Multifaceted roles of LhWRKY44 in promoting anthocyanin accumulation in Asiatic hybrid lilies (Lilium spp.)

Abstract

The Asiatic hybrid lily (Lilium spp.) is a horticultural crop with high commercial value and diverse anthocyanin pigmentation patterns. However, the regulatory mechanism underlying lily flower color has been largely unexplored. Here, we identified a WRKY transcription factor from lily tepals, LhWRKY44, whose expression was closely associated with anthocyanin accumulation. Functional verification indicated that LhWRKY44 positively regulated anthocyanin accumulation. LhWRKY44 physically interacted with LhMYBSPLATTER and directly bound to the LhMYBSPLATTER promoter, which enhanced the effect of the LhMYBSPLATTER-LhbHLH2 MBW complex activator on anthocyanin accumulation. Moreover, EMSA and dual-luciferase assays revealed that LhWRKY44 activated and bound to the promoters of gene LhF3H and the intracellular anthocyanin-related glutathione S-transferase gene LhGST. Interestingly, our further results showed that LhWRKY44 participated in light and drought-induced anthocyanin accumulation, and improved the drought tolerance in lily via activating stress-related genes. These results generated a multifaceted regulatory mechanism for the LhWRKY44-mediated enhancement by the environmental signal pathway of anthocyanin accumulation and expanded our understanding of the WRKY-mediated transcriptional regulatory hierarchy modulating anthocyanin accumulation in Asiatic hybrid lilies.

Acknowledgements

The authors thank the editor and the anonymous reviewers for their efforts to improve the manuscript. This study was supported by the National Natural Science Foundation of China (32172624, 32172612, 31672196) and the Programs for National Key R & D Plan (2019YFD1001002). We thank Prof. Nan Ma, Prof. Xia Cui, Prof. Allen A C, and Prof. Yuanwen Teng for the providing experimental plasmids and guidance, and we thank Prof. Ji Tian for providing apple calli materials.

Author contributions

J.M. and M.B. conceived and designed the experiments. M.B. and J.M. wrote the paper. M.B. performed the experiments. R.L., J.W., Y.Q., X.L., and Y.C. provided assistance. G.H., Y.Y., P.Y., and L.X. helped to revise the article. All authors read and approved the manuscript for submission.

Data availability

All relevant data in this study are provided in the article and its supplementary data files. The gene accession numbers used can be found in National Center for Biotechnology Information (NCBI) database, and they are listed in (Table S5, see online supplementary material).

Conflict of interest statement

The authors declare no competing interests.

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