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MsZEP, a novel zeaxanthin epoxidase gene from alfalfa (Medicago sativa), confers drought and salt tolerance in transgenic tobacco

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Abstract

Key message

The zeaxanthin epoxidase gene ( MsZEP ) was cloned and characterized from alfalfa and validated for its function of tolerance toward drought and salt stresses by heterologous expression in Nicotiana tabacum.

Abstract

Zeaxanthin epoxidase (ZEP) plays important roles in plant response to various environment stresses due to its functions in ABA biosynthetic and the xanthophyll cycle. To understand the expression characteristics and the biological functions of ZEP in alfalfa (Medicago sativa), a novel gene, designated as MsZEP (KM044311), was cloned, characterized and overexpressed in Nicotiana tabacum. The open reading frame of MsZEP contains 1992 bp nucleotides and encodes a 663-amino acid polypeptide. Amino acid sequence alignment indicated that deduced MsZEP protein was highly homologous to other plant ZEP sequences. Phylogenetic analysis showed that MsZEP was grouped into a branch with other legume plants. Real-time quantitative PCR revealed that MsZEP gene expression was clearly tissue-specific, and the expression levels were higher in green tissues (leaves and stems) than in roots. MsZEP expression decreased in shoots under drought, cold, heat and ABA treatment, while the expression levels in roots showed different trends. Besides, the results showed that nodules could up-regulate the MsZEP expression under non-stressful conditions and in the earlier stage of different abiotic stress. Heterologous expression of the MsZEP gene in N. tabacum could confer tolerance to drought and salt stress by affecting various physiological pathways, ABA levels and stress-responsive genes expression. Taken together, these results suggested that the MsZEP gene may be involved in alfalfa responses to different abiotic stresses and nodules, and could enhance drought and salt tolerance of transgenic tobacco by heterologous expression.

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Abbreviations

ABA:

Abscisic acid

Fv/Fm:

Maximum photochemical efficiency

GFP:

Green fluorescent protein

Gs:

Stomatal conductance

MDA:

Malonyldialdehyde

NA:

Alfalfa inoculated rhizobium

NN:

Alfalfa not-inoculated rhizobium

ORF:

Open reading frame

qRT-PCR:

Quantitative real-time PCR

RACE:

Rapid amplification of cDNA ends

RWC:

Relative water content

SOD:

Superoxide dismutase

Vx:

Violaxanthin

ZEP:

Zeaxanthin epoxidase

Zx:

Zeaxanthin

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Acknowledgments

This work was supported by the Project of National Natural Science Foundation of China (31372357, 31272490), the National Key Technology R&D Program in the 12th 5-year plan of China (2011BAD17B05) and the major project for Tibetan forage industry (Z2014C02N02).

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Authors and Affiliations

  1. Department of Grassland Science, College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, Shaanxi, China

    Zhiqiang Zhang, Yafang Wang, Leqin Chang, Tong Zhang, Jie An, Yushi Liu, Yuman Cao, Xia Zhao, Xuyang Sha, Tianming Hu & Peizhi Yang

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  1. Zhiqiang Zhang
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  2. Yafang Wang
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  3. Leqin Chang
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  4. Tong Zhang
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  5. Jie An
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  6. Yushi Liu
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  8. Xia Zhao
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  10. Tianming Hu
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  11. Peizhi Yang
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Correspondence to Tianming Hu or Peizhi Yang.

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Author contribution statement

Z.Q. Zhang performed the experiment, analyzed data and wrote the manuscript; Y.F. Wang cloned and analyzed the gene; L.Q. Chang, T. Zhang and J. An sampled the material and determined physiological indexes; Y.S. Liu and Y.M. Cao performed transgenic tobacco experiment; X. Zhao and X.Y. Sha performed qRT-PCR experiment; P.Z. Yang and T.M. Hu provided ideas, designed the research, and edited the manuscript; all authors read and approved the final manuscript.

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The authors declare that they have no conflict of interest.

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Communicated by L. Peña.

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Zhang, Z., Wang, Y., Chang, L. et al. MsZEP, a novel zeaxanthin epoxidase gene from alfalfa (Medicago sativa), confers drought and salt tolerance in transgenic tobacco. Plant Cell Rep 35, 439–453 (2016). https://doi.org/10.1007/s00299-015-1895-5

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