Physical mapping of DNA repetitive sequences to mitotic and meiotic chromosomes of Brassica oleracea var. alboglabra by fluorescence in situ hybridization

ARMSTRONG, SUSAN J; FRANSZ, PAUL; Marshall, David; JONES, GARETH H

doi:10.1038/sj.hdy.6884320

Cited by 19 publications

(25 citation statements)

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“…Robust molecular cytogenetic karyotypes established for B. rapa, B. oleracea, and B. napus: Numerous molecular cytogenetic studies have reported karyotypes in diploid and amphidiploid Brassica species on mitotic metaphase complements and meiotic prophase (pachytene) chromosomes (e.g., Armstrong et al 1998, Fukui et al 1998Snowdon et al 2002;Koo et al 2004;Lim et al 2005Lim et al , 2007Maluszynska and Hasterok 2005;Howell et al 2008;Kim et al 2009;Mun et al 2009). Among the crop species in the Brassicaceae, the B. rapa genetic, physical, and cytogenetic maps have begun to be integrated by BAC-FISH (Koo et al 2004;Lim et al 2005Lim et al , 2007Yang et al 2007;Kim et al 2009;Mun et al 2009;Xiong et al 2010).…”

Section: Discussionmentioning

confidence: 99%

“…Several karyotypes have been published for Brassica species on the basis of different staining methods, such as Giemsa staining, C-banding, CMA3/ 49-6-diamidino-2-phenylindole (DAPI) fluorescent staining, silver staining, and fluorescence in situ hybridization (FISH) with rDNA or pericentromeric tandem repeats (Olin-Faith and Hennen 1992;Maluszynska and Heslop-Harrison 1993;Cheng et al 1995;Snowdon et al 1997;Armstrong et al 1998;Fukui et al 1998;Hasterok et al 2001Hasterok et al , 2005Hasterok et al , 2006Howell et al 2002;Kulak et al 2002;Snowdon et al 2002;Ziolkowski and Sadowski 2002;Koo et al 2004;Maluszynska and Hasterok 2005;Lim et al 2005Lim et al , 2007. However, given the lack of specific chromosomal markers, even the best karyotypes developed could distinguish only six chromosomes in diploid B. rapa and three chromosomes in diploid B. oleracea.…”

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confidence: 99%

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Karyotype and Identification of All Homoeologous Chromosomes of AllopolyploidBrassica napusand Its Diploid Progenitors

2011

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Investigating recombination of homoeologous chromosomes in allopolyploid species is central to understanding plant breeding and evolution. However, examining chromosome pairing in the allotetraploid Brassica napus has been hampered by the lack of chromosome-specific molecular probes. In this study, we establish the identification of all homoeologous chromosomes of allopolyploid B. napus by using robust molecular cytogenetic karyotypes developed for the progenitor species Brassica rapa (A genome) and Brassica oleracea (C genome). The identification of every chromosome among these three Brassica species utilized genetically mapped bacterial artificial chromosomes (BACs) from B. rapa as probes for fluorescent in situ hybridization (FISH). With this BAC-FISH data, a second karyotype was developed using two BACs that contained repetitive DNA sequences and the ubiquitous ribosomal and pericentromere repeats. Using this diagnostic probe mix and a BAC that contained a C-genome repeat in two successive hybridizations allowed for routine identification of the corresponding homoeologous chromosomes between the A and C genomes of B. napus. When applied to the B. napus cultivar Stellar, we detected one chromosomal rearrangement relative to the parental karyotypes. This robust novel chromosomal painting technique will have biological applications for the understanding of chromosome pairing, homoeologous recombination, and genome evolution in the genus Brassica and will facilitate new applied breeding technologies that rely upon identification of chromosomes.

