A simple Agrobacterium tumefaciens-mediated transformation method for rapid transgene expression in Medicago truncatula root hairs
A simple Agrobacterium tumefaciens-mediated transformation method for rapid transgene expression in Medicago truncatula root hairs
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DOI：
10.1007/s11240-017-1323-3
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发表时间：
2017
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影响因子：
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通讯作者：
Céline Remblière;J. Fournier;Fernanda de Carvalho-Niebel;M. Chabaud
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作者：
Céline Remblière;J. Fournier;Fernanda de Carvalho-Niebel;M. Chabaud
蒺藜苜蓿是研究共生微生物和病原微生物相互作用的模式植物。虽然农杆菌介导的转化方法已经有很多种。truncatula，一个快速的根转化系统还没有用于这种模式植物。在这里，我们描述了一个简单的方法，快速转基因表达的根毛ofM。truncatula，使用与解除武装的hyperplasticA.根癌农杆菌菌株AGL 1。这种方法导致了各种GUS和荧光报告基因在M中的有效表达。蒺藜根的毛我们表明，共培养后2天，在位于根尖后0.5-2厘米的特定反应区的根毛中就检测到转基因表达。该方法可用于各种M。Truncatulagenotypes，特别适用于荧光融合蛋白亚细胞定位的快速研究。此外，在最初的共培养步骤中结合不同的农杆菌菌株有效地产生共转化的根毛，适合于在同一细胞中共定位不同的荧光融合蛋白。
Medicago truncatulais widely used as a model legume for symbiotic and pathogenic microbial interaction studies. Although a number ofAgrobacterium-mediated transformation methods have been developed forM. truncatula, a rapid root transformation system was not yet available for this model plant. Here, we describe an easy method for rapid transgene expression in root hairs ofM. truncatula, using young seedlings co-cultivated with the disarmed hypervirulentA. tumefaciensstrain AGL1. This method leads to efficient expression of various GUS and fluorescent reporters inM. truncatularoot hairs. We showed that transgene expression is detected as soon as 2 days following co-culture, in root hairs of a particular responsive zone lying 0.5–2 cm behind the root tip. This method can be used with a variety ofM. truncatulagenotypes, and is particularly useful for rapid investigation of the sub-cellular localization of fluorescent fusion proteins. Moreover, combining distinctAgrobacteriumstrains during the initial co-culture step efficiently generates co-transformed root hairs, suitable for co-localization of different fluorescent fusion proteins in the same cell.