5-Aminolevulinic acid improves cold resistance through regulation of SlMYB4/SlMYB88-SlGSTU43 module to scavenge reactive oxygen species in tomato

Abstract

Cold stress severely affects the growth and quality of tomato. 5-Aminolevulinic acid (ALA) can effectively improve tomato’s cold stress tolerance. In this study, a tomato glutathione S-transferase gene, SlGSTU43, was identified . Results showed that ALA strongly induced the expression of SlGSTU43 under cold stress . SlGSTU43-overexpressing lines showed increased resistance to cold stress through an enhanced ability to scavenge reactive oxygen species. On the contrary, slgstu43 mutant lines were sensitive to cold stress, and ALA did not improve their cold stress tolerance. Thus, SlGSTU43 is a key gene in the process of ALA improving tomato cold tolerance. Through yeast library screening, SlMYB4 and SlMYB88 were preliminarily identified as transcription factors that bind to the SlGSTU43 promoter. Electrophoretic mobility shift, yeast one-hybrid, dual luciferase, and chromatin immunoprecipitation assays experiments verified that SlMYB4 and SlMYB88 can bind to the SlGSTU43 promoter. Further experiments showed that SlMYB4 and SlMYB88 are involved in the process of ALA-improving tomato’s cold stress tolerance and they positively regulate the expression of SlGSTU43. The findings provide new insights into the mechanism by which ALA improves cold stress tolerance. SlGSTU43, as a valuable gene, could be added to the cold-responsive gene repository. Subsequently, it could be used in genetic engineering to enhance the cold tolerance of tomato.

Acknowledgements

We are grateful to Professor Mingjun Li (College of Horticulture, Northwest A&F University) for his help in paper writing, Miss Beibei He and Miss Yangyang Yuan (Horticulture Science Research Center, Northwest A&F University) for providing professional technical assistance with Fluorescence Microscope analysis. This work was supported by the National Key R&D Program of China (2019YFD1001902), the China Agriculture Research System (CARS-23-D06), and the Scientific & Technological Innovative Research Team of Shaanxi Province (2021TD-34).

Author contributions

Z.Z., T.L., and X.H. conceived and designed the experiments. Z.Z., L.Y, J.D., Y.Z., F.Z., Y.W., Y.G., Y.L., Y.W., T.L., Y.D., T.L., and X.H. performed experiments and analysed the data. Z.Z., T.L. and X.H. wrote the manuscript.

Data availability

The authors declare that all the data necessary to support the study's conclusions are included in the paper and the supplementary materials, or can be obtained upon request from the corresponding author.

Conflict of interest statement

The authors declare no competing commercial or financial interest.

Supplementary Data

Supplementary data is available at Horticulture Research online.

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