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Constitutive expression of a group 3 LEA protein from Medicago falcata (MfLEA3) increases cold and drought tolerance in transgenic tobacco

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MfLEA3 is involved in protection of catalase activity and confers multiple abiotic stress tolerance.

Abstract

Late embryogenesis abundant (LEA) proteins are involved in plant growth, development and abiotic stress tolerance. A member of group 3 LEA proteins from Medicago sativa subsp. falcata (L.) Arcang, MfLEA3, was investigated in the study. MfLEA3 transcript was induced in response to cold, dehydration, and abscisic acid (ABA), while the cold-induced transcript of MfLEA3 was blocked by pretreatment with inhibitor of ABA synthesis. Constitutive expression of MfLEA3 led to enhanced tolerance to cold, drought, and high-light stress in transgenic tobacco plants. Compared to accumulated reactive oxygen species (ROS) in the wild-type in response to treatments with low temperature, drought, and high light, ROS were not accumulated in transgenic plants. Superoxide dismutase, catalase (CAT), and ascorbate-peroxidase activities were increased in all plants after treatments with the above stresses, while higher CAT activity was maintained in transgenic plants compared with wild-type. However, transcript level of CAT-encoding genes including CAT1, CAT2, and CAT3 showed no significant difference between transgenic plants and wild-type, indicating that the higher CAT activity was not associated with its gene expression. ABA sensitivity and transcripts of several ABA and stress-responsive genes showed no difference between transgenic plant and wild-type, indicating that ABA signaling was not affected by constitutive expression of MfLEA3. The results suggest that MfLEA3 may be involved in the protection of CAT activity and confers multiple abiotic stress tolerance.

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Acknowledgements

The research was supported by the National Natural Science Foundation of China (31672481, 31701961), Chinese Agriculture Research System-Green Manure (CARS-22-G-04), and the Natural Science Foundation of Jiangsu Province (Grant no. BK20160728).

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Author notes

  1. Haifan Shi and Xueying He have contributed equally to the work.

Authors and Affiliations

  1. College of Grassland Science, Nanjing Agricultural University, Nanjing, 210095, China

    Haifan Shi & Zhenfei Guo

  2. State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangdong Engineering Research Center for Grassland Science, College of Life Sciences, South China Agricultural University, Guangzhou, 510642, China

    Xueying He, Yujuan Zhao & Shaoyun Lu

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  1. Haifan Shi
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Contributions

HS, XH, and JZ performed research and conducted data analyses. ZG and SL designed the research. HS and ZG wrote the manuscript.

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Correspondence to Zhenfei Guo.

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Communicated by Prakash P. Kumar.

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Supplementary file1 (PDF 86 kb)

Fig. S1.

Superoxide dismutase (SOD, a) and ascorbate-peroxidase (APX, b) activities in transgenic plants in comparison with the wild-type (WT) in response to abiotic stresses (PDF 1037 kb)

Fig. S2.

Analysis of transcript levels of NtDREB1 (a), NtDREB2 (b), NtDREB3 (c), NtDREB4 (d), NtERD10B (e), NtCOR15a (f), NtP5CS (g) in the transgenic tobacco plants in comparison with the wild-type (WT). The relative expression level was normalized to that of MfACTIN or MtACTIN. The same letter above the columns indicates no significant difference at P < 0.05. (PDF 3256 kb)

Fig. S3.

Analysis of ABA sensitivity in MfLEA3 transgenic plants in comparison with the wild-type. Seedling phenotypes (a) and shoot fresh weight (b) were recorded after 1-month of growing on 1/2 MS containing different concentrations of ABA. Roots phenotype (c) and root length (d) were recorded after ten-day of growing on 1/2 MS containing different concentrations of ABA. Three independent experiments were executed. Bar = 1 cm. Means of three biological replicates and standard errors are presented; the same letter above the column indicates no significant difference at P < 0.05 (PDF 318 kb)

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Shi, H., He, X., Zhao, Y. et al. Constitutive expression of a group 3 LEA protein from Medicago falcata (MfLEA3) increases cold and drought tolerance in transgenic tobacco. Plant Cell Rep 39, 851–860 (2020). https://doi.org/10.1007/s00299-020-02534-y

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