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Coexistence of NtCENH3 and two retrotransposons in tobacco centromeres.
Chromosome Res 2011; 19(5):591-605CR

Abstract

Although a centromeric DNA fragment of tobacco (Nicotiana tabacum), Nt2-7, has been reported, the overall structure of the centromeres remains unknown. To characterize the centromeric DNA sequences, we conducted a chromatin immunoprecipitation assay using anti-NtCENH3 antibody and chromatins isolated from two ancestral diploid species (Nicotiana sylvestris and Nicotiana tomentosiformis) of N. tabacum and isolated a 178-pb fragment, Nto1 from N. tomentosiformis, as a novel centromeric DNA. Fluorescence in situ hybridization (FISH) showed that Nto1 localizes on 24 out of 48 chromosomes in some cells of a BY-2 cell line. To identify the origins of the Nt2-7 and Nto1, a tobacco bacterial artificial chromosome (BAC) library was constructed from N. tabacum, and then screened by polymerase chain reaction (PCR) with primer sets designed from the Nt2-7 and Not1 DNA sequences. Twelve BAC clones were found to localize on the centromeric regions by FISH. We selected three BAC clones for sequencing and identified two centromeric retrotransposons, NtCR and NtoCR, the DNA sequences of which are similar to that of Nt2-7 and Nto1, respectively. Quantitative PCR analysis using coprecipitated DNA with anti-NtCENH3 clearly showed coexistence of NtCENH3 with both retrotransposons. These results indicate the possibility that these two retrotransposons act as centromeric DNA sequences in tobacco. NtoCR was found to be specific to N. tomentosiformis and T genome of N. tabacum, and a NtCR-like centromeric retrotransposon (TGRIV) exists in tomato. This specificity suggests that the times of amplification of these centromeric retrotransposons were different.

Authors+Show Affiliations

Institute of Plant Science and Resources, Okayama University, Kurashiki 710-0046, Japan. nagaki@rib.okayama-u.ac.jpNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

21626175

Citation

Nagaki, Kiyotaka, et al. "Coexistence of NtCENH3 and Two Retrotransposons in Tobacco Centromeres." Chromosome Research : an International Journal On the Molecular, Supramolecular and Evolutionary Aspects of Chromosome Biology, vol. 19, no. 5, 2011, pp. 591-605.
Nagaki K, Shibata F, Suzuki G, et al. Coexistence of NtCENH3 and two retrotransposons in tobacco centromeres. Chromosome Res. 2011;19(5):591-605.
Nagaki, K., Shibata, F., Suzuki, G., Kanatani, A., Ozaki, S., Hironaka, A., ... Murata, M. (2011). Coexistence of NtCENH3 and two retrotransposons in tobacco centromeres. Chromosome Research : an International Journal On the Molecular, Supramolecular and Evolutionary Aspects of Chromosome Biology, 19(5), pp. 591-605. doi:10.1007/s10577-011-9219-2.
Nagaki K, et al. Coexistence of NtCENH3 and Two Retrotransposons in Tobacco Centromeres. Chromosome Res. 2011;19(5):591-605. PubMed PMID: 21626175.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Coexistence of NtCENH3 and two retrotransposons in tobacco centromeres. AU - Nagaki,Kiyotaka, AU - Shibata,Fukashi, AU - Suzuki,Go, AU - Kanatani,Asaka, AU - Ozaki,Souichi, AU - Hironaka,Akiko, AU - Kashihara,Kazunari, AU - Murata,Minoru, Y1 - 2011/05/28/ PY - 2011/01/27/received PY - 2011/05/12/accepted PY - 2011/05/11/revised PY - 2011/6/1/entrez PY - 2011/6/1/pubmed PY - 2011/12/15/medline SP - 591 EP - 605 JF - Chromosome research : an international journal on the molecular, supramolecular and evolutionary aspects of chromosome biology JO - Chromosome Res. VL - 19 IS - 5 N2 - Although a centromeric DNA fragment of tobacco (Nicotiana tabacum), Nt2-7, has been reported, the overall structure of the centromeres remains unknown. To characterize the centromeric DNA sequences, we conducted a chromatin immunoprecipitation assay using anti-NtCENH3 antibody and chromatins isolated from two ancestral diploid species (Nicotiana sylvestris and Nicotiana tomentosiformis) of N. tabacum and isolated a 178-pb fragment, Nto1 from N. tomentosiformis, as a novel centromeric DNA. Fluorescence in situ hybridization (FISH) showed that Nto1 localizes on 24 out of 48 chromosomes in some cells of a BY-2 cell line. To identify the origins of the Nt2-7 and Nto1, a tobacco bacterial artificial chromosome (BAC) library was constructed from N. tabacum, and then screened by polymerase chain reaction (PCR) with primer sets designed from the Nt2-7 and Not1 DNA sequences. Twelve BAC clones were found to localize on the centromeric regions by FISH. We selected three BAC clones for sequencing and identified two centromeric retrotransposons, NtCR and NtoCR, the DNA sequences of which are similar to that of Nt2-7 and Nto1, respectively. Quantitative PCR analysis using coprecipitated DNA with anti-NtCENH3 clearly showed coexistence of NtCENH3 with both retrotransposons. These results indicate the possibility that these two retrotransposons act as centromeric DNA sequences in tobacco. NtoCR was found to be specific to N. tomentosiformis and T genome of N. tabacum, and a NtCR-like centromeric retrotransposon (TGRIV) exists in tomato. This specificity suggests that the times of amplification of these centromeric retrotransposons were different. SN - 1573-6849 UR - https://www.unboundmedicine.com/medline/citation/21626175/Coexistence_of_NtCENH3_and_two_retrotransposons_in_tobacco_centromeres_ L2 - https://doi.org/10.1007/s10577-011-9219-2 DB - PRIME DP - Unbound Medicine ER -