Measuring Meiotic Crossovers via Multi-Locus Genotyping of Single Pollen Grains in Barley
The detection of meiotic crossovers in crop plants currently relies on scoring DNA markers in a segregating population or cytological visualization. We investigated the feasibility of using flow-sorted haploid nuclei, Phi29 DNA polymerase-based whole-genome-amplification (WGA) and multi-locus KASP-g...
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| creator | Dreissig, Steven Fuchs, Jörg Cápal, Petr Kettles, Nicola Byrne, Ed Houben, Andreas |
| description | The detection of meiotic crossovers in crop plants currently relies on scoring DNA markers in a segregating population or cytological visualization. We investigated the feasibility of using flow-sorted haploid nuclei, Phi29 DNA polymerase-based whole-genome-amplification (WGA) and multi-locus KASP-genotyping to measure meiotic crossovers in individual barley pollen grains. To demonstrate the proof of concept, we used 24 gene-based physically mapped single nucleotide polymorphisms to genotype the WGA products of 50 single pollen nuclei. The number of crossovers per chromosome, recombination frequencies along chromosome 3H and segregation distortion were analysed and compared to a doubled haploid (DH) population of the same genotype. The number of crossovers and chromosome wide recombination frequencies show that this approach is able to produce results that resemble those obtained from other methods in a biologically meaningful way. Only the segregation distortion was found to be lower in the pollen population than in DH plants. |
| doi_str_mv | 10.1371/journal.pone.0137677 |
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We investigated the feasibility of using flow-sorted haploid nuclei, Phi29 DNA polymerase-based whole-genome-amplification (WGA) and multi-locus KASP-genotyping to measure meiotic crossovers in individual barley pollen grains. To demonstrate the proof of concept, we used 24 gene-based physically mapped single nucleotide polymorphisms to genotype the WGA products of 50 single pollen nuclei. The number of crossovers per chromosome, recombination frequencies along chromosome 3H and segregation distortion were analysed and compared to a doubled haploid (DH) population of the same genotype. The number of crossovers and chromosome wide recombination frequencies show that this approach is able to produce results that resemble those obtained from other methods in a biologically meaningful way. Only the segregation distortion was found to be lower in the pollen population than in DH plants.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0137677</identifier><identifier>PMID: 26356084</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Arabidopsis ; Arabidopsis thaliana ; Barley ; Biological effects ; Cell cycle ; Chromosomes, Plant ; Crossing over (Genetics) ; Crossing Over, Genetic ; Crossovers ; Deoxyribonucleic acid ; Distortion ; DNA ; DNA methylation ; DNA polymerase ; DNA, Plant ; DNA-directed DNA polymerase ; Enzymes ; Feasibility studies ; Genetic aspects ; Genetics ; Genome, Plant ; Genomes ; Genotyping ; Grains ; Haploidy ; Homogenization ; Hordeum - genetics ; Hordeum vulgare ; Loci ; Meiosis ; Meiosis - genetics ; Multilocus Sequence Typing ; Mutation ; Nuclei ; Nuclei (cytology) ; Physiological aspects ; Plants (botany) ; Pollen ; Pollen - genetics ; Quantitative Trait Loci ; Recombination ; Recombination, Genetic ; Single-nucleotide polymorphism</subject><ispartof>PloS one, 2015-09, Vol.10 (9), p.e0137677-e0137677</ispartof><rights>COPYRIGHT 2015 Public Library of Science</rights><rights>2015 Dreissig et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2015 Dreissig et al 2015 Dreissig