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Integrative analysis of DNA methylation, mRNAs, and small RNAs during maize embryo dedifferentiation

ABSTRACT: Background: Maize (Zea Mays) is an important model crop for transgenic studies. However, genetic transformation of maize requires embryonic calli derived from immature embryo, and the impact of utilizing tissue culture methods on the maize epigenome is poorly understood. Here, we generated whole-genome MeDIP-seq data examining DNA methylation in dedifferentiated and normal immature maize embryos. Results: We observed that most of the dedifferentiated embryos exhibited a methylation increase compared to normal embryos. Increased methylation at promoters was associated with down-regulated protein-coding gene expression; however, the correlation was not strong. Analysis of the callus and immature embryos indicated that the methylation increase was induced during induction of embryonic callus, suggesting phenotypic consequences may be caused by perturbations in genomic DNA methylation levels. The correlation between the 21-24nt small RNAs and DNA methylation regions were investigated but only a statistically significant correlation for 24nt small RNAs was observed. Conclusions: These data extend the significance of epigenetic changes during maize embryo callus formation, and the methylation changes might explain some of the observed embryonic callus variation in callus formation.

ORGANISM(S): Zea mays

PROVIDER: GSE84455 | GEO | 2017/05/10

SECONDARY ACCESSION(S): PRJNA329226

REPOSITORIES: GEO

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Proteome of Douglas-fir: a resource for genomic research

Project description:First Douglas fir proteomes by nLC-MS/MS from 12 different organs : root, stem, xylem, needle, bud, female and male flowers, immature and mature seed, immature and mature somatic embryos and callus.

2023-10-24 | PXD037815 | Pride

Differential small RNA expression upon induction and establishment of Tuxpeño maize embryogenic callus cultures

Project description:Purpose: Maize somatic embryogenesis is usually required to achieve genetic transformation and represents an important alternative in plant development. Although many embryogenesis-related genes have been studied in this model, the molecular mechanisms underlying cell dedifferentiation and further plant regeneration are not completely understood. Methods: Immature embryos smRNA profiles of 15-day-after-pollination (IE) and Embryogenic Callus from one (C1), four (C4), and ten months (C10) were generated by deep sequencing, using Illumina GAIIx. The sequence reads that passed quality filters were analyzed with two methods: Bowtie 1.1.2 and ShortStack 3.4. qRT–PCR validation for selected miRNAs was performed using SYBR Green assays. Results: We used high throughput sequencing to explore the sRNA populations during maize embryogenic callus induction and established subcultures from the Mexican cultivar VS-535, Tuxpeño landrace. We detected readjustments in 24 nt and 21-22 nt sRNA populations during the embryogenic callus establishment and maintenance. miRNAs related to stress response substantially increased upon callus proliferation establishment, correlating with a reduction in some of their target levels. On the other hand, while 24 nt-long hc-siRNAs derived from transposable retroelements transiently decreased in abundance during the embryogenic callus establishment, a population of 22 nt- hc-siRNAs increased. This was accompanied by reduction in transposon expression in the established callus subcultures. Conclusions: Stress- and development-related miRNAs are highly expressed upon maize EC callus induction and during maintenance subcultures, while miRNAs involved in hormone response only transiently increase during induction. The establishment of proliferative maize embryogenic callus is accompanied by important readjustments in the length of hc-siRNAs mapping to LTR retrotransposons, and their expression regulation.

2020-04-21 | GSE98055 | GEO

Transcriptome sequencing analysis of sorghum callus with various regeneration capacities

Project description:The applications of plant callus regeneration has been widely spreaded in agricultural improvement. By using immature sorghum embryos as explants, progress in successful genetic transformation has been made in sorghum. However, the underlying mechanism of callus differentiation is still largely unknown in sorghum. Here, we described three types of callus with different abilities of redifferentiation (Callus I-III), undergoing distinct induction from immature embryo in the variety of Hiro-1. In comparison to the non-embryonic Callus III who lost the ability of regeneration, the Callus I produced only some characterized adventitious roots and the embryonic Callus II is sufficient to regenerate whole plants. Genome-wide transcriptome profiles were performed to reveal the underlying mechenisms. The numbers of differentially expressed genes for the three types of callus vary from 5906 to 8029. Principal component analysis analysis demonstrated that gene expression patterns of Callus I and II were totally different from that of Callus III and differential leaves from Callus II, indicating that the compassions of Callus I and II provide clues for revealing regulations of regeneration in sorghum callus. Notably, KEGG and GO analysis showed that plant ribosome, lignin metabolic process, and metabolism of starch and sucrose are main processes that are associated with callus differentiation. Taken together, the results contributed the elucidation of molecular regulation in three types of callus with several regeneration abilities in sorghum.

