The Use of Zygotic Embryos as Explants for In Vitro Propagation: An Overview

Elhiti, Mohamed; Stasolla, Claudio

doi:10.1007/978-1-61737-988-8_17

Cited by 34 publications

(16 citation statements)

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“…Since the first reports of SE in the umbellifers Oenanthe aquatica and carrot by Harry Waris in 1957 (Krikorian and Simola 1999) and by Steward et al (1958), respectively, this regeneration pathway has become a model system for studying the physiological, biochemical, cellular and molecular mechanisms involved in the transition of somatic cells to the embryogenic state for many species (Elhiti and Stasolla 2011;Fehér et al 2003;Fehér 2015). Continuing the studies of P. edulis zygotic embryo-derived SE that were performed by Paim- Pinto et al (2011), Rosa et al (2015) and Rocha et al (2015), here we provide additional information based on an ultrastructural description of the events involved in embryogenic competence acquisition during the early stages of somatic embryo development.…”

Section: Discussionmentioning

confidence: 99%

Cellular and molecular changes associated with competence acquisition during passion fruit somatic embryogenesis: ultrastructural characterization and analysis of SERK gene expression

et al. 2015

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The integration of cellular and molecular data is essential for understanding the mechanisms involved in the acquisition of competence by plant somatic cells and the cytological changes that underlie this process. In the present study, we investigated the dynamics and fate of Passiflora edulis Sims cotyledon explants that were committed to somatic embryogenesis by characterizing the associated ultrastructural events and analysing the expression of a putative P. edulis ortholog of the Somatic Embryogenesis Receptor-like Kinase (SERK) gene. Embryogenic calli were obtained from zygotic embryo explants cultured on Murashige and Skoog medium supplemented with 2,4-dichlorophenoxyacetic acid and 6-benzyladenine. Callus formation was initiated by the division of cells derived from the protodermal and subprotodermal cells on the abaxial side of the cotyledons. The isodiametric protodermal cells of the cotyledon explants adopted a columnar shape and became meristematic at the onset of PeSERK expression, which was not initially detected in explant cells. Therefore, we propose that these changes represent the first observable steps towards the acquisition of a competent state within this regeneration system. PeSERK expression was limited to the early stages of somatic embryogenesis; the expression of this gene was confined to proembryogenic zones and was absent in the embryos after the globular stage. Our data also demonstrated that the dynamics of the mobilization of reserve compounds correlated with the differentiation of the embryogenic callus.

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Section: Discussionmentioning

confidence: 99%

Cellular and molecular changes associated with competence acquisition during passion fruit somatic embryogenesis: ultrastructural characterization and analysis of SERK gene expression

et al. 2015

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“…Here we demonstrated that Arabidopsis IZEs at the strictly defined late cotyledonary developmental stage can be induced to engage an alternative morphogenic pathway, shoot ORG, in addition to SE (Gaj, 2001). IZEs of other plant species have also been used successfully as explants for adventitious shoot induction (Elhiti and Stasolla, 2011).…”

Section: Discussionmentioning

confidence: 99%

“…IZE explants present high embryogenic potential (Gaj, 2011), and the culture conditions promoting induction of shoot ORG, the alternative morphogenic pathway, are already well defined in IZE culture (Elhiti and Stasolla, 2011). To assess the proposed in vitro system's usefulness for identifying the genetic mechanisms specific to the morphogenic pathway, we analyzed the expression patterns of LEC genes, key regulators of Arabidopsis embryogenesis, in IZE culture induced towards shoot ORG.…”

Section: Introductionmentioning

confidence: 99%

Immature Zygotic Embryo Cultures of Arabidopsis. A Model System for Molecular Studies on Morphogenic Pathways Induced In Vitro

