Ramos Javier, Bisseling Ton
Departamento de Nutrición Vegetal, Estación Experimental de Aula Dei, Consejo Superior de Investigaciones Científicas, Apdo 202, 50080 Zaragoza, Spain.
J Exp Bot. 2003 Oct;54(391):2245-50. doi: 10.1093/jxb/erg245. Epub 2003 Aug 8.
A new method for the isolation of root hairs from the model legume, Medicago truncatula, was developed. The procedure involves the propagation of detached roots on agar plates and the collection of root hairs by immersion in liquid nitrogen. Yields of up to 40 micro g of root hair protein were obtained from 50-100 root tips grown for 3 weeks on a single plate. The high purity of the root hair fraction was monitored by western blot analysis using an antibody to the pea epidermis specific protein PsRH2. Sequence analyses revealed that the protein homologous to PsRH2 in M. truncatula, MtRH2, is identical to the root protein MtPR10-1. The MtRH2 protein proved to be a useful endogenous marker to monitor root hair isolation since it is also specifically expressed in the root epidermis.
开发了一种从模式豆科植物蒺藜苜蓿中分离根毛的新方法。该程序包括在琼脂平板上培养离体根，并通过浸入液氮中来收集根毛。从在单个平板上生长3周的50 - 100个根尖中可获得高达40微克的根毛蛋白产量。使用针对豌豆表皮特异性蛋白PsRH2的抗体，通过蛋白质免疫印迹分析监测根毛部分的高纯度。序列分析表明，蒺藜苜蓿中与PsRH2同源的蛋白MtRH2与根蛋白MtPR10 - 1相同。由于MtRH2蛋白也在根表皮中特异性表达，因此它被证明是监测根毛分离的有用内源性标记物。
