MdWRKY9 overexpression confers intensive dwarfing in the M26 rootstock of apple by directly inhibiting brassinosteroid synthetase MdDWF4 expression
Dwarfing rootstocks enable high-density planting and are therefore highly desirable in modern apple (Malus domestica) production. M26 is a semi-dwarfing rootstock that is used worldwide, but identifying intensive dwarfing rootstock is a major goal of apple breeding programs. Herein, we show that MdW...
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| Veröffentlicht in: | The New phytologist 2018-02, Vol.217 (3), p.1086-1098 |
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| container_title | The New phytologist |
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| creator | Zheng, Xiaodong Zhao, Yu Shan, Dongqian Shi, Kun Wang, Lin Li, Qingtian Wang, Na Zhou, Jingzhe Yao, Junzhu Xue, Yuan Fang, Shuang Chu, Jinfang Guo, Yan Kong, Jin |
| description | Dwarfing rootstocks enable high-density planting and are therefore highly desirable in modern apple (Malus domestica) production. M26 is a semi-dwarfing rootstock that is used worldwide, but identifying intensive dwarfing rootstock is a major goal of apple breeding programs.
Herein, we show that MdWRKY9 mediates dwarfing by directly inhibiting the transcription of the brassinosteroid (BR) rate-limiting synthetase MdDWF4 and reducing BR production.
We found that the transcriptional factor MdWRKY9 is highly expressed in all tested dwarfing rootstocks. Transgenic lines of M26 rootstock overexpressing MdWRKY9 exhibit further dwarfing, which resulted from the reduced BR levels and was reversed via exogenous brassinolide treatment. Both an in vivo chromatin immunoprecipitation (ChIP) analysis and an in vitro electrophoretic mobility shift assay (EMSA) indicated that MdWRKY9 binds to the promoter of MdDWF4. Furthermore, MdWRKY9 repressed MdDWF4 expression in stable transgenic apple plants as determined by quantitative PCR. In addition, RNA-interfered expression of MdWRKY9 in transiently transformed apple calli led to a significant increase of MdDWF4, suggesting MdWRKY9 plays a critical role in regulating the expression of MdDWF4.
We report a novel dwarfing mechanism in perennial woody plants that involves WRKY-controlled BR production, and present a new dwarfing M26 rootstock for potential applications in apple production. |
| doi_str_mv | 10.1111/nph.14891 |
| format | Article |
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Herein, we show that MdWRKY9 mediates dwarfing by directly inhibiting the transcription of the brassinosteroid (BR) rate-limiting synthetase MdDWF4 and reducing BR production.
We found that the transcriptional factor MdWRKY9 is highly expressed in all tested dwarfing rootstocks. Transgenic lines of M26 rootstock overexpressing MdWRKY9 exhibit further dwarfing, which resulted from the reduced BR levels and was reversed via exogenous brassinolide treatment. Both an in vivo chromatin immunoprecipitation (ChIP) analysis and an in vitro electrophoretic mobility shift assay (EMSA) indicated that MdWRKY9 binds to the promoter of MdDWF4. Furthermore, MdWRKY9 repressed MdDWF4 expression in stable transgenic apple plants as determined by quantitative PCR. In addition, RNA-interfered expression of MdWRKY9 in transiently transformed apple calli led to a significant increase of MdDWF4, suggesting MdWRKY9 plays a critical role in regulating the expression of MdDWF4.
