A highly efficient maize nucellus protoplast system for transient gene expression and studying programmed cell death-related processes
Key message Conditions for the isolation and transfection of maize nucellus protoplasts were established. We demonstrated its utilization for protein expression, localization, protein–protein interaction, and the investigation of PCD-related processes. Plant protoplasts are an important and versatil...
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| Veröffentlicht in: | Plant cell reports 2015-07, Vol.34 (7), p.1239-1251 |
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| creator | Chen, Jiang Yi, Qiang Song, Qiaoheng Gu, Yong Zhang, Junjie Hu, Yufeng Liu, Hanmei Liu, Yinghong Yu, Guowu Huang, Yubi |
| description | Key message
Conditions for the isolation and transfection of maize nucellus protoplasts were established. We demonstrated its utilization for protein expression, localization, protein–protein interaction, and the investigation of PCD-related processes.
Plant protoplasts are an important and versatile cell system that is widely used in the analysis of gene characterization and diverse signaling pathways. Programmed cell death (PCD) occurs throughout the life of plants from embryogenesis to fertilization. The maize nucellus undergoes typical PCD during development of the embryo sac. The nucellus protoplast shows potential for use in research of PCD-related processes. No studies have reported previously the isolation and transfection of nucellus protoplasts. In this study, conditions for the isolation and transfection of maize nucellus protoplasts were established. The maize protoplast system can be used for protein expression, localization, and protein–protein interaction. We applied this system to investigate PCD-related processes. Quantitative real-time PCR analysis revealed that transient expression of MADS29 in the maize nucellus protoplast increases Cys-protease gene transcript level. In addition, β-glucuronidase and luciferase activity assays showed that MADS29 could enhance the promoter activities of the Cys-protease gene. Thus, we demonstrated the potential of a highly efficient maize nucellus protoplast system for transient gene expression and investigation of PCD-related processes. |
| doi_str_mv | 10.1007/s00299-015-1783-z |
| format | Article |
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Conditions for the isolation and transfection of maize nucellus protoplasts were established. We demonstrated its utilization for protein expression, localization, protein–protein interaction, and the investigation of PCD-related processes.
Plant protoplasts are an important and versatile cell system that is widely used in the analysis of gene characterization and diverse signaling pathways. Programmed cell death (PCD) occurs throughout the life of plants from embryogenesis to fertilization. The maize nucellus undergoes typical PCD during development of the embryo sac. The nucellus protoplast shows potential for use in research of PCD-related processes. No studies have reported previously the isolation and transfection of nucellus protoplasts. In this study, conditions for the isolation and transfection of maize nucellus protoplasts were established. The maize protoplast system can be used for protein expression, localization, and protein–protein interaction. We applied this system to investigate PCD-related processes. Quantitative real-time PCR analysis revealed that transient expression of MADS29 in the maize nucellus protoplast increases Cys-protease gene transcript level. In addition, β-glucuronidase and luciferase activity assays showed that MADS29 could enhance the promoter activities of the Cys-protease gene. Thus, we demonstrated the potential of a highly efficient maize nucellus protoplast system for transient gene expression and investigation of PCD-related processes.