• dr Elżbieta Wolny
Stanowisko: Adiunkt
Jednostka: Wydział Nauk Przyrodniczych
Adres: 40-032 Katowice, ul. Jagiellońska 28
Piętro: III
Numer pokoju: A-319
Telefon: (32) 2009 553
E-mail: elzbieta.wolny@us.edu.pl
Spis publikacji: Spis wg CINiBA
Spis publikacji: Spis wg OPUS
Scopus Author ID: 9939998300
Publikacje z bazy Scopus
2021
Piński, A.; Betekhtin, A.; Kwaśniewska, J.; Chajec, Ł.; Wolny, E. A.; Hasterok, R.
3,4‐dehydro‐l‐proline induces programmed cell death in the roots of brachypodium distachyon Journal Article
In: International Journal of Molecular Sciences, vol. 22, no. 14, 2021, ISSN: 16616596, (2).
@article{2-s2.0-85110027459,
title = {3,4‐dehydro‐l‐proline induces programmed cell death in the roots of brachypodium distachyon},
author = { A. Piński and A. Betekhtin and J. Kwaśniewska and Ł. Chajec and E.A. Wolny and R. Hasterok},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-85110027459&doi=10.3390%2fijms22147548&partnerID=40&md5=045642b6770419a71c99a63d6dd94f4c},
doi = {10.3390/ijms22147548},
issn = {16616596},
year = {2021},
date = {2021-01-01},
journal = {International Journal of Molecular Sciences},
volume = {22},
number = {14},
publisher = {MDPI},
abstract = {As cell wall proteins, the hydroxyproline‐rich glycoproteins (HRGPs) take part in plant growth and various developmental processes. To fulfil their functions, HRGPs, extensins (EXTs) in particular, undergo the hydroxylation of proline by the prolyl‐4‐hydroxylases. The activity of these enzymes can be inhibited with 3,4‐dehydro‐L‐proline (3;4‐DHP), which enables its application to reveal the functions of the HRGPs. Thus, to study the involvement of HRGPs in the development of root hairs and roots, we treated seedlings of Brachypodium distachyon with 250 μM, 500 μM, and 750 μM of 3,4‐DHP. The histological observations showed that the root epidermis cells and the cortex cells beneath them ruptured. The immunostaining experiments using the JIM20 antibody, which recognizes the EXT epitopes, demonstrated the higher abundance of this epitope in the control compared to the treated samples. The transmission electron microscopy analyses revealed morphological and ultrastructural features that are typical for the vacuolar‐type of cell death. Using the TUNEL test (terminal deoxynucleotidyl transferase dUTP nick end labelling), we showed an increase in the number of nuclei with damaged DNA in the roots that had been treated with 3,4‐DHP compared to the control. Finally, an analysis of two metacaspases’ gene activity revealed an increase in their expression in the treated roots. Altogether, our results show that inhibiting the prolyl‐4‐hydroxyl-ases with 3,4‐DHP results in a vacuolar‐type of cell death in roots, thereby highlighting the important role of HRGPs in root hair development and root growth. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.},
note = {2},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
As cell wall proteins, the hydroxyproline‐rich glycoproteins (HRGPs) take part in plant growth and various developmental processes. To fulfil their functions, HRGPs, extensins (EXTs) in particular, undergo the hydroxylation of proline by the prolyl‐4‐hydroxylases. The activity of these enzymes can be inhibited with 3,4‐dehydro‐L‐proline (3;4‐DHP), which enables its application to reveal the functions of the HRGPs. Thus, to study the involvement of HRGPs in the development of root hairs and roots, we treated seedlings of Brachypodium distachyon with 250 μM, 500 μM, and 750 μM of 3,4‐DHP. The histological observations showed that the root epidermis cells and the cortex cells beneath them ruptured. The immunostaining experiments using the JIM20 antibody, which recognizes the EXT epitopes, demonstrated the higher abundance of this epitope in the control compared to the treated samples. The transmission electron microscopy analyses revealed morphological and ultrastructural features that are typical for the vacuolar‐type of cell death. Using the TUNEL test (terminal deoxynucleotidyl transferase dUTP nick end labelling), we showed an increase in the number of nuclei with damaged DNA in the roots that had been treated with 3,4‐DHP compared to the control. Finally, an analysis of two metacaspases’ gene activity revealed an increase in their expression in the treated roots. Altogether, our results show that inhibiting the prolyl‐4‐hydroxyl-ases with 3,4‐DHP results in a vacuolar‐type of cell death in roots, thereby highlighting the important role of HRGPs in root hair development and root growth. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.
Wolny, E. A.; Skalska, A.; Brąszewska-Zalewska, A. J.; Mur, L. A. J.; Hasterok, R.
Defining the cell wall, cell cycle and chromatin landmarks in the responses of brachypodium distachyon to salinity Journal Article
In: International Journal of Molecular Sciences, vol. 22, no. 2, pp. 1-23, 2021, ISSN: 16616596, (12).
@article{2-s2.0-85100110960,
title = {Defining the cell wall, cell cycle and chromatin landmarks in the responses of brachypodium distachyon to salinity},
author = { E.A. Wolny and A. Skalska and A.J. Brąszewska-Zalewska and L.A.J. Mur and R. Hasterok},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-85100110960&doi=10.3390%2fijms22020949&partnerID=40&md5=aacee36bf01649a69d0bc62ec3e40d65},
doi = {10.3390/ijms22020949},
issn = {16616596},
year = {2021},
date = {2021-01-01},
journal = {International Journal of Molecular Sciences},
volume = {22},
number = {2},
pages = {1-23},
publisher = {MDPI AG},
abstract = {Excess salinity is a major stress that limits crop yields. Here, we used the model grass Brachypodium distachyon (Brachypodium) reference line Bd21 in order to define the key molecular events in the responses to salt during germination. Salt was applied either throughout the germination period (“salt stress”) or only after root emergence (“salt shock”). Germination was affected at ≥100 mM and root elongation at ≥75 mM NaCl. The expression of arabinogalactan proteins (AGPs), FLA1, FLA10, FLA11, AGP20 and AGP26, which regulate cell wall expansion (especially FLA11), were mostly induced by the “salt stress” but to a lesser extent by “salt shock”. Cytological assessment using two AGP epitopes, JIM8 and JIM13 indicated that “salt stress” increases the fluorescence signals in rhizodermal and exodermal cell wall. Cell division was suppressed at >75 mM NaCl. The cell cycle genes (CDKB1; CDKB2; CYCA3; CYCB1; WEE1) were induced by “salt stress” in a concentration-dependent manner but not CDKA, CYCA and CYCLIN-D4-1-RELATED. Under “salt shock”, the cell cycle genes were optimally expressed at 100 mM NaCl. These changes were consistent with the cell cycle arrest, possibly at the G1 phase. The salt-induced genomic damage was linked with the oxidative events via an increased glutathione accumulation. Histone acetylation and methylation and DNA methylation were visualized by immunofluorescence. Histone H4 acetylation at lysine 5 increased strongly whereas DNA methylation decreased with the application of salt. Taken together, we suggest that salt-induced oxidative stress causes genomic damage but that it also has epigenetic effects, which might modulate the cell cycle and AGP expression gene. Based on these landmarks, we aim to encourage functional genomics studies on the responses of Brachypodium to salt. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.},
note = {12},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Excess salinity is a major stress that limits crop yields. Here, we used the model grass Brachypodium distachyon (Brachypodium) reference line Bd21 in order to define the key molecular events in the responses to salt during germination. Salt was applied either throughout the germination period (“salt stress”) or only after root emergence (“salt shock”). Germination was affected at ≥100 mM and root elongation at ≥75 mM NaCl. The expression of arabinogalactan proteins (AGPs), FLA1, FLA10, FLA11, AGP20 and AGP26, which regulate cell wall expansion (especially FLA11), were mostly induced by the “salt stress” but to a lesser extent by “salt shock”. Cytological assessment using two AGP epitopes, JIM8 and JIM13 indicated that “salt stress” increases the fluorescence signals in rhizodermal and exodermal cell wall. Cell division was suppressed at >75 mM NaCl. The cell cycle genes (CDKB1; CDKB2; CYCA3; CYCB1; WEE1) were induced by “salt stress” in a concentration-dependent manner but not CDKA, CYCA and CYCLIN-D4-1-RELATED. Under “salt shock”, the cell cycle genes were optimally expressed at 100 mM NaCl. These changes were consistent with the cell cycle arrest, possibly at the G1 phase. The salt-induced genomic damage was linked with the oxidative events via an increased glutathione accumulation. Histone acetylation and methylation and DNA methylation were visualized by immunofluorescence. Histone H4 acetylation at lysine 5 increased strongly whereas DNA methylation decreased with the application of salt. Taken together, we suggest that salt-induced oxidative stress causes genomic damage but that it also has epigenetic effects, which might modulate the cell cycle and AGP expression gene. Based on these landmarks, we aim to encourage functional genomics studies on the responses of Brachypodium to salt. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.
Skalska, A.; Wolny, E. A.; Beckmann, M.; Doonan, J. H.; Hasterok, R.; Mur, L. A. J.
Allotetraploidization in brachypodium may have led to the dominance of one parent’s metabolome in germinating seeds Journal Article
In: Cells, vol. 10, no. 4, 2021, ISSN: 20734409.
@article{2-s2.0-85105054260,
title = {Allotetraploidization in brachypodium may have led to the dominance of one parent’s metabolome in germinating seeds},
author = { A. Skalska and E.A. Wolny and M. Beckmann and J.H. Doonan and R. Hasterok and L.A.J. Mur},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-85105054260&doi=10.3390%2fcells10040828&partnerID=40&md5=f2b582d61460aff5dc06392d44d8357d},
doi = {10.3390/cells10040828},
issn = {20734409},
year = {2021},
date = {2021-01-01},
journal = {Cells},
volume = {10},
number = {4},
publisher = {MDPI},
abstract = {Seed germination is a complex process during which a mature seed resumes metabolic activity to prepare for seedling growth. In this study, we performed a comparative metabolomic analysis of the embryo and endosperm using the community standard lines of three annual Brachypodium species, i.e., B. distachyon (Bd) and B. stacei (Bs) and their natural allotetraploid B. hybridum (BdBs) that has wider ecological range than the other two species. We explored how far the metabolomic impact of allotetraploidization would be observable as over-lapping changes at 4, 12, and 24 h after imbibition (HAI) with water when germination was initiated. Metabolic changes during germination were more prominent in Brachypodium embryos than in the endosperm. The embryo and endosperm metabolomes of Bs and BdBs were similar, and those of Bd were distinctive. The Bs and BdBs embryos showed increased levels of sugars and the tricarboxylic acid cycle compared to Bd, which could have been indicative of better nutrient mobilization from the endosperm. Bs and BdBs also showed higher oxalate levels that could aid nutrient transfer through altered cellular events. In Brachypodium endosperm, the thick cell wall, in addition to starch, has been suggested to be a source of nutrients to the embryo. Metabolites indicative of sugar metabolism in the endosperm of all three species were not prominent, suggesting that mobilization mostly occurred prior to 4 HAI. Hydroxycinnamic and monolignol changes in Bs and BdBs were consistent with cell wall remodeling that arose following the release of nutrients to the respective embryos. Amino acid changes in both the embryo and endosperm were broadly consistent across the species. Taking our data together, the formation of BdBs may have maintained much of the Bs metabolome in both the embryo and endosperm during the early stages of germination. In the embryo, this conserved Bs metabolome appeared to include an elevated sugar metabolism that played a vital role in germination. If these observations are confirmed in the future with more Brachypodium accessions, it would substantiate the dominance of the Bs metabolome in BdBs allotetraploidization and the use of metabolomics to suggest important adaptive changes. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Seed germination is a complex process during which a mature seed resumes metabolic activity to prepare for seedling growth. In this study, we performed a comparative metabolomic analysis of the embryo and endosperm using the community standard lines of three annual Brachypodium species, i.e., B. distachyon (Bd) and B. stacei (Bs) and their natural allotetraploid B. hybridum (BdBs) that has wider ecological range than the other two species. We explored how far the metabolomic impact of allotetraploidization would be observable as over-lapping changes at 4, 12, and 24 h after imbibition (HAI) with water when germination was initiated. Metabolic changes during germination were more prominent in Brachypodium embryos than in the endosperm. The embryo and endosperm metabolomes of Bs and BdBs were similar, and those of Bd were distinctive. The Bs and BdBs embryos showed increased levels of sugars and the tricarboxylic acid cycle compared to Bd, which could have been indicative of better nutrient mobilization from the endosperm. Bs and BdBs also showed higher oxalate levels that could aid nutrient transfer through altered cellular events. In Brachypodium endosperm, the thick cell wall, in addition to starch, has been suggested to be a source of nutrients to the embryo. Metabolites indicative of sugar metabolism in the endosperm of all three species were not prominent, suggesting that mobilization mostly occurred prior to 4 HAI. Hydroxycinnamic and monolignol changes in Bs and BdBs were consistent with cell wall remodeling that arose following the release of nutrients to the respective embryos. Amino acid changes in both the embryo and endosperm were broadly consistent across the species. Taking our data together, the formation of BdBs may have maintained much of the Bs metabolome in both the embryo and endosperm during the early stages of germination. In the embryo, this conserved Bs metabolome appeared to include an elevated sugar metabolism that played a vital role in germination. If these observations are confirmed in the future with more Brachypodium accessions, it would substantiate the dominance of the Bs metabolome in BdBs allotetraploidization and the use of metabolomics to suggest important adaptive changes. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.
