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  • 1
    ISSN: 1432-2048
    Keywords: Aegilops ; Chloroplast DNA ; DNA, chloroplast and nuclear ; Ploidy ; Triticum (chloroplast and nuclear genomes)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The possibility of estimating the proportion of chloroplast DNA (ctDNA) and nuclear DNA (nDNA) in nucleic-acid extracts by selective digestion with the methylation-sensitive restriction enzyme PstI, was tested using leaf extracts from Spinacia oleracea and Triticum aestivum. Values of ctDNA as percentage nDNA were estimated to be 14.58%±0.56 (SE) in S. oleracea leaves and 4.97%±0.36 (SE) in T. aestivum leaves. These estimates agree well with those already reported for the same type of leaf material. Selective digestion and quantitative dot-blot hybridisation were used to determine ctDNA as percentage nDNA in expanded leaf tissue from species of Triticum and Aegilops representing three levels of nuclear ploidy and six types of cytoplasm. No significant differences in leaf ctDNA content were detected: in the diploids the leaf ctDNA percentage ranged between 3.8% and 5.1%, and in the polyploids between 3.5% and 4.9%. Consequently, nuclear ploidy and nDNA amount were proportional to ctDNA amount (r(19)=0.935, P〉0.01) and hence to ctDNA copy number in the mature mesophyll cells of these species. There was a slight increase in ctDNA copy numbers per chloroplast at higher ploidy levels. The balance between numbers of nuclear and chloroplast genomes is discussed in relation to polyploidisation and to the nuclear control of ctDNA replication.
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  • 2
    ISSN: 1432-2242
    Keywords: Triticum ; Aegilops ; Chloroplast DNA ; Cytoplasmic inheritance ; Wheat evolution ; The B genome donor ; Restriction mapping ; Insertions ; Deletions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Restriction endonuclease analysis revealed interspecific and intraspecific variation between the chloroplast DNAs and therefore between the cytoplasms of 14 selected species of Triticum and Aegilops. Eleven distinct chloroplast DNA types were detected, the differences between them residing in the varied combination of a relatively few DNA alterations. The variation was simple enough for chloroplast DNA analysis to be used as a basis for the identification and classification of the Triticum and Aegilops cytoplasms. There was good agreement with the classification based on analysis of the phenotypic effects of the cytoplasm when combined with the T. aestivum nucleus in nuclear-cytoplasmic hybrids (Tsunewaki et al. 1976). There was however no correlation between specific chloroplast DNA alterations and any of the phenotypic effects known to be associated with specific cytoplasms. Although the diploid species examined included all those which have been suggested as possible donors of the cytoplasm and the B genome to T. aestivum, none of the chosen accessions belonged to the same cytoplasmic class as T. aestivum itself, except that of the tetraploid T. dicoccoides. Therefore, none of the diploid accessions analysed was the B genome donor. The analyses did however support several other suggestions which have been made concerning wheat ancestry. Scoring the different chloroplast DNA types according to the rarity of their banding patterns indicated that four of the eleven cytoplasms are of relatively recent origin. The DNA alterations most easily detectable by the limited comparison of the eleven Triticum/Aegilops chloroplast DNA types using only 4 endonucleases were insertions and deletions. These ranged between approximately 50 bp and 1,200 bp in size and most of them were clustered in 2 segments of the large single-copy region of the genome. Only two examples of the loss of restriction endonuclease sites through possible point mutations were observed. No variation was detected in the inverted repeat regions. Several of the deletions and insertions map close to known chloroplast protein genes, and there is also an indication that the more variable regions of the chloroplast genome may contain sequences which have allowed DNA recombination and rearrangement to occur.
    Type of Medium: Electronic Resource
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