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由茶麋子葡萄座腔菌Botryosphaeria dothidea引起的山核桃Carya cathayensis干腐病是山核桃栽培过程中的主要病害,该病菌表现明显的“潜伏侵染”特性。研究山核桃树体中干腐病菌的定量检测技术对山核桃干腐病的预测预报及科学防治具有重要的指导意义。根据茶麋子葡萄座腔菌的EF1α基因设计特异性引物,通过普通聚合酶链式反应(PCR)和实时荧光定量PCR(real-time PCR)扩增发现引物EFRT-F1/R1对山核桃干腐病病菌的特异性及扩增效率较高,可稳定扩增出230 bp的目标条带。应用此特异性引物建立的实时荧光定量PCR干腐病病菌检测方法能够定量检测出山核桃植株样品中的病菌含量,灵敏度要比普通PCR高100倍。
The dry rot of Carya cathayensis, a disease caused by the tea Botryosphaeria dothidea, is a major disease in the cultivation of pecans and shows significant “latent infection” characteristics. It is of great significance to study the quantitative detection technology of dry rot bacteria in the pecan tree for forecasting and scientific prevention and cure of dry rot disease in the pecan tree. According to the EF1α gene of Bacillus subtilis, specific primer was designed and EFRT-F1 / R1 was detected by ordinary polymerase chain reaction (PCR) and real-time PCR (Real-time PCR) The specificity of the rot pathogen and its high amplification efficiency can stably amplify the target band of 230 bp. Using this specific primer to establish a real-time fluorescence quantitative PCR dry rot bacteria detection method can quantitatively detect the bacteria in the pecan plant samples, the sensitivity of 100 times higher than the normal PCR.