2020
Gordon, S. P.; Contreras-Moreira, B.; Levy, J. J.; Djamei, A.; Czedik-Eysenberg, A.; Tartaglio, V. S.; Session, A.; Martin, J.; Cartwright, A.; Katz, A.; Singan, V. R.; Goltsman, E.; Barry, K.; Dinh-Thi, V. H.; Chalhoub, B.; Díaz-Pérez, A.; Sancho, R.; Lusinska, J.; Wolny, E. A.; Nibau, C.; Doonan, J. H.; Mur, L. A. J.; Plott, C.; Jenkins, J.; Hazen, S. P.; Lee, S. J.; Shu, S.; Goodstein, D.; Rokhsar, D.; Schmutz, J.; Hasterok, R.; Catalán, P.; Vogel, J. P.
Gradual polyploid genome evolution revealed by pan-genomic analysis of Brachypodium hybridum and its diploid progenitors Journal Article
In: Nature Communications, vol. 11, no. 1, 2020, ISSN: 20411723, (27).
@article{2-s2.0-85088698921,
title = {Gradual polyploid genome evolution revealed by pan-genomic analysis of Brachypodium hybridum and its diploid progenitors},
author = { S.P. Gordon and B. Contreras-Moreira and J.J. Levy and A. Djamei and A. Czedik-Eysenberg and V.S. Tartaglio and A. Session and J. Martin and A. Cartwright and A. Katz and V.R. Singan and E. Goltsman and K. Barry and V.H. Dinh-Thi and B. Chalhoub and A. Díaz-Pérez and R. Sancho and J. Lusinska and E.A. Wolny and C. Nibau and J.H. Doonan and L.A.J. Mur and C. Plott and J. Jenkins and S.P. Hazen and S.J. Lee and S. Shu and D. Goodstein and D. Rokhsar and J. Schmutz and R. Hasterok and P. Catalán and J.P. Vogel},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-85088698921&doi=10.1038%2fs41467-020-17302-5&partnerID=40&md5=a244ebf9c21dbfb16be4ae4a1e58bd83},
doi = {10.1038/s41467-020-17302-5},
issn = {20411723},
year = {2020},
date = {2020-01-01},
journal = {Nature Communications},
volume = {11},
number = {1},
publisher = {Nature Research},
abstract = {Our understanding of polyploid genome evolution is constrained because we cannot know the exact founders of a particular polyploid. To differentiate between founder effects and post polyploidization evolution, we use a pan-genomic approach to study the allotetraploid Brachypodium hybridum and its diploid progenitors. Comparative analysis suggests that most B. hybridum whole gene presence/absence variation is part of the standing variation in its diploid progenitors. Analysis of nuclear single nucleotide variants, plastomes and k-mers associated with retrotransposons reveals two independent origins for B. hybridum, ~1.4 and ~0.14 million years ago. Examination of gene expression in the younger B. hybridum lineage reveals no bias in overall subgenome expression. Our results are consistent with a gradual accumulation of genomic changes after polyploidization and a lack of subgenome expression dominance. Significantly, if we did not use a pan-genomic approach, we would grossly overestimate the number of genomic changes attributable to post polyploidization evolution. © 2020, This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply.},
note = {27},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Our understanding of polyploid genome evolution is constrained because we cannot know the exact founders of a particular polyploid. To differentiate between founder effects and post polyploidization evolution, we use a pan-genomic approach to study the allotetraploid Brachypodium hybridum and its diploid progenitors. Comparative analysis suggests that most B. hybridum whole gene presence/absence variation is part of the standing variation in its diploid progenitors. Analysis of nuclear single nucleotide variants, plastomes and k-mers associated with retrotransposons reveals two independent origins for B. hybridum, ~1.4 and ~0.14 million years ago. Examination of gene expression in the younger B. hybridum lineage reveals no bias in overall subgenome expression. Our results are consistent with a gradual accumulation of genomic changes after polyploidization and a lack of subgenome expression dominance. Significantly, if we did not use a pan-genomic approach, we would grossly overestimate the number of genomic changes attributable to post polyploidization evolution. © 2020, This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply.
2019
Lusinska, J.; Betekhtin, A.; López-Alvarez, D.; Catalán, P.; Jenkins, G.; Wolny, E. A.; Hasterok, R.
Comparatively barcoded chromosomes of brachypodium perennials tell the story of their karyotype structure and evolution Journal Article
In: International Journal of Molecular Sciences, vol. 20, no. 22, 2019, ISSN: 16616596, (7).
@article{2-s2.0-85074666863,
title = {Comparatively barcoded chromosomes of brachypodium perennials tell the story of their karyotype structure and evolution},
author = { J. Lusinska and A. Betekhtin and D. López-Alvarez and P. Catalán and G. Jenkins and E.A. Wolny and R. Hasterok},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-85074666863&doi=10.3390%2fijms20225557&partnerID=40&md5=1e87ac6eba426f4ef1f180cb67cf71f1},
doi = {10.3390/ijms20225557},
issn = {16616596},
year = {2019},
date = {2019-01-01},
journal = {International Journal of Molecular Sciences},
volume = {20},
number = {22},
publisher = {MDPI AG},
abstract = {The Brachypodium genus is an informative model system for studying grass karyotype organization. Previous studies of a limited number of species and reference chromosomes have not provided a comprehensive picture of the enigmatic phylogenetic relationships in the genus. Comparative chromosome barcoding, which enables the reconstruction of the evolutionary history of individual chromosomes and their segments, allowed us to infer the relationships between putative ancestral karyotypes of extinct species and extant karyotypes of current species. We used over 80 chromosome-specific BAC (bacterial artificial chromosome) clones derived from five reference chromosomes of B. distachyon as probes against the karyotypes of twelve accessions representing five diploid and polyploid Brachypodium perennials. The results showed that descending dysploidy is common in Brachypodium and occurs primarily via nested chromosome fusions. Brachypodium distachyon was rejected as a putative ancestor for allotetraploid perennials and B. stacei for B. mexicanum. We propose two alternative models of perennial polyploid evolution involving either the incorporation of a putative x = 5 ancestral karyotype with different descending dysploidy patterns compared to B. distachyon chromosomes or hybridization of two x = 9 ancestors followed by genome doubling and descending dysploidy. Details of the karyotype structure and evolution in several Brachypodium perennials are revealed for the first time. © 2019 by the authors. Licensee MDPI, Basel, Switzerland.},
note = {7},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
The Brachypodium genus is an informative model system for studying grass karyotype organization. Previous studies of a limited number of species and reference chromosomes have not provided a comprehensive picture of the enigmatic phylogenetic relationships in the genus. Comparative chromosome barcoding, which enables the reconstruction of the evolutionary history of individual chromosomes and their segments, allowed us to infer the relationships between putative ancestral karyotypes of extinct species and extant karyotypes of current species. We used over 80 chromosome-specific BAC (bacterial artificial chromosome) clones derived from five reference chromosomes of B. distachyon as probes against the karyotypes of twelve accessions representing five diploid and polyploid Brachypodium perennials. The results showed that descending dysploidy is common in Brachypodium and occurs primarily via nested chromosome fusions. Brachypodium distachyon was rejected as a putative ancestor for allotetraploid perennials and B. stacei for B. mexicanum. We propose two alternative models of perennial polyploid evolution involving either the incorporation of a putative x = 5 ancestral karyotype with different descending dysploidy patterns compared to B. distachyon chromosomes or hybridization of two x = 9 ancestors followed by genome doubling and descending dysploidy. Details of the karyotype structure and evolution in several Brachypodium perennials are revealed for the first time. © 2019 by the authors. Licensee MDPI, Basel, Switzerland.
Borowska-Zuchowska, N.; Robaszkiewicz, E.; Wolny, E. A.; Betekhtin, A.; Hasterok, R.
Ribosomal DNA loci derived from Brachypodium stacei are switched off for major parts of the life cycle of Brachypodium hybridum Journal Article
In: Journal of Experimental Botany, vol. 70, no. 3, pp. 871-883, 2019, ISSN: 00220957, (3).
@article{2-s2.0-85061128284,
title = {Ribosomal DNA loci derived from Brachypodium stacei are switched off for major parts of the life cycle of Brachypodium hybridum},
author = { N. Borowska-Zuchowska and E. Robaszkiewicz and E.A. Wolny and A. Betekhtin and R. Hasterok},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-85061128284&doi=10.1093%2fjxb%2fery425&partnerID=40&md5=a123ef59aae852b182e62c863f89ff7c},
doi = {10.1093/jxb/ery425},
issn = {00220957},
year = {2019},
date = {2019-01-01},
journal = {Journal of Experimental Botany},
volume = {70},
number = {3},
pages = {871-883},
publisher = {Oxford University Press},
abstract = {Nucleolar dominance is an epigenetic phenomenon that occurs in some plant and animal allopolyploids and hybrids, whereby only one ancestral set of 35S rRNA genes retains the ability to form the nucleolus while the rDNA loci derived from the other progenitor are transcriptionally silenced. There is substantial evidence that nucleolar dominance is regulated developmentally. This study focuses upon the establishment and/or maintenance of nucleolar dominance during different stages of development in the model grass allotetraploid Brachypodium hybridum. Fluorescence in situ hybridization with a 25S rDNA probe to cells in three-dimensional cytogenetic preparations showed that nucleolar dominance is present not only in root meristematic and differentiated cells of this species, but also in male meiocytes at prophase I, tetrads of microspores, and different embryonic tissues. The inactive state of Brachypodium stacei-originated rDNA loci was confirmed by silver staining. Only B. distachyon-derived 35S rDNA loci formed nucleoli in the aforementioned tissues, whereas B. stacei-like loci remained highly condensed and thus transcriptionally suppressed. The establishment of nucleolar dominance during earlier stages of B. hybridum embryo development cannot be ruled out. However, we propose that gradual pseudogenization of B. stacei-like loci in the evolution of the allotetraploid seems to be more likely. © 2018 The Author(s).},
note = {3},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Nucleolar dominance is an epigenetic phenomenon that occurs in some plant and animal allopolyploids and hybrids, whereby only one ancestral set of 35S rRNA genes retains the ability to form the nucleolus while the rDNA loci derived from the other progenitor are transcriptionally silenced. There is substantial evidence that nucleolar dominance is regulated developmentally. This study focuses upon the establishment and/or maintenance of nucleolar dominance during different stages of development in the model grass allotetraploid Brachypodium hybridum. Fluorescence in situ hybridization with a 25S rDNA probe to cells in three-dimensional cytogenetic preparations showed that nucleolar dominance is present not only in root meristematic and differentiated cells of this species, but also in male meiocytes at prophase I, tetrads of microspores, and different embryonic tissues. The inactive state of Brachypodium stacei-originated rDNA loci was confirmed by silver staining. Only B. distachyon-derived 35S rDNA loci formed nucleoli in the aforementioned tissues, whereas B. stacei-like loci remained highly condensed and thus transcriptionally suppressed. The establishment of nucleolar dominance during earlier stages of B. hybridum embryo development cannot be ruled out. However, we propose that gradual pseudogenization of B. stacei-like loci in the evolution of the allotetraploid seems to be more likely. © 2018 The Author(s).
2018
Wolny, E. A.; Betekhtin, A.; Rojek-Jelonek, M.; Brąszewska-Zalewska, A. J.; Lusinska, J.; Hasterok, R.
Germination and the early stages of seedling development in brachypodium distachyon Journal Article
In: International Journal of Molecular Sciences, vol. 19, no. 10, 2018, ISSN: 16616596, (31).
