MdAIL5 overexpression promotes apple adventitious shoot regeneration by regulating hormone signaling and activating the expression of shoot development-related genes

Abstract

Adventitious shoot (AS) regeneration is a significant factor in the genetic transformation of horticultural plants. It is also a noteworthy approach to their vegetative propagation. AS regeneration remains highly dependent on the genotype or maturity of explants. We here found that the AS regeneration abilities of apple leaves were positively correlated with MdAIL5 expression. MdAIL5 overexpression dramatically increased AS regeneration efficiency. Notably, MdAIL5 overexpression could restore the AS formation ability of explants to a certain extent, which was lost with an increase in maturity. Endogenous hormone detection revealed that MdAIL5 overexpression changed the contents of auxin, cytokinin (CK), and other hormones in apple leaves. Transcriptome analysis revealed that many genes related to auxin, CK, and brassinolide signaling pathways were significantly and differentially expressed between MdAIL5-overexpressing transgenic apple and wild-type apple plants. Yeast one-hybrid assays, the electrophoretic mobility shift assay, and the dual-luciferase reporter assay revealed that MdAIL5 directly binds to MdARF9 and MdHB14 promoters and positively affects their expression. We here established a model of MdAIL5 regulating AS formation, which acts as a theoretical basis for facilitating genotype- or explant maturity-independent AS regeneration in the future.

Acknowledgements

This work was financially supported by the China Agriculture Research System (Grant No. CARS-27), the National Natural Science Foundation of China (Grant No. 32202463), and the Agricultural Science and Technology Innovation Program (Grant No. CAAS-ASTIP-2021-RIP-02). We would like to thank Prof. Zhihong Zhang (Shenyang Agricultural University, Shenyang, Liaoning) for providing tissue-cultured ‘GL-3’ plants. We would like to thank Prof. Jialong Yao (New Zealand Institute for Plant and Food Research Limited, Mount Albert Research Centre, Auckland, New Zealand) for providing the pGreenII-0800-LUC plasmid. We would like to thank Prof. Wei Li (China Agricultural University, Beijing) for providing pJG4-5 and pLacZi plasmids.

Author contributions

C.X.Z., X.L.H., P.H.C., L.Y.Z. and K.L. conceived and planned the research. C.X.Z. and X.L.H. supervised the research. K.L., Z.L.L., A.Y., J.D.Y., P.H.C., X.L.H., G.P.Y. and C.X.Z. performed the experiments. K.L. and X.L.H. conducted data analysis. K.L. wrote the manuscript. C.X.Z. and X.L.H. edited the manuscript. All authors have read and agreed to the final version of the manuscript.

Data availability

The data used to support the study findings are included within the article.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary data

Supplementary data is available at Horticulture Research online.

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