Characterization and expression analysis of waxy alleles in barley accessions
Granule Bound Starch Synthase I (GBSS I) encoded by the waxy gene plays an important role in accumulating amylose during the development of starch granules in barley. In this study, we isolated and characterized waxy alleles of three waxy (GSHO 908, GSHO 1828 and NA 40) and two non-waxy barley acces...
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| Veröffentlicht in: | Genetica 2013-06, Vol.141 (4-6), p.227-238 |
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| creator | Ma, Jian Jiang, Qian-Tao Zhao, Quan-Zhi Zhao, Shan Lan, Xiu-Jin Dai, Shou-Fen Lu, Zhen-Xiang Liu, Chunji Wei, Yu-Ming Zheng, You-Liang |
| description | Granule Bound Starch Synthase I (GBSS I) encoded by the waxy gene plays an important role in accumulating amylose during the development of starch granules in barley. In this study, we isolated and characterized waxy alleles of three waxy (GSHO 908, GSHO 1828 and NA 40) and two non-waxy barley accessions (PI 483237 and CIho 15773), estimated the expression patterns of waxy genes via Real-time quantitative PCR (RT-qPCR), investigated promoter activity by analyzing promoter-GUS expression, and examined possible effects of waxy alleles on starch granule morphology in barley accessions by scanning electron microscopy (SEM). A 193-bp insertion in intron 1, a 15-bp insertion in the coding region, and some single nucleotide polymorphic sites were detected in the waxy barley accessions. In addition, a 397-bp deletion containing the TATA box, transcription starting point, exon 1 and partial intron 1 were also identified in the waxy barley accessions. RT-qPCR analysis showed that waxy accessions had lower waxy expression levels than those of non-waxy accessions. Transient expression assays showed that GUS activity driven by the 1,029-bp promoter of the non-waxy accessions was stronger than that driven by the 822-bp promoter of the waxy accessions. SEM revealed no apparent differences of starch granule morphology between waxy and non-waxy accessions. Our results showed that the 397-bp deletion identified in the waxy barley accessions is likely responsible for the reduction of waxy transcript, leading to lower concentrations of GBSS I protein thus lower amylose content. |
| doi_str_mv | 10.1007/s10709-013-9721-x |
| format | Article |
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In this study, we isolated and characterized waxy alleles of three waxy (GSHO 908, GSHO 1828 and NA 40) and two non-waxy barley accessions (PI 483237 and CIho 15773), estimated the expression patterns of waxy genes via Real-time quantitative PCR (RT-qPCR), investigated promoter activity by analyzing promoter-GUS expression, and examined possible effects of waxy alleles on starch granule morphology in barley accessions by scanning electron microscopy (SEM). A 193-bp insertion in intron 1, a 15-bp insertion in the coding region, and some single nucleotide polymorphic sites were detected in the waxy barley accessions. In addition, a 397-bp deletion containing the TATA box, transcription starting point, exon 1 and partial intron 1 were also identified in the waxy barley accessions. RT-qPCR analysis showed that waxy accessions had lower waxy expression levels than those of non-waxy accessions. Transient expression assays showed that GUS activity driven by the 1,029-bp promoter of the non-waxy accessions was stronger than that driven by the 822-bp promoter of the waxy accessions. SEM revealed no apparent differences of starch granule morphology between waxy and non-waxy accessions. Our results showed that the 397-bp deletion identified in the waxy barley accessions is likely responsible for the reduction of waxy transcript, leading to lower concentrations of GBSS I protein thus lower amylose content.</description><identifier>ISSN: 0016-6707</identifier><identifier>EISSN: 1573-6857</identifier><identifier>DOI: 10.1007/s10709-013-9721-x</identifier><identifier>PMID: 23690246</identifier><language>eng</language><publisher>Dordrecht: Springer-Verlag</publisher><subject>Alleles ; Amylose - chemistry ; Animal Genetics and Genomics ; Barley ; Biomedical and Life Sciences ; Carbohydrate Metabolism - genetics ; Evolutionary Biology ; Gene Expression ; Gene Order ; Genes, Plant ; Hordeum - genetics ; Hordeum - metabolism ; Hordeum vulgare ; Human Genetics ; Life Sciences ; Microbial Genetics and Genomics ; Nucleotide Motifs ; Plant Genetics and Genomics ; polymerase chain reaction ; Polymorphism, Genetic ; Promoter Regions, Genetic ; scanning electron microscopy ; Sequence Deletion ; single nucleotide polymorphism ; Starch ; Starch - ultrastructure ; starch granules ; starch synthase ; Starch Synthase - chemistry ; Starch Synthase - genetics ; TATA box ; Waxes</subject><ispartof>Genetica, 2013-06, Vol.141 (4-6), p.227-238</ispartof><rights>Springer Science+Business Media Dordrecht 2013</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c429t-136c798045daec9a9eacc5615d1a529fa3aa7a6df83f2c29866d973ef3940a413</citedby><cites>FETCH-LOGICAL-c429t-136c798045daec9a9eacc5615d1a529fa3aa7a6df83f2c29866d973ef3940a413</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://link.springer.com/content/pdf/10.1007/s10709-013-9721-x$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s10709-013-9721-x$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>314,780,784,27923,27924,41487,42556,51318</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23690246$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Ma, Jian</creatorcontrib><creatorcontrib>Jiang, Qian-Tao</creatorcontrib><creatorcontrib>Zhao, Quan-Zhi</creatorcontrib><creatorcontrib>Zhao, Shan</creatorcontrib><creatorcontrib>Lan, Xiu-Jin</creatorcontrib><creatorcontrib>Dai, Shou-Fen</creatorcontrib><creatorcontrib>Lu, Zhen-Xiang</creatorcontrib><creatorcontrib>Liu, Chunji</creatorcontrib><creatorcontrib>Wei, Yu-Ming</creatorcontrib><creatorcontrib>Zheng, You-Liang</creatorcontrib><title>Characterization and expression analysis of waxy alleles in barley accessions</title><title>Genetica</title><addtitle>Genetica</addtitle><addtitle>Genetica</addtitle><description>Granule Bound Starch Synthase I (GBSS I) encoded by the waxy gene plays an important role in accumulating amylose during the development of starch granules in barley. In this study, we isolated and characterized waxy alleles of three waxy (GSHO 908, GSHO 1828 and NA 40) and two non-waxy barley accessions (PI 483237 and CIho 15773), estimated the expression patterns of waxy genes via Real-time quantitative PCR (RT-qPCR), investigated promoter activity by analyzing promoter-GUS expression, and examined possible effects of waxy alleles on starch granule morphology in barley accessions by scanning electron microscopy (SEM). A 193-bp insertion in intron 1, a 15-bp insertion in the coding region, and some single nucleotide polymorphic sites were detected in the waxy barley accessions. In addition, a 397-bp deletion containing the TATA box, transcription starting point, exon 1 and partial intron 1 were also identified in the waxy barley accessions. RT-qPCR analysis showed that waxy accessions had lower waxy expression levels than those of non-waxy accessions. Transient expression assays showed that GUS activity driven by the 1,029-bp promoter of the non-waxy accessions was stronger than that driven by the 822-bp promoter of the waxy accessions. SEM revealed no apparent differences of starch granule morphology between waxy and non-waxy accessions. Our results showed that the 397-bp deletion identified in the waxy barley accessions is likely responsible for the reduction of waxy transcript, leading to lower concentrations of GBSS I protein thus lower amylose content.</description><subject>Alleles</subject><subject>Amylose - chemistry</subject><subject>Animal Genetics and Genomics</subject><subject>Barley</subject><subject>Biomedical and Life Sciences</subject><subject>Carbohydrate Metabolism - genetics</subject><subject>Evolutionary Biology</subject><subject>Gene Expression</subject><subject>Gene Order</subject><subject>Genes, Plant</subject><subject>Hordeum - genetics</subject><subject>Hordeum - metabolism</subject><subject>Hordeum vulgare</subject><subject>Human Genetics</subject><subject>Life Sciences</subject><subject>Microbial Genetics and Genomics</subject><subject>Nucleotide Motifs</subject><subject>Plant Genetics and Genomics</subject><subject>polymerase chain reaction</subject><subject>Polymorphism, Genetic</subject><subject>Promoter Regions, Genetic</subject><subject>scanning electron microscopy</subject><subject>Sequence Deletion</subject><subject>single nucleotide polymorphism</subject><subject>Starch</subject><subject>Starch - ultrastructure</subject><subject>starch granules</subject><subject>starch synthase</subject><subject>Starch Synthase - chemistry</subject><subject>Starch Synthase - genetics</subject><subject>TATA 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and expression analysis of waxy alleles in barley accessions</title><author>Ma, Jian ; Jiang, Qian-Tao ; Zhao, Quan-Zhi ; Zhao, Shan ; Lan, Xiu-Jin ; Dai, Shou-Fen ; Lu, Zhen-Xiang ; Liu, Chunji ; Wei, Yu-Ming ; Zheng, You-Liang</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c429t-136c798045daec9a9eacc5615d1a529fa3aa7a6df83f2c29866d973ef3940a413</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2013</creationdate><topic>Alleles</topic><topic>Amylose - chemistry</topic><topic>Animal