Chromosome dynamics visualized with an anti-centromeric histone H3 antibody in Allium.

Due to the ease with which chromosomes can be observed, the Allium species, and onion in particular, have been familiar materials employed in cytogenetic experiments in biology. In this study, centromeric histone H3 (CENH3)-coding cDNAs were identified in four Allium species (onion, welsh onion, gar...

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Autores principales:	Kiyotaka Nagaki, Maki Yamamoto, Naoki Yamaji, Yasuhiko Mukai, Minoru Murata
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/b61929a3792f47948a5fe36e4b693b67
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spelling	oai:doaj.org-article:b61929a3792f47948a5fe36e4b693b672021-11-18T08:05:57ZChromosome dynamics visualized with an anti-centromeric histone H3 antibody in Allium.1932-620310.1371/journal.pone.0051315https://doaj.org/article/b61929a3792f47948a5fe36e4b693b672012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23236469/?tool=EBIhttps://doaj.org/toc/1932-6203Due to the ease with which chromosomes can be observed, the Allium species, and onion in particular, have been familiar materials employed in cytogenetic experiments in biology. In this study, centromeric histone H3 (CENH3)-coding cDNAs were identified in four Allium species (onion, welsh onion, garlic and garlic chives) and cloned. Anti-CENH3 antibody was then raised against a deduced amino acid sequence of CENH3 of welsh onion. The antibody recognized all CENH3 orthologs of the Allium species tested. Immunostaining with the antibody enabled clear visualization of chromosome behavior during mitosis in the species. Furthermore, three-dimensional (3D) observation of mitotic cell division was achieved by subjecting root sections to immunohistochemical techniques. The 3D dynamics of the cells and position of cell-cycle marker proteins (CENH3 and α-tubulin) were clearly revealed by immunohistochemical staining with the antibodies. The immunohistochemical analysis made it possible to establish an overview of the location of dividing cells in the root tissues. This breakthrough in technique, in addition to the two centromeric DNA sequences isolated from welsh onion by chromatin immuno-precipitation using the antibody, should lead to a better understanding of plant cell division. A phylogenetic analysis of Allium CENH3s together with the previously reported plant CENH3s showed two separate clades for monocot species tested. One clade was made from CENH3s of the Allium species with those of Poaceae species, and the other from CENH3s of a holocentric species (Luzula nivea). These data may imply functional differences of CENH3s between holocentric and monocentric species. Centromeric localization of DNA sequences isolated from welsh onion by chromatin immuno-precipitation (ChIP) using the antibody was confirmed by fluorescence in situ hybridization and ChIP-quantitative PCR.Kiyotaka NagakiMaki YamamotoNaoki YamajiYasuhiko MukaiMinoru MurataPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 12, p e51315 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Kiyotaka Nagaki Maki Yamamoto Naoki Yamaji Yasuhiko Mukai Minoru Murata Chromosome dynamics visualized with an anti-centromeric histone H3 antibody in Allium.
description	Due to the ease with which chromosomes can be observed, the Allium species, and onion in particular, have been familiar materials employed in cytogenetic experiments in biology. In this study, centromeric histone H3 (CENH3)-coding cDNAs were identified in four Allium species (onion, welsh onion, garlic and garlic chives) and cloned. Anti-CENH3 antibody was then raised against a deduced amino acid sequence of CENH3 of welsh onion. The antibody recognized all CENH3 orthologs of the Allium species tested. Immunostaining with the antibody enabled clear visualization of chromosome behavior during mitosis in the species. Furthermore, three-dimensional (3D) observation of mitotic cell division was achieved by subjecting root sections to immunohistochemical techniques. The 3D dynamics of the cells and position of cell-cycle marker proteins (CENH3 and α-tubulin) were clearly revealed by immunohistochemical staining with the antibodies. The immunohistochemical analysis made it possible to establish an overview of the location of dividing cells in the root tissues. This breakthrough in technique, in addition to the two centromeric DNA sequences isolated from welsh onion by chromatin immuno-precipitation using the antibody, should lead to a better understanding of plant cell division. A phylogenetic analysis of Allium CENH3s together with the previously reported plant CENH3s showed two separate clades for monocot species tested. One clade was made from CENH3s of the Allium species with those of Poaceae species, and the other from CENH3s of a holocentric species (Luzula nivea). These data may imply functional differences of CENH3s between holocentric and monocentric species. Centromeric localization of DNA sequences isolated from welsh onion by chromatin immuno-precipitation (ChIP) using the antibody was confirmed by fluorescence in situ hybridization and ChIP-quantitative PCR.
format	article
author	Kiyotaka Nagaki Maki Yamamoto Naoki Yamaji Yasuhiko Mukai Minoru Murata
author_facet	Kiyotaka Nagaki Maki Yamamoto Naoki Yamaji Yasuhiko Mukai Minoru Murata
author_sort	Kiyotaka Nagaki
title	Chromosome dynamics visualized with an anti-centromeric histone H3 antibody in Allium.
title_short	Chromosome dynamics visualized with an anti-centromeric histone H3 antibody in Allium.
title_full	Chromosome dynamics visualized with an anti-centromeric histone H3 antibody in Allium.
title_fullStr	Chromosome dynamics visualized with an anti-centromeric histone H3 antibody in Allium.
title_full_unstemmed	Chromosome dynamics visualized with an anti-centromeric histone H3 antibody in Allium.
title_sort	chromosome dynamics visualized with an anti-centromeric histone h3 antibody in allium.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/b61929a3792f47948a5fe36e4b693b67
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_version_	1718422252910804992

Chromosome dynamics visualized with an anti-centromeric histone H3 antibody in Allium.

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