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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 84 (1992), S. 26-32 
    ISSN: 1432-2242
    Keywords: Lysine ; Rice ; Mutant ; Protein ; β-glucanases
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A rice mutant with unique protein expression/ transport properties has been established as cells in liquid suspension and partially characterized. Mutants were originally recovered from anther calli grown for three cycles at inhibitory levels of lysine + threonine and one cycle of S-(2-aminoethyl)cysteine. Cell suspension cultures were started from high lysine-containing seeds regenerated from the inhibitor selections. Cultures of the mutant produce 2 times as much protein per unit weight as is produced by the control. Significant portions of the proteins are exported from the cells into the surrounding medium. The mutant also has 20% greater lysine content in the exported protein than the control. This cell suspension line should be particularly useful for biochemical and molecular studies on protein synthesis and processing phenomena in cereals.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 92 (1996), S. 255-262 
    ISSN: 1432-2242
    Keywords: Key words Rice  ;  β-1 ; 3 glucanases  ;  Mutant  ;  Protein export  ;  Extracellular  ;  Cell culture  ;  Stress  ;  Pathogenesis-related
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  Mutant rice cells (Oryza sativaL.) grown in liquid suspension cultures exported greater quantities of protein and β-glucanases than controls. These mutants were isolated from anther calli resistant to 1 mM lysine plus threonine (LT), regenerated and reestablished as cell suspension cultures from seeds. Cellular protein levels are genetically conditioned, and the levels of extracellular proteins and enzyme activities are inversely related to that of the cellular portions. The rechallenge of cells with 1 mM LT inhibited the expression of both β1,3-glucanases and β-1,4-glucosidases but had no significant effect upon the levels of chitinase activity. Mutant cells were more sensitive than controls to stress caused by exogenous LT. In general, under exogenous LT stress the mutant/control ratio for extracellular glucanases increased as the assay conditions were changed from a basic to an acidic pH. The specific activity of β-glucanases was highest in media and lowest in cells. Both the mutant and control cells exported β-glucanases into the suspension medium, but the level of activity in media was greater in that in which the mutant was suspended. The export was probably modulated by the internal protein levels which were highest in mutant cells without LT. Seedlings from mutants with enhanced lysine also had enhanced acidic β-glucanase activity.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Theoretical and applied genetics 92 (1996), S. 255-262 
    ISSN: 1432-2242
    Keywords: Rice ; β-1,3 glucanases ; Mutant ; Protein export ; Extracellular ; Cell culture ; Stress ; Pathogenesis-related
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Mutant rice cells (Oryza sativa L.) grown in liquid suspension cultures exported greater quantities of protein and β-glucanases than controls. These mutants were isolated from anther calli resistant to 1 mM lysine plus threonine (LT), regenerated and reestablished as cell suspension cultures from seeds. Cellular protein levels are genetically conditioned, and the levels of extracellular proteins and enzyme activities are inversely related to that of the cellular portions. The rechallenge of cells with 1 mM LT inhibited the expression of both β-1,3-glucanases and β-1,4-glucosidases but had no significant effect upon the levels of chitinase activity. Mutant cells were more sensitive than controls to stress caused by exogenous LT. In general, under exogenous LT stress the mutant/control ratio for extracellular glucanases increased as the assay conditions were changed from a basic to an acidic pH. The specific activity of βglucanases was highest in media and lowest in cells. Both the mutant and control cells exported β-glucanases into the suspension medium, but the level of activity in media was greater in that in which the mutant was suspended. The export was probably modulated by the internal protein levels which were highest in mutant cells without LT. Seedlings from mutants with enhanced lysine also had enhanced acidic β-glucanase activity.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-2242
    Keywords: Oryza sativa L. ; Biochemical selections ; Analogs ; Mutants ; Seed size ; Opaque
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Lysine is a limiting amino acid for optimal nutritional quality in rice grain. In vitro selections using inhibitory levels of lysine plus threonine or s-aminoethylcysteine allow the predictable recovery of variants with elevated levels of lysine and protein. These methods may generate useful starting germplasm for plant breeders. This study was conducted to define the genetics of lysine mutants in progeny from crosses of mutants derived from cells cultured in vitro in the presence of inhibitory levels of lysine plus threonine and s-(2-aminoethyl)-cysteine. In vitro selections produce a wide range of mutants, including endosperm mutants with elevated lysine and protein levels as well as mutants for high and low seed weights. Mutants were analyzed for lysine content by the endosperm half-seed method in which the halves without the embryo were ground and acid hydrolyzed for amino acid determinations. The halves with the embryos were preserved for later germination. In two different F2 populations derived from a cross of a selected mutant x M-101, a parental marker, there was an inverse relationship between seed weight and percent lysine in endosperm protein (R2 0.52 and 0.56). The F2 segregation patterns show that elevated lysine is inherited as a recessive gene and that increased lysine is correlated with decreased seed size. F3 and F4 data provide evidence for the transmission of high lysine genes to advanced germplasm in rice. This work supports our earlier conclusions that high lysine phenotypes can be recovered predictably from in vitro selections. The elevated lysine phenotypes are frequently, but not exclusively, associated with opaque seed. Some segregants from crosses produced increased lysine in plants with near normal seed weight and good fertility.
    Type of Medium: Electronic Resource
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