The long non-coding RNA MSTRG.32189-PcmiR399b-PcUBC24 module regulates phosphate accumulation and disease resistance to Botryosphaeria dothidea in pear

Abstract

Pear ring rot disease (Botryosphaeria dothidea) is a significant threat to the healthy development of the pear industry. Recent research has identified the functional role of long non-coding RNAs (lncRNAs) in various biological processes of plants. The role of lncRNAs in the pear defense response remains unknown. In this study, transcriptome sequencing was used to analyze lncRNAs in pear stem infected with B. dothidea. It identified 3555 lncRNAs, of which 286 were significantly differentially expressed. GO and KEGG analyses showed that cis- and trans-regulated target genes were enriched in multiple disease resistance-related pathways. More specifically, MSTRG.32189, predicted as an endogenous target mimic (eTM), was significantly down-regulated in response to B. dothidea infection, and was confirmed to inhibit the cleavage effect of PcmiR399b on PcUBC24. OE-MSTRG.32189 transgenic Arabidopsis exhibited lower Pi content and weaker disease resistance to Botrytis cinerea compared with wild type. In pear callus, overexpression of MSTRG.32189 negatively regulated PcmiR399b, which decreased Pi content and reduced disease resistance. Overexpressing PcmiR399b in pear callus exhibited the opposite effects compared with OE-MSTRG.32189. Overexpression and knockout of PcUBC24 further clarified that PcUBC24 negatively regulates Pi content and disease resistance to B. dothidea infection. Furthermore, the ROS levels and expressions of disease resistance pathway-related genes were regulated by the MSTRG.32189-PcmiR399b-PcUBC24 module in transgenic pear callus, which contributed to disease resistance. Overall, our results demonstrated the role of lncRNAs in the pear defense response, revealing that the MSTRG.32189-PcmiR399b-PcUBC24 module regulates phosphate accumulation and disease resistance to B. dothidea infection in pear.

Acknowledgements

This work was financially supported by grants from the National Natural Science Foundation of China (grant 31972321) and the Earmarked Fund for China Agriculture Research System (grant CARS-28). We thank Prof. Chunying Kang and Prof. Qiang Xu from Huazhong Agricultural University for providing vectors of pK7WG2D and pKSE401.

Author contributions

L.W. supervised the research. L.W. and Y.Y. conceived the project and designed the experiments. Y.Y. and S.L. conducted most of the experiments. Y.Y. analyzed the data and wrote the article. Y.H., X.Z., and Y.L. contributed to pear callus transformation. L.W. revised the manuscript. X.H., N.H., and G.W. provided suggestions on the manuscript.

Data availability

Sequence data generated during this work can be found in the GenBank database. Sequences of MSTRG.32189 and PcUBC24 CDS have been uploaded to GenBank under accession numbers PP104288 and PP104289, respectively. RNA-seq raw reads used for pear lncRNA analysis in this study have been uploaded to NCBI under accession number PRJNA1072809.

Conflict of interest statement

L.W., Y.Y., S.L., Y.H., N.H., and G.W. have filed a provisional patent application related to this work.

Supplementary data

Supplementary data are available at Horticulture Research online.

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