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Discovery of microRNAs and transcript targets related to witches’ broom disease in Paulownia fortunei by high-throughput sequencing and degradome approach

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Abstract

Paulownia witches’ broom (PaWB) caused by the phytoplasma is a devastating disease of Paulownia trees. It has caused heavy yield losses to Paulownia production worldwide. However, knowledge of the transcriptional and post-transcriptional regulation of gene expression by microRNAs (miRNAs), especially miRNAs responsive to PaWB disease stress, is still rudimentary. In this study, to identify miRNAs and their transcript targets that are responsive to PaWB disease stress, six sequencing libraries were constructed from healthy (PF), PaWB-infected (PFI), and PaWB-infected, 20 mg L−1 methyl methane sulfonate-treated (PFI20) P. fortunei seedlings. As a result, 95 conserved miRNAs belonging to 18 miRNA families, as well as 122 potential novel miRNAs, were identified. Most of them were found to be a response to PaWB disease-induced stress, and the expression levels of these miRNAs were validated by quantitative real-time PCR analysis. The study simultaneously identified 109 target genes from the P. fortunei for 14 conserved miRNA families and 24 novel miRNAs by degradome sequencing. Furthermore, the functions of the miRNA targets were annotated based on Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analysis. The results presented here provide the groundwork for further analysis of miRNAs and target genes responsive to the PaWB disease stress, and could be also useful for addressing new questions to better understand the mechanisms of plant infection by phytoplasma in the future.

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Acknowledgments

This study was funded by the National Natural Science Foundation of China (Grant Nos. 30271082, 30571496, U1204309), by the Outstanding Talents Project of Henan Province (Grant No. 122101110700), by the Transformation Project of the National Agricultural Scientific and Technological Achievement of China (Grant No. 2012GB2D000271), and by the Science and Technology Innovation Team Project of Zhengzhou City, China (Grant No. 121PCXTD515).

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Authors and Affiliations

  1. Institute of Paulownia, Henan Agricultural University, 95 Wenhua Road, Jinsui District, Zhengzhou, 450002, Henan, People’s Republic of China

    Suyan Niu, Guoqiang Fan, Minjie Deng, Zhenli Zhao, Enkai Xu & Lin Cao

  2. College of Forestry, Henan Agricultural University, 95 Wenhua Road, Jinsui District, Zhengzhou, 450002, Henan, People’s Republic of China

    Suyan Niu, Guoqiang Fan, Minjie Deng, Zhenli Zhao, Enkai Xu & Lin Cao

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  1. Suyan Niu
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  2. Guoqiang Fan
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  3. Minjie Deng
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  4. Zhenli Zhao
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  5. Enkai Xu
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Correspondence to Guoqiang Fan.

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Communicated by S. Hohmann.

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Niu, S., Fan, G., Deng, M. et al. Discovery of microRNAs and transcript targets related to witches’ broom disease in Paulownia fortunei by high-throughput sequencing and degradome approach. Mol Genet Genomics 291, 181–191 (2016). https://doi.org/10.1007/s00438-015-1102-y

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