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Overexpression of an alfalfa (Medicago sativa) gene, MsDUF, negatively impacted seed germination and response to osmotic stress in transgenic tobacco

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Abstract

Many stress-responsive genes have been identified in alfalfa (Medicago sativa L.). The function of these genes, however, are mostly not understood. We reported previously a novel stress-responsive gene, MsDUF, from alfalfa that was up-regulated under drought stress. In the present study, we examined its function by overexpressing the gene in Nicotiana tabacum. We found that overexpression of MsDUF reduced seed vigor and germination percentage under normal conditions or osmotic stress. The reduced seed vigor and germination was associated with an increased ABA content in the overexpressor seeds. Further analysis revealed that overexpression of MsDUF resulted in up-regulation of transcript levels of ABA biosynthesis genes (ZEP, NCED1 and NCED6) in the seeds. Compared with wild type, MsDUF-overexpression seedlings displayed significantly lower chlorophyll content and reduced soluble sugar content under normal conditions. MDA content was significantly higher in MsDUF-overexpressors compared to wild type under ABA treatment, while soluble sugar content and peroxidase activities were significantly lower in MsDUF-overexpressors. Our results suggest that MsDUF may act as a negative regulator in controlling seed vigor and responses to osmotic stress in plants.

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Abbreviations

ABA:

Abscisic acid

GA:

Gibberellic acid

GI:

Germination index

GP:

Germination percentage

SVI:

Seed vigor index

PCR:

Polymerase chain reaction

qRT-PCR:

Quantitative real-time PCR

DUF:

Domains of unknown function

WT:

Wild type

OS:

Osmotic stress

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Acknowledgements

This work was supported by the Project of National Natural Science Foundation of China (Grant Nos. 31572456, 31601987), the major Project for Tibetan forage industry (2016), and China Agriculture Research System (Grant No. CARS-35-40).

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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

      Yafang Wang, Zhiqiang Zhang, Houmei Liu, Yunru An, Bo Han, Leqin Chang, Tianming Hu & Peizhi Yang

    2. College of Grassland Resources and Environment, Inner Mongolia Agricultural University, Hohhot, 010011, Inner Mongolia, China

      Zhiqiang Zhang

    3. Department of Biology and Microbiology, South Dakota State University, Brookings, SD, 57007, USA

      Yajun Wu

    4. College of Animal Science and Technology, Yunnan Agriculture University, Kunming, 650201, China

      Bo Han

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    1. Yafang Wang
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    Contributions

    YW and ZZ performed the whole experiment. YW and YW analyzed the data and wrote the manuscript. BH provided the transgenic tobacco seeds. HL, YA and LC participated in the gene expression, antioxidant enzyme and soluble sugar measurement. PY and TH proposed the ideas, designed the experiment, and edited the manuscript. All authors read and approved the final manuscript.

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    Correspondence to Tianming Hu or Peizhi Yang.

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    Additional information

    Communicated by Sergio J. Ochatt.

    Yafang Wang and Zhiqiang Zhang have contributed equally to this work.

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    Wang, Y., Zhang, Z., Liu, H. et al. Overexpression of an alfalfa (Medicago sativa) gene, MsDUF, negatively impacted seed germination and response to osmotic stress in transgenic tobacco. Plant Cell Tiss Organ Cult 132, 525–534 (2018). https://doi.org/10.1007/s11240-017-1348-7

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