Abstract
Heat stress can induce the cultured microspores into embryogenesis. In this study, whole genome bisulphite sequencing was employed to study global DNA methylation variations after short-term heat shock (STHS) treatments in cultured microspores of Brassica napus cv. Topas. Our results indicated that treatment on cultured Topas microspores at 32 °C for 6 h triggered DNA hypomethylation, particularly in the CG and CHG contexts. And the total number of T32 (Topas 32 °C for 6 h) vs. T0 (Topas 0 h) differentially methylated region-related genes (DRGs) was approximately two-fold higher than that of T18 (Topas 18 °C for 6 h) vs. T0 DRGs, which suggested that 32 °C might be a more intense external stimulus than 18 °C resulting in more changes in the DNA methylation status of cultured microspores. Additionally, 32 °C treatment for 6 h led to increased CHG differential methylations of transposons (DMTs), which were mainly constituted by overlaps between the hypomethylated differentially methylated regions (hypo-DMRs) and transposon elements (TEs). Further analysis demonstrated that the DRGs and their paralogs exhibited differential methylated/demethylated patterns. To summarize, the present study is the first methylome analysis of cultured microspores in response to STHS and may provide valuable information on the roles of DNA methylation in heat response.
Highlights
IntroductionHeat pre-treatment is one of the significant stresses for microspore embryogenesis (ME) induction[7,15], few studies have examined whether DNA methylation changes in cultured microspores after short-term heat shock (STHS) treatment
Involved in the control of cell growth during heat stress in tobacco BY-2 cells[2]
Our results showed that the T32 vs. T0 and T32 vs. T18 DMRs were mainly comprised of hypo-DMRs rather than hyper-DMRs (Fig. 3C and Supplementary Data S1)
Summary
Heat pre-treatment is one of the significant stresses for ME induction[7,15], few studies have examined whether DNA methylation changes in cultured microspores after STHS treatment. We tried to employ genome-wide bisulphite sequencing (GWBS) to decipher global DNA methylation variations after 32 °C and 18 °C treatments for 6 h in cultured microspores of B. napus cv. Our results revealed that 32 °C heat treatment for 6 h was sufficient to induce global DNA hypomethylation in cultured Topas microspores. 32 °C might be a more intense external stimulus than 18 °C generating more changes in the DNA methylation status of cultured microspores. The present study is the first methylome analysis of cultured microspores in response to STHS and may provide valuable information on the roles of DNA methylation in heat response
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