@article{2-s2.0-85054137635,
title = {Germination and the early stages of seedling development in brachypodium distachyon},
author = { E.A. Wolny and A. Betekhtin and M. Rojek-Jelonek and A.J. Brąszewska-Zalewska and J. Lusinska and R. Hasterok},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-85054137635&doi=10.3390%2fijms19102916&partnerID=40&md5=6bc3a0bce97cc709e1970b6a859568af},
doi = {10.3390/ijms19102916},
issn = {16616596},
year = {2018},
date = {2018-01-01},
journal = {International Journal of Molecular Sciences},
volume = {19},
number = {10},
publisher = {MDPI AG},
abstract = {Successful germination and seedling development are crucial steps in the growth of a new plant. In this study, we investigated the course of the cell cycle during germination in relation to grain hydration in the model grass Brachypodium distachyon (Brachypodium) for the first time. Flow cytometry was performed to monitor the cell cycle progression during germination and to estimate DNA content in embryo tissues. The analyses of whole zygotic embryos revealed that the relative DNA content was 2C, 4C, 8C, and 16C. Endoreplicated nuclei were detected in the scutellum and coleorhiza cells, whereas the rest of the embryo tissues only had nuclei with a 2C and 4C DNA content. This study was accompanied by a spatiotemporal profile analysis of the DNA synthetic activity in the organs of Brachypodium embryos during germination using EdU labelling. Upon imbibition, nuclear DNA replication was initiated in the radicle within 11 h and subsequently spread towards the plumule. The first EdU-labelled prophases were observed after 14 h of imbibition. Analysis of selected genes that are involved in the regulation of the cell cycle, such as those encoding cyclin-dependent kinases and cyclins, demonstrated an increase in their expression profiles. © 2018 by the authors. Licensee MDPI, Basel, Switzerland.},
note = {31},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Successful germination and seedling development are crucial steps in the growth of a new plant. In this study, we investigated the course of the cell cycle during germination in relation to grain hydration in the model grass Brachypodium distachyon (Brachypodium) for the first time. Flow cytometry was performed to monitor the cell cycle progression during germination and to estimate DNA content in embryo tissues. The analyses of whole zygotic embryos revealed that the relative DNA content was 2C, 4C, 8C, and 16C. Endoreplicated nuclei were detected in the scutellum and coleorhiza cells, whereas the rest of the embryo tissues only had nuclei with a 2C and 4C DNA content. This study was accompanied by a spatiotemporal profile analysis of the DNA synthetic activity in the organs of Brachypodium embryos during germination using EdU labelling. Upon imbibition, nuclear DNA replication was initiated in the radicle within 11 h and subsequently spread towards the plumule. The first EdU-labelled prophases were observed after 14 h of imbibition. Analysis of selected genes that are involved in the regulation of the cell cycle, such as those encoding cyclin-dependent kinases and cyclins, demonstrated an increase in their expression profiles. © 2018 by the authors. Licensee MDPI, Basel, Switzerland.
Lusinska, J.; Majka, J.; Betekhtin, A.; Susek, K.; Wolny, E. A.; Hasterok, R.
Chromosome identification and reconstruction of evolutionary rearrangements in Brachypodium distachyon, B. stacei and B. hybridum Journal Article
In: Annals of Botany, vol. 122, no. 3, pp. 445-459, 2018, ISSN: 03057364, (22).
@article{2-s2.0-85054793804,
title = {Chromosome identification and reconstruction of evolutionary rearrangements in Brachypodium distachyon, B. stacei and B. hybridum},
author = { J. Lusinska and J. Majka and A. Betekhtin and K. Susek and E.A. Wolny and R. Hasterok},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-85054793804&doi=10.1093%2faob%2fmcy086&partnerID=40&md5=502a5a8f625a9f3465e1961f5b82860d},
doi = {10.1093/aob/mcy086},
issn = {03057364},
year = {2018},
date = {2018-01-01},
journal = {Annals of Botany},
volume = {122},
number = {3},
pages = {445-459},
publisher = {Oxford University Press},
abstract = {Background and Aims The Brachypodium genus represents a useful model system to study grass genome organization. Palaeogenomic analyses (e.g. Murat F; Armero A; Pont C; Klopp C; Salse J. 2017. Reconstructing the genome of the most recent common ancestor of flowering plants. Nature Genetics49: 490-496) have identified polyploidization and dysploidy as the prime mechanisms driving the diversity of plant karyotypes and nested chromosome fusions (NCFs) crucial for shaping grass chromosomes. This study compares the karyotype structure and evolution in B. distachyon (genome Bd), B. stacei (genome Bs) and in their putative allotetraploid B. hybridum (genomes BdBs). Methods Brachypodium chromosomes were measured and identified using multicolour fluorescence in situ hybridization (mcFISH). For higher resolution, comparative chromosome barcoding was developed using sets of low-repeat, physically mapped B. distachyon-derived bacterial artificial chromosome (BAC) clones. Key Results All species had rather small chromosomes, and essentially all in the Bs genome were morphometrically indistinguishable. Seven BACs combined with two rDNA-based probes provided unambiguous and reproducible chromosome discrimination. Comparative chromosome barcoding revealed NCFs that contributed to the reduction in the x = 12 chromosome number that has been suggested for the intermediate ancestral grass karyotype. Chromosome Bd3 derives from two NCFs of three ancestral chromosomes (Os2; Os8; Os10). Chromosome Bs6 shows an ancient Os8/Os10 NCF, whilst Bs4 represents Os2 only. Chromosome Bd4 originated from a descending dysploidy that involves two NCFs of Os12, Os9 and Os11. The specific distribution of BACs along Bs9 and Bs5, in both B. stacei and B. hybridum, suggests a Bs genome-specific Robertsonian rearrangement. Conclusions mcFISH-based karyotyping identifies all chromosomes in Brachypodium annuals. Comparative chromosome barcoding reveals rearrangements responsible for the diverse organization of Bd and Bs genomes and provides new data regarding karyotype evolution since the split of the two diploids. The fact that no chromosome rearrangements were observed in B. hybridum compared with the karyotypes of its phylogenetic ancestors suggests prolonged genome stasis after the formation of the allotetraploid. © 2018 The Author(s).},
note = {22},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Background and Aims The Brachypodium genus represents a useful model system to study grass genome organization. Palaeogenomic analyses (e.g. Murat F; Armero A; Pont C; Klopp C; Salse J. 2017. Reconstructing the genome of the most recent common ancestor of flowering plants. Nature Genetics49: 490-496) have identified polyploidization and dysploidy as the prime mechanisms driving the diversity of plant karyotypes and nested chromosome fusions (NCFs) crucial for shaping grass chromosomes. This study compares the karyotype structure and evolution in B. distachyon (genome Bd), B. stacei (genome Bs) and in their putative allotetraploid B. hybridum (genomes BdBs). Methods Brachypodium chromosomes were measured and identified using multicolour fluorescence in situ hybridization (mcFISH). For higher resolution, comparative chromosome barcoding was developed using sets of low-repeat, physically mapped B. distachyon-derived bacterial artificial chromosome (BAC) clones. Key Results All species had rather small chromosomes, and essentially all in the Bs genome were morphometrically indistinguishable. Seven BACs combined with two rDNA-based probes provided unambiguous and reproducible chromosome discrimination. Comparative chromosome barcoding revealed NCFs that contributed to the reduction in the x = 12 chromosome number that has been suggested for the intermediate ancestral grass karyotype. Chromosome Bd3 derives from two NCFs of three ancestral chromosomes (Os2; Os8; Os10). Chromosome Bs6 shows an ancient Os8/Os10 NCF, whilst Bs4 represents Os2 only. Chromosome Bd4 originated from a descending dysploidy that involves two NCFs of Os12, Os9 and Os11. The specific distribution of BACs along Bs9 and Bs5, in both B. stacei and B. hybridum, suggests a Bs genome-specific Robertsonian rearrangement. Conclusions mcFISH-based karyotyping identifies all chromosomes in Brachypodium annuals. Comparative chromosome barcoding reveals rearrangements responsible for the diverse organization of Bd and Bs genomes and provides new data regarding karyotype evolution since the split of the two diploids. The fact that no chromosome rearrangements were observed in B. hybridum compared with the karyotypes of its phylogenetic ancestors suggests prolonged genome stasis after the formation of the allotetraploid. © 2018 The Author(s).
Betekhtin, A.; Milewska-Hendel, A.; Chajec, Ł.; Rojek-Jelonek, M.; Nowak, K.; Kwaśniewska, J.; Wolny, E. A.; Kurczyńska, E. U.; Hasterok, R.
5-Azacitidine induces cell death in a tissue culture of brachypodium distachyon Journal Article
In: International Journal of Molecular Sciences, vol. 19, no. 6, 2018, ISSN: 16616596, (14).
@article{2-s2.0-85048889878,
title = {5-Azacitidine induces cell death in a tissue culture of brachypodium distachyon},
author = { A. Betekhtin and A. Milewska-Hendel and Ł. Chajec and M. Rojek-Jelonek and K. Nowak and J. Kwaśniewska and E.A. Wolny and E.U. Kurczyńska and R. Hasterok},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-85048889878&doi=10.3390%2fijms19061806&partnerID=40&md5=72a6b817774ef0346f00f6530b07a6ac},
doi = {10.3390/ijms19061806},
issn = {16616596},
year = {2018},
date = {2018-01-01},
journal = {International Journal of Molecular Sciences},
volume = {19},
number = {6},
publisher = {MDPI AG},
abstract = {Morphological and histological observations revealed that, at a concentration of 50 µM, 5-azacitidine (5-azaC) totally inhibited the induction of embryogenic masses (EM), while the cultivation of explants (zygotic embryos; ZEs) in the presence of 5 µM of 5-azaC led to the formation of a callus with EM in 10% of the cases. Transmission electron microscopy (TEM) analyzes revealed the presence of the morphological and ultrastructural features that are typical for the vacuolar type of cell death in the callus cells that were treated. A TUNEL assay confirmed the presence of DNA double-strand breaks for the callus cells that had been treated with both 5 and 50 µM 5-azaC concentrations. Analysis of the gene expression of selected cell death markers demonstrated a reduced expression of metacaspase, protein executer 1 (EX1), and thioredoxin (TRX) in the callus cells that had been treated compared to the control culture. The strongest increase in the gene activity was characteristic for glutathione S-transferase (GST). Our studies also included an analysis of the distribution of some arabinogalactan proteins (AGPs) and extensin epitopes, which can be used as markers of cells that are undergoing death in a Brachypodium distachyon tissue culture. © 2018 by the authors. Licensee MDPI, Basel, Switzerland.},
note = {14},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Morphological and histological observations revealed that, at a concentration of 50 µM, 5-azacitidine (5-azaC) totally inhibited the induction of embryogenic masses (EM), while the cultivation of explants (zygotic embryos; ZEs) in the presence of 5 µM of 5-azaC led to the formation of a callus with EM in 10% of the cases. Transmission electron microscopy (TEM) analyzes revealed the presence of the morphological and ultrastructural features that are typical for the vacuolar type of cell death in the callus cells that were treated. A TUNEL assay confirmed the presence of DNA double-strand breaks for the callus cells that had been treated with both 5 and 50 µM 5-azaC concentrations. Analysis of the gene expression of selected cell death markers demonstrated a reduced expression of metacaspase, protein executer 1 (EX1), and thioredoxin (TRX) in the callus cells that had been treated compared to the control culture. The strongest increase in the gene activity was characteristic for glutathione S-transferase (GST). Our studies also included an analysis of the distribution of some arabinogalactan proteins (AGPs) and extensin epitopes, which can be used as markers of cells that are undergoing death in a Brachypodium distachyon tissue culture. © 2018 by the authors. Licensee MDPI, Basel, Switzerland.
2017
Wolny, E. A.; Brąszewska-Zalewska, A. J.; Kroczek, D.; Hasterok, R.
In: Protoplasma, vol. 254, no. 5, pp. 2045-2052, 2017, ISSN: 0033183X, (8).
@article{2-s2.0-85013775595,
title = {Histone H3 and H4 acetylation patterns are more dynamic than those of DNA methylation in Brachypodium distachyon embryos during seed maturation and germination},
author = { E.A. Wolny and A.J. Brąszewska-Zalewska and D. Kroczek and R. Hasterok},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-85013775595&doi=10.1007%2fs00709-017-1088-x&partnerID=40&md5=8beaeb9cef0bb0cad966271c94b002e0},
doi = {10.1007/s00709-017-1088-x},
issn = {0033183X},
year = {2017},
date = {2017-01-01},
journal = {Protoplasma},
volume = {254},
number = {5},
pages = {2045-2052},
publisher = {Springer-Verlag Wien},
abstract = {The transition of seeds from a dry to a metabolically active state requires significant changes in both the spatial and temporal patterns of gene expression, and this transcriptional reprogramming involves various modifications of the chromatin structure. There are several factors that can greatly influence the structure of chromatin, one of which is the chemical modifications of histone proteins and DNA itself. In this study, we analysed the distribution of three epigenetic markers, i.e. acetylation of histone H4 (H4K16ac) and histone H3 (H3K18ac) as well as DNA methylation (5mC) in Brachypodium distachyon embryos during the four stages of seed development—maturation, desiccation (quiescence), imbibition and germination. Our results indicate that both H4K16ac and H3K18ac are at a very high level in embryos during seed imbibition, but that the patterns of DNA methylation are considerably more stable in embryos during seed development. © 2017, The Author(s).},
note = {8},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
The transition of seeds from a dry to a metabolically active state requires significant changes in both the spatial and temporal patterns of gene expression, and this transcriptional reprogramming involves various modifications of the chromatin structure. There are several factors that can greatly influence the structure of chromatin, one of which is the chemical modifications of histone proteins and DNA itself. In this study, we analysed the distribution of three epigenetic markers, i.e. acetylation of histone H4 (H4K16ac) and histone H3 (H3K18ac) as well as DNA methylation (5mC) in Brachypodium distachyon embryos during the four stages of seed development—maturation, desiccation (quiescence), imbibition and germination. Our results indicate that both H4K16ac and H3K18ac are at a very high level in embryos during seed imbibition, but that the patterns of DNA methylation are considerably more stable in embryos during seed development. © 2017, The Author(s).