Kraut¹,

Wójcikowska²,

Ledwoń³

et al. 2011

Acta Biologica Cracoviensia Series Botanica

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To understand the molecular mechanism controlling in vitro plant morphogenesis, a culture system enabling induction of alternative morphogenic pathways (somatic embryogenesis, SE; shoot organogenesis, ORG) in a well defined population of somatic cells is needed. Arabidopsis is the most useful model plant for genomic studies, but a system in which SE or ORG can be induced alternatively in the same type of explant has not been proposed. Immature zygotic embryos (IZEs) of Arabidopsis provide the only explants with embryogenic potential, and have been recommended for studying mechanisms of SE induced in vitro. This study was aimed at defining culture conditions promoting induction of alternative morphogenic pathways: shoot ORG in IZE explants. The established protocol involves pretreatment of IZE explants with liquid auxin-rich callus induction (CIM) medium, followed by subculture on solid cytokinin-rich shoot induction medium (SIM). The method enables efficient shoot induction in Columbia (Col-0) and Wassilewskija (Ws), genotypes commonly used in molecular studies. During 3 weeks of culture up to 90% of Col-0 and 70% of Ws explants regenerated shoots via an indirect morphogenic pathway. We analyzed the qRT-PCR expression patterns of the LEC (LEC1, LEC2 and FUS3) genes, the key regulators of Arabidopsis embryogenesis, in the IZE explants induced to promote shoot ORG. The sharp decline of LEC expression on SIM medium confirmed that culture of Arabidopsis IZE explants enables experimental manipulation of the morphogenic response of somatic cells. A scheme illustrating various in vitro morphogenic responses of IZEs in relation to hormonal treatment is presented.K Ke ey y w wo or rd ds s: : Arabidopsis, immature zygotic embryos, in vitro morphogenesis, shoot regeneration.

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“…In many plants zygotic embryos provide explants with high SE potential, although their developmental stage allowing for embryogenic transition varies considerably between species (Elhiti and Stasolla 2011). Mature zygotic embryos (MZEs) are efficient in SE induction in some species (Prakash and Gurumurthi 2010;Chen et al 2010;You et al 2011;Pinto et al 2011), mainly in cereals (Sharma et al 2005); but immature zygotic embryos (IZEs) have been most frequently used to start embryogenic culture in plants of various taxonomic groups (Gaj 2004).…”

Section: Introductionmentioning

confidence: 99%

Expression of seed storage product genes (CRA1 and OLEO4) in embryogenic cultures of somatic tissues of Arabidopsis

Gliwicka

Nowak

Cieśla

et al. 2011

Plant Cell Tiss Organ Cult

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In Arabidopsis, immature zygotic embryos (IZEs) at the late cotyledon stage provide the only explants that can be used to induce in vitro somatic embryogenesis (SE) with high efficiency. The most conspicuous characteristic of SE-competent IZEs is the accumulation of seed storage reserves (SSR), as proteins and lipids. In order to elucidate an assumed role of these compounds in the mechanisms involved in tissue capacity for SE, the genes encoding the main seed storage protein, cruciferin (CRU-CIFERIN1, CRA1) and the lipid body-related protein oleosin (OLEOSIN4, OLEO4), were studied. Significantly higher transcriptional activity of both genes, CRA1 and OLEO4, in embryogenic than in non-embryogenic cultures, were indicated. However, their activity under in vitro culture were found not to be induced by auxin treatment or LEC2 expression, and were unspecific for SE induction. In addition, the results on mutants severely impaired in SE response indicated that high activity of SSR genes in explant tissue is not sufficient for SE induction. On the other hand, the cra1 and oleo4 insertional mutants were found to be defective in their capacity for SE. In addition, it was found that the mutants displayed lower tolerance to high salinity and osmotic stress. Altogether, the results suggest an indirect influence of SSR genes on the embryogenic capacity of cultured tissues possibly via improvement of cell response to stress imposed in vitro.

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The Use of Zygotic Embryos as Explants for In Vitro Propagation: An Overview

Cited by 34 publications

References 100 publications

Cellular and molecular changes associated with competence acquisition during passion fruit somatic embryogenesis: ultrastructural characterization and analysis of SERK gene expression

Cellular and molecular changes associated with competence acquisition during passion fruit somatic embryogenesis: ultrastructural characterization and analysis of SERK gene expression

Immature Zygotic Embryo Cultures of Arabidopsis. A Model System for Molecular Studies on Morphogenic Pathways Induced In Vitro

Expression of seed storage product genes (CRA1 and OLEO4) in embryogenic cultures of somatic tissues of Arabidopsis

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