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  • 1
    ISSN: 1432-2242
    Keywords: Key words Perennial ryegrass (Lolium perenne)  ; Cytoplasmic male sterility  ;  Mitochondrial DNA  ; atp9  ;  Chimaeric open reading frame
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract  The most striking difference between the mtDNAs of the fertile L. perenne line LPSB21 and the male-sterile line CMS9B290, is the presence in the former and the absence in the latter of a 5.6-kb HindIII fragment. This difference between fertile and sterile lines was the starting point for a detailed molecular analysis of the mitochondrial genome in the region spanning the 5.6-kb HindIII fragment in fertile L. perenne and the corresponding region in CMS9B290. Restriction mapping and Southern-blot analyses indicated that rearrangement of the mitochondrial genome consistent with a deletion/insertion event had occurred in the sterile line. Nucleotide-sequence analysis of the rearranged region in CMS9B290 revealed the presence of (1) a novel chimaeric gene, orf-C9, comprising the first six codons of atp9 fused to a further 118 codons of an unknown sequence and (2) a truncated version of an open reading frame, orf-L , originally identified in LPSB21 mtDNA. Northern-blot analysis confirmed the absence of orf-L transcripts and the presence of orf-C9 transcripts in the mtRNA of CMS9B290.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-2242
    Keywords: Diagnostic probe ; Mitochondrial DNA (mtDNA) ; Cytoplasmic male sterility (CMS) ; Lolium perenne
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Analysis of reciprocal crosses between nonrestoring fertile genotypes and restored male-sterile genotypes of Lolium perenne confirmed the cytoplasmic nature of the sterility trait. This prompted a search for a molecular probe that could be used to distinguish between fertile and cytoplasmic male-sterile (CMS) cytoplasms. We describe the identification and cloning of a 4.5-kb BamHI-HindIII restriction fragment from the mtDNA of the CMS line. The cloned fragment (pCMS45) failed to hybridise to sequences in the mtDNA of fertile lines and was thus capable of unambiguously distinguishing between fertile and CMS cytoplasms. The use of pCMS45 as a diagnostic probe provided a simple test for positive identification of young non-flowering plants carrying the CMS cytoplasm and also permitted confirmation at the molecular level of the maternal transmission of the CMS trait suggested by the genetic data.
    Type of Medium: Electronic Resource
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