Molecular biology of somatic embryogenesis in conifers

Dong, Jin-Zhuo; Dunstan, David I.

doi:10.1007/978-94-017-2311-4_3

Cited by 19 publications

(14 citation statements)

References 163 publications

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“…LEA proteins are produced in abundance during late embryo development and their expression is linked to the acquisition of desiccation tolerance, although their function is not yet clear (Wise and Tunnacliffe 2004). Further, many LEA genes are induced by abscisic acid (ABA) and their level of expression tracks endogenous ABA levels (Bray 1997), a phenomenon that has also been observed during conifer embryogenesis (Dong and Dunstan 2000). The experiment with the LEA-like gene of P. pinaster was included in this study to demonstrate that our calculation method generates results that are consistent with results gained using other methods to assay gene expression.…”

Section: Discussionmentioning

confidence: 99%

Evaluation of control transcripts in real-time RT-PCR expression analysis during maritime pine embryogenesis

Gonçalves

Cairney

Marôco

et al. 2005

Planta

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In order to determine the suitability of reference or housekeeping genes as internal controls in real-time reverse transcriptase PCR (RT-PCR) assays for quantification of target mRNAs, we studied the levels of expression of four candidate reference genes in maritime pine by real-time RT-PCR. The expression levels obtained for glyceraldehyde-3-phosphate-dehydrogenase, 18S ribosomal RNA, eukaryotic translation initiation factor eIF4AII and ubiquitin in nine stages of embryo development revealed that none of the genes tested proved to be suitable as an internal control. Copy number quantification of the four transcripts showed an average relative variation of seven fold. We propose that the combination of a precise method for RNA quantification, internal controls for monitoring RT reaction and PCR efficiency and a robust external standard curve can guarantee a reliable absolute quantification of mRNA transcripts in real time RT-PCR. This approach may avoid the controversy in the use of housekeeping genes and may assume special significance in tissues undergoing developmental changes.

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Section: Discussionmentioning

confidence: 99%

Evaluation of control transcripts in real-time RT-PCR expression analysis during maritime pine embryogenesis

Gonçalves

Cairney

Marôco

et al. 2005

Planta

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“…In the second phase embryos pass through the typical stages of zygotic embryogenesis, i.e., globular, heart-shaped and torpedo-shaped stages in dicots, globular, scutellar (transition), and coleoptilar stages in monocots, or globular, early cotyledonary and late cotyledonary embryos in conifers (Gray et al 1995;Toonen and de Vries 1996;Dong and Dunstan 2000).…”

Section: Changes In Endogenous Hormone Contents During Expression Of Sementioning

confidence: 99%

“…Those works that involve molecular and genetic studies of hormonal regulation will not be discussed here (further information on this particular topic can be found in Dudits et al 1995;Dodeman et al 1997;Dong and Dunstan 2000;Thorpe 2000;Stasolla et al 2002;Phillips 2004;von Arnold et al 2005, among other sources). Whenever possible, review articles will be employed to avoid citing huge amounts of literature that could be found elsewhere.…”

Section: Introductionmentioning

confidence: 99%

Involvement of Plant Hormones and Plant Growth Regulators on in vitro Somatic Embryogenesis

2005

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In spite of the importance attained by somatic embryogenesis and of the many studies that have been conducted on this developmental process, there are still many aspects that are not fully understood. Among those features, the involvement of plant hormones and plant growth regulators on determining the conversion of somatic onto embryogenic tissues, and on allowing progression and maturation of somatic embryos, are far away from being completely comprehended. Part of these difficulties relies on the frequent appearance of contradictory results when studying the effect of a particular stimulus over a specific stage in somatic embryogenesis. Recent progress achieved on understanding the interaction between exogenously added plant growth regulators over the concentration of endogenous hormones, together with the involvement of sensitivity of the tissues to particular hormone groups, might help clarifying the occurrence of divergent patterns in somatic embryogenesis, and in tissue culture in general. The aspects described above, emphasizing on the effect of the concentration of plant hormones and of the addition of plant growth regulators during the different phases of somatic embryogenesis, will be reviewed in this paper. Citations will be limited to review articles as much as possible and to individual articles only in those cases in which very specific or recent information is presented.Abbreviations: 2,4-D -2

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“…The differing nature of somatic and zygotic embryos (ZEs) may cause deficient maturation. It has been suggested that high-quality SEs most closely resemble their zygotic counterparts in morphological, molecular and biochemical respects (Flinn et al 1993;Dong and Dunstan 2000). We previously demonstrated that accumulation of Em-like transcripts during SE maturation positively correlates with improved quality and germination capacity (Š underlíková and Wilhelm 2002).…”

Section: Introductionmentioning

confidence: 96%

Isolation and characterization of an embryo-specific Em-like gene of pedunculate oak (Quercus robur L.) and its temporal and spatial expression patterns during somatic and zygotic embryo development

Šunderlíková

Salaj

Matušíková

et al. 2008

Trees

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The oak Em gene, QrEm, was isolated from Quercus robur L. zygotic embryos in RT-PCR using degenerate primers. The full-length cDNA was obtained by RACE. Sequence analyses revealed a high degree of similarity to previously identified Em-like genes from both monocots and dicots. Northern blot analyses showed that QrEm gene is expressed in zygotic embryos during the mid-maturation phase, with a decline following towards the end of seed development. In somatic embryos, abundant transcript accumulation was observed after treatment with 6% sorbitol or after partial desiccation. In situ hybridization experiments displayed similar tissue-specific expression patterns in both zygotic and somatic embryos, predominantly in the meristematic regions of root and shoot apices as well as provascular tissues, whereas almost no hybridization signal was found in the cotyledons. Wounding, drought and cold all failed to induce expression of the QrEm gene in vegetative tissue, suggesting that the QrEm gene is embryo-specific and its expression is developmentally regulated.

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Molecular biology of somatic embryogenesis in conifers

Cited by 19 publications

References 163 publications

Evaluation of control transcripts in real-time RT-PCR expression analysis during maritime pine embryogenesis

Evaluation of control transcripts in real-time RT-PCR expression analysis during maritime pine embryogenesis

Involvement of Plant Hormones and Plant Growth Regulators on in vitro Somatic Embryogenesis

Isolation and characterization of an embryo-specific Em-like gene of pedunculate oak (Quercus robur L.) and its temporal and spatial expression patterns during somatic and zygotic embryo development

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