Physical mapping resources for large plant genomes: radiation hybrids for wheat D-genome progenitor <it>Aegilops tauschii</it>

<p>Abstract</p> <p>Background</p> <p>Development of a high quality reference sequence is a daunting task in crops like wheat with large (~17Gb), highly repetitive (>80%) and polyploid genome. To achieve complete sequence assembly of such genomes, development of a hig...

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Main Authors: Kumar Ajay, Simons Kristin, Iqbal Muhammad J, de Jiménez Monika, Bassi Filippo M, Ghavami Farhad, Al-Azzam Omar, Drader Thomas, Wang Yi, Luo Ming-Cheng, Gu Yong Q, Denton Anne, Lazo Gerard R, Xu Steven S, Dvorak Jan, Kianian Penny MA, Kianian Shahryar F
Format: Article
Language:English
Published: BMC 2012-11-01
Series:BMC Genomics
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Online Access:http://www.biomedcentral.com/1471-2164/13/597
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author Kumar Ajay
Simons Kristin
Iqbal Muhammad J
de Jiménez Monika
Bassi Filippo M
Ghavami Farhad
Al-Azzam Omar
Drader Thomas
Wang Yi
Luo Ming-Cheng
Gu Yong Q
Denton Anne
Lazo Gerard R
Xu Steven S
Dvorak Jan
Kianian Penny MA
Kianian Shahryar F
author_facet Kumar Ajay
Simons Kristin
Iqbal Muhammad J
de Jiménez Monika
Bassi Filippo M
Ghavami Farhad
Al-Azzam Omar
Drader Thomas
Wang Yi
Luo Ming-Cheng
Gu Yong Q
Denton Anne
Lazo Gerard R
Xu Steven S
Dvorak Jan
Kianian Penny MA
Kianian Shahryar F
author_sort Kumar Ajay
collection DOAJ
description <p>Abstract</p> <p>Background</p> <p>Development of a high quality reference sequence is a daunting task in crops like wheat with large (~17Gb), highly repetitive (>80%) and polyploid genome. To achieve complete sequence assembly of such genomes, development of a high quality physical map is a necessary first step. However, due to the lack of recombination in certain regions of the chromosomes, genetic mapping, which uses recombination frequency to map marker loci, alone is not sufficient to develop high quality marker scaffolds for a sequence ready physical map. Radiation hybrid (RH) mapping, which uses radiation induced chromosomal breaks, has proven to be a successful approach for developing marker scaffolds for sequence assembly in animal systems. Here, the development and characterization of a RH panel for the mapping of D-genome of wheat progenitor <it>Aegilops tauschii</it> is reported<it>.</it></p> <p>Results</p> <p>Radiation dosages of 350 and 450 Gy were optimized for seed irradiation of a synthetic hexaploid (AABBDD) wheat with the D-genome of <it>Ae. tauschii</it> accession AL8/78. The surviving plants after irradiation were crossed to durum wheat (AABB), to produce pentaploid RH<sub>1</sub>s (AABBD), which allows the simultaneous mapping of the whole D-genome. A panel of 1,510 RH<sub>1</sub> plants was obtained, of which 592 plants were generated from the mature RH<sub>1</sub> seeds, and 918 plants were rescued through embryo culture due to poor germination (<3%) of mature RH<sub>1</sub> seeds. This panel showed a homogenous marker loss (2.1%) after screening with SSR markers uniformly covering all the D-genome chromosomes. Different marker systems mostly detected different lines with deletions. Using markers covering known distances, the mapping resolution of this RH panel was estimated to be <140kb. Analysis of only 16 RH lines carrying deletions on chromosome 2D resulted in a physical map with <it>cM/cR</it> ratio of 1:5.2 and 15 distinct bins. Additionally, with this small set of lines, almost all the tested ESTs could be mapped. A set of 399 most informative RH lines with an average deletion frequency of ~10% were identified for developing high density marker scaffolds of the D-genome.</p> <p>Conclusions</p> <p>The RH panel reported here is the first developed for any wild ancestor of a major cultivated plant species. The results provided insight into various aspects of RH mapping in plants, including the genetically effective cell number for wheat (for the first time) and the potential implementation of this technique in other plant species. This RH panel will be an invaluable resource for mapping gene based markers, developing a complete marker scaffold for the whole genome sequence assembly, fine mapping of markers and functional characterization of genes and gene networks present on the D-genome.</p>
