Molecular Cloning and Functional Characterization of a Salt Tolerance- Associated Gene IbNFU1 from Sweetpotato

doi:10.1016/S2095-3119(13)60202-6

Journal of Integrative Agriculture ›› 2013, Vol. 12 ›› Issue (1): 27-35.DOI: 10.1016/S2095-3119(13)60202-6

Molecular Cloning and Functional Characterization of a Salt Tolerance- Associated Gene IbNFU1 from Sweetpotato

WANG Lian-jun, HE Shao-zhen, ZHAI Hong, LIU De-gao, WANGYan-nan , LIU Qing-chang

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

收稿日期:2011-08-11 出版日期:2013-01-01 发布日期:2013-01-24
通讯作者: Correspondence LIU Qing-chang, Tel/Fax: +86-10-62733710, E-mail: liuqc@cau.edu.cn
基金资助:
This work was supported by the China Agricultural Research System (Sweetpotato), the National High-Tech Res e a rch and Deve lopment Program of Chin a (2009AA10Z102), the National Transgenic Plants Project of China (2009ZX08009-064B), and the National Natural Science Foundation of China (31071468).

Molecular Cloning and Functional Characterization of a Salt Tolerance- Associated Gene IbNFU1 from Sweetpotato

WANG Lian-jun, HE Shao-zhen, ZHAI Hong, LIU De-gao, WANGYan-nan , LIU Qing-chang

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

Received:2011-08-11 Online:2013-01-01 Published:2013-01-24
Contact: Correspondence LIU Qing-chang, Tel/Fax: +86-10-62733710, E-mail: liuqc@cau.edu.cn
Supported by:
This work was supported by the China Agricultural Research System (Sweetpotato), the National High-Tech Res e a rch and Deve lopment Program of Chin a (2009AA10Z102), the National Transgenic Plants Project of China (2009ZX08009-064B), and the National Natural Science Foundation of China (31071468).

摘要/Abstract

摘要： Iron-sulfur cluster biosynthesis involving the nitrogen fixation (Nif) proteins has been proposed as a general mechanism acting in various organisms. NifU-like protein may play an important role in protecting plants against abiotic and biotic stresses. Based on the EST sequence selected from salt-stressed suppression subtractive hybridization (SSH) cDNA library constructed with a salt-tolerant mutant LM79, a NFU gene, termed IbNFU1, was cloned from sweetpotato (Ipomoea batatas (L.) Lam.) via rapid amplification of cDNA ends (RACE). The cDNA sequence of 1 117 bp contained an 846 bp open reading frame encoding a 281 amino acids polypeptide with a molecular weight of 30.5 kDa and an isoelectric point (pI) of 5.12. IbNFU1 gene contained a conserved Cys-X-X-Cys motif in C-terminal of the iron-sulfur cluster domain. The deduced amino acid sequence had 66.08 to 71.99% sequence identity to NFU genes reported in Arabidopsis thaliana, Eucalyptus grandis and Vitis vinifera. Real-time quantitative PCR analysis revealed that the expression level of IbNFU1 gene was significantly higher in the roots of the mutant LM79 compared to the wild-type Lizixiang. Transgenic tobacco (cv. Wisconsin 38) plants expressing IbNFU1 gene exhibited significantly higher salt tolerance compared to the untransformed control plants. It is proposed that IbNFU1 gene has an important function for salt tolerance of plants.

关键词: cloning , IbNFU1 gene , Ipomoea batatas (L.) Lam. , salt tolerance

Abstract: Iron-sulfur cluster biosynthesis involving the nitrogen fixation (Nif) proteins has been proposed as a general mechanism acting in various organisms. NifU-like protein may play an important role in protecting plants against abiotic and biotic stresses. Based on the EST sequence selected from salt-stressed suppression subtractive hybridization (SSH) cDNA library constructed with a salt-tolerant mutant LM79, a NFU gene, termed IbNFU1, was cloned from sweetpotato (Ipomoea batatas (L.) Lam.) via rapid amplification of cDNA ends (RACE). The cDNA sequence of 1 117 bp contained an 846 bp open reading frame encoding a 281 amino acids polypeptide with a molecular weight of 30.5 kDa and an isoelectric point (pI) of 5.12. IbNFU1 gene contained a conserved Cys-X-X-Cys motif in C-terminal of the iron-sulfur cluster domain. The deduced amino acid sequence had 66.08 to 71.99% sequence identity to NFU genes reported in Arabidopsis thaliana, Eucalyptus grandis and Vitis vinifera. Real-time quantitative PCR analysis revealed that the expression level of IbNFU1 gene was significantly higher in the roots of the mutant LM79 compared to the wild-type Lizixiang. Transgenic tobacco (cv. Wisconsin 38) plants expressing IbNFU1 gene exhibited significantly higher salt tolerance compared to the untransformed control plants. It is proposed that IbNFU1 gene has an important function for salt tolerance of plants.

Key words: cloning , IbNFU1 gene , Ipomoea batatas (L.) Lam. , salt tolerance

WANG Lian-jun, HE Shao-zhen, ZHAI Hong, LIU De-gao, WANGYan-nan , LIU Qing-chang. Molecular Cloning and Functional Characterization of a Salt Tolerance- Associated Gene IbNFU1 from Sweetpotato[J]. Journal of Integrative Agriculture, 2013, 12(1): 27-35.

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Molecular Cloning and Functional Characterization of a Salt Tolerance- Associated Gene IbNFU1 from Sweetpotato

Molecular Cloning and Functional Characterization of a Salt Tolerance- Associated Gene IbNFU1 from Sweetpotato

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