Agroinjection of tomato fruits. A tool for rapid functional analysis of transgenes directly in fruit
Agroinjection of tomato fruits. A tool for rapid functional analysis of transgenes directly in fruit
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DOI：
10.1104/pp.105.068221
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发表时间：
2006-01-01
期刊：
影响因子：
7.4
通讯作者：
Granell, A
中科院分区：
文献类型：
--
作者：
Orzaez, D;Mirabel, S;Granell, A
外源基因在植物组织中的瞬时表达是植物生物技术的一个有价值的工具。为了缩短水果基因功能分析的时间，我们开发了一种可应用于番茄（Solanum lycopersicum cv Micro Tom）水果的瞬时方法。结果发现，通过果实花柱顶端注射农杆菌培养物导致果实完全浸润。这种渗透方法被称为水果农杆菌注射，使 35S 花椰菜花叶病毒驱动的 β-葡萄糖醛酸酶和黄色荧光蛋白在水果中瞬时表达高水平，在胎盘周围表达水平较高，在果皮中表达水平中等。水果农杆菌注射液的有效性在三个案例研究中进行了分析：(1) 拟南芥 (Arabidopsis thaliana) 启动子的热休克调节，(2) 重组 IgA 抗体的生产作为分子农业的一个例子，以及 (3) 病毒诱导的胡萝卜素生物合成途径基因沉默。在所有三个实例中，该技术都被证明是在水果中快速表达转基因的有效工具。
Transient expression of foreign genes in plant tissues is a valuable tool for plant biotechnology. To shorten the time for gene functional analysis in fruits, we developed a transient methodology that could be applied to tomato (Solanum lycopersicum cv Micro Tom) fruits. It was found that injection of Agrobacterium cultures through the fruit stylar apex resulted in complete fruit infiltration. This infiltration method, named fruit agroinjection, rendered high levels of 35S Cauliflower mosaic virus-driven beta-glucuronidase and yellow fluorescence protein transient expression in the fruit, with higher expression levels around the placenta and moderate levels in the pericarp. Usefulness of fruit agroinjection was assayed in three case studies: (1) the heat shock regulation of an Arabidopsis (Arabidopsis thaliana) promoter, (2) the production of recombinant IgA antibodies as an example of molecular farming, and (3) the virus-induced gene silencing of the carotene biosynthesis pathway. In all three instances, this technology was shown to be efficient as a tool for fast transgene expression in fruits.