2015
Wolny, E. A.; Brąszewska-Zalewska, A. J.; Kroczek, D.; Hasterok, R.
In situ analysis of epigenetic modifications in the chromatin of Brachypodium distachyon embryos Journal Article
In: Plant Signaling and Behavior, vol. 10, no. 5, pp. 1-2, 2015, ISSN: 15592316, (1).
@article{2-s2.0-84944031628,
title = {In situ analysis of epigenetic modifications in the chromatin of Brachypodium distachyon embryos},
author = { E.A. Wolny and A.J. Brąszewska-Zalewska and D. Kroczek and R. Hasterok},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-84944031628&doi=10.1080%2f15592324.2015.1011948&partnerID=40&md5=3b979188a97b567a2cf78e22419d2247},
doi = {10.1080/15592324.2015.1011948},
issn = {15592316},
year = {2015},
date = {2015-01-01},
journal = {Plant Signaling and Behavior},
volume = {10},
number = {5},
pages = {1-2},
publisher = {Taylor and Francis Inc.},
abstract = {Epigenetic modifications of the chromatin structure are crucial for many biological processes and act on genes during the development and germination of seeds. The spatial distribution of 3 epigenetic markers, i.e. H4K5ac, H3K4me2 and H3K4me1 was investigated in ‘matured,’ ‘dry,’ ‘imbibed” and ‘germinating’ embryos of a model grass, Brachypodium. Our results indicate that the patterns of epigenetic modification differ in the various types of tissues of embryos that were analyzed. Such a tissue-specific manner of these modifications may be linked to the switch of the gene expression profiles in various organs of the developing embryo. © 2015 Taylor & Francis Group, LLC.},
note = {1},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Epigenetic modifications of the chromatin structure are crucial for many biological processes and act on genes during the development and germination of seeds. The spatial distribution of 3 epigenetic markers, i.e. H4K5ac, H3K4me2 and H3K4me1 was investigated in ‘matured,’ ‘dry,’ ‘imbibed” and ‘germinating’ embryos of a model grass, Brachypodium. Our results indicate that the patterns of epigenetic modification differ in the various types of tissues of embryos that were analyzed. Such a tissue-specific manner of these modifications may be linked to the switch of the gene expression profiles in various organs of the developing embryo. © 2015 Taylor & Francis Group, LLC.
2014
Słomka, A.; Wolny, E. A.; Kuta, E.
Viola tricolor (Violaceae) is a karyologically unstable species Journal Article
In: Plant Biosystems, vol. 148, no. 4, pp. 602-608, 2014, ISSN: 11263504, (6).
@article{2-s2.0-84922158000,
title = {Viola tricolor (Violaceae) is a karyologically unstable species},
author = { A. Słomka and E.A. Wolny and E. Kuta},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-84922158000&doi=10.1080%2f11263504.2013.788576&partnerID=40&md5=86cf4b7a0894b0c1d9e0206ea8c2e489},
doi = {10.1080/11263504.2013.788576},
issn = {11263504},
year = {2014},
date = {2014-01-01},
journal = {Plant Biosystems},
volume = {148},
number = {4},
pages = {602-608},
publisher = {Taylor and Francis Ltd.},
abstract = {Viola tricolor is a pseudometallophyte covering heavy-metal-polluted and non-polluted areas. The species is a member of the evolutionarily young sect. Melanium of Viola. In this study, we sought to determine whether the karyotype of V. tricolor is stable with respect to chromosome structure or is altered depending on environmental conditions (heavy-metal-polluted vs. non-polluted areas). We established the karyotypes of plant material originating from a Zakopane meadow (non-metallicolous population) and from the Bukowno mine waste heap (metallicolous population), showing evident interpopulation differentiation in chromosome type (2M+20 m+2sm+2st vs. 18 m+8sm), in the number, size, and distribution of rDNA loci (25S and 5S), and also in chromosome mutations, mainly fission of chromosomes into acentric fragments and translocation of the fragments. Variable numbers of both 25S and 5S rDNA loci were distributed at different positions of the chromosomes and not on specific pairs of chromosomes. The results clearly indicate that the karyotype of V. tricolor results from the unstable genetic structure of the species. This character, typical for relatively young evolutionary groups, proves its membership to the Melanium section considered to be young within the genus Viola. © 2013, © 2013 Societá Botanica Italiana.},
note = {6},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Viola tricolor is a pseudometallophyte covering heavy-metal-polluted and non-polluted areas. The species is a member of the evolutionarily young sect. Melanium of Viola. In this study, we sought to determine whether the karyotype of V. tricolor is stable with respect to chromosome structure or is altered depending on environmental conditions (heavy-metal-polluted vs. non-polluted areas). We established the karyotypes of plant material originating from a Zakopane meadow (non-metallicolous population) and from the Bukowno mine waste heap (metallicolous population), showing evident interpopulation differentiation in chromosome type (2M+20 m+2sm+2st vs. 18 m+8sm), in the number, size, and distribution of rDNA loci (25S and 5S), and also in chromosome mutations, mainly fission of chromosomes into acentric fragments and translocation of the fragments. Variable numbers of both 25S and 5S rDNA loci were distributed at different positions of the chromosomes and not on specific pairs of chromosomes. The results clearly indicate that the karyotype of V. tricolor results from the unstable genetic structure of the species. This character, typical for relatively young evolutionary groups, proves its membership to the Melanium section considered to be young within the genus Viola. © 2013, © 2013 Societá Botanica Italiana.
Wolny, E. A.; Brąszewska-Zalewska, A. J.; Hasterok, R.
Spatial distribution of epigenetic modifications in Brachypodium distachyon embryos during seed maturation and germination Journal Article
In: PLoS ONE, vol. 9, no. 7, 2014, ISSN: 19326203, (18).
@article{2-s2.0-84904103572,
title = {Spatial distribution of epigenetic modifications in Brachypodium distachyon embryos during seed maturation and germination},
author = { E.A. Wolny and A.J. Brąszewska-Zalewska and R. Hasterok},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-84904103572&doi=10.1371%2fjournal.pone.0101246&partnerID=40&md5=73dbd091faa86c9270cf0ac270eeb31e},
doi = {10.1371/journal.pone.0101246},
issn = {19326203},
year = {2014},
date = {2014-01-01},
journal = {PLoS ONE},
volume = {9},
number = {7},
publisher = {Public Library of Science},
abstract = {Seed development involves a plethora of spatially and temporally synchronised genetic and epigenetic processes. Although it has been shown that epigenetic mechanisms, such as DNA methylation and chromatin remodelling, act on a large number of genes during seed development and germination, to date the global levels of histone modifications have not been studied in a tissue-specific manner in plant embryos. In this study we analysed the distribution of three epigenetic markers, i.e. H4K5ac, H3K4me2 and H3K4me1 in 'matured', 'dry' and 'germinating' embryos of a model grass, Brachypodium distachyon (Brachypodium). Our results indicate that the abundance of these modifications differs considerably in various organs and tissues of the three types of Brachypodium embryos. Embryos from matured seeds were characterised by the highest level of H4K5ac in RAM and epithelial cells of the scutellum, whereas this modification was not observed in the coleorhiza. In this type of embryos H3K4me2 was most evident in epithelial cells of the scutellum. In 'dry' embryos H4K5ac was highest in the coleorhiza but was not present in the nuclei of the scutellum. H3K4me1 was the most elevated in the coleoptile but absent from the coleorhiza, whereas H3K4me2 was the most prominent in leaf primordia and RAM. In embryos from germinating seeds H4K5ac was the most evident in the scutellum but not present in the coleoptile, similarly H3K4me1 was the highest in the scutellum and very low in the coleoptile, while the highest level of H3K4me2 was observed in the coleoptile and the lowest in the coleorhiza. The distinct patterns of epigenetic modifications that were observed may be involved in the switch of the gene expression profiles in specific organs of the developing embryo and may be linked with the physiological changes that accompany seed desiccation, imbibition and germination. © 2014 Wolny et al.},
note = {18},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Seed development involves a plethora of spatially and temporally synchronised genetic and epigenetic processes. Although it has been shown that epigenetic mechanisms, such as DNA methylation and chromatin remodelling, act on a large number of genes during seed development and germination, to date the global levels of histone modifications have not been studied in a tissue-specific manner in plant embryos. In this study we analysed the distribution of three epigenetic markers, i.e. H4K5ac, H3K4me2 and H3K4me1 in 'matured', 'dry' and 'germinating' embryos of a model grass, Brachypodium distachyon (Brachypodium). Our results indicate that the abundance of these modifications differs considerably in various organs and tissues of the three types of Brachypodium embryos. Embryos from matured seeds were characterised by the highest level of H4K5ac in RAM and epithelial cells of the scutellum, whereas this modification was not observed in the coleorhiza. In this type of embryos H3K4me2 was most evident in epithelial cells of the scutellum. In 'dry' embryos H4K5ac was highest in the coleorhiza but was not present in the nuclei of the scutellum. H3K4me1 was the most elevated in the coleoptile but absent from the coleorhiza, whereas H3K4me2 was the most prominent in leaf primordia and RAM. In embryos from germinating seeds H4K5ac was the most evident in the scutellum but not present in the coleoptile, similarly H3K4me1 was the highest in the scutellum and very low in the coleoptile, while the highest level of H3K4me2 was observed in the coleoptile and the lowest in the coleorhiza. The distinct patterns of epigenetic modifications that were observed may be involved in the switch of the gene expression profiles in specific organs of the developing embryo and may be linked with the physiological changes that accompany seed desiccation, imbibition and germination. © 2014 Wolny et al.
Idziak-Helmcke, D.; Hazuka, I.; Poliwczak, B.; Wiszynska, A.; Wolny, E. A.; Hasterok, R.
Insight into the karyotype evolution of Brachypodium species using comparative chromosome barcoding Journal Article
In: PLoS ONE, vol. 9, no. 3, 2014, ISSN: 19326203, (26).
