Whole Mount in situ Localization of miRNAs and mRNAs During Somatic Embryogenesis in Arabidopsis

Wójcik, Anna; Mosiołek, Magdalena; Karcz, J; Nodine, Michael D.; Gaj, Małgorzata D.

doi:10.3389/fpls.2018.01277

Cited by 12 publications

(8 citation statements)

References 58 publications

(80 reference statements)

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“…However, most of the reporter lines monitor the gene promoter activity rather than localize the corresponding gene transcript. Hence, a novel method of the whole-mount in situ hybridization (WISH) was recently proposed to analyze the spatiotemporal pattern of miRNAs in SE-induced tissue [84].…”

Section: Auxin-related Mirnas Fine-tune the Genetic Network That Contmentioning

confidence: 99%

Epigenetic Regulation of Auxin-Induced Somatic Embryogenesis in Plants

Wójcikowska

Wójcik

Gaj

2020

IJMS

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Somatic embryogenesis (SE) that is induced in plant explants in response to auxin treatment is closely associated with an extensive genetic reprogramming of the cell transcriptome. The significant modulation of the gene transcription profiles during SE induction results from the epigenetic factors that fine-tune the gene expression towards embryogenic development. Among these factors, microRNA molecules (miRNAs) contribute to the post-transcriptional regulation of gene expression. In the past few years, several miRNAs that regulate the SE-involved transcription factors (TFs) have been identified, and most of them were involved in the auxin-related processes, including auxin metabolism and signaling. In addition to miRNAs, chemical modifications of DNA and chromatin, in particular the methylation of DNA and histones and histone acetylation, have been shown to shape the SE transcriptomes. In response to auxin, these epigenetic modifications regulate the chromatin structure, and hence essentially contribute to the control of gene expression during SE induction. In this paper, we describe the current state of knowledge with regard to the SE epigenome. The complex interactions within and between the epigenetic factors, the key SE TFs that have been revealed, and the relationships between the SE epigenome and auxin-related processes such as auxin perception, metabolism, and signaling are highlighted.

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Section: Auxin-related Mirnas Fine-tune the Genetic Network That Contmentioning

confidence: 99%

Epigenetic Regulation of Auxin-Induced Somatic Embryogenesis in Plants

Wójcikowska

Wójcik

Gaj

2020

IJMS

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“…MmuMiR124 was chosen as it has no plant homolog and no complementary sequences were detected in the Arabidopsis genome (Ghosh Dastidar et al . 2016; Wójcik et al . 2018).…”

Section: Resultsmentioning

confidence: 99%

“…The 3' UTR of this CLV3 transgene was modified to contain a site complementary to mouse microRNA MmuMiR124. MmuMiR124 was chosen as it has no plant homolog and no complementary sequences were detected in the Arabidopsis genome (Ghosh Dastidar et al 2016;Wójcik et al 2018). This recombinant construct, pCLV3::CLV3:MmuMiR124'::eCLV3, is hereafter abbreviated to 'C' (Figure 1b).…”

Section: Phloem-expressed Artificial Mirna Influences Gene Expression In Sam Stem Cellsmentioning

confidence: 99%

small RNA can move long distances through plant vasculature to influence gene expression in shoot apical meristems

