Flow Cytometric Analysis and Chromosome Sorting of Barley (Hordeum vulgare L.)

Lee, Jai‐Heon; Arumuganathan, K.; Chung, Youngjin; Kim, Kee‐Young; Chung, Won-Bok; Bae, Kae-Sun; Kim, Doh-Hoon; Chung, Dae-Soo; Kwon, Oh Chang

doi:10.1007/s10059-000-0619-y

Cited by 10 publications

(4 citation statements)

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“…Examples of the typical flow karyotypes for different human cell lines are shown in Figure 9A. In all cases the conventional spectrum comparable with the data published [Dean, 1985; Lee et al, 2000] has been obtained and, in each case, the spectrum contains chromosome aggregates (clumps) as expected on the right‐hand part (Fig. 9A, brackets).…”

Section: Resultssupporting

confidence: 80%

“…Nonrandom chromosomal associations on the mitotic ring are reported for some cell types [Kuwano et al, 1992; Nagele et al, 1995]. It was supposed that small chromosomes were more likely to be involved in association [Van den Engh et al, 1984; Lee et al, 2000]. In contrast to this supposition, the clump formation in our experiments does not depend on the chromosome length.…”

Section: Discussioncontrasting

confidence: 64%

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Evidence for the existence of satellite DNA‐containing connection between metaphase chromosomes

Kuznetsova

Enukashvily

Noniashvili

et al. 2007

J of Cellular Biochemistry

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Physical connections between mitotic chromosomes have been reported previously. It was assumed that the interchromosome connection was based on the DNA-protein thread. However, the data about DNA sequences and protein component in the thread is fragmentary. We demonstrated on the mouse cultured cell line and prematurely condensed chromosomes that: (a) all four mouse satellite DNA fragments (major and minor satellite, mouse satellite 3 (MS3) and mouse satellite 4 (MS4)) were involved in the thread formation; (b) MS4 was involved in the thread to the least extent among all the other fragments; (c) telomere was never a member of the thread; (d) the thread was synthesized at a late G(2) phase; (e) RNA helicase p68 and CENP-B were among the protein components of the interchromosome connection. It was shown by FACS analysis that in mouse and human cell lines: (1) the flow karyotype spectrums were never free from chromosome aggregates; (2) chromosome association did not depend on the chromosome length and each chromosome was free to associate with the other.

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Section: Resultssupporting

confidence: 80%

Section: Discussioncontrasting

confidence: 64%

Evidence for the existence of satellite DNA‐containing connection between metaphase chromosomes

Kuznetsova

Enukashvily

Noniashvili

et al. 2007

J of Cellular Biochemistry

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“…In a majority of plant species analyzed so far, differences in relative DNA content, or DNA base content among the chromosomes, are too small to permit discrimination of all chromosomes using a single DNA fluorochrome. Unfortunately, bivariate flow karyotyping after simultaneous chromosome staining with AT-and GC-binding fluorochromes, which is a standard in human flow karyotyping [67], does not lead to a significant improvement [68,69] and has been rarely used. An alternative, and to date the only successful method for bivariate flow karyotyping in plants, has been to fluorescently label a repetitive DNA sequence which is abundant and non-homogenously distributed across the chromosomes of the karyotype.…”

Section: Bivariate Flow Karyotypingmentioning

confidence: 99%

Chromosome analysis and sorting

et al. 2021

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Flow cytometric analysis and sorting of plant mitotic chromosomes has been mastered by only a few laboratories worldwide. Yet, it has been contributing significantly to progress in plant genetics, including the production of genome assemblies and the cloning of important genes. The dissection of complex genomes by flow sorting into the individual chromosomes that represent small parts of the genome reduces DNA sample complexity and streamlines projects relying on molecular and genomic techniques. Whereas flow cytometric analysis, that is, chromosome classification according to fluorescence and light scatter properties, is an integral part of any chromosome sorting project, it has rarely been used on its own due to lower resolution and sensitivity as compared to other cytogenetic methods. To perform chromosome analysis and sorting, commercially available electrostatic droplet sorters are suitable. However, in order to resolve and purify chromosomes of interest the instrument must offer high resolution of optical signals as well as stability during long runs. The challenge is thus not the instrumentation, but the adequate sample preparation. The sample must be a suspension of intact mitotic metaphase chromosomes and the protocol, which includes the induction of cell cycle synchrony, accumulation of dividing cells at metaphase, and release of undamaged chromosomes, is time consuming and laborious and needs to be performed very carefully. Moreover, in addition to fluorescent staining chromosomal DNA, the protocol may include specific labelling of DNA repeats to facilitate discrimination of particular chromosomes. This review introduces the applications of chromosome sorting in plants, and discusses in detail sample preparation, chromosome analysis and sorting to achieve the highest purity in flowsorted fractions, and their suitability for downstream applications.

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“…List of Triticeae species in which flow cytometric chromosome sorting has been reported (adapted fromDoležel et al (2014)) . (1999);Lee et al (2000);Suchánková et al (2006);Mayer et al (2009Mayer et al ( , 2011 …”

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confidence: 99%

Genomics of Wild Relatives and Alien Introgressions

Rey

Molnár

Doležel

2015

Alien Introgression in Wheat

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Alien introgression breeding is an attractive approach to recover genetic variation that was lost during wheat domestication and breeding. New alleles and genes may be introduced from wild relatives from the tribe Triticeae, which exhibit large genetic variation and many potentially useful traits. Although a range of wheat-alien introgression lines has been developed, apart from the 1BL.1RS translocation, only a few commercial wheat cultivars benefitted from alien introgression. This is a consequence of poor knowledge of genome structure of wild donors, limited ability to control chromosome behavior during meiosis in interspecific hybrids and introgression lines, difficulties in eliminating unwanted chromatin transferred simultaneously with genes of interest, as well as a lack of tools permitting large-scale production and characterization of introgression lines. Recent advances in molecular and flow cytogenetics and genomics are bound to change the situation. New insights into the meiotic recombination raise the hopes for the ability to control its frequency and distribution. The progress in comparative genome analysis provides clues about the genome collinearity between wild species and wheat, making it possible to assess chances for chromosome recombination and predict its outcomes. Genomics tools enable massive and high-resolution screening of hybrids and their progenies and characterize their genomes, including the development of markers linked to traits of interest. Until recently, little attention has been paid to the function of introgressed genes and their interaction with the host genome. However, also this has been changing and all these achievements make the breeding of improved wheat cultivars using wild germplasm a realistic goal. Keywords (separated by "-") Wheat breeding-Alien gene function-Chromosome flow sorting-Chromosome genomics-Epigenetic modifications-Genome sequencing-Interspecific hybridization-Molecular cytogenetics-Molecular markers AUTHOR QUERIES

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Flow Cytometric Analysis and Chromosome Sorting of Barley (Hordeum vulgare L.)

Cited by 10 publications

References 0 publications

Evidence for the existence of satellite DNA‐containing connection between metaphase chromosomes

Evidence for the existence of satellite DNA‐containing connection between metaphase chromosomes

Chromosome analysis and sorting

Genomics of Wild Relatives and Alien Introgressions

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