Cloning and Functional Analysis of Lycopene ε-Cyclase (IbLCYe) Gene from Sweetpotato, Ipomoea batata

来源 :Journal of Integrative Agriculture | 被引量 : 0次 | 上传用户:jiahong222
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This paper reported firstly successful cloning of lycopene ε-cyclase(IbLCYe) gene from sweetpotato,Ipomoea batatas(L.) Lam.Using rapid amplification of cDNA ends(RACE),IbLCYe gene was cloned from sweetpotato cv.Nongdafu 14 with high carotenoid content.The 1 805 bp cDNA sequence of IbLCYe gene contained a 1 236 bp open reading frame(ORF) encoding a 411 amino acids polypeptide with a molecular weight of 47 kDa and an isoelectric point(pI) of 6.95.IbLCYe protein contained one potential lycopene ε-cyclase domain and one potential FAD(flavinadenine dinucleotide)/NAD(P)(nicotinamide adenine dinucleotide phosphate)-binding domain,indicating that this protein shares the typical characteristics of LCYe proteins.The gDNA of IbLCYe gene was 4 029 bp and deduced to contain 5 introns and 6 exons.Real-time quantitative PCR analysis revealed that the expression level of IbLCYe gene was significantly higher in the storage roots of Nongdafu 14 than those in the leaves and stems.Transgenic tobacco(cv.Wisconsin 38) expressing IbLCYe gene accumulated significantly more β-carotene compared to the untransformed control plants.These results showed that IbLCYe gene has an important function for the accumulation of carotenoids of sweetpotato. This paper reported successful cloning of lycopene ε-cyclase (IbLCYe) gene from sweetpotato, Ipomoea batatas (L.) Lam. Using rapid amplification of cDNA ends (RACE), IbLCYe gene was cloned from sweetpotato cv.Nongdafu 14 with high carotenoid content The 1 805 bp cDNA sequence of the IbLCYe gene contained a 1 236 bp open reading frame (ORF) encoding a 411 amino acids polypeptide with a molecular weight of 47 kDa and an isoelectric point (pI) of 6.95.IbLCYe protein contained one potential lycopene ε-cyclase domain and one potential FAD (flavinadenine dinucleotide) / NAD (P) (nicotinamide adenine dinucleotide phosphate) -binding domain, indicating that this protein shares the typical characteristics of LCYe proteins. The gDNA of IbLCYe gene was 4 029 bp and deduced to contain 5 introns and 6 exons. Real-time quantitative PCR analysis revealed that the expression level of IbLCYe gene was significantly higher in the storage roots of Nongdafu 14 than those in the leaves and stems. Transgenic tobacco (cv. Wisconsin 38) expressing IbLCYe genes accumulated significantly more β-carotene compared to the untransformed control plants. These results showed that IbLCYe genes have an important function for the accumulation of carotenoids of sweetpotato.
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