</description><identifier>ISSN: 0721-7714</identifier><identifier>EISSN: 1432-203X</identifier><identifier>DOI: 10.1007/s00299-015-1783-z</identifier><identifier>PMID: 25786591</identifier><language>eng</language><publisher>Berlin/Heidelberg: Springer Berlin Heidelberg</publisher><subject>Apoptosis ; Biomedical and Life Sciences ; Biotechnology ; Cell Biology ; Cell death ; Corn ; Embryonic growth stage ; Gene Expression ; Gene Expression Regulation, Plant ; Genes, Plant ; Life Sciences ; Original Paper ; Osmotic Pressure ; Plant Biochemistry ; Plant Proteins - metabolism ; Plant Sciences ; Plants, Genetically Modified ; Pollination ; Promoter Regions, Genetic - genetics ; Protein Binding ; Protein Interaction Maps ; Protein Transport ; Protoplasts - metabolism ; Real-Time Polymerase Chain Reaction ; Subcellular Fractions - metabolism ; Transfection ; Zea mays ; Zea mays - cytology ; Zea mays - genetics</subject><ispartof>Plant cell reports, 2015-07, Vol.34 (7), p.1239-1251</ispartof><rights>Springer-Verlag Berlin Heidelberg 2015</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c541t-c60fe15b9c2b345615a6acec0d90b56cba731e3c261e14f3fcf04afa9e3120863</citedby><cites>FETCH-LOGICAL-c541t-c60fe15b9c2b345615a6acec0d90b56cba731e3c261e14f3fcf04afa9e3120863</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/s00299-015-1783-z$$EPDF$$P50$$Gspringer$$H</linktopdf><linktohtml>$$Uhttps://link.springer.com/10.1007/s00299-015-1783-z$$EHTML$$P50$$Gspringer$$H</linktohtml><link.rule.ids>315,781,785,27928,27929,41492,42561,51323</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25786591$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Chen, Jiang</creatorcontrib><creatorcontrib>Yi, Qiang</creatorcontrib><creatorcontrib>Song, Qiaoheng</creatorcontrib><creatorcontrib>Gu, Yong</creatorcontrib><creatorcontrib>Zhang, Junjie</creatorcontrib><creatorcontrib>Hu, Yufeng</creatorcontrib><creatorcontrib>Liu, Hanmei</creatorcontrib><creatorcontrib>Liu, Yinghong</creatorcontrib><creatorcontrib>Yu, Guowu</creatorcontrib><creatorcontrib>Huang, Yubi</creatorcontrib><title>A highly efficient maize nucellus protoplast system for transient gene expression and studying programmed cell death-related processes</title><title>Plant cell reports</title><addtitle>Plant Cell Rep</addtitle><addtitle>Plant Cell Rep</addtitle><description>Key message
Conditions for the isolation and transfection of maize nucellus protoplasts were established. We demonstrated its utilization for protein expression, localization, protein–protein interaction, and the investigation of PCD-related processes.
Plant protoplasts are an important and versatile cell system that is widely used in the analysis of gene characterization and diverse signaling pathways. Programmed cell death (PCD) occurs throughout the life of plants from embryogenesis to fertilization. The maize nucellus undergoes typical PCD during development of the embryo sac. The nucellus protoplast shows potential for use in research of PCD-related processes. No studies have reported previously the isolation and transfection of nucellus protoplasts. In this study, conditions for the isolation and transfection of maize nucellus protoplasts were established. The maize protoplast system can be used for protein expression, localization, and protein–protein interaction. We applied this system to investigate PCD-related processes. Quantitative real-time PCR analysis revealed that transient expression of MADS29 in the maize nucellus protoplast increases Cys-protease gene transcript level. In addition, β-glucuronidase and luciferase activity assays showed that MADS29 could enhance the promoter activities of the Cys-protease gene. Thus, we demonstrated the potential of a highly efficient maize nucellus protoplast system for transient gene expression and investigation of PCD-related processes.</description><subject>Apoptosis</subject><subject>Biomedical and Life Sciences</subject><subject>Biotechnology</subject><subject>Cell Biology</subject><subject>Cell death</subject><subject>Corn</subject><subject>Embryonic growth stage</subject><subject>Gene Expression</subject><subject>Gene Expression Regulation, Plant</subject><subject>Genes, Plant</subject><subject>Life Sciences</subject><subject>Original Paper</subject><subject>Osmotic Pressure</subject><subject>Plant Biochemistry</subject><subject>Plant Proteins - metabolism</subject><subject>Plant Sciences</subject><subject>Plants, Genetically Modified</subject><subject>Pollination</subject><subject>Promoter