We report a novel dwarfing mechanism in perennial woody plants that involves WRKY-controlled BR production, and present a new dwarfing M26 rootstock for potential applications in apple production.</description><identifier>ISSN: 0028-646X</identifier><identifier>EISSN: 1469-8137</identifier><identifier>DOI: 10.1111/nph.14891</identifier><identifier>PMID: 29165808</identifier><language>eng</language><publisher>England: New Phytologist Trust</publisher><subject>Brassinolide ; brassinosteroid ; Breeding ; Chromatin ; DNA ; dwarfing ; Electrophoretic mobility ; Gene expression ; Immunoprecipitation ; M26 ; Malus domestica ; MdDWF4 ; MdWRKY9 ; Nucleic acids ; Nucleotide sequence ; PCR ; Plant breeding ; Planting density ; Ribonucleic acid ; RNA ; Rootstocks ; Transcription ; Transgenic plants ; Woody plants</subject><ispartof>The New phytologist, 2018-02, Vol.217 (3), p.1086-1098</ispartof><rights>Copyright © 2018 New Phytologist Trust</rights><rights>2017 The Authors. New Phytologist © 2017 New Phytologist Trust</rights><rights>2017 The Authors. New Phytologist © 2017 New Phytologist Trust.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/90018539$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/90018539$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>314,776,780,799,1411,1427,27903,27904,45553,45554,46388,46812,57996,58229</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/29165808$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zheng, Xiaodong</creatorcontrib><creatorcontrib>Zhao, Yu</creatorcontrib><creatorcontrib>Shan, Dongqian</creatorcontrib><creatorcontrib>Shi, Kun</creatorcontrib><creatorcontrib>Wang, Lin</creatorcontrib><creatorcontrib>Li, Qingtian</creatorcontrib><creatorcontrib>Wang, Na</creatorcontrib><creatorcontrib>Zhou, Jingzhe</creatorcontrib><creatorcontrib>Yao, Junzhu</creatorcontrib><creatorcontrib>Xue, Yuan</creatorcontrib><creatorcontrib>Fang, Shuang</creatorcontrib><creatorcontrib>Chu, Jinfang</creatorcontrib><creatorcontrib>Guo, Yan</creatorcontrib><creatorcontrib>Kong, Jin</creatorcontrib><title>MdWRKY9 overexpression confers intensive dwarfing in the M26 rootstock of apple by directly inhibiting brassinosteroid synthetase MdDWF4 expression</title><title>The New phytologist</title><addtitle>New Phytol</addtitle><description>Dwarfing rootstocks enable high-density planting and are therefore highly desirable in modern apple (Malus domestica) production. M26 is a semi-dwarfing rootstock that is used worldwide, but identifying intensive dwarfing rootstock is a major goal of apple breeding programs.
Herein, we show that MdWRKY9 mediates dwarfing by directly inhibiting the transcription of the brassinosteroid (BR) rate-limiting synthetase MdDWF4 and reducing BR production.
We found that the transcriptional factor MdWRKY9 is highly expressed in all tested dwarfing rootstocks. Transgenic lines of M26 rootstock overexpressing MdWRKY9 exhibit further dwarfing, which resulted from the reduced BR levels and was reversed via exogenous brassinolide treatment. Both an in vivo chromatin immunoprecipitation (ChIP) analysis and an in vitro electrophoretic mobility shift assay (EMSA) indicated that MdWRKY9 binds to the promoter of MdDWF4. Furthermore, MdWRKY9 repressed MdDWF4 expression in stable transgenic apple plants as determined by quantitative PCR. In addition, RNA-interfered expression of MdWRKY9 in transiently transformed apple calli led to a significant increase of MdDWF4, suggesting MdWRKY9 plays a critical role in regulating the expression of MdDWF4.
