Mandal B, Csinos A S, Martinez-Ochoa N, Pappu H R
Department of Plant Pathology, University of Georgia, Coastal Plain Experiment Station, Tifton, GA 31793, USA.
J Virol Methods. 2008 Apr;149(1):195-8. doi: 10.1016/j.jviromet.2007.12.007. Epub 2008 Feb 13.
A rapid and efficient method of inoculation for Tomato spotted wilt tospovirus (TSWV) was achieved by applying the inoculum with a device consisting of a spray gun, an atomizer and a CO2-powered sprayer. The inoculum contained infected leaf sap prepared in 0.1M phosphate buffer, pH 7.0, 0.2% sodium sulfite and 0.01 M 2-mercaptoethanol (1g: 10 ml) and 1% each of Celite 545 and Carborundum 320 grit. The spray application of chilled inoculum at the rate of 1.1 ml/plant and at an air pressure of 4.1 bar resulted in systemic infection nearly to a 100% of the tobacco (Nicotiana tabacum) plants inoculated. The inoculation procedure was successfully applied to two other important host species of TSWV, peanut (Arachis hypogaea) and tomato (Lycopersicon esculentum), where 75.0-100% and 72.2-91.6% plants developed systemic infection, respectively. The approach facilitated a much faster inoculation of test plants with TSWV as it was estimated to be about 50 times quicker (depending on the plant species) than the hand inoculation. The procedure is suitable for rapid and simultaneous inoculation of a large number of test plants with TSWV and should facilitate screening of germplasm and breeding lines for virus resistance.
通过使用一种由喷枪、雾化器和二氧化碳驱动喷雾器组成的装置来施加接种物，实现了一种快速高效的番茄斑萎病毒（TSWV）接种方法。接种物包含在pH 7.0的0.1M磷酸盐缓冲液、0.2%亚硫酸钠和0.01M 2-巯基乙醇（1g:10 ml）中制备的感染叶汁，以及各1%的硅藻土545和320目金刚砂。以1.1 ml/株的速率和4.1巴的气压喷雾施用冷却的接种物，导致接种的烟草（Nicotiana tabacum）植株几乎100%发生系统感染。该接种程序成功应用于TSWV的另外两种重要寄主植物，花生（Arachis hypogaea）和番茄（Lycopersicon esculentum），分别有75.0 - 100%和72.2 - 91.6%的植株发生系统感染。该方法使TSWV对试验植物的接种速度大大加快，据估计比手工接种快约50倍（取决于植物种类）。该程序适用于用TSWV对大量试验植物进行快速同时接种，应有助于筛选种质和抗病育种系。
