Cloning and Characterization of a Salt Tolerance-Associated Gene Encoding Trehalose-6-Phosphate Synthase in Sweetpotato

doi:10.1016/S2095-3119(13)60534-1

Journal of Integrative Agriculture ›› 2014, Vol. 13 ›› Issue (8): 1651-1661.DOI: 10.1016/S2095-3119(13)60534-1

Cloning and Characterization of a Salt Tolerance-Associated Gene Encoding Trehalose-6-Phosphate Synthase in Sweetpotato

JIANG Tao, ZHAI Hong, WANG Fei-bing, ZHOU Hua-nan, SI Zeng-zhi, 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:2013-04-12 出版日期:2014-08-01 发布日期:2014-08-02
通讯作者: LIU Qing-chang, Tel/Fax: +86-10-62733710, E-mail: liuqc@cau.edu.cn
基金资助:
This work was supported by the National Natural Science Foundation of China (31271777), the China Agriculture Research System (CARS-11, Sweetpotato), the National High-Tech R&D Program of China (2012AA101204), and the Beijing Key Discipline Program, China.

Cloning and Characterization of a Salt Tolerance-Associated Gene Encoding Trehalose-6-Phosphate Synthase in Sweetpotato

JIANG Tao, ZHAI Hong, WANG Fei-bing, ZHOU Hua-nan, SI Zeng-zhi, 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:2013-04-12 Online:2014-08-01 Published:2014-08-02
Contact: LIU Qing-chang, Tel/Fax: +86-10-62733710, E-mail: liuqc@cau.edu.cn
Supported by:
This work was supported by the National Natural Science Foundation of China (31271777), the China Agriculture Research System (CARS-11, Sweetpotato), the National High-Tech R&D Program of China (2012AA101204), and the Beijing Key Discipline Program, China.

摘要/Abstract

摘要： Trehalose plays an important role in metabolic regulation and abiotic stress tolerance in a variety of organisms. In plants, its biosynthesis is catalyzed by two key enzymes: trehalose-6-phosphate synthase (TPS) and trehalose-6-phosphate phosphatase (TPP). In the present study, a TPS gene, named IbTPS, was first isolated from sweetpotato (Ipomoea batatas (L.) Lam.) cv. Lushu 3 by rapid amplification of cDNA ends (RACE). The open reading frame (ORF) contained 2 580 nucleotides encoding 859 amino acids with a molecular weight of 97.433 kDa and an isoelectric point (pI) of 5.7. The deduced amino acid sequence showed high identities with TPS of other plants. Real-time quantitative PCR analysis revealed that the expression level of IbTPS gene was significantly higher in stems of Lushu 3 than in its leaves and roots. Subcellular localization analysis in onion epidermal cells indicated that IbTPS gene was located in the nucleus. Transgenic tobacco (cv. Wisconsin 38) plants over-expressing IbTPS gene exhibited significantly higher salt tolerance compared with the control plant. Trehalose and proline content was found to be significantly more accumulated in transgenic tobacco plants than in the wild-type and several stress tolerance related genes were up-regulated. These results suggest that IbTPS gene may enhance salt tolerance of plants by increasing the amount of treahalose and proline and regulating the expression of stress tolerance related genes.

关键词: cloning , IbTPS gene , Ipomoea batatas (L.) Lam. , salt tolerance

Abstract: Trehalose plays an important role in metabolic regulation and abiotic stress tolerance in a variety of organisms. In plants, its biosynthesis is catalyzed by two key enzymes: trehalose-6-phosphate synthase (TPS) and trehalose-6-phosphate phosphatase (TPP). In the present study, a TPS gene, named IbTPS, was first isolated from sweetpotato (Ipomoea batatas (L.) Lam.) cv. Lushu 3 by rapid amplification of cDNA ends (RACE). The open reading frame (ORF) contained 2 580 nucleotides encoding 859 amino acids with a molecular weight of 97.433 kDa and an isoelectric point (pI) of 5.7. The deduced amino acid sequence showed high identities with TPS of other plants. Real-time quantitative PCR analysis revealed that the expression level of IbTPS gene was significantly higher in stems of Lushu 3 than in its leaves and roots. Subcellular localization analysis in onion epidermal cells indicated that IbTPS gene was located in the nucleus. Transgenic tobacco (cv. Wisconsin 38) plants over-expressing IbTPS gene exhibited significantly higher salt tolerance compared with the control plant. Trehalose and proline content was found to be significantly more accumulated in transgenic tobacco plants than in the wild-type and several stress tolerance related genes were up-regulated. These results suggest that IbTPS gene may enhance salt tolerance of plants by increasing the amount of treahalose and proline and regulating the expression of stress tolerance related genes.

Key words: cloning , IbTPS gene , Ipomoea batatas (L.) Lam. , salt tolerance

JIANG Tao, ZHAI Hong, WANG Fei-bing, ZHOU Hua-nan, SI Zeng-zhi, HE Shao-zhen , LIU Qing-chang. Cloning and Characterization of a Salt Tolerance-Associated Gene Encoding Trehalose-6-Phosphate Synthase in Sweetpotato[J]. Journal of Integrative Agriculture, 2014, 13(8): 1651-1661.

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Cloning and Characterization of a Salt Tolerance-Associated Gene Encoding Trehalose-6-Phosphate Synthase in Sweetpotato

Cloning and Characterization of a Salt Tolerance-Associated Gene Encoding Trehalose-6-Phosphate Synthase in Sweetpotato

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