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  • 1
    ISSN: 1573-5028
    Keywords: ADP-glucose pyrophosphorylase ; barley ; endosperm ; PCR ; cDNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Several cDNAs encoding the small and large subunit of ADP-glucose pyrophosphorylase (AGP) were isolated from total RNA of the starchy endosperm, roots and leaves of barley by polymerase chain reaction (PCR). Sets of degenerate oligonucleotide primers, based on previously published conserved amino acid sequences of plant AGP, were used for synthesis and amplification of the cDNAs. For either the endosperm, roots and leaves, the restriction analysis of PCR products (ca. 550 nucleotides each) has revealed heterogeneity, suggesting presence of three transcripts for AGP in the endosperm and roots, and up to two AGP transcripts in the leaf tissue. Based on the derived amino acid sequences, two clones from the endosperm, beps and bepl, were identified as coding for the small and large subunit of AGP, respectively, while a leaf transcript (blpl) encoded the putative large subunit of AGP. There was about 50% identity between the endosperm clones, and both of them were about 60% identical to the leaf cDNA. Northern blot analysis has indicated that beps and bepl are expressed in both the endosperm and roots, while blpl is detectable only in leaves. Application of the PCR technique in studies on gene structure and gene expression of plant AGP is discussed.
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 18 (1992), S. 1195-1198 
    ISSN: 1573-5028
    Keywords: abscisic acid ; barley ; endosperm ; seed protein
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1573-5028
    Keywords: barley ; endosperm coenocyte ; differential screening ; modified aleurone cells ; nucellus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The cereal endosperm develops from a coenocyte to a cellular storage organ through formation of nucleo-cytoplasmic domains and cell wall deposition in the interzones between these domains. During its early stages, the endosperm develops in close contact with nucellus, the sporophytic tissue which gives rise to the megagametophyte. Owing to the positioning of the two tissues deeply within the ovary, neither cell types have been easily accessible for molecular studies. In this paper we report for the first time the cloning of molecular markers for the barley endosperm coenocyte and the nucellus. The novel END1 and NUC1 cDNAs were isolated by differential screening of a cDNA library from 5 DAP (days after pollination) ovaries using a positive probe from hand-dissected embryo sacs with adhering nucellus and testa cell layers, and a negative probe from pericarp. In situ and northern blot hybridization data show that END1 transcripts are asymmetrically distributed in teh endosperm coenocyte limited to an area over the nucellar projection. In the cellular endosperm, END1 transcripts are present in modified aleurone cells and a few layers of ventral starchy endosperm cells. The second clone, NUC1, hybridizes to transcripts in the nucellus before fertilization and in autolyzing nucellus cells after fertilization. At later stages, after the disappearance of nucellus, NUC1 transcripts are present in the nucellar epidermis and in the lateral cells of the nucellar projection. This work provide tools for future elucidation of the genes specifying endosperm histogenesis.
    Type of Medium: Electronic Resource
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