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  • 1
    ISSN: 1432-2242
    Keywords: Wheat ; Anther-derived doubled haploids ; Chloroplast DNA ; Mitochondrial DNA ; Restriction patterns
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Chloroplast and mitochondrial compartments of a parental line of wheat (Triticum aestivum L., cv. ‘Moisson’) and its anther-derived doubled haploid lines have been analyzed and compared on the basis of their DNA restriction patterns. The results obtained show that no noticeable difference can be detected between doubled haploid lines and parental line at the level of ctDNA and mtDNA organization. It may be concluded that in vitro culture by itself does not systematically generate a cytoplasmic variation in germ cells.
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  • 2
    ISSN: 1432-2242
    Keywords: Ribosomal DNA ; Restriction fragment length polymorphism ; Anther-derived doubled haploids ; Gametoclonal variation ; Wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Restriction fragment length polymorphism of the wheat nuclear ribosomal DNA has been studied in several steps of a breeding scheme, including parental genotypes, F1 hybrid, F9 generation, and anther-derived doubled haploid lines obtained from F9. Ribosomal DNA represents a suitable molecular marker in following segregation and possible divergency from expected homozygosity after haploidization of an androgenetic process. It has been shown to undergo variations among the first cycle-doubled haploid lines in the relative amount of two different sizes of ribosomal DNA repeat units. The specificity and peculiar properties of the plant system used allowed us to assign an intrachromosomal location (short arm of the chromosomes 1B, 1R or 6B) to several ribosomal DNA repeat units that differ by the length of their nontranscribed spacer region.
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  • 3
    ISSN: 1432-2242
    Keywords: Wheat ; Somatic tissue culture ; Mitochondrial DNA ; Chondriome variability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have previously shown that the mitochondrial genome of long-term tissue cultures prepared from immature embryos of several varieties of cultivated wheat underwent variety-specific rearrangements resulting from either changes in the relative amounts of subgenomic components or from the appearance of novel genomic configurations. In the present work, both categories of rearrangements were studied in long-term tissue cultures initiated from other explants (shoot meristem, young leaf base, young root tip, immature inflorescence) of the same wheat variety (Chinese Spring) and were compared to those previously obtained with immature embryo cultures. Two main patterns of reorganization were found in a region of the mitochondrial genome known to be hypervariable in structure. In addition, some of the novel subgenomic configurations were obviously organ/tissue-specific whereas others were present in more than one type of organ. In several instances, the age of culture was found to determine the degree of mitochondrial DNA rearrangement. The data presented in this study strengthen the hypothesis of an association between a particular organization of the mitochondrial genome in tissue culture and its regeneration capacity.
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  • 4
    ISSN: 1432-2242
    Keywords: Wheat ; Somatic tissue culture ; Regenerated plants ; Mitochondrial DNA ; Chondriome variability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have previously shown that tissue cultures derived from various explants of the wheat variety Chinese Spring exhibit organ/tissue-specific changes in the organization of their mitochondrial genome. The aim of this work was to study the influence of passage out of in-vitro culture, and subsequent plant regeneration, on the in vitro “induced” reorganization of this genome. In all cases but one, subgenomic configurations present in both the donor parent and the tissue culture were evident, in corresponding regenerated plants. The presence, in regenerated plants, of subgenomic configurations found in tissue culture but undetectable in the donor parent appeared to be both timeand organ/tissue-dependent. Moreover, when present, these novel organizations were not systematically found in all regenerated plants. Finally, novel subgenomic configurations were specifically detected after passage out of in-vitro culture. As these results were obtained from a single plant variety, they clearly confirm the extreme plasticity of mitochondrial genome structure in response to in-vitro culture.
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  • 5
    ISSN: 1432-2242
    Keywords: Wheat ; Anther-derived doubled haploids ; Gametoclonal variation ; Ribosomal DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The organization of the nuclear ribosomal DNA from a parental line of wheat (Triticum aestivum L., cv. ‘César’) and its anther-derived first cycle and second cycle doubled haploid lines has been analyzed by DNA-DNA molecular hybridization. Restricted DNA has been probed by three subclones of wheat nuclear ribosomal DNA covering the entire repeat unit. No significant difference was detected in the extent of methylation of ribosomal DNA of the doubled haploid lines with respect to the parental line. On the other hand, a variation has been found in the organization of the nontranscribed spacer region of ribosomal DNA of the first cycle doubled haploid line. This variation remains stable after a second cycle of in vitro androgenesis. However, one out of five second cycle doubled haploid lines so far tested showed an additional hybridization band present in the parental line but lacking in the first cycle doubled haploid line.
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  • 6
    ISSN: 1432-2242
    Keywords: Mitochondrial DNA ; Chondriome variability ; In vitro culture ; Plant regeneration ; Wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Plants have been regenerated from short-and long-term in vitro somatic tissue cultures made from immature embryos of the hexaploid wheat cultivar “Chinese Spring”. The mitochondrial genome organization of each regenerated plantlet was studied, after one selfing, by probing Sal I-restricted total DNA with cloned Sal I fragments of wheat mitochondrial DNA derived from a segment of the genome, which displays marked structural changes in response to in vitro culture. Short-term in vitro cultures give rise to regenerated plants whose mitochondrial genome organization is either close to that of the parental cultivar or to that of embryogenic callus cultures, except for a single plant which has an organization resembling that of short-term non-embryogenic cultures. In contrast, all but one of the plants regenerated from long-term cultures exhibited a mitochondrial genome organization similar to that of long-term nonembryogenic cultures. In addition, extra labelled bands were detected in some of the regenerated plants with two of the probes used. These results emphasize the importance of the duration of the in vitro step preceding the regeneration process: the longer it is, the higher the probability is of obtaining mitochondrial DNA variability in regenerated plants. Furthermore, since increasing the duration of the in vitro stetp results in the production of regenerated plants with a mitochondrial genome organization resembling that of non-embryogenic tissue cultures, the question is thus raised as to whether regeneration from long-term cultures is suitable for use in plant breeding.
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  • 7
    ISSN: 1432-2242
    Keywords: Wheat ; Somatic tissue culture ; Regeneration ; Reciprocal crosses ; Mitochondrial DNA variability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The transmission of a structurally-hypervariable fraction of the mitochondrial genome has been studied in 42 F1 progenies obtained from reciprocal crosses between self-pollinated alloplasmic wheat plants regenerated after long-term somatic embryogenesis. This fraction of the genome is maternally and stoichiometrically inherited. In contrast, some additional restriction fragments specific to regenerated plants display a more complex mode of sexual transmission: one of the additional fragments was stoichiometrically and systematically inherited whereas two others were detected only in certain F1 hybrids. Assuming that the detection, by Southern analysis, of such a fragment in regenerated plants is due to the amplification of a pre-existing substoichiometric molecule generated by the activation of a rare recombination event, our results suggest that the probability of detecting a novel fragment in the F1 hybrids could be determined by the length of the repeated sequence at which recombination occurs.
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