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doi:10.1534/genetics.105.048363
A more recent version of this article appeared on January 1, 2006.
REGULAR RESEARCH PAPERS |
Sister chromatids are often incompletely cohesed in meristematic and endopolyploid interphase nuclei of Arabidopsis thaliana
Veit Schubert 1, Marco Klatte 1, Ales Pecinka 1, Armin Meister 1, Zuzana Jasencakova 2 and Ingo Schubert 1*
1 Institute of Plant Genetics and Crop Plant Research (IPK)
2 Dulbecco Telethon Institute of Genetics & Biophysics CNR
* To whom correspondence should be addressed. E-mail: schubert{at}ipk-gatersleben.de.
Submitted on July 16, 2005
Revised on September 3, 2005
Accepted on 5 September 2005
We analysed whether sister chromatids are continuously cohesed in meristematic and endopolyploid Arabidopsis interphase nuclei by studying sister chromatid alignment at various chromosomal positions. FISH with individual BACs to flow-sorted 4C root and leaf nuclei frequently yielded more than two hybridization signals indicating incomplete or lacking sister chromatid alignment. Up to 100% of 8C, 16C and 32C nuclei showed no sister chromatid alignment at defined positions. Simultaneous FISH with BACs from different chromosomal positions revealed more frequent sister chromatid alignment in terminal than in mid arm positions. Centromeric positions were mainly aligned up to a ploidy level of 16C but became separated or dispersed in 32C nuclei. DNA hypomethylation (of the whole genome) and transcriptional activity (at FWA gene position) did not impair sister chromatid alignment. Only 6.1% of 4C leaf nuclei showed sister chromatid separation of entire chromosome 1 top arm territories. Homozygous transgenic tandem repeat (lac operator) arrays showing somatic homologous pairing more often than average euchromatic loci did not promote an increased frequency of sister chromatid alignment. The high frequency of separated sister chromatid arm positions in >4C nuclei suggests that sister chromatid cohesion is variable, dynamic and not obligatory along the entire chromosome arms in meristematic and differentiated Arabidopsis nuclei.
Key Words: Arabidopsis thaliana, fluorescent in situ hybridization, meristematic and endopolyploid nuclei, nuclear organization, sister chromatid cohesion
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