcLEC1 Gene Transformation Into Arabidopsis Thaliana

Posted on:2016-03-22Degree:MasterType:Thesis
Country:ChinaCandidate:Y L ChenFull Text:PDF
GTID:2180330482981930Subject:Biochemistry and Molecular Biology
Abstract/Summary:PDF Full Text Request
Pistacia chinensis Bunge is a kind of important woody oil plant, its seed oil can be as raw material for biodiesel refining, indicating that P. chinensis is a greatly potential biomass energy tree species. Analysis of P. chinensis seed transcriptome with high-throughput sequencing technology, is conducive to the identification of fatty acid biosynthesis related genes, base on which the key transcription factor genes were cloned and transformed into Arabidopsis for functional identification. The study would be advantageous to the variety improvement and utilization of P. chinensis, of which the obtained results summarized as follows:1. RNA was extracted from P. chinensis seeds for cDNA library construction and transcriptome high-throughput sequencing. For the result,5.2G data was produced and assembled into 209,559 transcripts, from which a total of 127,545 unigenes were identified and were annotated with the public databases.Then the specific unigenes related to the oil accumulation were determined.2. Base on the analysis of transcriptome sequencing, the transcription factor gene PcWRI1 related to the regulation of seed oil accumulation was cloned. Then the eukaryotic expression vector of PcWRI1 and PcLEC1 gene that cloned in previous studies was constructed respectively, and transformed into Arabidopsis thaliana mediated by Agrobacterium.3. The phenotypic observation and seed oil detection was performed on the homozygous generations of transgenic A. thaliana plants with PcWRI1 and PcLECl gene obtained by molecular detection and Chi square test.4. The tissue-specific expression of PcWRI1 and PcLEC1 gene and period-specific expression of oil accumulation related genes in the transgenic A. thaliana plants was analyzed by using the Real-time fluorescent quantitative PCR. The result showed that the PcWRI1 gene has a high expression level in the later period of pod development, but a low expression level observed in the early period of pod development, as well as in flower and vegetative organs, and PcLEC1 gene has a high expression level not only in the development pod, but also in the vegetative organs of transgenic A. thaliana plants. For the oil accumulation related genes, some of the key enzyme genes involved in oil biosynthesis showed up-regulated expression in development pods of transgenic A. thaliana with PcWRIl and PcLEC1 gene.The study above could lay an important foundation for the further in-depth identification of PcWRI1 and PcLEC1 gene function and for its application in genetic improvement of Pistacia chinensis seed oil content.
Keywords/Search Tags:Pistacia Chinensis Bunge, transcriptome sequencing, PcLEC1, PcWRI1, genetic transformation, expression analysis
PDF Full Text Request
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