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  • 1
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Plant, cell & environment 20 (1997), S. 0 
    ISSN: 1365-3040
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A mutational approach was taken to identify genes required for low-temperature growth of the chilling-tolerant plant Arabidopsis thaliana. The screen identified mutants that were specifically compromised in their ability to grow at 5°C but were indistinguishable from wild type when grown at 22°C. The populations screened were mutated either by ethyl methanesulphonate or by T-DNA insertion. In both cases symptoms at 5°C included chlorosis, reduced growth, necrosis and death. This diversity of phenotypes demonstrates roles for chilling-tolerance responses in such diverse processes as organdie biogenesis, cell metabolism and cell and organ development. Co-segregation analysis on the first five mutants isolated from the T-DNA lines indicated that in three of them, pfc1, pfc2 and sop1, the chilling phenotype is the result of T-DNA insertion in a gene required for chilling tolerance rather than the creation of a temperature-conditional mutation in an essential housekeeping gene. This identification of T-DNA tagged alleles will facilitate cloning of the PFC1, PFC2 and SOP1 loci and allow for the biochemical and molecular genetic characterization of these chilling-tolerance genes and the proteins that they encode.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-2048
    Keywords: Avena (phytochrome) ; Immunoprecipitation ; Monoclonal antibody ; Phytochrome from green tissue ; Western blotting (immunoblot analysis)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A method is described for the extraction of phytochrome from chlorophyllous shoots of Avena sativa L. Poly(ethyleneimine) and salt fractionation are used to reduce chlorophyll and to increase the phytochrome concentration sufficiently to permit spectral and immunochemical analyses. The phototransformation difference spectrum of this phytochrome is distinct from that of phytochrome from etiolated shoots in that the maximum in the red region of the difference spectrum is shifted about 15 nm to a shorter wavelength. Immunochemical probing of electroblotted proteins (Western blotting), using a method sensitive to 50 pg, demonstrates the presence of two polypeptides in green tissue that bind antiphytochrome antibodies: a predominant species with a relative molecular mass (Mr) of 118000 and a lesser-abundant 124000-Mr polypeptide. Under nondenaturing conditions all of the 124000-Mr species is immunoprecipitable, but the 118000-Mr species remains in the supernatant. Peptide mapping and immunochemical analysis with monoclonal antibodies show that the 118000-Mr species has structural features that differ from etiolated-oat phytochrome. Mixing experiments show that these structural differences are intrinsic to the molecular species from these two tissues rather than being the result of post-homogenization modifications or interfering substances in the green-tissue extracts. Together the data indicate that the phytochrome that predominates in green-tissue has a polypeptide distinct from the well-characterized molecule from etiolated tissue.
    Type of Medium: Electronic Resource
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