Regions, Genetic - genetics</subject><subject>Protein Binding</subject><subject>Protein Interaction Maps</subject><subject>Protein Transport</subject><subject>Protoplasts - metabolism</subject><subject>Real-Time Polymerase Chain Reaction</subject><subject>Subcellular Fractions - metabolism</subject><subject>Transfection</subject><subject>Zea mays</subject><subject>Zea mays - cytology</subject><subject>Zea mays - genetics</subject><issn>0721-7714</issn><issn>1432-203X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNqFkU-L1TAUxYMoznP0A7iRgBs30dykTZrlMIx_YMCNgruSpjd9Hdr0maTgex_Az23qG0UEcRXI-Z1zk3sIeQ78NXCu3yTOhTGMQ81AN5KdHpAdVFIwweWXh2THtQCmNVQX5ElKd5wXUavH5ELUulG1gR35fkX347CfjhS9H92IIdPZjiekYXU4TWuih7jk5TDZlGk6powz9UukOdqQfuIDBqT47RAxpXEJ1Iaeprz2xzEMm3mIdp6xp1sc7dHmPYs42VyuiuqKC9NT8sjbKeGz-_OSfH578-n6Pbv9-O7D9dUtc3UFmTnFPULdGSc6WdUKaqusQ8d7w7tauc5qCSidUIBQeemd55X11qAEwRslL8mrc26Z_HXFlNt5TNvDbMBlTS3osiNTSYD_o6ppSgmgZEFf_oXeLWsM5SOFMkaA1EYUCs6Ui0tKEX17iONs47EF3m59tuc-29Jnu_XZnornxX3y2pUd_nb8KrAA4gykIoUB4x-j_5n6A6x6rmA</recordid><startdate>20150701</startdate><enddate>20150701</enddate><creator>Chen, Jiang</creator><creator>Yi, Qiang</creator><creator>Song, Qiaoheng</creator><creator>Gu, Yong</creator><creator>Zhang, Junjie</creator><creator>Hu, Yufeng</creator><creator>Liu, Hanmei</creator><creator>Liu, Yinghong</creator><creator>Yu, Guowu</creator><creator>Huang, Yubi</creator><general>Springer Berlin Heidelberg</general><general>Springer Nature B.V</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>3V.</scope><scope>7QL</scope><scope>7T5</scope><scope>7T7</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</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>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>20150701</creationdate><title>A highly efficient maize nucellus protoplast system for transient gene expression and studying programmed cell death-related processes</title><author>Chen, Jiang ; Yi, Qiang ; Song, Qiaoheng ; Gu, Yong ; Zhang, Junjie ; Hu, Yufeng ; Liu, Hanmei ; Liu, Yinghong ; Yu, Guowu ; Huang, Yubi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c541t-c60fe15b9c2b345615a6acec0d90b56cba731e3c261e14f3fcf04afa9e3120863</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Apoptosis</topic><topic>Biomedical and Life Sciences</topic><topic>Biotechnology</topic><topic>Cell Biology</topic><topic>Cell death</topic><topic>Corn</topic><topic>Embryonic growth stage</topic><topic>Gene Expression</topic><topic>Gene Expression Regulation, Plant</topic><topic>Genes, Plant</topic><topic>Life Sciences</topic><topic>Original Paper</topic><topic>Osmotic Pressure</topic><topic>Plant Biochemistry</topic><topic>Plant Proteins - metabolism</topic><topic>Plant Sciences</topic><topic>Plants, Genetically Modified</topic><topic>Pollination</topic><topic>Promoter Regions, Genetic - genetics</topic><topic>Protein Binding</topic><topic>Protein Interaction Maps</topic><topic>Protein Transport</topic><topic>Protoplasts - metabolism</topic><topic>Real-Time Polymerase Chain Reaction</topic><topic>Subcellular Fractions - metabolism</topic><topic>Transfection</topic><topic>Zea mays</topic><topic>Zea mays - cytology</topic><topic>Zea mays - genetics</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Chen, Jiang</creatorcontrib><creatorcontrib>Yi, Qiang</creatorcontrib><creatorcontrib>Song, Qiaoheng</creatorcontrib><creatorcontrib>Gu, Yong</creatorcontrib><creatorcontrib>Zhang, Junjie</creatorcontrib><creatorcontrib>Hu, Yufeng</creatorcontrib><creatorcontrib>Liu, Hanmei</creatorcontrib><creatorcontrib>Liu, Yinghong</creatorcontrib><creatorcontrib>Yu, Guowu</creatorcontrib><creatorcontrib>Huang, Yubi</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Immunology Abstracts</collection><collection>Industrial and Applied Microbiology Abstracts (Microbiology A)</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Agricultural Science Collection</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Biology Database (Alumni Edition)</collection><collection>Medical Database (Alumni Edition)</collection><collection>ProQuest Pharma Collection</collection><collection>Technology Research Database</collection><collection>ProQuest SciTech