@article{2-s2.0-84899832143,
title = {Insight into the karyotype evolution of Brachypodium species using comparative chromosome barcoding},
author = { D. Idziak-Helmcke and I. Hazuka and B. Poliwczak and A. Wiszynska and E.A. Wolny and R. Hasterok},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-84899832143&doi=10.1371%2fjournal.pone.0093503&partnerID=40&md5=f792b899dc05809b408cc66f5118db9d},
doi = {10.1371/journal.pone.0093503},
issn = {19326203},
year = {2014},
date = {2014-01-01},
journal = {PLoS ONE},
volume = {9},
number = {3},
publisher = {Public Library of Science},
abstract = {Paleogenomic studies based on bioinformatic analyses of DNA sequences have enabled unprecedented insight into the evolution of grass genomes. They have revealed that nested chromosome fusions played an important role in the divergence of modern grasses. Nowadays, studies on karyotype evolution based on the sequence analysis can also be effectively complemented by the fine-scale cytomolecular approach. In this work, we studied the karyotype evolution of small genome grasses using BAC-FISH based comparative chromosome barcoding in four Brachypodium species: diploid B. distachyon (2n = 10) and B. sylvaticum (2n = 18), diploid (2n = 18) and allopolyploid (2n = 28) B. pinnatum as well as B. phoenicoides (2n = 28). Using BAC clones derived from the B. distachyon genomic libraries for the chromosomes Bd2 and Bd3, we identified the descending dysploidy events that were common for diploids with x = 9 and B. distachyon as well as two nested chromosome fusions that were specific only for B. distachyon. We suggest that dysploidy events that are shared by different lineages of the genus had already appeared in their common ancestor. We also show that additional structural rearrangements, such as translocations and duplications, contributed to increasing genome diversification in the species analysed. No chromosomes structured exactly like Bd2 and Bd3 were found in B. pinnatum (2n = 28) and B. phoenicoides. The structure of Bd2 and Bd3 homeologues belonging to the two genomes in the allopolyploids resembled the structure of their counterparts in the 2n = 18 diploids. These findings reinforce the hypothesis which excludes B. distachyon as a potential parent for Eurasian perennial Brachypodium allopolyploids. Our cytomolecular data elucidate some mechanisms of the descending dysploidy in monocots and enable reconstructions of the evolutionary events which shaped the extant karyotypes in both the genus Brachypodium and in grasses as a whole. © 2014 Idziak et al.},
note = {26},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Paleogenomic studies based on bioinformatic analyses of DNA sequences have enabled unprecedented insight into the evolution of grass genomes. They have revealed that nested chromosome fusions played an important role in the divergence of modern grasses. Nowadays, studies on karyotype evolution based on the sequence analysis can also be effectively complemented by the fine-scale cytomolecular approach. In this work, we studied the karyotype evolution of small genome grasses using BAC-FISH based comparative chromosome barcoding in four Brachypodium species: diploid B. distachyon (2n = 10) and B. sylvaticum (2n = 18), diploid (2n = 18) and allopolyploid (2n = 28) B. pinnatum as well as B. phoenicoides (2n = 28). Using BAC clones derived from the B. distachyon genomic libraries for the chromosomes Bd2 and Bd3, we identified the descending dysploidy events that were common for diploids with x = 9 and B. distachyon as well as two nested chromosome fusions that were specific only for B. distachyon. We suggest that dysploidy events that are shared by different lineages of the genus had already appeared in their common ancestor. We also show that additional structural rearrangements, such as translocations and duplications, contributed to increasing genome diversification in the species analysed. No chromosomes structured exactly like Bd2 and Bd3 were found in B. pinnatum (2n = 28) and B. phoenicoides. The structure of Bd2 and Bd3 homeologues belonging to the two genomes in the allopolyploids resembled the structure of their counterparts in the 2n = 18 diploids. These findings reinforce the hypothesis which excludes B. distachyon as a potential parent for Eurasian perennial Brachypodium allopolyploids. Our cytomolecular data elucidate some mechanisms of the descending dysploidy in monocots and enable reconstructions of the evolutionary events which shaped the extant karyotypes in both the genus Brachypodium and in grasses as a whole. © 2014 Idziak et al.
2013
Inda, L. A.; Wolny, E. A.
Fluorescent in situ hybridization of the ribosomal RNA genes (5S and 35S) in the genus Lolium: Lolium canariense, the missing link with Festuca? Journal Article
In: Anales del Jardin Botanico de Madrid, vol. 70, no. 1, pp. 97-102, 2013, ISSN: 02111322, (5).
@article{2-s2.0-84884524306,
title = {Fluorescent in situ hybridization of the ribosomal RNA genes (5S and 35S) in the genus Lolium: Lolium canariense, the missing link with Festuca?},
author = { L.A. Inda and E.A. Wolny},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-84884524306&doi=10.3989%2fajbm.2329&partnerID=40&md5=400cb57013414635bc11d4de3b4f6629},
doi = {10.3989/ajbm.2329},
issn = {02111322},
year = {2013},
date = {2013-01-01},
journal = {Anales del Jardin Botanico de Madrid},
volume = {70},
number = {1},
pages = {97-102},
abstract = {Two groups of taxa can be distinguished within the genus Lolium L. based on the pollination system, chromosome size, chromosomal location of nrDNA (5S and 35S (18S-5.8S-26S)] and nrDNA phylogeny. The first group includes self-pollinated taxa (L. temulentum; L. persicum and L. remotum), whereas the second group comprises cross-pollinated species (L. perenne; L. multiflorum and L. rigidum). Here we describe that the autogamous species have two 5S sites and four 35S sites in their genome. Two of the 35S sites are present in the chromosomes containing the 5S regions. The allogamous taxa possess two 5S rDNA sites and 6-10 35S sites per genome, depending on the species. Two of these regions (35S) may also be present in the chromosomes bearing 5S sites. Our study also demonstrates that Lolium canariense shows a distinctive pattern. It has two 5S and four 35S sites. In this case, the 35S loci are located in different chromosomes than the 5S. This cytogenetic pattern is consistent with that of Festuca pratensis. Thus, despite being allogamous, Lolium canariense does not entirely fit in either of the groups defined for the genus Lolium. The physical mapping of the nrDNA regions in L. canariense is different, and resembles that of F. pratensis, suggesting that this Macaronesian Lolium could be intermediate between Festuca and Lolium. © 2013.},
note = {5},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Two groups of taxa can be distinguished within the genus Lolium L. based on the pollination system, chromosome size, chromosomal location of nrDNA (5S and 35S (18S-5.8S-26S)] and nrDNA phylogeny. The first group includes self-pollinated taxa (L. temulentum; L. persicum and L. remotum), whereas the second group comprises cross-pollinated species (L. perenne; L. multiflorum and L. rigidum). Here we describe that the autogamous species have two 5S sites and four 35S sites in their genome. Two of the 35S sites are present in the chromosomes containing the 5S regions. The allogamous taxa possess two 5S rDNA sites and 6-10 35S sites per genome, depending on the species. Two of these regions (35S) may also be present in the chromosomes bearing 5S sites. Our study also demonstrates that Lolium canariense shows a distinctive pattern. It has two 5S and four 35S sites. In this case, the 35S loci are located in different chromosomes than the 5S. This cytogenetic pattern is consistent with that of Festuca pratensis. Thus, despite being allogamous, Lolium canariense does not entirely fit in either of the groups defined for the genus Lolium. The physical mapping of the nrDNA regions in L. canariense is different, and resembles that of F. pratensis, suggesting that this Macaronesian Lolium could be intermediate between Festuca and Lolium. © 2013.
Brąszewska-Zalewska, A. J.; Wolny, E. A.; Smialek, L.; Hasterok, R.
Tissue-Specific Epigenetic Modifications in Root Apical Meristem Cells of Hordeum vulgare Journal Article
In: PLoS ONE, vol. 8, no. 7, 2013, ISSN: 19326203, (25).
@article{2-s2.0-84881192851,
title = {Tissue-Specific Epigenetic Modifications in Root Apical Meristem Cells of Hordeum vulgare},
author = { A.J. Brąszewska-Zalewska and E.A. Wolny and L. Smialek and R. Hasterok},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-84881192851&doi=10.1371%2fjournal.pone.0069204&partnerID=40&md5=0031f46a741a0280b8b236fbf4964b49},
doi = {10.1371/journal.pone.0069204},
issn = {19326203},
year = {2013},
date = {2013-01-01},
journal = {PLoS ONE},
volume = {8},
number = {7},
abstract = {Epigenetic modifications of chromatin structure are essential for many biological processes, including growth and reproduction. Patterns of DNA and histone modifications have recently been widely studied in many plant species, although there is virtually no data on the spatial and temporal distribution of epigenetic markers during plant development. Accordingly, we have used immunostaining techniques to investigate epigenetic modifications in the root apical meristem of Hordeum vulgare. Histone H4 acetylation (H4K5ac), histone H3 dimethylation (H3K4me2; H3K9me2) and DNA methylation (5mC) patterns were established for various root meristem tissues. Distinct levels of those modifications were visualised in the root cap, epidermis, cortex and vascular tissues. The lateral root cap cells seem to display the highest level of H3K9me2 and 5mC. In the epidermis, the highest level of 5mC and H3K9me2 was detected in the nuclei from the boundary of the proximal meristem and the elongation zone, while the vascular tissues were characterized by the highest level of H4K5ac. Some of the modified histones were also detectable in the cytoplasm in a highly tissue-specific manner. Immunolocalisation of epigenetic modifications of chromatin carried out in this way, on longitudinal or transverse sections, provides a unique topographic context within the organ, and will provide some answers to the significant biological question of tissue differentiation processes during root development in a monocotyledon plant species. © 2013 Braszewska-Zalewska et al.},
note = {25},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Epigenetic modifications of chromatin structure are essential for many biological processes, including growth and reproduction. Patterns of DNA and histone modifications have recently been widely studied in many plant species, although there is virtually no data on the spatial and temporal distribution of epigenetic markers during plant development. Accordingly, we have used immunostaining techniques to investigate epigenetic modifications in the root apical meristem of Hordeum vulgare. Histone H4 acetylation (H4K5ac), histone H3 dimethylation (H3K4me2; H3K9me2) and DNA methylation (5mC) patterns were established for various root meristem tissues. Distinct levels of those modifications were visualised in the root cap, epidermis, cortex and vascular tissues. The lateral root cap cells seem to display the highest level of H3K9me2 and 5mC. In the epidermis, the highest level of 5mC and H3K9me2 was detected in the nuclei from the boundary of the proximal meristem and the elongation zone, while the vascular tissues were characterized by the highest level of H4K5ac. Some of the modified histones were also detectable in the cytoplasm in a highly tissue-specific manner. Immunolocalisation of epigenetic modifications of chromatin carried out in this way, on longitudinal or transverse sections, provides a unique topographic context within the organ, and will provide some answers to the significant biological question of tissue differentiation processes during root development in a monocotyledon plant species. © 2013 Braszewska-Zalewska et al.
Wolny, E. A.; Fidyk, W.; Hasterok, R.
Karyotyping of Brachypodium pinnatum (2n = 18) chromosomes using cross-species BAC-FISH Journal Article
In: Genome, vol. 56, no. 4, pp. 239-243, 2013, ISSN: 08312796, (10).
@article{2-s2.0-84878227807,
title = {Karyotyping of Brachypodium pinnatum (2n = 18) chromosomes using cross-species BAC-FISH},
author = { E.A. Wolny and W. Fidyk and R. Hasterok},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-84878227807&doi=10.1139%2fgen-2013-0012&partnerID=40&md5=aa498c8d9b7beadf868a5ece165d2678},
doi = {10.1139/gen-2013-0012},
issn = {08312796},
year = {2013},
date = {2013-01-01},
journal = {Genome},
volume = {56},
number = {4},
pages = {239-243},
abstract = {Identification of individual chromosomes in a complement is usually a difficult task in the case of most plant species, especially for those with small, numerous, and morphologically uniform chromosomes. In this paper, we demonstrate that the landmarks produced by cross-species fluorescence in situ hybridisation (FISH) of Brachypodium distachyon derived bacterial artificial chromosome (BAC) clones can be used for discrimination of Brachypodium pinnatum (2n = 18) chromosomes. Selected sets of clones were hybridised in several sequential experiments performed on exactly the same chromosome spreads, using reprobing of cytological preparations. Analysis of the morphometric features of B. pinnatum chromosomes was performed to establish their total length, the position of centromeres, and the position of BAC-based landmarks in relation to the centromere, thereby enabling their effective karyotyping, which is a prerequisite for more complex study of the grass genome structure and evolution at the cytomolecular level. © 2013 Published by NRC Research Press.},
note = {10},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Identification of individual chromosomes in a complement is usually a difficult task in the case of most plant species, especially for those with small, numerous, and morphologically uniform chromosomes. In this paper, we demonstrate that the landmarks produced by cross-species fluorescence in situ hybridisation (FISH) of Brachypodium distachyon derived bacterial artificial chromosome (BAC) clones can be used for discrimination of Brachypodium pinnatum (2n = 18) chromosomes. Selected sets of clones were hybridised in several sequential experiments performed on exactly the same chromosome spreads, using reprobing of cytological preparations. Analysis of the morphometric features of B. pinnatum chromosomes was performed to establish their total length, the position of centromeres, and the position of BAC-based landmarks in relation to the centromere, thereby enabling their effective karyotyping, which is a prerequisite for more complex study of the grass genome structure and evolution at the cytomolecular level. © 2013 Published by NRC Research Press.
2012
Breda, E.; Wolny, E. A.; Hasterok, R.
Intraspecific polymorphism of ribosomal DNA loci number and morphology in Brachypodium pinnatum and Brachypodium sylvaticum Journal Article
In: Cellular and Molecular Biology Letters, vol. 17, no. 4, pp. 526-541, 2012, ISSN: 14258153, (6).