We report a novel dwarfing mechanism in perennial woody plants that involves WRKY-controlled BR production, and present a new dwarfing M26 rootstock for potential applications in apple production.</description><subject>Brassinolide</subject><subject>brassinosteroid</subject><subject>Breeding</subject><subject>Chromatin</subject><subject>DNA</subject><subject>dwarfing</subject><subject>Electrophoretic mobility</subject><subject>Gene expression</subject><subject>Immunoprecipitation</subject><subject>M26</subject><subject>Malus domestica</subject><subject>MdDWF4</subject><subject>MdWRKY9</subject><subject>Nucleic acids</subject><subject>Nucleotide sequence</subject><subject>PCR</subject><subject>Plant breeding</subject><subject>Planting density</subject><subject>Ribonucleic acid</subject><subject>RNA</subject><subject>Rootstocks</subject><subject>Transcription</subject><subject>Transgenic plants</subject><subject>Woody plants</subject><issn>0028-646X</issn><issn>1469-8137</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2018</creationdate><recordtype>article</recordtype><recordid>eNpdkc1u1DAUhS0EotPCggcAWWLDJq3_4p8lammLaAEhUGEV2Y7DeMjYwfa05Dl4YTydUiTuxlf2d8617gHgGUaHuNZRmJaHmEmFH4AFZlw1ElPxECwQIrLhjH_dA_s5rxBCquXkMdgjCvNWIrkAvy_7q0_vvikYr11yv6bkcvYxQBvD4FKGPhQXsr92sL_RafDhe72CZengJeEwxVhyifYHjAPU0zQ6aGbY--RsGedKLr3xZSsySVfjEHNxKfoe5jlUk6JzNepPrk4Z_Df8CXg06DG7p3fnAfhy-ubz8Xlz8eHs7fHri2ZFKceN1VJKaoShhGHVW2GQdlbQ3mKDUGuYYINWhmJruaaCEkIGQpzppcBcakIPwKud75Tiz43LpVv7bN046uDiJndYccE4IVRW9OV_6CpuUqi_q5QUrGUSbw1f3FEbs3Z9NyW_1mnu_q67Akc74MaPbr5_x6jb5tjVHLvbHLv3H89vm6p4vlOs6p7TvUIhhGVLFf0DEJqbZQ</recordid><startdate>20180201</startdate><enddate>20180201</enddate><creator>Zheng, Xiaodong</creator><creator>Zhao, Yu</creator><creator>Shan, Dongqian</creator><creator>Shi, Kun</creator><creator>Wang, Lin</creator><creator>Li, Qingtian</creator><creator>Wang, Na</creator><creator>Zhou, Jingzhe</creator><creator>Yao, Junzhu</creator><creator>Xue, Yuan</creator><creator>Fang, Shuang</creator><creator>Chu, Jinfang</creator><creator>Guo, Yan</creator><creator>Kong, Jin</creator><general>New Phytologist Trust</general><general>Wiley Subscription Services, Inc</general><scope>NPM</scope><scope>7QO</scope><scope>7SN</scope><scope>8FD</scope><scope>C1K</scope><scope>F1W</scope><scope>FR3</scope><scope>H95</scope><scope>L.G</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>20180201</creationdate><title>MdWRKY9 overexpression confers intensive dwarfing in the M26 rootstock of apple by directly inhibiting brassinosteroid synthetase MdDWF4 expression</title><author>Zheng, Xiaodong ; Zhao, Yu ; Shan, Dongqian ; Shi, Kun ; Wang, Lin ; Li, Qingtian ; Wang, Na ; Zhou, Jingzhe ; Yao, Junzhu ; Xue, Yuan ; Fang, Shuang ; Chu, Jinfang ; Guo, Yan ; Kong, Jin</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-j3361-ca8883b7b32419dc7b0aec73dc1b005b474fa9b31cc6a373222f22ebd87168a23</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2018</creationdate><topic>Brassinolide</topic><topic>brassinosteroid</topic><topic>Breeding</topic><topic>Chromatin</topic><topic>DNA</topic><topic>dwarfing</topic><topic>Electrophoretic mobility</topic><topic>Gene expression</topic><topic>Immunoprecipitation</topic><topic>M26</topic><topic>Malus domestica</topic><topic>MdDWF4</topic><topic>MdWRKY9</topic><topic>Nucleic acids</topic><topic>Nucleotide sequence</topic><topic>PCR</topic><topic>Plant breeding</topic><topic>Planting density</topic><topic>Ribonucleic acid</topic><topic>RNA</topic><topic>Rootstocks</topic><topic>Transcription</topic><topic>Transgenic plants</topic><topic>Woody plants</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Zheng, Xiaodong</creatorcontrib><creatorcontrib>Zhao, Yu</creatorcontrib><creatorcontrib>Shan, Dongqian</creatorcontrib><creatorcontrib>Shi, Kun</creatorcontrib><creatorcontrib>Wang, Lin</creatorcontrib><creatorcontrib>Li, Qingtian</creatorcontrib><creatorcontrib>Wang, Na</creatorcontrib><creatorcontrib>Zhou, Jingzhe</creatorcontrib><creatorcontrib>Yao, Junzhu</creatorcontrib><creatorcontrib>Xue, Yuan</creatorcontrib><creatorcontrib>Fang, Shuang</creatorcontrib><creatorcontrib>Chu, Jinfang</creatorcontrib><creatorcontrib>Guo, Yan</creatorcontrib><creatorcontrib>Kong, Jin</creatorcontrib><collection>PubMed</collection><collection>Biotechnology