Genetics and Genomics</topic><topic>Barley</topic><topic>Biomedical and Life Sciences</topic><topic>Carbohydrate Metabolism - genetics</topic><topic>Evolutionary Biology</topic><topic>Gene Expression</topic><topic>Gene Order</topic><topic>Genes, Plant</topic><topic>Hordeum - genetics</topic><topic>Hordeum - metabolism</topic><topic>Hordeum vulgare</topic><topic>Human Genetics</topic><topic>Life Sciences</topic><topic>Microbial Genetics and Genomics</topic><topic>Nucleotide Motifs</topic><topic>Plant Genetics and Genomics</topic><topic>polymerase chain reaction</topic><topic>Polymorphism, Genetic</topic><topic>Promoter Regions, Genetic</topic><topic>scanning electron microscopy</topic><topic>Sequence Deletion</topic><topic>single nucleotide polymorphism</topic><topic>Starch</topic><topic>Starch - ultrastructure</topic><topic>starch granules</topic><topic>starch synthase</topic><topic>Starch Synthase - chemistry</topic><topic>Starch Synthase - genetics</topic><topic>TATA box</topic><topic>Waxes</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Ma, Jian</creatorcontrib><creatorcontrib>Jiang, Qian-Tao</creatorcontrib><creatorcontrib>Zhao, Quan-Zhi</creatorcontrib><creatorcontrib>Zhao, Shan</creatorcontrib><creatorcontrib>Lan, Xiu-Jin</creatorcontrib><creatorcontrib>Dai, Shou-Fen</creatorcontrib><creatorcontrib>Lu, 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Edition</collection><collection>Genetics Abstracts</collection><jtitle>Genetica</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Ma, Jian</au><au>Jiang, Qian-Tao</au><au>Zhao, Quan-Zhi</au><au>Zhao, Shan</au><au>Lan, Xiu-Jin</au><au>Dai, Shou-Fen</au><au>Lu, Zhen-Xiang</au><au>Liu, Chunji</au><au>Wei, Yu-Ming</au><au>Zheng, You-Liang</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Characterization and expression analysis of waxy alleles in barley accessions</atitle><jtitle>Genetica</jtitle><stitle>Genetica</stitle><addtitle>Genetica</addtitle><date>2013-06-01</date><risdate>2013</risdate><volume>141</volume><issue>4-6</issue><spage>227</spage><epage>238</epage><pages>227-238</pages><issn>0016-6707</issn><eissn>1573-6857</eissn><abstract>Granule Bound Starch Synthase I (GBSS I) encoded by the waxy gene plays an important role in accumulating amylose during the development of starch granules in barley. In this study, we isolated and characterized waxy alleles of three waxy (GSHO 908, GSHO 1828 and NA 40) and two non-waxy barley accessions (PI 483237 and CIho 15773), estimated the expression patterns of waxy genes via Real-time quantitative PCR (RT-qPCR), investigated promoter activity by analyzing promoter-GUS expression, and examined possible effects of waxy alleles on starch granule morphology in barley accessions by scanning electron microscopy (SEM). A 193-bp insertion in intron 1, a 15-bp insertion in the coding region, and some single nucleotide polymorphic sites were detected in the waxy barley accessions. In addition, a 397-bp deletion containing the TATA box, transcription starting point, exon 1 and partial intron 1 were also identified in the waxy barley accessions. RT-qPCR analysis showed that waxy accessions had lower waxy expression levels than those of non-waxy accessions. Transient expression assays showed that GUS activity driven by the 1,029-bp promoter of the non-waxy accessions was stronger than that driven by the 822-bp promoter of the waxy accessions. SEM revealed no apparent differences of starch granule morphology between waxy and non-waxy accessions. Our results showed that the 397-bp deletion identified in the waxy barley accessions is likely responsible for the reduction of waxy transcript, leading to lower concentrations of GBSS I protein thus lower amylose content.</abstract><cop>Dordrecht</cop><pub>Springer-Verlag</pub><pmid>23690246</pmid><doi>10.1007/s10709-013-9721-x</doi><tpages>12</tpages></addata></record> |
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| subjects | Alleles Amylose - chemistry Animal Genetics and Genomics Barley Biomedical and Life Sciences Carbohydrate Metabolism - genetics Evolutionary Biology Gene Expression Gene Order Genes, Plant Hordeum - genetics Hordeum - metabolism Hordeum vulgare Human Genetics Life Sciences Microbial Genetics and Genomics Nucleotide Motifs Plant Genetics and Genomics polymerase chain reaction Polymorphism, Genetic Promoter Regions, Genetic scanning electron microscopy Sequence Deletion single nucleotide polymorphism Starch Starch - ultrastructure starch granules starch synthase Starch Synthase - chemistry Starch Synthase - genetics TATA box Waxes |
| title | Characterization and expression analysis of waxy alleles in barley accessions |
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