Identification of genes related to Paulownia witches' broom by AFLP and MSAP.

Xibing Cao,Yanpeng Dong,Minjie Deng,Zhenli Zhao,Guoqiang Fan

doi:10.3390/ijms150814669

Xibing Cao, Yanpeng Dong + Show 3 more

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https://doi.org/10.3390/ijms150814669

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Abstract

DNA methylation is believed to play important roles in regulating gene expression in plant growth and development. Paulownia witches’ broom (PaWB) infection has been reported to be related to gene expression changes in paulownia plantlets. To determine whether DNA methylation is associated with gene expression changes in response to phytoplasma, we investigated variations in genomic DNA sequence and methylation in PaWB plantlets treated with methyl methane sulfonate (MMS) using amplified fragment length polymorphism (AFLP) and methylation-sensitive amplification polymorphism (MSAP) techniques, respectively. The results indicated that PaWB seedings recovered a normal morphology after treatment with more than 15 mg·L−1 MMS. PaWB infection did not cause changes of the paulownia DNA sequence at the AFLP level; However, DNA methylation levels and patterns were altered. Quantitative real-time PCR (qRT-PCR) showed that three of the methylated genes were up-regulated and three were down-regulated in the MMS-treated PaWB plantlets that had regained healthy morphology. These six genes might be involved in transcriptional regulation, plant defense, signal transduction and energy. The possible roles of these genes in PaWB are discussed. The results showed that changes of DNA methylation altered gene expression levels, and that MSAP might help identify genes related to PaWB.

Highlights

Epigenetic inheritance refers to phenotypic modifications in the absence of changes in the DNA sequence, and DNA methylation is one of the main epigenetic modifications
The methyl methane sulfonate (MMS)-treated Paulownia witches’ broom (PaWB) plantlets regained a healthy morphology
Both the PaWB plantlets treated with 30 and 45 mg·L−1 MMS changed into healthy morphology, while only very tiny axillary buds were found in the PaWB plantlets treated with mg·L−1 MMS (Table S1)

Summary

Introduction

Epigenetic inheritance refers to phenotypic modifications in the absence of changes in the DNA sequence, and DNA methylation is one of the main epigenetic modifications. DNA methylation is associated with many biological processes, including transcriptional silencing of transgenes, regulation of gene expression, and genomic imprinting [2,3,4], and has become the focus of many studies. It was reported that DNA methylation regulated gene expression under biotic stress; for example, DNA methylation was found to induce post-transcriptional gene silencing in tomato infected with the Tomato yellow leaf curl Sardinia virus [5], and DNA methylation changed the transcript levels of genes in response to Burkholderia phytofirmans strain PsJN in infected potato plants [6]. Paulownia is one of the fastest growing shade trees in the world The wood of this tree possesses several valuable characteristics, such as decay-resistance, strength, specific gravity, and fiber length, making it useful for furniture, pulp, and paper production. It has a prosperous root system, which makes it a good candidate for growing together with crops for efficient use of resources

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