Minow

Coneva

Lesy

et al. 2021

Preprint

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In plants, small RNA (sRNA) can regulate gene expression via post transcriptional gene silencing (PTGS) or through RNA-directed DNA methylation (RdDM) leading to transcriptional gene silencing (TGS). sRNA is mobile throughout the plant, with movement occurring short distances from cell-to-cell as well as long distances through the vasculature via phloem trafficking. The range of long-distance sRNA mediated signaling from the vasculature to the shoot apical meristem (SAM) is not clear. To investigate this, two independent transgenic approaches were used to examine trafficking of phloem-expressed sRNA to the SAM in Arabidopsis thaliana. First, the phloem companion-cell specific promoter SUC2 was used to drive expression of an inverted repeat complementary to FLOWERING LOCUS D (FD), a flowering time regulator expressed exclusively in the SAM. In a separate experiment, the SUC2 promoter was used to express an artificial microRNA (aMiR) designed to target a synthetic CLAVATA3 (CLV3) target in the SAM stem cells. Both systems provide evidence of a phloem-to-SAM sRNA communication axis connecting distal regions of the plant to the stem cells of the SAM, which ultimately gives rise to all shoot tissues, including gametes. Thus, phloem-to-SAM sRNA movement defines an important link between sRNA expressed in distal regions of the plant and the growing shoot. Importantly, phloem-to-SAM sRNA trafficking may allow somatic sRNA to direct SAM RdDM, fixing transient sRNA expression events into stable epigenetic changes.

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“…In recent years, SE has been employed in plant species as a propagation tool in place of seed reproduction ( Ipekci and Gozukirmizi, 2003 ; Li et al, 2012 ), especially for plants with extremely low reproduction rates or maintaining agronomic traits. To date, the process and mechanism of SE in model plants such as Arabidopsis have been well elucidated ( Kadokura et al, 2018 ; Wojcik et al, 2018 ); however, the widespread application of in vitro embryogenesis is limited to the low responsiveness of many plant species and genotypes, especially in woody plants.…”

Section: Discussionmentioning

confidence: 99%

Direct and Indirect Somatic Embryogenesis Induction in Camellia oleifera Abel

et al. 2021

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Camellia oleifera Abel. is an important woody oil species; however, the shortage of rapid and industrialized seedling culture is a large constraint on the development of the tea oil industry. Somatic embryogenesis (SE) is one of the main powerful biotechnological tools for plant mass regeneration, but the largely unknown SE in C. oleifera limits the scale production of clonal plants. In this study, we described a high-efficiency SE system via direct and indirect pathways in C. oleifera and investigated the effect of genotype, explant age and phytohormones on SE. In the direct pathway, somatic embryos were highly induced from immature seeds 220 days after full blossom, and the development of embryoids was achieved with a combination of 0.19 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.05 mg/L thidiazuron (TDZ). In the indirect pathway, embryogenic calli were induced from the same explants in medium containing 1.5 mg/L 2,4-D, while 0.75 mg/L 2,4-D treatment led to high proliferation rates for embryogenic calli. The addition of 0.19 mg/L 2,4-D alone stimulated the production of globular embryos while causing a 75% loss of the induction rate in the heart embryo stage. Upon transfer of the globular embryos to phytohormone-free medium, an optimal induction rate of 62.37% from globular embryos to cotyledonary embryos was obtained. These data suggest that the subsequent differentiation process after the globular embryo stage in ISE is more similar to an endogenous phytohormones-driven process. Mature embryos germinated to produce intact plantlets on half-strength MS basal medium with a regeneration rate of 63.67%. Histological analysis confirmed the vascular bundle isolation of embryoids from the mother tissue. We further studied the different varieties and found that there were no significant genotype differences for SE induction efficiency in C. oleifera. Thus, we established a high-efficiency induction system for direct and indirect somatic embryogenesis (ISE) in C. oleifera and regenerated intact plantlets via SE, not organogenesis. ISE has a more complicated induction and regulatory mechanism than direct somatic embryogenesis. The improved protocol of SE would benefit mass propagation and genetic manipulation in C. oleifera.

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Whole Mount in situ Localization of miRNAs and mRNAs During Somatic Embryogenesis in Arabidopsis

Cited by 12 publications

References 58 publications

Epigenetic Regulation of Auxin-Induced Somatic Embryogenesis in Plants

Epigenetic Regulation of Auxin-Induced Somatic Embryogenesis in Plants

small RNA can move long distances through plant vasculature to influence gene expression in shoot apical meristems

Direct and Indirect Somatic Embryogenesis Induction in Camellia oleifera Abel

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