@article{2-s2.0-84873267734,
title = {Intraspecific polymorphism of ribosomal DNA loci number and morphology in Brachypodium pinnatum and Brachypodium sylvaticum},
author = { E. Breda and E.A. Wolny and R. Hasterok},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-84873267734&doi=10.2478%2fs11658-012-0025-4&partnerID=40&md5=28d2364b5c42eb384ee1a12ab80631e5},
doi = {10.2478/s11658-012-0025-4},
issn = {14258153},
year = {2012},
date = {2012-01-01},
journal = {Cellular and Molecular Biology Letters},
volume = {17},
number = {4},
pages = {526-541},
publisher = {BioMed Central Ltd.},
abstract = {The genus Brachypodium has become the target of extensive cytomolecular studies since one of its representatives, B. distachyon, has been accepted as a model plant for temperate cereals and forage grasses. Recent preliminary studies suggested that intraspecific rDNA polymorphism can occur in at least two members of the genus, B. sylvaticum and B. pinnatum, so the aim of this study was to further analyse this phenomenon. FISH with 25S rDNA and 5S rDNA probes was performed on somatic metaphase chromosomes, supplemented by the silver staining technique which distinguishes transcriptionally active from inactive 18S-5. 8S-25S rDNA loci. The number, size and chromosomal distribution of 5S rDNA loci were very constant: two loci were invariably observed in all studied diploid accessions of both species, while four 5S rDNA loci were present in the tetraploid B. pinnatum. In contrast to 5S rDNA loci, those of the 35S rDNA were more variable. Two or three loci were observed in the diploid B. pinnatum and four in tetraploid accessions. In chromosome complements of B. sylvaticum accessions from two to six 35S rDNA sites were detected. Regardless of total rDNA locus number, only two were transcriptionally active in diploid accessions of both species, while two or four were active in the tetraploid B. pinnatum. Additionally, the fluorescent CMA/DAPI banding method was used to identify the relation between rDNA sites and CMA+ bands. It was revealed that the number and chromosomal distribution of CMA+ bands are in congruence only with 35S rDNA loci which gave strong FISH signals. © 2012 Versita Warsaw and Springer-Verlag Wien.},
note = {6},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
The genus Brachypodium has become the target of extensive cytomolecular studies since one of its representatives, B. distachyon, has been accepted as a model plant for temperate cereals and forage grasses. Recent preliminary studies suggested that intraspecific rDNA polymorphism can occur in at least two members of the genus, B. sylvaticum and B. pinnatum, so the aim of this study was to further analyse this phenomenon. FISH with 25S rDNA and 5S rDNA probes was performed on somatic metaphase chromosomes, supplemented by the silver staining technique which distinguishes transcriptionally active from inactive 18S-5. 8S-25S rDNA loci. The number, size and chromosomal distribution of 5S rDNA loci were very constant: two loci were invariably observed in all studied diploid accessions of both species, while four 5S rDNA loci were present in the tetraploid B. pinnatum. In contrast to 5S rDNA loci, those of the 35S rDNA were more variable. Two or three loci were observed in the diploid B. pinnatum and four in tetraploid accessions. In chromosome complements of B. sylvaticum accessions from two to six 35S rDNA sites were detected. Regardless of total rDNA locus number, only two were transcriptionally active in diploid accessions of both species, while two or four were active in the tetraploid B. pinnatum. Additionally, the fluorescent CMA/DAPI banding method was used to identify the relation between rDNA sites and CMA+ bands. It was revealed that the number and chromosomal distribution of CMA+ bands are in congruence only with 35S rDNA loci which gave strong FISH signals. © 2012 Versita Warsaw and Springer-Verlag Wien.
2011
Słomka, A.; Siwińska, D.; Wolny, E. A.; Kellner, K.; Kuta, E.
Influence of a heavy-metal-polluted environment on Viola tricolor genome size and chromosome number Journal Article
In: Acta Biologica Cracoviensia Series Botanica, vol. 53, no. 1, pp. 7-15, 2011, ISSN: 00015296, (9).
@article{2-s2.0-80053355540,
title = {Influence of a heavy-metal-polluted environment on Viola tricolor genome size and chromosome number},
author = { A. Słomka and D. Siwińska and E.A. Wolny and K. Kellner and E. Kuta},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-80053355540&doi=10.2478%2fv10182-011-0001-8&partnerID=40&md5=58fedcffca784a953938a587fb28df9c},
doi = {10.2478/v10182-011-0001-8},
issn = {00015296},
year = {2011},
date = {2011-01-01},
journal = {Acta Biologica Cracoviensia Series Botanica},
volume = {53},
number = {1},
pages = {7-15},
abstract = {Intraspecific changes in genome size and chromosome number lead to divergence and species evolution. Heavy metals disturb the cell cycle and cause mutations. Areas contaminated by heavy metals (metalliferous sites) are places where microevolutionary processes accelerate; very often only a few generations are enough for a new genotype to arise. This study, which continues our long-term research on Viola tricolor (Violaceae), a species occurring on both metalliferous (Zn; Pb; Cd; Cu) and non-metalliferous soils in Western and Central Europe, is aimed at determining the influence of environments polluted with heavy metals on genome size and karyological variability. The genome size of V. tricolor ranged from 3.801 to 4.203 pg, but the differences between metallicolous and non-metallicolous populations were not statistically significant. Altered chromosome numbers were significantly more frequent in material from the polluted sites than from the non-polluted sites (43% versus 28%). Besides the standard chromosome number (2n = 26), aneuploid cells with lower (2n = 18-25) or higher (2n = 27; 28) chromosome numbers were found in plants from both types of site, but polyploid (2n = 42) cells were observed only in plants from the metalliferous locality. The lack of correlation between chromosome variability in root meristematic cells and genome size estimated from peduncle cells can be attributed to elimination of somatic mutations in generative meristem, producing chromosome-stable non-meristematic tissues in the peduncle. © Polish Academy of Sciences and Jagiellonian University, Cracow 2011.},
note = {9},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Intraspecific changes in genome size and chromosome number lead to divergence and species evolution. Heavy metals disturb the cell cycle and cause mutations. Areas contaminated by heavy metals (metalliferous sites) are places where microevolutionary processes accelerate; very often only a few generations are enough for a new genotype to arise. This study, which continues our long-term research on Viola tricolor (Violaceae), a species occurring on both metalliferous (Zn; Pb; Cd; Cu) and non-metalliferous soils in Western and Central Europe, is aimed at determining the influence of environments polluted with heavy metals on genome size and karyological variability. The genome size of V. tricolor ranged from 3.801 to 4.203 pg, but the differences between metallicolous and non-metallicolous populations were not statistically significant. Altered chromosome numbers were significantly more frequent in material from the polluted sites than from the non-polluted sites (43% versus 28%). Besides the standard chromosome number (2n = 26), aneuploid cells with lower (2n = 18-25) or higher (2n = 27; 28) chromosome numbers were found in plants from both types of site, but polyploid (2n = 42) cells were observed only in plants from the metalliferous locality. The lack of correlation between chromosome variability in root meristematic cells and genome size estimated from peduncle cells can be attributed to elimination of somatic mutations in generative meristem, producing chromosome-stable non-meristematic tissues in the peduncle. © Polish Academy of Sciences and Jagiellonian University, Cracow 2011.
Idziak-Helmcke, D.; Betekhtin, A.; Wolny, E. A.; Susek, K.; Wright, J.; Febrer, M.; Bevan, M. W.; Jenkins, G.; Hasterok, R.
Painting the chromosomes of Brachypodium-current status and future prospects Journal Article
In: Chromosoma, vol. 120, no. 5, pp. 469-479, 2011, ISSN: 00095915, (46).
@article{2-s2.0-80054097796,
title = {Painting the chromosomes of Brachypodium-current status and future prospects},
author = { D. Idziak-Helmcke and A. Betekhtin and E.A. Wolny and K. Susek and J. Wright and M. Febrer and M.W. Bevan and G. Jenkins and R. Hasterok},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-80054097796&doi=10.1007%2fs00412-011-0326-9&partnerID=40&md5=e58f0c0baf758f71c389d6c6eb612290},
doi = {10.1007/s00412-011-0326-9},
issn = {00095915},
year = {2011},
date = {2011-01-01},
journal = {Chromosoma},
volume = {120},
number = {5},
pages = {469-479},
abstract = {Chromosome painting is one of the most powerful and spectacular tools of modern molecular cytogenetics, enabling complex analyses of nuclear genome structure and evolution. For many years, this technique was restricted to the study of mammalian chromosomes, as it failed to work in plant genomes due mainly to the presence of large amounts of repetitive DNA common to all the chromosomes of the complement. The availability of ordered, chromosome-specific BAC clones of Arabidopsis thaliana containing relatively little repetitive genomic DNA enabled the first chromosome painting in dicotyledonous plants. Here, we show for the first time chromosome painting in three different cytotypes of a monocotyledonous plant-the model grass, Brachypodium distachyon. Possible directions of further detailed studies are proposed, such as the evolution of grass karyotypes, the behaviour of meiotic chromosomes, and the analysis of chromosome distribution at interphase. © 2011 The Author(s).},
note = {46},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Chromosome painting is one of the most powerful and spectacular tools of modern molecular cytogenetics, enabling complex analyses of nuclear genome structure and evolution. For many years, this technique was restricted to the study of mammalian chromosomes, as it failed to work in plant genomes due mainly to the presence of large amounts of repetitive DNA common to all the chromosomes of the complement. The availability of ordered, chromosome-specific BAC clones of Arabidopsis thaliana containing relatively little repetitive genomic DNA enabled the first chromosome painting in dicotyledonous plants. Here, we show for the first time chromosome painting in three different cytotypes of a monocotyledonous plant-the model grass, Brachypodium distachyon. Possible directions of further detailed studies are proposed, such as the evolution of grass karyotypes, the behaviour of meiotic chromosomes, and the analysis of chromosome distribution at interphase. © 2011 The Author(s).
Wolny, E. A.; Susek, K.; Hasterok, R.; Langdon, T.
Compact genomes and complex evolution in the genus Brachypodium Journal Article
In: Chromosoma, vol. 120, no. 2, pp. 199-212, 2011, ISSN: 00095915, (32).
@article{2-s2.0-79955523870,
title = {Compact genomes and complex evolution in the genus Brachypodium},
author = { E.A. Wolny and K. Susek and R. Hasterok and T. Langdon},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-79955523870&doi=10.1007%2fs00412-010-0303-8&partnerID=40&md5=69284c1a124962d5ffd5895e0bd45961},
doi = {10.1007/s00412-010-0303-8},
issn = {00095915},
year = {2011},
date = {2011-01-01},
journal = {Chromosoma},
volume = {120},
number = {2},
pages = {199-212},
abstract = {The temperate annual grass Brachypodium distachyon is a diploid species with a chromosome base number of 5. It is strikingly different from other Eurasian species of the genus, which are perennial and often polyploid, with the diploids typically having base numbers of 8 or 9. Previously, phylogenies indicated that B. distachyon split from the other species early in the evolution of the genus, while its genome sequence revealed that extensive synteny on a chromosomal scale had been maintained with rice, a tropical grass with a base number of 12. Here we show evidence that B. distachyon may have a homoploid origin, involving ancestral interspecific hybridisation, although it does not appear to be a component of any of the perennial Eurasian allopolyploids. Using a cytogenetic approach, we show that dysploidy in Brachypodium has not followed a simple progression. © 2010 Springer-Verlag.},
note = {32},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
The temperate annual grass Brachypodium distachyon is a diploid species with a chromosome base number of 5. It is strikingly different from other Eurasian species of the genus, which are perennial and often polyploid, with the diploids typically having base numbers of 8 or 9. Previously, phylogenies indicated that B. distachyon split from the other species early in the evolution of the genus, while its genome sequence revealed that extensive synteny on a chromosomal scale had been maintained with rice, a tropical grass with a base number of 12. Here we show evidence that B. distachyon may have a homoploid origin, involving ancestral interspecific hybridisation, although it does not appear to be a component of any of the perennial Eurasian allopolyploids. Using a cytogenetic approach, we show that dysploidy in Brachypodium has not followed a simple progression. © 2010 Springer-Verlag.
2010
Febrer, M.; Goicoechea, J. L.; Wright, J.; McKenzie, N.; Song, X.; Lin, J.; Collura, K.; Wissotski, M.; Yu, Y.; Ammiraju, J. S. S.; Wolny, E. A.; Idziak-Helmcke, D.; Betekhtin, A.; Kudrna, D.; Hasterok, R.; Wing, R. A.; Bevan, M. W.
An integrated physical, genetic and cytogenetic map of Brachypodium distachyon, a model system for grass research Journal Article
In: PLoS ONE, vol. 5, no. 10, 2010, ISSN: 19326203, (41).
