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Soybean late embryogenesis abundant protein GmLEA4 interacts with GmCaM1, enhancing seed vigor in transgenic Arabidopsis under high temperature and humidity stress

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Abstract

Calmodulin (CaM) as a ubiquitous Ca2+-binding protein is involved in multiple signaling processes expressed in all the eukaryotic cells, while the late embryogenesis abundant (LEA) proteins are a large family of hydrophilic proteins that accumulate in developing seeds to ensure protection against water deficit stress. In our previous study, GmLEA4 was screened as a candidate GmCaM1-interacting protein by yeast two-hybrid assay while GmCaM1 was found to be highly related to the formation of seed vigor under high temperature and humidity (HTH) stress in soybean. In the present study, GmLEA4 was localized on the cell membrane and nucleus. The interaction between GmCaM1 and GmLEA4 was further confirmed by yeast two-hybrid assay, bimolecular fluorescence complementation (BiFC) and GST pull down. GmLEA4 and GmCaM1 showed higher expression levels in seeds than in other tissues and were involved in response to HTH stress. The overexpression of GmCaM1 and GmLEA4 in Arabidopsis raised the ROS scavenging ability in seeds and improved seed vitality under HTH stress. Our results indicated that GmLEA4 interacts with GmCaM1, maybe participating in seed vigor formation under HTH stress in soybean.

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Abbreviations

BiFC:

Bimolecular fluorescence complementation

CaM:

Calmodulin

CV:

Cultivar

LEA4:

Late embryogenesis abundant 4

GFP:

Green fluorescent protein

HTH:

High temperature and humidity

HT:

High temperature

HH:

High humidity

ORF:

Open reading frame

PCR:

Polymerase chain reaction

qRT-PCR:

Quantitative reverse transcription PCR

ROS:

Reactive oxygen species

YFP:

Yellow fluorescent protein

WT:

Wild type

CAT:

Catalase

SOD:

Superoxide

POD:

Peroxidase

MDA:

Malondialdehyde

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Acknowledgements

The authors gratefully acknowledge the partial financial support from the project supported by the National Natural Science Foundation of China (31671772, and 31971996) and the Ministry of Science and Technology of China (2018YFD0100905) for this research.

Funding

Funding was provided National Natural Science Foundation of China (Grant nos. 31671772, 31971996), Ministry of Science and Technology of the People's Republic of China (Grant no. 2018YFD0100905).

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Authors and Affiliations

  1. State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing, 210095, China

    Yingzi Shen, Jiaping Wei, Yali Zhou, Yajing Zhu, Sushuang Liu, Yilong Wang & Hao Ma

  2. Department of Life Science and Health, Huzhou University, Huzhou, 313000, China

    Sushuang Liu

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  1. Yingzi Shen
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  2. Jiaping Wei
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Contributions

The work presented here was carried out in collaboration among all the authors. YS carried out most of the laboratory experiments and prepared preliminary manuscript; JW, YZ, YJZ, SL, and YW carried out the seed germination and enzyme activities assay; HM designed the experiments and corrected the manuscript. All the authors have read and approved the final manuscript.

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Correspondence to Hao Ma.

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Communicated by Zhong-Hua Chen.

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10725_2022_930_MOESM1_ESM.jpg

Supplementary file1 Supplemental Fig S1. Effects of HT, HH and HTH stresses on plants of single gene transformed Arabidopsis lines and WT plants. a, Illustration of 3-week-old WT and single gene transformed Arabidopsis lines (C3 and L6) under the control [23 °C/20 °C, 12 h/12 h (light/dark) and 70% RH], HT stress [40 °C/20 °C, 12 h/12 h (light/dark) and 70% RH], HH stress [23 °C/20 °C, 12 h/12 h (light/dark) and 100% RH] and HTH stress [40 °C/20 °C, 12 h/12 h (light/dark) and 100% RH] for 2 d, respectively. b, SOD activity, CAT activity, POD activity and MDA content in leaves of GmCaM1 transformed Arabidopsis lines (C1, C2 and C3), respectively; c, SOD activity, CAT activity, POD activity and MDA content in leaves of GmLEA4 transformed Arabidopsis lines (L4, L5 and L6), respectively; d, e, Staining of H2O2 in the leaves of transgenic Arabidopsis lines (C3 and L6) and WT plants under control, HT, HH and HTH stresses, respectively. Values shown are mean ± SD from three biological replicates (**p < 0.01). (JPG 409 kb)

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Shen, Y., Wei, J., Zhou, Y. et al. Soybean late embryogenesis abundant protein GmLEA4 interacts with GmCaM1, enhancing seed vigor in transgenic Arabidopsis under high temperature and humidity stress. Plant Growth Regul 99, 583–595 (2023). https://doi.org/10.1007/s10725-022-00930-w

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