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Hokkaido University Collection of Scholarly and Academic Papers >
Graduate School of Agriculture / Faculty of Agriculture >
Peer-reviewed Journal Articles, etc >
Microperforated leaf blotting on polyvinylidene difluoride and nylon membranes to analyze spatial distribution of endogenous and viral gene expression in plant leaves
| 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)
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Submitter: 中原 健二
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