An efficient protocol for perennial ryegrass mesophyll protoplast isolation and transformation, and its application on interaction study between LpNOL and LpNYC1

dc.contributor.authorYu, Guohuien
dc.contributor.authorCheng, Qiangen
dc.contributor.authorXie, Zhenien
dc.contributor.authorXu, Binen
dc.contributor.authorHuang, Bingruen
dc.contributor.authorZhao, Bingyu Y.en
dc.contributor.departmentSchool of Plant and Environmental Sciencesen
dc.date.accessioned2017-08-03T20:11:17Zen
dc.date.available2017-08-03T20:11:17Zen
dc.date.issued2017-06-05en
dc.date.updated2017-08-03T10:59:49Zen
dc.description.abstractBackground Perennial ryegrass (Lolium perenne L.) is an important temperate grass used for turf and forage purposes. With the increasing accumulation of genomic and transcriptomic data of perennial ryegrass, an efficient protoplast and transient gene expression protocol is highly desirable for in vivo gene functional studies in its homologous system. Results In this report, a highly efficient protoplast isolation (5.6 × 107 protoplasts per gram of leaf material) and transient expression (plasmid transformation efficiency at 55.2%) was developed and the detailed protocol presented. Using this protocol, the subcellular locations of two ryegrass proteins were visualized in chloroplasts and nuclei, respectively, and protein–protein interaction between two chlorophyll catabolic enzymes (LpNOL and LpNYC1) was recorded in its homologous system for the first time. Conclusions This efficient protoplast isolation and transformation protocol is sufficient for studies on protein subcellular localization and protein–protein interaction, and shall be suitable for many other molecular biology applications where the mesophyll protoplast system is desirable in perennial ryegrass.en
dc.description.versionPublished versionen
dc.format.mimetypeapplication/pdfen
dc.identifier.citationPlant Methods. 2017 Jun 05;13(1):46en
dc.identifier.doihttps://doi.org/10.1186/s13007-017-0196-0en
dc.identifier.urihttp://hdl.handle.net/10919/78651en
dc.language.isoenen
dc.rightsCreative Commons Attribution 4.0 Internationalen
dc.rights.holderThe Author(s)en
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/en
dc.titleAn efficient protocol for perennial ryegrass mesophyll protoplast isolation and transformation, and its application on interaction study between LpNOL and LpNYC1en
dc.title.serialPlant Methodsen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten
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