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The few-celled microspore embryo transcriptome of Brassica napus

ABSTRACT: Haploid embryos can be induced from cultured immature pollen following a stress treatment. In Brassica napus (B. napus), application of the histone/lysine deacetylase (HDAC/KDAC) inhibitor trichostatin A (TSA) to pollen cultures enhances the production of differentiated embryos and embryogenic callus when applied together with heat stress (Li et al., 2014). To identify genes associated with the induction of B. napus haploid embryogenesis, we employed fluorescence-activated cell sorting on microspore cultures of the LEAFY COTYLEDON1 embryo reporter line (LEC1:LEC1-GFP, Li et al., 2014) to select for embryogenic structures developing in culture, followed by RNA-sequencing. Our transcriptome analysis of the 3-day-old enriched embryo populations and in vitro cultured pollen revealed different regulatory pathways underlying the early development of embryogenic structures. Comparing FACS-sorted embryogenic transcriptomes from cultures treated with heat stress combined with either a low or a high concentration of TSA allowed us to assign transcription profiles to embryogenic structures that have a low or high potential to develop into histodifferentiated embryos.

ORGANISM(S): Brassica napus

PROVIDER: GSE207576 | GEO | 2025/08/18

REPOSITORIES: GEO

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Project description:Somatic embryogenesis (SE) exemplifies the unique developmental plasticity of plant cells. The regulatory processes, including epigenetic modifications controlling embryogenic reprogramming of cell transcriptome, have just started to be revealed. To identify the genes of histone acetylation-regulated expression in SE, we analyzed global transcriptomes of Arabidopsis explants undergoing embryogenic induction in response to treatment with histone deacetylase inhibitor, trichostatin A (TSA). The TSA-induced and auxin (2,4-dichlorophenoxyacetic acid; 2,4-D)-induced transcriptomes were compared. RNA-seq results revealed the similarities of the TSA- and auxin-induced transcriptomic responses that involve extensive deregulation, mostly repression, of the majority of genes. Within the differentially expressed genes (DEGs), we identified the master regulators (transcription factors TF) of SE, genes involved in biosynthesis, signaling, and polar transport of auxin and NITRILASE-encoding genes of the function in indole-3-acetic acid (IAA) biosynthesis. TSA-upregulated TF genes of essential functions in auxin-induced SE, included LEC1/LEC2, FUS3, AGL15, MYB118, PHB, PHV, PLTs, and WUS/WOXs. The TSA-induced transcriptome revealed also extensive upregulation of stress-related genes, including those related to stress hormone biosynthesis. In line with transcriptomic data, TSA-induced explants accumulated salicylic acid (SA) and abscisic acid (ABA), suggesting the role of histone acetylation (Hac) in regulating stress hormone-related responses during SE induction. Since mostly the adaxial side of cotyledon explant contributes to SE induction, we also identified organ polarity-related genes responding to TSA treatment, including AIL7/PLT7, RGE1, LBD18, 40, HB32, CBF1, and ULT2. Analysis of the relevant mutants supported the role of polarity-related genes in SE induction. The study results provide a step forward in deciphering the epigenetic network controlling embryogenic transition in somatic cells of plants.

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The few-celled microspore embryo transcriptome of Brassica napus

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