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Application of flow cytometry for the study of plant genomes

100%
Dolezel J.
|
|
issue 3
285-302
EN
During the last fifteen years, flow cytometry and sorting greatly contributed to the improvement of our knowledge of plant genome structure and function. This paper reviews the applications of flow cytometry for the analysis of isolated nuclei and chromosomes. Because of its speed, precision and convenience, this method of analysis of the nuclear DNA content finds an enormous number of applications which cover basic research, breeding and production. The results obtained with chromosome analysis and sorting indicate that the technique might greatly simplify the analysis of plant genomes at the molecular level.
2

Karyological characterization of sugar beet gynogenetic lines cultured in vitro

63%
Svirshchevskaya A., Dolezel J.
Journal of Applied Genetics
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2001
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vol. 42
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issue 1
21-32
EN
Flow cytometry was used to screen ploidy levels in 47 cultured in vitro sugar beet gynogenetic lines of various origin and age, obtained after plant regeneration from unfertilized ovules. When donor plants were diploid, the majority of regenerants were found to have cells with 1C, 2C and 4C relative DNA content (mainly haploid and diploid) and there were large differences in the rate of spontaneous in vitro chromosome doubling between individual homozygous lines. Six ovule-derived lines regenerated from fertile and sterile diploid donors of forty-five lines were solid diploids from the very early stages of their in vitro cultivation, and thus could not be classified as doubled haploids. In the case of tetraploid donor plants, the gynogenetic regenerants demonstrated 2x-ploidy level. The results obtained in chimeric plants with both haploid and diploid cells indicated the possibility to overcome mixoploidy by their re-cultivation through generative shoot tip culture. The flow cytometry method confirmed data obtained by conventional microscopic chromosome counting in dividing leaf cells and was found very useful for screening of a large number of regenerants and for characterizing the process of in vitro gynogenetic lines formation in sugar beet.
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