标题:Evaluation of parameters affecting Agrobacterium-mediated transient expression in citrus

作者:Li, Fang Dai, Su-ming Deng, Zi-niu Li, Da-zhi Long, Gui-you Li, Na Li, Yi Gentile, Alexandra

作者单位:National Center for Citrus Improvement Changsha/Horticulture and Landscape College, Hunan Agricultural University, Changsha 410128, P.R.China

摘要:Agrobacterium-mediated transient expression assays are a convenient alternative to stable expression because they are simple, easy to perform, and achieve gene expression rapidly. This study investigated the factors affecting transient gene expression efficiency in citrus by observing the cryo-sectioning of leaf samples under a laser confocal microscope. These factors included the composition of the infiltration buffer, the Agrobacterium cell density, the leaf development stage, the incubation temperature, and plant genotype. The highest transient expression level of yellow fluorescent protein (YFP) was detected in Mexican lime (Citrus aurantifolia) on the third day after the intermediate-aged leaves were infiltrated with the improved infiltration buffer 1 (15 mmol L-1 2-(N-morpholino) ethanesulfonic acid, 10 mmol L-1 MgCl2, and 200 pmol L-1 acetosyringone), which had an optical density of 0.8 and was incubated at 22 degrees C. Additionally, this transient expression assay was applied to other citrus genotypes.\' Of note, trifoliate orange (Poncirus trifoliate) and kumquat (Fortunella obovate) had higher expression efficiency than other six genotypes of the Citrus genus. Our study provides research basis for the selection of optimization strategies in transient gene expression and improves the method for available genome investigation in citrus.

关键词:citrus, Agrobacterium, transient expression efficiency, yellow fluorescent prote

文献注录:Li, Fang Dai, Su-ming Deng, Zi-niu Li, Da-zhi Long, Gui-you Li, Na Li, Yi Gentile, Alexandra. Evaluation of parameters affecting Agrobacterium-mediated transient expression in citrus [J]. Journal of Integrative Agriculture. 2016, 3: 572-579.

报/刊名:Journal of Integrative Agriculture》,发表于2016 / 第 3 期。

文献类型: [J] (文献级别:SCI刊物

页码: 572-579 页 / 共8

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