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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell reports 15 (1996), S. 516-521 
    ISSN: 1432-203X
    Keywords: conifer ; Picea glauca ; RNA extraction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A simple and efficient procedure suitable for extraction of high-quality RNA from cultured conifer tissues, somatic embryos, zygotic embryos, needles, stem and root tissues was developed. It produced from 100 μg up to 700 μg total RNA per gram tissue dependent on the types of tissues used. RNA quality was estimated by spectrophotometry, agarose gel electrophoresis, in vitro translation of mRNA, cDNA synthesis and Northern blot analysis. The method also worked well with Arabidopsis thaliana and tobacco tissues.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-2048
    Keywords: Key words: Abscisic acid ; Gene expression ; lea genes ; Picea (somatic embryogenesis) ; Somatic embryo development
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract. Five abscisic acid (ABA)-responsive cDNAs that were identified by differential screening of a white spruce [Picea glauca (Moench) Voss] cotyledonary somatic embryo cDNA library are characterized. Three of the cDNAs (PgEMB12, 14, 15) are predicted to encode homologues of different late-embryogenesis-abundant (LEA) proteins, a further two (PgEMB 5 and 23) share little similarity to any known DNA or protein sequences. When suspension cultures were fed 1.5 × 105 M S-(+)-ABA and assessed for transcript abundance over 48 h, gene expression corresponding to each lea-like gene was inducible and was evident during the 48-h period. Conversely, transcripts were at a very low abundance when suspensions were fed R-(−)-ABA. When suspensions were treated with a range of S-(+)-ABA and R-(−)-ABA concentrations (10−7–10−4 M), expression was induced at S-(+)-ABA concentrations of 10−5 M or above. Expression corresponding to PgEMB5 and PgEMB23 indicates less specificity for ABA stereostructure, with similar expression patterns being observed for either enantiomer during the 48 h after feeding. The two corresponding genes also appear more responsive to ABA concentration than the predicted lea genes, within the range examined (10−7–10−4 M).
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-2048
    Keywords: Abscisic acid ; Endochitinase ; Fungal elicitation ; β-1,3-Glucanase ; Picea glauca ; Somatic embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Two cDNAs isolated from white spruce [Picea glauca (Moench) Voss] somatic embryos, are predicted to encode a basic class IV chitinase and a β-1,3-glucanase, respectively corresponding to genesPgChi-1 andPgGlu-1. Each represents a multigene family in spruce. Transcripts homologous toPgChi-1 orPgGlu-1 genes were highly abundant in embryogenic tissues and gradually decreased after tissues were placed on abscisic acid-containing maturation medium, with lowest abundance in globular embryos. Transcripts related toPgGlu-1 became highly abundant again in early cotyledonary embryos but decreased thereafter, whereas transcripts related toPgChi-1 were also highly abundant in late cotyledonary embryos and plantlets in vitro; transcripts were either low (PgChi-1) or were not detectable (PgGlu-1) in needles. Wounding, drying and flooding stresses enhancedPgChi-1-andPgGlu-1-related gene expression. Fungal cell wall suspension enhancedPgGlu-1-related transcript accumulation, but reducedPgChi-1-related transcript abundance within 24 h.PgChi-1 andPgGlu-1 and their homologues may have roles in plant defense, and possibly developmental roles during spruce somatic embryo maturation.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-2048
    Keywords: Abscisic acid ; Embryo abundant gene ; Gene expression ; Polyethylene glycol ; Picea ; Somatic embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Embryogenic tissues of white spruce [Picea glauca (Moench) Voss] remain in an early developmental stage while cultured on 2,4-dichlorophenoxyacetic acid and N6-benzyladenine, but develop to cotyledonary embryos when these phytohormones are replaced by abscisic acid. Twenty-eight cDNAs were isolated from cotyledonary embryos by differential screening against immature embryo and non-embryonic tissues. Temporal expression patterns of these cDNAs during ABA-stimulated somatic embryo development were observed. This showed that clones could be allocated to various groups, including embryo-abundant, embryo-maturation-induced, and those whose expression was modulated during embryo development, germination or in non-embryogenic tissues. Expression corresponding to these cDNA clones showed that there were various responses to exogenous ABA or polyethylene glycol during a period of 48 h. Analyses of DNA and predicted amino acid sequence revealed that 12 of 28 cDNA clones had no known homologues, while others were predicted to encode different late-embryogenesis-abundant proteins, early methionine-labelled proteins, storage proteins, heat-shock proteins, glycine-rich cell wall protein, metallothionein-like protein and some other metabolic enzymes.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-2048
    Keywords: Abscisic acid ; Somatic embryogenesis ; Gene expression ; Heat shock protein ; Picea ; Polyethylene glycol
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Three cDNAs (PgEMB22, 27 and 29) predicted to encode low-molecular-weight (LMW) heat-shock proteins (HSPs) were cloned and characterized from white spruce [Picea glauca (Moench) Voss] somatic embryo tissues by differentially screening a cotyledonary embryo cDNA library. Clone PgEMB22 is predicted to encode a putative mitochondria-localized LMW HSP, and PgEMB27 and 29 are predicted to encode different cytoplasmic class II LMW HSPs, although they share 84.7% identity within DNA coding regions and 83.0% identity for predicted proteins. They are developmentally regulated during somatic embryo development and subsequent embryo germination, in addition they show strong response to heat-shock stress. Transcripts of the two kinds of hsp genes could be detected in embryogenic tissues before induction of embryo maturation, but subsequently increased, being most abundant at late embryo stages. Gene expression levels were very low or not detectable in germinated plantlets or needle tissues from older plants. Abscisic acid and polyethylene glycol, stimulators for spruce embryo maturation, could also induce the hsp genes.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1573-5028
    Keywords: embryo-abundant cDNAs ; gene expression ; gymnosperm ; Picea glauca ; somatic embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Six somatic embryogenesis-associated cDNAs (PgEMB2, 6, 7, 8, 24 and 34) from white spruce (Picea glauca (Moench) Voss) somatic embryos have been characterized. Transcript accumulation during somatic embryo development and subsequent germination related to these genes, indicated that they were developmentally regulated. The transcripts related to clones PgEMB2, 6, 24 and 34 were also detected during zygotic embryo development, but transcripts of clones PgEMB7 and 8 were not. PgEMB24 had a similar gene expression pattern to spruce Em-like late embryo abundant (lea) gene, but other clones had no similarities in gene expression to either spruce lea-like or storage protein genes. Abscisic acid, a stimulator for spruce somatic embryo maturation, did not obviously affect gene expression corresponding to these cDNAs. The predicted proteins are distinguishable from known LEA proteins based on analyses of hydropathy plots, amino acid compositions and deduced protein structures. The similarities of the spruce cDNAs, and protein sequences predicted from these cDNAs, to other sequence data are described.
    Type of Medium: Electronic Resource
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