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Cloning and Expression Characterization of the Athocyanin Biosynthesis Key Genes in Eggplant (solanum Melongena L.)

Author: LiXiang
Tutor: ChenHuoYing
School: Shanghai Jiaotong University
Course: Horticulture
Keywords: Eggplant Anthocyanin Clone Dihydroflavonol 4 - reductase Anthocyanin 5-O Glycosyltransferase
CLC: S641.1
Type: Master's thesis
Year: 2011
Downloads: 214
Quote: 0
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Abstract


Eggplant is an important vegetable crops. In recent years, increasingly in-depth studies on eggplant anthocyanins, type of eggplant anthocyanins, the antioxidant effect has an in-depth study. Therefore, further research eggplant anthocyanin biosynthesis path, looking for differences in anthocyanin content between the the eggplant different resources mechanism to explore in depth the health effects of eggplant, improve eggplant breeding quality of the work provided a theoretical basis. We used two phenotypic differences obvious eggplant resources phenotype as the experimental material for the green stems and white flowers, ginkgo Yunnan round white eggplant purple stems, purple flowers, purple fruit Yunnan purple long eggplant and phenotype. Discovered by studying 14 of anthocyanin synthesis related gene expression in Yunnan Purple long eggplant, and Yunnan round white eggplant commodity mature peel, dihydroflavonol 4 - reductase the gene (SmDFR) and the anthocyanin 5 -O glucoside transferase gene (Sm5GT), differential expression. Then the the eggplant in SmDFR and Sm5GT gene full-length cDNA clone. The length of the cDNA of the SmDFR for 1503bp, the coding region of 1149bp from 87-1235, the deduced 383 amino acids. Yunnan round white eggplant SmDFR (YRWSmDFR) compared to the nucleotide sequence in Yunnan Purple long eggplant SmDFR (YLPSmDFR) two-locus mutation, the nucleotide sequence of the 632 mutations to synonymous mutations, 792 mutations were missense mutations, making YRWSmDFR amino acid sequence from lysine to glutamic acid 236. The the SmDFR deduced amino acid sequence having a typical NADPH binding motif and substrate binding motif, in determining the activity of the 145 points, SmDFR aspartate. Expression of SmDFR Purple long eggplant in Yunnan and Yunnan round white eggplant has a great difference. Yunnan purple long eggplant purple organization YLPSmDFR expression to be significantly greater than Yunnan round white eggplant same organization YRWSmDFR. Through southern hybridization, SmDFR The Purple long eggplant in Yunnan and Yunnan round white eggplant are single copy. The the YLPSmDFR and YRWSmDFR prokaryotic expression activity of the the detection fusion protein dihydro kaempferol dihydro quercetin dihydromyricetin, three substrates. YLPSmDFR and YRWSmDFR the activity dihydromyricetin greater than dihydro-quercetin and dihydro kaempferol, at the same time YLPSmDFR For activity dihydromyricetin To significantly greater than YRWSmDFR. These results indicate that, SmDFR may cause eggplant anthocyanins difference key genes and YRWSmDFR the mutation at 236 may further weaken the enzyme activity. Sm5GT length of the cDNA is 1789bp, and the coding region of 1413bp from 67-1479, the deduced 470 amino acids. The deduced amino acid sequence has a typical the 5GT substrate binding motif. Expression of all detected Sm5GT Purple long eggplant in Yunnan and Yunnan round white eggplant, but the amount of expression in Yunnan purple a long eggplant purple petals, and commodities purple peel slightly larger than the of Yunnan round white eggplant. This result indicates that Sm5GT may have a wider range of substrates.

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CLC: > Agricultural Sciences > Gardening > Vegetable gardening > Solanaceous > Eggplant
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