Research Abstracts</collection><collection>Ecology Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ASFA: Aquatic Sciences and Fisheries Abstracts</collection><collection>Engineering Research Database</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) 1: Biological Sciences & Living Resources</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Professional</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>The New phytologist</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Zheng, Xiaodong</au><au>Zhao, Yu</au><au>Shan, Dongqian</au><au>Shi, Kun</au><au>Wang, Lin</au><au>Li, Qingtian</au><au>Wang, Na</au><au>Zhou, Jingzhe</au><au>Yao, Junzhu</au><au>Xue, Yuan</au><au>Fang, Shuang</au><au>Chu, Jinfang</au><au>Guo, Yan</au><au>Kong, Jin</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>MdWRKY9 overexpression confers intensive dwarfing in the M26 rootstock of apple by directly inhibiting brassinosteroid synthetase MdDWF4 expression</atitle><jtitle>The New phytologist</jtitle><addtitle>New Phytol</addtitle><date>2018-02-01</date><risdate>2018</risdate><volume>217</volume><issue>3</issue><spage>1086</spage><epage>1098</epage><pages>1086-1098</pages><issn>0028-646X</issn><eissn>1469-8137</eissn><abstract>Dwarfing rootstocks enable high-density planting and are therefore highly desirable in modern apple (Malus domestica) production. M26 is a semi-dwarfing rootstock that is used worldwide, but identifying intensive dwarfing rootstock is a major goal of apple breeding programs.
Herein, we show that MdWRKY9 mediates dwarfing by directly inhibiting the transcription of the brassinosteroid (BR) rate-limiting synthetase MdDWF4 and reducing BR production.
We found that the transcriptional factor MdWRKY9 is highly expressed in all tested dwarfing rootstocks. Transgenic lines of M26 rootstock overexpressing MdWRKY9 exhibit further dwarfing, which resulted from the reduced BR levels and was reversed via exogenous brassinolide treatment. Both an in vivo chromatin immunoprecipitation (ChIP) analysis and an in vitro electrophoretic mobility shift assay (EMSA) indicated that MdWRKY9 binds to the promoter of MdDWF4. Furthermore, MdWRKY9 repressed MdDWF4 expression in stable transgenic apple plants as determined by quantitative PCR. In addition, RNA-interfered expression of MdWRKY9 in transiently transformed apple calli led to a significant increase of MdDWF4, suggesting MdWRKY9 plays a critical role in regulating the expression of MdDWF4.
We report a novel dwarfing mechanism in perennial woody plants that involves WRKY-controlled BR production, and present a new dwarfing M26 rootstock for potential applications in apple production.</abstract><cop>England</cop><pub>New Phytologist Trust</pub><pmid>29165808</pmid><doi>10.1111/nph.14891</doi><tpages>13</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | The New phytologist, 2018-02, Vol.217 (3), p.1086-1098 |
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| source | Wiley Online Library Journals Frontfile Complete; Jstor Complete Legacy; Wiley Free Content; EZB-FREE-00999 freely available EZB journals |
| subjects | Brassinolide brassinosteroid Breeding Chromatin DNA dwarfing Electrophoretic mobility Gene expression Immunoprecipitation M26 Malus domestica MdDWF4 MdWRKY9 Nucleic acids Nucleotide sequence PCR Plant breeding Planting density Ribonucleic acid RNA Rootstocks Transcription Transgenic plants Woody plants |
| title | MdWRKY9 overexpression confers intensive dwarfing in the M26 rootstock of apple by directly inhibiting brassinosteroid synthetase MdDWF4 expression |
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