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small RNA-mediated regulation during Zea mays (maize) Somatic Embryogenesis [RNA-Seq]

Project description:We report the role of sRNAs populations during the induction of callus tissues from VS-535 maize embryos displaying contrasting in vitro embryogenic potential; characterized through Next-generation sequencing (NGS). We conclude that the Embryogenic Response during Maize Somatic Embryogenesis induction is closely related to sRNAs regulation and depends on the developmental stage of the explant.

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small RNA-mediated regulation during Zea mays (maize) Somatic Embryogenesis [smRNA-Seq]

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Cytological, biochemical and molecular events related to the embryogenic state in Douglas-fir

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2019-05-17 | PXD011176 | Pride

The maize methylome modulates mRNA splicing and reveals widespread paramutation guided by small RNA.

Project description:Methylation of chromosomal DNA in animals and plants is a fundamental mechanism of epigenetic regulation, and the maize genome, with its diverse complement of transposons and repeats, is a paradigm for transgenerational mechanisms such as paramutation and imprinting. We have determined the genome-wide cytosine methylation map of two maize inbred lines, B73 and Mo17, at high coverage and at single nucleotide resolution. Transposon methylation is highest in CG (65%) and CHG (50%) contexts (where H = A, C or T), while methylation in CHH (5%) contexts is guided by 24nt small interfering RNA (siRNA), and not by 21-22nt siRNA. We have found that CG (8%) methylation seems to deter insertion of Mutator transposons into exons, while CHH and CHG methylation at splice donor and acceptor sites strongly inhibits RNA splicing. Methylation differences between parents are inherited in recombinant inbred lines, but methylation switches, guided by siRNA, are widespread and persist for up to 8 generations. These differences influence splicing, and recurrent switching suggest that paramutation is much more common than previously supposed, and may contribute to heterosis. Our results provide a comprehensive high resolution resource for maize genome methylation, as well as a map of recurrent transgenerational epigenetic shifts (paramutation) in the two most commonly used inbred maize lines. Genome-wide cytosine methylation map in 2 maize strains by bisulfite sequencing, and RNA and small RNA profiles in the same tissue using Illumina platform.

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Genome-wide methylation profile of mitochondrial DNA across bovine preimplantation development

Project description:This study characterized variations in the methylation profile of mitochondrial DNA (mtDNA) during initial bovine embryo development and correlated the presence of methylation with mtDNA transcription. Bovine oocytes were obtained from abattoir ovaries and submitted to in vitro culture procedures. Oocytes and embryos were collected at various stages (immature oocyte, IM; mature oocyte, MII; zygote, ZY; 4-cells, 4C; 16-cells, 16C and blastocysts, BL). Total DNA (including mtDNA) was used for Whole Genome Enzymatic Methyl Sequencing and for quantification of mtDNA copy number. Extracted RNA was used for quantification of mitochondrial transcripts (ND6, CYTB, tRNA-Phe and tRNA-Gln) using Droplet Digital PCR. The number of mtDNA copies per oocyte/embryo was found to be similar, while methylation levels in mtDNA varied among stages. Higher total methylation levels were found mainly at 4C and 16C. In specific gene regions, higher methylation levels were also observed at 4C and 16C (ND6, CYTB and tRNA-Phe), as well as an inverse correlation with the quantity of transcripts for these regions. This is a first description of epigenetic changes occurring in mtDNA during early embryonic development. Our results indicate that methylation might regulate the mtDNA transcription at a local level, particularly around the time of embryonic genome activation.

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The maize methylome modulates mRNA splicing and reveals widespread paramutation guided by small RNA.

2013-06-03 | GSE39232 | GEO

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2020-08-03 | GSE140969 | GEO

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