@article{2-s2.0-78149437473,
title = {An integrated physical, genetic and cytogenetic map of Brachypodium distachyon, a model system for grass research},
author = { M. Febrer and J.L. Goicoechea and J. Wright and N. McKenzie and X. Song and J. Lin and K. Collura and M. Wissotski and Y. Yu and J.S.S. Ammiraju and E.A. Wolny and D. Idziak-Helmcke and A. Betekhtin and D. Kudrna and R. Hasterok and R.A. Wing and M.W. Bevan},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-78149437473&doi=10.1371%2fjournal.pone.0013461&partnerID=40&md5=d1ddbfdfca8fbfda0316627d6007d582},
doi = {10.1371/journal.pone.0013461},
issn = {19326203},
year = {2010},
date = {2010-01-01},
journal = {PLoS ONE},
volume = {5},
number = {10},
abstract = {The pooid subfamily of grasses includes some of the most important crop, forage and turf species, such as wheat, barley and Lolium. Developing genomic resources, such as whole-genome physical maps, for analysing the large and complex genomes of these crops and for facilitating biological research in grasses is an important goal in plant biology. We describe a bacterial artificial chromosome (BAC)-based physical map of the wild pooid grass Brachypodium distachyon and integrate this with whole genome shotgun sequence (WGS) assemblies using BAC end sequences (BES). The resulting physical map contains 26 contigs spanning the 272 Mb genome. BES from the physical map were also used to integrate a genetic map. This provides an independent vaildation and confirmation of the published WGS assembly. Mapped BACs were used in Fluorescence In Situ Hybridisation (FISH) experiments to align the integrated physical map and sequence assemblies to chromosomes with high resolution. The physical, genetic and cytogenetic maps, integrated with whole genome shotgun sequence assemblies, enhance the accuracy and durability of this important genome sequence and will directly facilitate gene isolation. © 2010 Febrer et al.},
note = {41},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
The pooid subfamily of grasses includes some of the most important crop, forage and turf species, such as wheat, barley and Lolium. Developing genomic resources, such as whole-genome physical maps, for analysing the large and complex genomes of these crops and for facilitating biological research in grasses is an important goal in plant biology. We describe a bacterial artificial chromosome (BAC)-based physical map of the wild pooid grass Brachypodium distachyon and integrate this with whole genome shotgun sequence (WGS) assemblies using BAC end sequences (BES). The resulting physical map contains 26 contigs spanning the 272 Mb genome. BES from the physical map were also used to integrate a genetic map. This provides an independent vaildation and confirmation of the published WGS assembly. Mapped BACs were used in Fluorescence In Situ Hybridisation (FISH) experiments to align the integrated physical map and sequence assemblies to chromosomes with high resolution. The physical, genetic and cytogenetic maps, integrated with whole genome shotgun sequence assemblies, enhance the accuracy and durability of this important genome sequence and will directly facilitate gene isolation. © 2010 Febrer et al.
2009
Wolny, E. A.; Hasterok, R.
Comparative cytogenetic analysis of the genomes of the model grass Brachypodium distachyon and its close relatives Journal Article
In: Annals of Botany, vol. 104, no. 5, pp. 873-881, 2009, ISSN: 03057364, (54).
@article{2-s2.0-70349587942,
title = {Comparative cytogenetic analysis of the genomes of the model grass Brachypodium distachyon and its close relatives},
author = { E.A. Wolny and R. Hasterok},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-70349587942&doi=10.1093%2faob%2fmcp179&partnerID=40&md5=44169818ca6e826ec9ec1625450a9209},
doi = {10.1093/aob/mcp179},
issn = {03057364},
year = {2009},
date = {2009-01-01},
journal = {Annals of Botany},
volume = {104},
number = {5},
pages = {873-881},
abstract = {Background and AimsBrachypodium is a small genus of temperate grasses that comprises 12-15 species. Brachypodium distachyon is now well established as a model species for temperate cereals and forage grasses. In contrast to B. distachyon, other members of the genus have been poorly investigated at the chromosome level or not at all.MethodsTwenty accessions comprising six species and two subspecies of Brachypodium were analysed cytogenetically. Measurements of nuclear genome size were made by flow cytometry. Chromosomal localization of 18-5·8-25S rDNA and 5S rDNA loci was performed by dual-colour fluorescence in situ hybridization (FISH) on enzymatically digested root-tip meristematic cells. For comparative phylogenetic analyses genomic in situ hybridization (GISH) applied to somatic chromosome preparations was used.Key ResultsAll Brachypodium species examined have rather small genomes and chromosomes. Their chromosome numbers and genome sizes vary from 2n = 10 and 0·631 pg/2C in B. distachyon to 2n = 38 and 2·57 pg/2C in B. retusum, respectively. Genotypes with 18 and 28 chromosomes were found among B. pinnatum accessions. GISH analysis revealed that B. pinnatum with 28 chromosomes is most likely an interspecific hybrid between B. distachyon (2n = 10) and B. pinnatum (2n = 18). Two other species, B. phoenicoides and B. retusum, are also allopolyploids and B. distachyon or a close relative seems to be one of their putative ancestral species. In chromosomes of all species examined the 45S rDNA loci are distally distributed whereas loci for 5S rDNA are pericentromeric. ConclusionsThe increasing significance of B. distachyon as a model grass emphasizes the need to understand the evolutionary relationships in the genus Brachypodium and to ensure consistency in the biological nomenclature of its species. Modern molecular cytogenetic techniques such as FISH and GISH are suitable for comparative phylogenetic analyses and may provide informative chromosome- and/or genome-specific landmarks.},
note = {54},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Background and AimsBrachypodium is a small genus of temperate grasses that comprises 12-15 species. Brachypodium distachyon is now well established as a model species for temperate cereals and forage grasses. In contrast to B. distachyon, other members of the genus have been poorly investigated at the chromosome level or not at all.MethodsTwenty accessions comprising six species and two subspecies of Brachypodium were analysed cytogenetically. Measurements of nuclear genome size were made by flow cytometry. Chromosomal localization of 18-5·8-25S rDNA and 5S rDNA loci was performed by dual-colour fluorescence in situ hybridization (FISH) on enzymatically digested root-tip meristematic cells. For comparative phylogenetic analyses genomic in situ hybridization (GISH) applied to somatic chromosome preparations was used.Key ResultsAll Brachypodium species examined have rather small genomes and chromosomes. Their chromosome numbers and genome sizes vary from 2n = 10 and 0·631 pg/2C in B. distachyon to 2n = 38 and 2·57 pg/2C in B. retusum, respectively. Genotypes with 18 and 28 chromosomes were found among B. pinnatum accessions. GISH analysis revealed that B. pinnatum with 28 chromosomes is most likely an interspecific hybrid between B. distachyon (2n = 10) and B. pinnatum (2n = 18). Two other species, B. phoenicoides and B. retusum, are also allopolyploids and B. distachyon or a close relative seems to be one of their putative ancestral species. In chromosomes of all species examined the 45S rDNA loci are distally distributed whereas loci for 5S rDNA are pericentromeric. ConclusionsThe increasing significance of B. distachyon as a model grass emphasizes the need to understand the evolutionary relationships in the genus Brachypodium and to ensure consistency in the biological nomenclature of its species. Modern molecular cytogenetic techniques such as FISH and GISH are suitable for comparative phylogenetic analyses and may provide informative chromosome- and/or genome-specific landmarks.
2007
Khattak, G. S. S.; Wolny, E. A.; Saeed, I.
In: Pakistan Journal of Botany, vol. 39, no. 5, pp. 1511-1515, 2007, ISSN: 05563321, (7).
@article{2-s2.0-47249088758,
title = {Detection of ribosomal DNA sites in chickpea (Cicer arietinum L.) and mungbean (Vigna radiata (L.) wiltzek) by fluorescence in situ hybridization},
author = { G.S.S. Khattak and E.A. Wolny and I. Saeed},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-47249088758&partnerID=40&md5=441229411ce36efda58cd5cfb9fffd7c},
issn = {05563321},
year = {2007},
date = {2007-01-01},
journal = {Pakistan Journal of Botany},
volume = {39},
number = {5},
pages = {1511-1515},
abstract = {Fluorescence in situ hybridization (FISH) technique was used to detect rDNA sites in chickpea and mungbean. In chickpea, the rDNA sites were detected on three pairs of chromosomes. Among these three pairs of chromosomes, one pair exhibited both 25S rDNA and 5S rDNA sites, while in case of other two pairs of chromosomes the 25S rDNA and 5S rDNA sites were located separately on separate pair of chromosomes. In case of mungbean, rDNA sites were found on four pairs of chromosomes. Each 25S rDNA and 5S rDNA had separate sites on two pairs of chromosomes. Active Nucleolus Organizer Regions (NORs) of both the crops were detected through silver staining technique. One and two pairs of chromosomes were detected active for NORs in chickpea and in mungbean, respectively.},
note = {7},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Fluorescence in situ hybridization (FISH) technique was used to detect rDNA sites in chickpea and mungbean. In chickpea, the rDNA sites were detected on three pairs of chromosomes. Among these three pairs of chromosomes, one pair exhibited both 25S rDNA and 5S rDNA sites, while in case of other two pairs of chromosomes the 25S rDNA and 5S rDNA sites were located separately on separate pair of chromosomes. In case of mungbean, rDNA sites were found on four pairs of chromosomes. Each 25S rDNA and 5S rDNA had separate sites on two pairs of chromosomes. Active Nucleolus Organizer Regions (NORs) of both the crops were detected through silver staining technique. One and two pairs of chromosomes were detected active for NORs in chickpea and in mungbean, respectively.
2006
Hasterok, R.; Marasek, A.; Donnison, I. S.; Armstead, I.; Thomas, A.; King, I. P.; Wolny, E. A.; Idziak-Helmcke, D.; Draper, J.; Jenkins, G.
In: Genetics, vol. 173, no. 1, pp. 349-362, 2006, ISSN: 00166731, (104).
@article{2-s2.0-33744489394,
title = {Alignment of the genomes of Brachypodium distachyon and temperate cereals and grasses using bacterial artificial chromosome landing with fluorescence in situ hybridization},
author = { R. Hasterok and A. Marasek and I.S. Donnison and I. Armstead and A. Thomas and I.P. King and E.A. Wolny and D. Idziak-Helmcke and J. Draper and G. Jenkins},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-33744489394&doi=10.1534%2fgenetics.105.049726&partnerID=40&md5=fe1c6cb39d5cf5a72551bb8ff32a8943},
doi = {10.1534/genetics.105.049726},
issn = {00166731},
year = {2006},
date = {2006-01-01},
journal = {Genetics},
volume = {173},
number = {1},
pages = {349-362},
abstract = {As part of an initiative to develop Brachypodium distachyon as a genomic "bridge" species between rice and the temperate cereals and grasses, a BAC library has been constructed for the two diploid (2n = 2x = 10) genotypes, ABR1 and ABR5. The library consists of 9100 clones, with an approximate average insert size of 88 kb, representing 2.22 genome equivalents. To validate the usefulness of this species for comparative genomics and gene discovery in its larger genome relatives, the library was screened by PCR using primers designed on previously mapped rice and Poaceae sequences. Screening indicated a degree of synteny between these species and B. distachyon, which was confirmed by fluorescent in situ hybridization of the marker-selected BACs (BAC landing) to the 10 chromosome arms of the karyotype, with most of the BACs hybridizing as single loci on known chromosomes. Contiguous BACs colocalized on individual chromosomes, thereby confirming the conservation of genome synteny and proving that B. distachyon has utility as a temperate grass model species alternative to rice. Copyright © 2006 by the Genetics Society of America.},
note = {104},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
As part of an initiative to develop Brachypodium distachyon as a genomic "bridge" species between rice and the temperate cereals and grasses, a BAC library has been constructed for the two diploid (2n = 2x = 10) genotypes, ABR1 and ABR5. The library consists of 9100 clones, with an approximate average insert size of 88 kb, representing 2.22 genome equivalents. To validate the usefulness of this species for comparative genomics and gene discovery in its larger genome relatives, the library was screened by PCR using primers designed on previously mapped rice and Poaceae sequences. Screening indicated a degree of synteny between these species and B. distachyon, which was confirmed by fluorescent in situ hybridization of the marker-selected BACs (BAC landing) to the 10 chromosome arms of the karyotype, with most of the BACs hybridizing as single loci on known chromosomes. Contiguous BACs colocalized on individual chromosomes, thereby confirming the conservation of genome synteny and proving that B. distachyon has utility as a temperate grass model species alternative to rice. Copyright © 2006 by the Genetics Society of America.
Hasterok, R.; Wolny, E. A.; Hosiawa, M.; Kowalczyk, M.; Kulak-Ksiazczyk, S.; Ksiązczyk, T.; Heneen, W. K.; Małuszyńska, J.
Comparative analysis of rDNA distribution in chromosomes of various species of Brassicaceae Journal Article
In: Annals of Botany, vol. 97, no. 2, pp. 205-216, 2006, ISSN: 03057364, (114).
