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Microperforated leaf blotting on polyvinylidene difluoride and nylon membranes to analyze spatial distribution of endogenous and viral gene expression in plant leaves

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Please use this identifier to cite or link to this item:http://hdl.handle.net/2115/67241

Title: Microperforated leaf blotting on polyvinylidene difluoride and nylon membranes to analyze spatial distribution of endogenous and viral gene expression in plant leaves
Authors: Murakami, Taiki Browse this author
Tayama, Ryou Browse this author
Nakahara, Kenji S Browse this author →KAKEN DB
Keywords: leaf protein distribution
leaf RNA distribution
leaf microperforation
Issue Date: Sep-2016
Publisher: Springer Japan
Journal Title: Journal of General Plant Pathology
Volume: 82
Issue: 5
Start Page: 254
End Page: 260
Publisher DOI: 10.1007/s10327-016-0671-x
Abstract: Leaf blotting to detect proteins and investigate their spatial distribution in leaves has so far mainly been used to detect viral coat proteins that accumulate abundantly in infected leaves, but rarely to detect endogenous plant proteins. We improved the method for detecting endogenous proteins. We found that microperforating leaves with bundled pins before blotting, then pressing leaves with a rolling pin onto polyvinylidene difluoride (PVDF) membranes enabled even blotting of sap. This microperforated leaf blotting (mPLB) was also suitable for use with nylon membranes to detect leaf RNA. The mPLB revealed that accumulation of two endogenous proteins, calmodulin-like rgs-CaM and actin, was respectively positively and negatively associated with that of viral coat protein in tobacco leaves infected with cucumber mosaic virus (CMV). When a tobacco plant primed with benzothiadiazole was inoculated with CMV, mPLB showed that the infection was restricted to some areas of the leaf, and that in these areas the mRNA encoding tobacco pathogenesis-related protein 1, an indicator of salicylic acid-mediated immune responses, was induced. These results demonstrate the effectiveness of mPLB for investigating the spatial distribution of endogenous and viral gene expression in leaves.
Rights: The final publication is available at link.springer.com
Type: article (author version)
URI: http://hdl.handle.net/2115/67241
Appears in Collections:農学院・農学研究院 (Graduate School of Agriculture / Faculty of Agriculture) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 中原 健二

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