et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c692t-704f469ede261244544028238ba039b4162bf2e4fe0e8cf7a82c39b12c92abc3</citedby><cites>FETCH-LOGICAL-c692t-704f469ede261244544028238ba039b4162bf2e4fe0e8cf7a82c39b12c92abc3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4565660/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4565660/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,860,881,2096,2915,23845,27901,27902,53766,53768,79343,79344</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/26356084$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Lichten, Michael</contributor><creatorcontrib>Dreissig, Steven</creatorcontrib><creatorcontrib>Fuchs, Jörg</creatorcontrib><creatorcontrib>Cápal, Petr</creatorcontrib><creatorcontrib>Kettles, Nicola</creatorcontrib><creatorcontrib>Byrne, Ed</creatorcontrib><creatorcontrib>Houben, Andreas</creatorcontrib><title>Measuring Meiotic Crossovers via Multi-Locus Genotyping of Single Pollen Grains in Barley</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>The detection of meiotic crossovers in crop plants currently relies on scoring DNA markers in a segregating population or cytological visualization. We investigated the feasibility of using flow-sorted haploid nuclei, Phi29 DNA polymerase-based whole-genome-amplification (WGA) and multi-locus KASP-genotyping to measure meiotic crossovers in individual barley pollen grains. To demonstrate the proof of concept, we used 24 gene-based physically mapped single nucleotide polymorphisms to genotype the WGA products of 50 single pollen nuclei. The number of crossovers per chromosome, recombination frequencies along chromosome 3H and segregation distortion were analysed and compared to a doubled haploid (DH) population of the same genotype. The number of crossovers and chromosome wide recombination frequencies show that this approach is able to produce results that resemble those obtained from other methods in a biologically meaningful way. Only the segregation distortion was found to be lower in the pollen population than in DH plants.</description><subject>Arabidopsis</subject><subject>Arabidopsis thaliana</subject><subject>Barley</subject><subject>Biological effects</subject><subject>Cell cycle</subject><subject>Chromosomes, Plant</subject><subject>Crossing over (Genetics)</subject><subject>Crossing Over, Genetic</subject><subject>Crossovers</subject><subject>Deoxyribonucleic acid</subject><subject>Distortion</subject><subject>DNA</subject><subject>DNA methylation</subject><subject>DNA polymerase</subject><subject>DNA, Plant</subject><subject>DNA-directed DNA polymerase</subject><subject>Enzymes</subject><subject>Feasibility studies</subject><subject>Genetic aspects</subject><subject>Genetics</subject><subject>Genome, Plant</subject><subject>Genomes</subject><subject>Genotyping</subject><subject>Grains</subject><subject>Haploidy</subject><subject>Homogenization</subject><subject>Hordeum - genetics</subject><subject>Hordeum vulgare</subject><subject>Loci</subject><subject>Meiosis</subject><subject>Meiosis - genetics</subject><subject>Multilocus Sequence Typing</subject><subject>Mutation</subject><subject>Nuclei</subject><subject>Nuclei (cytology)</subject><subject>Physiological aspects</subject><subject>Plants (botany)</subject><subject>Pollen</subject><subject>Pollen - genetics</subject><subject>Quantitative Trait Loci</subject><subject>Recombination</subject><subject>Recombination, Genetic</subject><subject>Single-nucleotide polymorphism</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><sourceid>DOA</sourceid><recordid>eNqNk1Fv0zAQxyMEYmPwDRBEQkLw0GI7jpO8II0KSqVWQ2xC4slynXPryrU7O6not8dZs6lBe0B5cHT-3d--v--S5DVGY5wV-NPGtd4KM945C2MUQ6woniTnuMrIiBGUPT35P0tehLBBKM9Kxp4nZ4RlOUMlPU9-L0CE1mu7ShegXaNlOvEuBLcHH9K9FumiNY0ezZ1sQzoF65rDrqOdSq_jaiD94YwBm0690Dak2qZfhDdweJk8U8IEeNWvF8nNt683k--j-dV0NrmcjySrSDMqEFWUVVADYZhQmlOKSEmycilQVi0pZmSpCFAFCEqpClESGeOYyIqIpcwukrdH2Z1xgfemBI4LjKqSRPVIzI5E7cSG77zeCn_gTmh-F3B-xYWPhRvgGaqWSEmFVZVTVdOyjteKVtWsyhAmZdT63J_WLrdQS7CNF2YgOtyxes1Xbs9pznLGUBT40At4d9tCaPhWBwnGCAuuvbs3zrMip0VE3_2DPl5dT61