Cloning And Characterization Of A Candidate Gene For The Dormancy Of Peanut

Peanut planting in our country is common, but its dormancy will affect the quality and yield of peanut, which brings some problems to agricultural production, so the research on the dormancy of peanut has important significance. The research on the dormancy of peanut is limited to the classical genetics and the changes of endogenous substances. Howerver, there were only few research reports regarding its molecular biology. In this study, the gene oleosin2 was obtained from the peanut genome by PCR technology, and constructed over-expression vector for this gene. At the same time, 3 candidate gene were cloned by RT-PCR, which may be related to dormancy. We successfully constructed over-expression vector and subcellular localization vector for the relative gene. The main results of this study includes:1. The oleosin2 gene was cloned form the peanut "Y-1" genome by PCR technology, and constructed the over-expression vector p Cambia2300EC-oleosin2. Through Agrobacterium infection transformed into peanut, by PCR obtained transgenic seed, and carried out germination experiment and quality analysis, found that the gene was not related to dormancy, but was related to oli. When the gene is over expressed in the peanut, the oil content of the peanut is improved.2. In this study, a novel Ah ABI3 gene was cloned from the mature seed of peanut by RT-PCR method. The Ah ABI3 gene was predicted to contain an open reading frame of 2313 bp which encoded 770 amino acids. As shown by homology comparison analysis, it had high similarity between Ah ABI3 and ABI3 from other selected plants(such as Arabidopsis and Cicer arietinum). Phylogenetic analysis indicated that Ah ABI3 was in closest relationship with ABI3 from Cicer arietinum. We successfully constructed over-expression vector and subcellular localization vector for this gene. The over expression vector was constructed and transformed into Arabidopsis thaliana by Agrobacterium mediated transformation, and get positive seedings, which provided a foundation for further research of Ah ABI3 gene function.3. In this study, a novel AhNCED1 gene was cloned from the seed of peanut by RT-PCR method. The Ah NCED1 gene was predicted to contain an open reading frame of 1806 bp which encoded 601 amino acids.Its encoded protein had the molecular weight as 66.8 k Da and p I as 8.49. As shown by homology comparison analysis, it had high similarity between Ah NCED1 and NCED from other selected plants(such as Arabidopsis and Stylosanthes guianensis). Phylogenetic analysis indicated that Ah NCED1 was in closest relationship with NCED from Stylosanthes guianensis. We successfully constructed subcellular localization and over-express vector for this gene, which provided a foundation for further research of Ah NCED1 gene function.4. In this study, a novel Ah CYP707A1 gene was cloned from the seed of peanut by RT-PCR method. The Ah CYP707A1 gene was predicted to contain an open reading frame of 1401 bp which encoded 466 amino acids.The results showed that the molecular weight as 53.2 k Da and p I as 9.29. As shown by homology comparison analysis, it had high similarity between Ah CYP707A1 and ABA 8’-hydroxylase from other selected plants(such as Arabidopsis and Medicago truncatula). Phylogenetic analysis indicated that Ah CYP707A1 was in closest relationship with CYP707 A from Medicago truncatula. We successfully constructed over-express and subcellular localization vector for this gene, which provided a foundation for further research of Ah CYP707A1 gene function.