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Cloning and functional validation of molybdenum cofactor sulfurase gene from Ammopiptanthus nanus

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Abstract

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The molybdenum cofactor sulfurase gene ( AnMCSU ) was cloned from xerophytic desert plant Ammopiptanthus nanus and validated for its function of tolerance toward abiotic stresses by heterologous expression in Arabidopsis thaliana.

Abstract

Molybdenum cofactor sulfurase participates in catalyzing biosynthesis of abscisic acid, which plays a crucial role in the response of plants to abiotic stresses. In this study, we cloned molybdenum cofactor sulfurase gene (AnMCSU) from a super-xerophytic desert plant, Ammopiptanthus nanus, by using rapid amplification of cDNA ends method. This gene has a total length of 2544 bp, with a 5′- and a 3′-untranslated region of 167 and 88 bp, and an open reading frame of 2289 bp, which encodes an 84.85 kDa protein of 762 amino acids. The putative amino acid sequence shares high homology and conserved amino acid residues crucial for the function of molybdenum cofactor sulfurases with other leguminous species. The encoded protein of the AnMCSU gene was located in the cytoplasm by transient expression in Nicotiana benthamiana. The result of real-time quantitative PCR showed that the expression of the AnMCSU gene was induced by heat, dehydration, high salt stresses, and ABA induction, and inhibited by cold stress. The heterologous expression of the AnMCSU gene significantly enhanced the tolerance of Arabidopsis thaliana to high salt, cold, osmotic stresses, and abscisic acid induction. All these results suggest that the AnMCSU gene might play a crucial role in the adaptation of A. nanus to abiotic stress and has potential to be applied to transgenic improvement of commercial crops.

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Abbreviations

ABA:

Abscisic acid

MoCo:

Molybdenum cofactor

MCSU:

Molybdenum cofactor sulfurase

cDNA:

Complementary DNA

RACE:

Rapid amplification of cDNA ends

qRT-PCR:

Quantitative real-time PCR

ORF:

Open reading frame

UTR:

Untranslated region

eGFP:

Enhanced green fluorescent protein

CaMV:

Cauliflower mosaic virus

OCS:

Octopine synthetase

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Acknowledgments

This work was supported by the National Key Science and Technology Special Project (2014ZX08003-004) and the National Natural Science Foundation of China (31071433). The authors thank the technical support from the Key Laboratory of Biology and Genetic Improvement of Maize in Southwest Region and the anonymous reviewers for their critical reading and modification suggestions.

Conflict of interest

The authors declare that they have no conflict of interest.

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    Authors and Affiliations

    1. Maize Research Institute, Sichuan Agricultural University, Chengdu, 611130, Sichuan, People’s Republic of China

      Hao Qiang Yu, Yuan Yuan Zhang, Tai Ming Yong, Shu Feng Zhou, Feng Ling Fu & Wan Chen Li

    2. Faculty of Plant Science, Tarim University, Alar, 843300, Xinjiang, People’s Republic of China

      Yan Ping Liu

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    1. Hao Qiang Yu
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    2. Yuan Yuan Zhang
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    3. Tai Ming Yong
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    Correspondence to Feng Ling Fu or Wan Chen Li.

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    Communicated by Q. Zhao.

    H. Q. Yu and Y. Y. Zhang contributed equally to this work.

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    Yu, H.Q., Zhang, Y.Y., Yong, T.M. et al. Cloning and functional validation of molybdenum cofactor sulfurase gene from Ammopiptanthus nanus . Plant Cell Rep 34, 1165–1176 (2015). https://doi.org/10.1007/s00299-015-1775-z

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