ELEBb5eK5shPll7QzklRlR40foeJXw1bL2EFKx_gg4eMgITIN_GlWog2Bz65__j979WvIvj9h1yBMsw7OtI12NgxBegRl144e1IPJGPFuAO7d4N0A8H4AYtqb0wd6SLrv-Owvygv_MQ</recordid><startdate>20150910</startdate><enddate>20150910</enddate><creator>Dreissig, Steven</creator><creator>Fuchs, Jörg</creator><creator>Cápal, Petr</creator><creator>Kettles, Nicola</creator><creator>Byrne, Ed</creator><creator>Houben, Andreas</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>IOV</scope><scope>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QO</scope><scope>7RV</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TG</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>KL.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PDBOC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope></search><sort><creationdate>20150910</creationdate><title>Measuring Meiotic Crossovers via Multi-Locus Genotyping of Single Pollen Grains in Barley</title><author>Dreissig, Steven ; Fuchs, Jörg ; Cápal, Petr ; Kettles, Nicola ; Byrne, Ed ; Houben, Andreas</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c692t-704f469ede261244544028238ba039b4162bf2e4fe0e8cf7a82c39b12c92abc3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Arabidopsis</topic><topic>Arabidopsis thaliana</topic><topic>Barley</topic><topic>Biological effects</topic><topic>Cell cycle</topic><topic>Chromosomes, Plant</topic><topic>Crossing over (Genetics)</topic><topic>Crossing Over, Genetic</topic><topic>Crossovers</topic><topic>Deoxyribonucleic acid</topic><topic>Distortion</topic><topic>DNA</topic><topic>DNA methylation</topic><topic>DNA polymerase</topic><topic>DNA, Plant</topic><topic>DNA-directed DNA polymerase</topic><topic>Enzymes</topic><topic>Feasibility studies</topic><topic>Genetic aspects</topic><topic>Genetics</topic><topic>Genome, Plant</topic><topic>Genomes</topic><topic>Genotyping</topic><topic>Grains</topic><topic>Haploidy</topic><topic>Homogenization</topic><topic>Hordeum - 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We investigated the feasibility of using flow-sorted haploid nuclei, Phi29 DNA polymerase-based whole-genome-amplification (WGA) and multi-locus KASP-genotyping to measure meiotic crossovers in individual barley pollen grains. To demonstrate the proof of concept, we used 24 gene-based physically mapped single nucleotide polymorphisms to genotype the WGA products of 50 single pollen nuclei. The number of crossovers per chromosome, recombination frequencies along chromosome 3H and segregation distortion were analysed and compared to a doubled haploid (DH) population of the same genotype. The number of crossovers and chromosome wide recombination frequencies show that this approach is able to produce results that resemble those obtained from other methods in a biologically meaningful way. Only the segregation distortion was found to be lower in the pollen population than in DH plants.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>26356084</pmid><doi>10.1371/journal.pone.0137677</doi><oa>free_for_read</oa></addata></record> |
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| subjects | Arabidopsis Arabidopsis thaliana Barley Biological effects Cell cycle Chromosomes, Plant Crossing over (Genetics) Crossing Over, Genetic Crossovers Deoxyribonucleic acid Distortion DNA DNA methylation DNA polymerase DNA, Plant DNA-directed DNA polymerase Enzymes Feasibility studies Genetic aspects Genetics Genome, Plant Genomes Genotyping Grains Haploidy Homogenization Hordeum - genetics Hordeum vulgare Loci Meiosis Meiosis - genetics Multilocus Sequence Typing Mutation Nuclei Nuclei (cytology) Physiological aspects Plants (botany) Pollen Pollen - genetics Quantitative Trait Loci Recombination Recombination, Genetic Single-nucleotide polymorphism |
| title | Measuring Meiotic Crossovers via Multi-Locus Genotyping of Single Pollen Grains in Barley |
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