@article{2-s2.0-31744436819,
title = {Comparative analysis of rDNA distribution in chromosomes of various species of Brassicaceae},
author = { R. Hasterok and E.A. Wolny and M. Hosiawa and M. Kowalczyk and S. Kulak-Ksiazczyk and T. Ksiązczyk and W.K. Heneen and J. Małuszyńska},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-31744436819&doi=10.1093%2faob%2fmcj031&partnerID=40&md5=0d82c13868701a373afa8f6633a37d79},
doi = {10.1093/aob/mcj031},
issn = {03057364},
year = {2006},
date = {2006-01-01},
journal = {Annals of Botany},
volume = {97},
number = {2},
pages = {205-216},
abstract = {• Background and Aims: The Brassicaceae family encompasses numerous species of great agronomic importance, belonging to such genera, as Brassica, Raphanus, Sinapis and Armoracia. Many of them are characterized by extensive intraspecific diversity of phenotypes. The present study focuses on the polymorphism of number, appearance and chromosomal localization of ribosomal DNA (rDNA) sites and, when possible, in relation to polyploidy, in 42 accessions of Brassica species and ten accessions of Diplotaxis, Eruca, Raphanus and Sinapis species. • Methods: Chromosomal localization of ribosomal DNA was carried out using dual colour fluorescence in situ hybridization (FISH) with 5S rDNA and 25S rDNA sequences as probes on enzymatically digested root-tip meristematic cells. • Key Results: Loci for 5S and 18S-5.8S-25S rDNA were determined for the first time in six taxa, and previously unreported rDNA constellations were described in an additional 12 accessions. FISH revealed frequent polymorphism in number, appearance and chromosomal localization of both 5S and 25S rDNA sites. This phenomenon was most commonly observed in the A genome of Brassica, where it involves exclusively pericentromeric sites of 5S and 25S rRNA genes. The intraspecific polymorphism was between subspecies/varieties or within a variety or cultivar (i.e. interindividual). • Conclusions: The number of rDNA sites can differ up to 5-fold in species with the same chromosome number. In addition to the eight previously reported chromosomal types with ribosomal genes, three new variant types are described. The extent of polymorphism is genome dependent. Comparing the A, B and C genomes revealed the highest rDNA polymorphism in the A genome. The loci carrying presumably inactive ribosomal RNA genes are particularly prone to polymorphism. It can also be concluded that there is no obvious polyploidization-related tendency to reduce the number of ribosomal DNA loci in the allotetraploid species, when compared with their putative diploid progenitors. The observed differences are rather caused by the prevailing polymorphism within the diploids and allotetraploids. This would make it difficult to predict expected numbers of rDNA loci in natural polyploids. © The Author 2005. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved.},
note = {114},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
• Background and Aims: The Brassicaceae family encompasses numerous species of great agronomic importance, belonging to such genera, as Brassica, Raphanus, Sinapis and Armoracia. Many of them are characterized by extensive intraspecific diversity of phenotypes. The present study focuses on the polymorphism of number, appearance and chromosomal localization of ribosomal DNA (rDNA) sites and, when possible, in relation to polyploidy, in 42 accessions of Brassica species and ten accessions of Diplotaxis, Eruca, Raphanus and Sinapis species. • Methods: Chromosomal localization of ribosomal DNA was carried out using dual colour fluorescence in situ hybridization (FISH) with 5S rDNA and 25S rDNA sequences as probes on enzymatically digested root-tip meristematic cells. • Key Results: Loci for 5S and 18S-5.8S-25S rDNA were determined for the first time in six taxa, and previously unreported rDNA constellations were described in an additional 12 accessions. FISH revealed frequent polymorphism in number, appearance and chromosomal localization of both 5S and 25S rDNA sites. This phenomenon was most commonly observed in the A genome of Brassica, where it involves exclusively pericentromeric sites of 5S and 25S rRNA genes. The intraspecific polymorphism was between subspecies/varieties or within a variety or cultivar (i.e. interindividual). • Conclusions: The number of rDNA sites can differ up to 5-fold in species with the same chromosome number. In addition to the eight previously reported chromosomal types with ribosomal genes, three new variant types are described. The extent of polymorphism is genome dependent. Comparing the A, B and C genomes revealed the highest rDNA polymorphism in the A genome. The loci carrying presumably inactive ribosomal RNA genes are particularly prone to polymorphism. It can also be concluded that there is no obvious polyploidization-related tendency to reduce the number of ribosomal DNA loci in the allotetraploid species, when compared with their putative diploid progenitors. The observed differences are rather caused by the prevailing polymorphism within the diploids and allotetraploids. This would make it difficult to predict expected numbers of rDNA loci in natural polyploids. © The Author 2005. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved.
2005
Hasterok, R.; Ksiązczyk, T.; Wolny, E. A.; Małuszyńska, J.
FISH and GISH analysis of Brassica genomes Journal Article
In: Acta Biologica Cracoviensia Series Botanica, vol. 47, no. 1, pp. 185-192, 2005, ISSN: 00015296, (21).
@article{2-s2.0-28944441289,
title = {FISH and GISH analysis of Brassica genomes},
author = { R. Hasterok and T. Ksiązczyk and E.A. Wolny and J. Małuszyńska},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-28944441289&partnerID=40&md5=a5ca35de39bf658c5c304b07d941005b},
issn = {00015296},
year = {2005},
date = {2005-01-01},
journal = {Acta Biologica Cracoviensia Series Botanica},
volume = {47},
number = {1},
pages = {185-192},
abstract = {Fluorescence and genomic in situ hybridization (FISH and GISH) methods were used for discrimination of Brassica genomes. The three diploid and three allotetraploid species of Brassica, known as the "U-triangle," represent an attractive model for molecular and cytological analysis of genome changes during phylogeny in the genus Brassica. The use of genomic DNA probes enabled unambiguous discrimination of the ancestral genomes in B. juncea and B. carinata, and was only partially successful inB. napus. GISH signals in all genomes were localized predominantly in pericentromeric regions of chromosomes. Simultaneous application of genomic and ribosomal DNA probes in multicolor GISH and FISH allowed identification of a significant number of chromosomes in the B. juncea complement. The study also revealed that species of Brassica possess Arabidopsis-type telomeric repeats which in all genomes occupied exclusively terminal, that is, telomeric, locations of chromosomes. © Polish Academy of Sciences, 2005.},
note = {21},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Fluorescence and genomic in situ hybridization (FISH and GISH) methods were used for discrimination of Brassica genomes. The three diploid and three allotetraploid species of Brassica, known as the "U-triangle," represent an attractive model for molecular and cytological analysis of genome changes during phylogeny in the genus Brassica. The use of genomic DNA probes enabled unambiguous discrimination of the ancestral genomes in B. juncea and B. carinata, and was only partially successful inB. napus. GISH signals in all genomes were localized predominantly in pericentromeric regions of chromosomes. Simultaneous application of genomic and ribosomal DNA probes in multicolor GISH and FISH allowed identification of a significant number of chromosomes in the B. juncea complement. The study also revealed that species of Brassica possess Arabidopsis-type telomeric repeats which in all genomes occupied exclusively terminal, that is, telomeric, locations of chromosomes. © Polish Academy of Sciences, 2005.
Hasterok, R.; Wolny, E. A.; Kulak, S.; Zdziechiewicz, A.; Małuszyńska, J.; Heneen, W. K.
Molecular cytogenetic analysis of Brassica rapa-Brassica oleracea var. alboglabra monosomic addition lines Journal Article
In: Theoretical and Applied Genetics, vol. 111, no. 2, pp. 196-205, 2005, ISSN: 00405752, (32).
@article{2-s2.0-22844431646,
title = {Molecular cytogenetic analysis of Brassica rapa-Brassica oleracea var. alboglabra monosomic addition lines},
author = { R. Hasterok and E.A. Wolny and S. Kulak and A. Zdziechiewicz and J. Małuszyńska and W.K. Heneen},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-22844431646&doi=10.1007%2fs00122-005-1942-7&partnerID=40&md5=21f499c1e9ea7ecb86be9b4f1acc8c7e},
doi = {10.1007/s00122-005-1942-7},
issn = {00405752},
year = {2005},
date = {2005-01-01},
journal = {Theoretical and Applied Genetics},
volume = {111},
number = {2},
pages = {196-205},
abstract = {Interspecific alien chromosome addition lines can be very useful for gene mapping and studying chromosome homoeology between closely related species. In this study we demonstrate a simple but robust manner of identifying individual C-genome chromosomes (C5; C8 and C9) in the A-genome background through the simultaneous use of 5S and 25S ribosomal probes on mitotic and meiotic chromosomes of three different Brassica rapa-B. oleracea var. alboglabra monosomic addition lines. Sequential silver staining and fluorescence in situ hybridisation indicated that 18S-5.8S-25S rRNA genes on the additional chromosome C9 are expressed in the A-genome background. Meiotic behaviour of the additional chromosomes was studied in pollen mother cells at diakinesis and metaphase I. In all of the addition lines the alien chromosome was most frequently observed as a univalent. The alien chromosome C5, which carries an intercalary 5S rDNA locus, occasionally formed trivalents that involved either rDNA- or non rDNA-carrying chromosomes from the A genome. In the case of chromosomes C8 and C9, the most frequently observed intergenomic associations involved the regions occupied by 18S-5.8S-25S ribosomal RNA genes. It is possible that not all such associations represent true pairing but are remnants of nucleolar associations from the preceding interphase. Variations in the numbers and distribution of 5S and 25S rDNA sites between cultivars of B. oleracea, B. oleracea var. alboglabra and B. rapa are discussed. © Springer-Verlag 2005.},
note = {32},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Interspecific alien chromosome addition lines can be very useful for gene mapping and studying chromosome homoeology between closely related species. In this study we demonstrate a simple but robust manner of identifying individual C-genome chromosomes (C5; C8 and C9) in the A-genome background through the simultaneous use of 5S and 25S ribosomal probes on mitotic and meiotic chromosomes of three different Brassica rapa-B. oleracea var. alboglabra monosomic addition lines. Sequential silver staining and fluorescence in situ hybridisation indicated that 18S-5.8S-25S rRNA genes on the additional chromosome C9 are expressed in the A-genome background. Meiotic behaviour of the additional chromosomes was studied in pollen mother cells at diakinesis and metaphase I. In all of the addition lines the alien chromosome was most frequently observed as a univalent. The alien chromosome C5, which carries an intercalary 5S rDNA locus, occasionally formed trivalents that involved either rDNA- or non rDNA-carrying chromosomes from the A genome. In the case of chromosomes C8 and C9, the most frequently observed intergenomic associations involved the regions occupied by 18S-5.8S-25S ribosomal RNA genes. It is possible that not all such associations represent true pairing but are remnants of nucleolar associations from the preceding interphase. Variations in the numbers and distribution of 5S and 25S rDNA sites between cultivars of B. oleracea, B. oleracea var. alboglabra and B. rapa are discussed. © Springer-Verlag 2005.
2003
Małuszyńska, J.; Kwaśniewska, J.; Wolny, E. A.
Chromosomal aberrations in Crepis capillaris cells detected by FISH Journal Article
In: Folia Histochemica et Cytobiologica, vol. 41, no. 2, pp. 101-104, 2003, ISSN: 02398508, (11).
@article{2-s2.0-0037247227,
title = {Chromosomal aberrations in Crepis capillaris cells detected by FISH},
author = { J. Małuszyńska and J. Kwaśniewska and E.A. Wolny},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-0037247227&partnerID=40&md5=2892c6d31a0977bc3dcb1151b6ce5ef9},
issn = {02398508},
year = {2003},
date = {2003-01-01},
journal = {Folia Histochemica et Cytobiologica},
volume = {41},
number = {2},
pages = {101-104},
publisher = {Polish Histochemical and Cytochemical Society},
abstract = {Crepis capillaris (2n=6) is an excellent plant for the assay of chromosome aberrations after mutagenic treatment. It has simple karyotype: three pairs of morphologically distinct and relatively large chromosomes. The frequency of structural chromosome aberrations and micronuclei in root meristem cells has been used for evaluation of the genotoxicity of chemicals and environmental pollutants. The introduction of fluorescence in situ hybridization method allows more detailed detection and localization of chromosomal rearrangements not only in mitotic but also in interphase nuclei. We demonstrate a few examples of the detection of chromosomal aberrations using rDNA and telomeric sequences as probes for in situ hybridization to C. capillaris chromosomes.},
note = {11},
keywords = {},
pubstate = {published},
tppubtype = {article}
}
Crepis capillaris (2n=6) is an excellent plant for the assay of chromosome aberrations after mutagenic treatment. It has simple karyotype: three pairs of morphologically distinct and relatively large chromosomes. The frequency of structural chromosome aberrations and micronuclei in root meristem cells has been used for evaluation of the genotoxicity of chemicals and environmental pollutants. The introduction of fluorescence in situ hybridization method allows more detailed detection and localization of chromosomal rearrangements not only in mitotic but also in interphase nuclei. We demonstrate a few examples of the detection of chromosomal aberrations using rDNA and telomeric sequences as probes for in situ hybridization to C. capillaris chromosomes.