Cloning and Functional Analysis of Lycopene ε-Cyclase (IbLCYe) Gene from Sweetpotato, Ipomoea batatas (L.) Lam.

doi:10.1016/S2095-3119(13)60299-3

Journal of Integrative Agriculture ›› 2013, Vol. 12 ›› Issue (5): 773-780.DOI: 10.1016/S2095-3119(13)60299-3

Cloning and Functional Analysis of Lycopene ε-Cyclase (IbLCYe) Gene from Sweetpotato, Ipomoea batatas (L.) Lam.

YU Ling, ZHAI Hong, CHEN Wei, HE Shao-zhen , LIU Qing-chang

Beijing Key Laboratory of Crop Genetic Improvement/Laboratory of Crop Heterosis and Utilization, Ministry of Education/College of Agriculture and Biotechnology, China Agricultural University, Beijing 100193, P.R.China

收稿日期:2012-06-06 出版日期:2013-05-01 发布日期:2013-05-01
通讯作者: Correspondence LIU Qing-chang, Tel: +86-10-62733710, E-mail: liuqc@cau.edu.cn
基金资助:
This work was supported by the China Agriculture Research System (Sweetpotato) and the National High-Tech Research and Development Project of China (2011AA100607 and 2012AA101204).

Cloning and Functional Analysis of Lycopene ε-Cyclase (IbLCYe) Gene from Sweetpotato, Ipomoea batatas (L.) Lam.

YU Ling, ZHAI Hong, CHEN Wei, HE Shao-zhen , LIU Qing-chang

Beijing Key Laboratory of Crop Genetic Improvement/Laboratory of Crop Heterosis and Utilization, Ministry of Education/College of Agriculture and Biotechnology, China Agricultural University, Beijing 100193, P.R.China

Received:2012-06-06 Online:2013-05-01 Published:2013-05-01
Contact: Correspondence LIU Qing-chang, Tel: +86-10-62733710, E-mail: liuqc@cau.edu.cn
Supported by:
This work was supported by the China Agriculture Research System (Sweetpotato) and the National High-Tech Research and Development Project of China (2011AA100607 and 2012AA101204).

摘要/Abstract

摘要： 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.

关键词: IbLCYe gene , cloning , functional analysis , carotenoid , Ipomoea batatas (L.) Lam.

Abstract: 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.

Key words: IbLCYe gene , cloning , functional analysis , carotenoid , Ipomoea batatas (L.) Lam.

YU Ling, ZHAI Hong, CHEN Wei, HE Shao-zhen , LIU Qing-chang. Cloning and Functional Analysis of Lycopene ε-Cyclase (IbLCYe) Gene from Sweetpotato, Ipomoea batatas (L.) Lam.[J]. Journal of Integrative Agriculture, 2013, 12(5): 773-780.

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Cloning and Functional Analysis of Lycopene ε-Cyclase (IbLCYe) Gene from Sweetpotato, Ipomoea batatas (L.) Lam.

Cloning and Functional Analysis of Lycopene ε-Cyclase (IbLCYe) Gene from Sweetpotato, Ipomoea batatas (L.) Lam.

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