Collection</collection><collection>ProQuest Natural Science Collection</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>Agricultural & Environmental Science Collection</collection><collection>ProQuest Central Essentials</collection><collection>Biological Science Collection</collection><collection>Proquest Central</collection><collection>Natural Science Collection</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Engineering Research Database</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Central Student</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>SciTech Premium Collection</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>ProQuest Biological Science Collection</collection><collection>Agricultural Science Database</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Medical Database</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biological Science Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Plant cell reports</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Chen, Jiang</au><au>Yi, Qiang</au><au>Song, Qiaoheng</au><au>Gu, Yong</au><au>Zhang, Junjie</au><au>Hu, Yufeng</au><au>Liu, Hanmei</au><au>Liu, Yinghong</au><au>Yu, Guowu</au><au>Huang, Yubi</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A highly efficient maize nucellus protoplast system for transient gene expression and studying programmed cell death-related processes</atitle><jtitle>Plant cell reports</jtitle><stitle>Plant Cell Rep</stitle><addtitle>Plant Cell Rep</addtitle><date>2015-07-01</date><risdate>2015</risdate><volume>34</volume><issue>7</issue><spage>1239</spage><epage>1251</epage><pages>1239-1251</pages><issn>0721-7714</issn><eissn>1432-203X</eissn><abstract>Key message
Conditions for the isolation and transfection of maize nucellus protoplasts were established. We demonstrated its utilization for protein expression, localization, protein–protein interaction, and the investigation of PCD-related processes.
Plant protoplasts are an important and versatile cell system that is widely used in the analysis of gene characterization and diverse signaling pathways. Programmed cell death (PCD) occurs throughout the life of plants from embryogenesis to fertilization. The maize nucellus undergoes typical PCD during development of the embryo sac. The nucellus protoplast shows potential for use in research of PCD-related processes. No studies have reported previously the isolation and transfection of nucellus protoplasts. In this study, conditions for the isolation and transfection of maize nucellus protoplasts were established. The maize protoplast system can be used for protein expression, localization, and protein–protein interaction. We applied this system to investigate PCD-related processes. Quantitative real-time PCR analysis revealed that transient expression of MADS29 in the maize nucellus protoplast increases Cys-protease gene transcript level. In addition, β-glucuronidase and luciferase activity assays showed that MADS29 could enhance the promoter activities of the Cys-protease gene. Thus, we demonstrated the potential of a highly efficient maize nucellus protoplast system for transient gene expression and investigation of PCD-related processes.</abstract><cop>Berlin/Heidelberg</cop><pub>Springer Berlin Heidelberg</pub><pmid>25786591</pmid><doi>10.1007/s00299-015-1783-z</doi><tpages>13</tpages></addata></record> |
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| subjects | Apoptosis Biomedical and Life Sciences Biotechnology Cell Biology Cell death Corn Embryonic growth stage Gene Expression Gene Expression Regulation, Plant Genes, Plant Life Sciences Original Paper Osmotic Pressure Plant Biochemistry Plant Proteins - metabolism Plant Sciences Plants, Genetically Modified Pollination Promoter Regions, Genetic - genetics Protein Binding Protein Interaction Maps Protein Transport Protoplasts - metabolism Real-Time Polymerase Chain Reaction Subcellular Fractions - metabolism Transfection Zea mays Zea mays - cytology Zea mays - genetics |
| title | A highly efficient maize nucellus protoplast system for transient gene expression and studying programmed cell death-related processes |
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