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spelling doaj.art-ce70ae732a8a4bc2975db1e31cea66042022-12-22T01:45:04ZengBMCBMC Genomics1471-21642012-11-0113159710.1186/1471-2164-13-597Physical mapping resources for large plant genomes: radiation hybrids for wheat D-genome progenitor <it>Aegilops tauschii</it>Kumar AjaySimons KristinIqbal Muhammad Jde Jiménez MonikaBassi Filippo MGhavami FarhadAl-Azzam OmarDrader ThomasWang YiLuo Ming-ChengGu Yong QDenton AnneLazo Gerard RXu Steven SDvorak JanKianian Penny MAKianian Shahryar F<p>Abstract</p> <p>Background</p> <p>Development of a high quality reference sequence is a daunting task in crops like wheat with large (~17Gb), highly repetitive (>80%) and polyploid genome. To achieve complete sequence assembly of such genomes, development of a high quality physical map is a necessary first step. However, due to the lack of recombination in certain regions of the chromosomes, genetic mapping, which uses recombination frequency to map marker loci, alone is not sufficient to develop high quality marker scaffolds for a sequence ready physical map. Radiation hybrid (RH) mapping, which uses radiation induced chromosomal breaks, has proven to be a successful approach for developing marker scaffolds for sequence assembly in animal systems. Here, the development and characterization of a RH panel for the mapping of D-genome of wheat progenitor <it>Aegilops tauschii</it> is reported<it>.</it></p> <p>Results</p> <p>Radiation dosages of 350 and 450 Gy were optimized for seed irradiation of a synthetic hexaploid (AABBDD) wheat with the D-genome of <it>Ae. tauschii</it> accession AL8/78. The surviving plants after irradiation were crossed to durum wheat (AABB), to produce pentaploid RH<sub>1</sub>s (AABBD), which allows the simultaneous mapping of the whole D-genome. A panel of 1,510 RH<sub>1</sub> plants was obtained, of which 592 plants were generated from the mature RH<sub>1</sub> seeds, and 918 plants were rescued through embryo culture due to poor germination (<3%) of mature RH<sub>1</sub> seeds. This panel showed a homogenous marker loss (2.1%) after screening with SSR markers uniformly covering all the D-genome chromosomes. Different marker systems mostly detected different lines with deletions. Using markers covering known distances, the mapping resolution of this RH panel was estimated to be <140kb. Analysis of only 16 RH lines carrying deletions on chromosome 2D resulted in a physical map with <it>cM/cR</it> ratio of 1:5.2 and 15 distinct bins. Additionally, with this small set of lines, almost all the tested ESTs could be mapped. A set of 399 most informative RH lines with an average deletion frequency of ~10% were identified for developing high density marker scaffolds of the D-genome.</p> <p>Conclusions</p> <p>The RH panel reported here is the first developed for any wild ancestor of a major cultivated plant species. The results provided insight into various aspects of RH mapping in plants, including the genetically effective cell number for wheat (for the first time) and the potential implementation of this technique in other plant species. This RH panel will be an invaluable resource for mapping gene based markers, developing a complete marker scaffold for the whole genome sequence assembly, fine mapping of markers and functional characterization of genes and gene networks present on the D-genome.</p>http://www.biomedcentral.com/1471-2164/13/597<it>Aegilops tauschii</it>Genetically effective cell numberPhysical mappingRadiation hybrid mappingRepeat DNA junction markerWheat
spellingShingle Kumar Ajay
Simons Kristin
Iqbal Muhammad J
de Jiménez Monika
Bassi Filippo M
Ghavami Farhad
Al-Azzam Omar
Drader Thomas
Wang Yi
Luo Ming-Cheng
Gu Yong Q
Denton Anne
Lazo Gerard R
Xu Steven S
Dvorak Jan
Kianian Penny MA
Kianian Shahryar F
Physical mapping resources for large plant genomes: radiation hybrids for wheat D-genome progenitor <it>Aegilops tauschii</it>
BMC Genomics
<it>Aegilops tauschii</it>
Genetically effective cell number
Physical mapping
Radiation hybrid mapping
Repeat DNA junction marker
Wheat
title Physical mapping resources for large plant genomes: radiation hybrids for wheat D-genome progenitor <it>Aegilops tauschii</it>
title_full Physical mapping resources for large plant genomes: radiation hybrids for wheat D-genome progenitor <it>Aegilops tauschii</it>
title_fullStr Physical mapping resources for large plant genomes: radiation hybrids for wheat D-genome progenitor <it>Aegilops tauschii</it>
title_full_unstemmed Physical mapping resources for large plant genomes: radiation hybrids for wheat D-genome progenitor <it>Aegilops tauschii</it>
title_short Physical mapping resources for large plant genomes: radiation hybrids for wheat D-genome progenitor <it>Aegilops tauschii</it>
title_sort physical mapping resources for large plant genomes radiation hybrids for wheat d genome progenitor it aegilops tauschii it
topic <it>Aegilops tauschii</it>
Genetically effective cell number
Physical mapping
Radiation hybrid mapping
Repeat DNA junction marker
Wheat
url http://www.biomedcentral.com/1471-2164/13/597
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