Authors
Yiming Wang, Jingni Wu, Sang Gon Kim, Sun Tae Kim, and Kyu Young Kang
Summary
We developed a protocol for in planta gene expression analyses in sliced rice sheath cells by modifying and optimizing biolistic particle bombardment techniques. By obtaining thin sections (400 um) of rice sheath cells, auto-fluorescence tissues from chlorophyll were eliminated. This system was validated through the localization of marker genes specifically expressed in nuclei, plasma membranes, and vacuolar membranes. Additionally, high transformation efficiencies of approximately 30% were achieved. Therefore, this protocol provides a new and rapid method for transient gene expression assays in rice. Protein secretion was examined in rice sheath cells using predicted secretory proteins from rice blast fungus, indicating that this method is practical for host plant-microbe interaction studies. The transient expression protocol established here is well suited for protein localization, secretion, and host-pathogen protein interaction studies in rice. A typical experiment can be completed in 3 days.Further details
The protocol was published on Protocol Exchange on 28 June 2013. To see the entire protocol, click on the source link.Advertisement
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