1、A highly efficient rice green tissue A highly efficient rice green tissue protoplast system for transient protoplast system for transient gene expression and studying gene expression and studying light/chloroplast-related processeslight/chloroplast-related processes水稻原生質(zhì)體分離與GFP，BiFC等方法研究意義研究意義植物原生質(zhì)體

2、是常見的與生理相關(guān)的細(xì)胞組織，可用于分析基因的功能特性，有重要意義。在水稻研究中，有關(guān)懸浮培養(yǎng)細(xì)胞和黃化苗原生質(zhì)體研究較多，但綠色組織原生質(zhì)體研究較少。綠色植物原生質(zhì)體成功用于植物與激素、代謝物及環(huán)境因素相關(guān)的信號轉(zhuǎn)導(dǎo)的研究。簡化并提高了基因短暫表達(dá)的系統(tǒng)，并結(jié)合蛋白質(zhì)免疫印染、點(diǎn)位及蛋白質(zhì)-蛋白質(zhì)相關(guān)試驗。主要內(nèi)容主要內(nèi)容 Here, we report a simplified method for isolating protoplasts from normally cultivated young rice green tissue without the need for unne

3、cessary chemicals and a vacuum device. 主要內(nèi)容主要內(nèi)容Transfections of the generated protoplasts with plasmids of a wide range of sizes (4.5-13 kb) and co-transfections with multiple plasmids achieved impressively high efficiencies and allowed evaluations by 1) protein immunoblotting analysis, 2) subcellul

4、ar localization assays, 3) protein-protein interaction analysis.圖片圖片 GFP Chl Merged BrightGFP Chl Merged Bright問題7頁：The upper panel contains low power imagesshowing high co-expression efficiency peroxisomes過氧化物酶體研究結(jié)果研究結(jié)果 Importantly, the rice green tissue protoplasts were photosynthetically active and sensitive to the retrograde plastid signaling inducer norflurazon (NF). Transient expression of the GFP-tagged light-related transcription factor OsGLK1 markedly upregulated transcript levels of the endogeneous photosynthetic genes OsLhcb1, OsLhcp, GADPH and RbcS, which were