烟草苗床期普通花叶病(TMV)组织印迹检测技术研究

Study on Tissue Blotting Detection Technology of Tobacco Mosaic Virus (TMV) in Tobacco Seedbed Period

摘要: 本文探讨了烟草苗床期TMV组织印迹检测技术，包括第一抗血清最适浓度的确定、第一抗血清和第一抗血清最佳温度及时间的优化、烟苗最佳取样部位的选择、感染烟草普通花叶病毒后的发病时间、不同品种烟草花叶病毒发病情况检测等。结果表明：第一抗血清最适浓度以1000倍效果较理想；用第一抗血清稀释1000倍，第二抗血清稀释6000倍，样品在37℃恒温振荡器60 r/min封闭状态下，与第一抗血清连接1 h，37℃恒温震荡，与第二抗血清连接1 h，37℃恒温震荡，效果最好；烟苗最佳取样部位的选择为叶柄中部；接种后第1天烟苗茎的各部位都有病毒积累，茎尖病毒积累出现较早，病毒接种第6天烟苗各个部位都检测到病毒的积累；试验了KRK26、K326、云烟87、红大、云烟97等5个品种，检测的5个样品均被TMV侵染，云烟97感染TMV的感病率最高，K326感病率最低。

Abstract: Tobacco mosaic virus (TMV) tissue blotting detection technology in the seedling period of tobacco was discussed, including the determination of the most suitable concentration of the first antiserum, optimization of optimal temperature and time of first antiserum and second antiserum, selection of the best sampling parts of tobacco seedling, time of onset after infection with tobacco Mosaic virus and detection of the incidence of tobacco Mosaic virus of different varieties. The results show that the optimum concentration of the first antiserum was 1000 times. The first antiserum was diluted 1000 times, the second antiserum was diluted 6000 times. Samples in 37℃ thermostat oscillator 60 r/min under the closed state, connected to the first antiserum 1 h, 37℃ constant temperature oscillation, connected with the second antiserum 1 h, 37℃ constant temperature oscillation, have the best effect. The best sampling site of tobacco seedling is the middle of petiole. On the first day after inoculation, virus accumulation occurred in all parts of the stem of tobacco seedling, stem-tip virus accumulation occurred earlier. Accumulation of virus was detected in all parts of the tobacco seedling on the 6th day after inoculation. Five varieties including KRK26, K326, Yunyan 87, Hongda and Yunyan 97 were tested. All 5 samples tested were infected by TMV. The incidence rate of TMV infection was highest in Yunyan 97 and lowest in K326.