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

Karyotype and Identification of All Homoeologous Chromosomes of AllopolyploidBrassica napusand Its Diploid Progenitors

2011

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“…Westar and N-o-1, a DH line derived from Westar (provided by John Innes Centre, Norwich, UK). Methods for staging and fixing anthers have been described previously (Armstrong et al 1998;Howell et al 2002). Slides were prepared from these anthers following enzyme digestion (Armstrong et al 1998;Howell et al 2002) but cytohelicase was omitted from the enzyme mixture and the digestion time was extended to 2 hr.…”

Section: Methodsmentioning

confidence: 99%

A and C Genome Distinction and Chromosome Identification inBrassica napusby Sequential Fluorescencein SituHybridization and Genomicin SituHybridization

et al. 2008

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The two genomes (A and C) of the allopolyploid Brassica napus have been clearly distinguished using genomic in situ hybridization (GISH) despite the fact that the two extant diploids, B. rapa (A, n ¼ 10) and B. oleracea (C, n ¼ 9), representing the progenitor genomes, are closely related. Using DNA from B. oleracea as the probe, with B. rapa DNA and the intergenic spacer of the B. oleracea 45S rDNA as the block, hybridization occurred on 9 of the 19 chromosome pairs along the majority of their length. The pattern of hybridization confirms that the two genomes have remained distinct in B. napus line DH12075, with no significant genome homogenization and no large-scale translocations between the genomes. Fluorescence in situ hybridization (FISH)-with 45S rDNA and a BAC that hybridizes to the pericentromeric heterochromatin of several chromosomes-followed by GISH allowed identification of six chromosomes and also three chromosome groups. Our procedure was used on the B. napus cultivar Westar, which has an interstitial reciprocal translocation. Two translocated segments were detected in pollen mother cells at the pachytene stage of meiosis. Using B. oleracea chromosome-specific BACs as FISH probes followed by GISH, the chromosomes involved were confirmed to be A7 and C6.

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“…Slide preparations and FISH: Slide preparations were made from individual anthers and root tips, adjusting the digestion time as necessary (Armstrong et al 1998;Howell et al 2002). Methods for the slide pretreatment, hybridization, and posthybridization procedures for FISH have been described previously (Howell et al 2002).…”

Section: Methodsmentioning

confidence: 99%

Evolution of Neo-Sex Chromosomes inSilene diclinis

2009

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A small cluster of dioecious species in the plant genus Silene has evolved chromosomal sex determination and sex chromosomes relatively recently, within the last 10 million years (MY). Five dioecious Silene species (section Elisanthe) are very closely related (1-2 MY of divergence) and it was previously thought that all five have similar sex chromosomes. Here we demonstrate that in one of these species, Silene diclinis, the sex chromosomes have been significantly rearranged, resulting in the formation of neo-sex chromosomes. Fluorescence in situ hybridization with genic and repetitive probes revealed that in S. diclinis a reciprocal translocation has occurred between the ancestral Y chromosome and an autosome, resulting in chromosomes designated Y1 and Y2. Both Y1 and Y2 chromosomes are male specific. Y1 pairs with the X chromosome and with the autosome (the neo-X), which cosegregates with X. Y2 pairs only with the neo-X, forming a chain X-Y1-neo-X-Y2 in male meiosis. Despite very recent formation of the neo-sex chromosomes in S. diclinis, they are present in all surveyed individuals throughout the species range. Evolution of neo-sex chromosomes may be the cause of partial reproductive isolation of this species and could have been the isolating mechanism that drove speciation of S. diclinis.

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Physical mapping of DNA repetitive sequences to mitotic and meiotic chromosomes of Brassica oleracea var. alboglabra by fluorescence in situ hybridization

Cited by 19 publications

References 21 publications

Karyotype and Identification of All Homoeologous Chromosomes of AllopolyploidBrassica napusand Its Diploid Progenitors

Karyotype and Identification of All Homoeologous Chromosomes of AllopolyploidBrassica napusand Its Diploid Progenitors

A and C Genome Distinction and Chromosome Identification inBrassica napusby Sequential Fluorescencein SituHybridization and Genomicin SituHybridization

Evolution of Neo-Sex Chromosomes inSilene diclinis

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