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Cuyacot, Abigail Rubiato,Won, So Youn,Park, Sang Kun,Sohn, Seong-Han,Lee, Jungho,Kim, Jung Sun,Kim, Hyun Hee,Lim, Ki-Byung,Hwang, Yoon-Jung Elsevier 2016 Scientia horticulturae Vol.198 No.-
<P><B>Abstract</B></P> <P>The sequencing of <I>Chrysanthemum boreale</I> genome is now in progress. However, the repetitive DNA sequences that dominates greatly in the genome have not yet been elucidated in <I>Chrysanthemum</I>. Therefore, we performed fluorescence <I>in situ</I> hybridization (FISH) by using the mitotic metaphase chromosome of <I>C. boreale</I> with well-identified repeats and C<SUB>0</SUB>t DNA as probes. The most conserved repeats, rDNAs, as well as telomeric repeats were first observed. Three pairs of 45S rDNA sites were located at the termini of the short arms of chromosomes 3, 7, and 8, while one pair of 5S rDNA signals was observed in the middle of the long arm of chromosome 4. <I>Arabidopsis</I> -type telomeric repeats colocalizing with 45S rDNA sites, were detected at the ends of both chromosome arms. Next, on the basis of DNA reassociation kinetics, three types of C<SUB>0</SUB>t DNA that exhibited different degrees of repetitive nature were fractionated and used as FISH probes to map the repetitive sequences. Signals from all C<SUB>0</SUB>t DNAs were successfully observed, but their coverage on the chromosomes was different for C<SUB>0</SUB>t-1, C<SUB>0</SUB>t-10, and C<SUB>0</SUB>t-100. C<SUB>0</SUB>t-1 FISH signals showed intensity in the telomeric region and were dispersed on both chromosome arms, except for distal regions. C<SUB>0</SUB>t-10 signals were observed almost in all parts of the chromosome, with greater intensity around pericentromeric regions. Bright C<SUB>0</SUB>t-100 signals were observed throughout the chromosome excluding some distal parts. C<SUB>0</SUB>t FISH signals covered the rDNAs and telomeric repeats, thus signifying their repetitive attributes. The relative proportions of repetitive DNA sequences in <I>C. boreale</I> genome is discussed.</P> <P><B>Highlights</B></P> <P> <UL> <LI> rDNAs (5S and 45S), telomeric repeats and C<SUB>0</SUB>t DNAs in <I>Chrysanthemum boreale</I> chromosome are localized. </LI> <LI> FISH karyotype of <I>C. boreale</I> is clarified. </LI> <LI> 45S rDNA loci show colocalization with <I>Arabidopsis</I> -type telomeric repeats. </LI> <LI> Repetitive DNAs (C<SUB>0</SUB>t DNA) of <I>C. boreale</I> localize predominantly in pericentromeric and telomeric regions. </LI> <LI> Relative proportions of repetitive sequences in <I>C. boreale</I> genome is discussed. </LI> </UL> </P>
Abigail Rubiato Cuyacot,황윤정,임기병,김현희 한국원예학회 2017 Horticulture, Environment, and Biotechnology Vol.58 No.5
The aim of this study was to gain insight into the basic genetics of Chrysanthemum zawadskii Herbich ssp. lucidum (NAK.) Y. Lee, a Chrysanthemum subspecies from Ulleung Island, South Korea, by investigating its karyotypicand cytogenetic characteristics, and to contribute to the karyosystematic study of the genus Chrysanthemum (tribe:Anthemideae). Fluorescence in situ hybridization (FISH) was used to characterize the ploidy level, chromosomeconstitution, and distribution of 5S and 18S ribosomal DNA (rDNA) and Arabidopsis-type telomere repeats (TTTAGGG)nin C. zawadskii ssp. lucidum chromosomes. The somatic metaphase chromosome number of C. zawadskii ssp. lucidumwas tetraploid (2n = 4x = 36). Signals from four 5S rDNA loci were detected in the middle of the long arms ofsome chromosomes, whereas signals from eight 18S rDNA loci were located at the termini of the short arms. The tworDNA markers localized to different chromosomes with no co-localization. The Arabidopsis-type telomeric repeats,however, co-localized with 18S rDNA sites at the ends of both chromosomal arms. This co-localization of telomericsequences and 18S rDNA has also been observed in other Chrysanthemum species with various levels of ploidy,indicating that it is a general feature of the genus. DAPI (4′,6-diamidino-2-phenylindole) bands incidentally detectedafter FISH are also illustrated. These preliminary data contribute towards understanding the karyotypic trends androle of polyploidy in the evolution of the genus Chrysanthemum.
C0t Analysis of Chrysanthemum boreale: the Realization of its Genome Characteristics
Abigail Rubiato Cuyacot,So Youn Won,Sang Kun Park,Seong-Han Sohn,Ki-Byung Lim,Hyun Hee Kim,Franklin Hinosa Mancia,Yoon-Jung Hwang 한국육종학회 2015 한국육종학회 심포지엄 Vol.2015 No.07
In the genus Chrysanthemum, repetitive DNA sequences, the dominant part of a genome, are still to be elucidated. To explore the matter, the present study applied fluorescent in situ hybridization (FISH) to the mitotic metaphase chromosome of Chrysanthemum boreale with C0t DNA as probes. Based on DNA re-assotiation kinetics, three kinds of C0t DNA exhibiting different degrees of repetitive nature were fractionated and used as FISH probes to map the repetitive sequences. Signals from all C0t DNAs were successfully observed but their coverage on the chromosomes was different among C0t-1, C0t-10, and C0t-100. C0t-1 FISH signals resulted to have its intensity on the telomeric region and were also dispersed on both chromosome arms except for some distal regions. In C0t-10, signals were observed in all parts of the chromosome with greater intensity around pericentromeric regions. FISH with C0t-100 DNA was observed in bright signals all over the chromosome. Signals of C0t FISH found in this study covered the regions where ribosomal DNAs and telomeric repeats of C. boreale have been distributed (previous report), thus signifying their repetitive attributes. The present results could enhance the efficiency of studying genomes, chromosomes and repetitive sequences of C. boreale and subsequently hasten the realization of the genetic scheme of Chrysanthemum.