摘要
猕猴桃溃疡病是严重危害猕猴桃产业发展的重要细菌性病害,由于致病菌潜伏期不表现病症,常错过最佳的防治时期。为实现猕猴桃溃疡病田间无症带菌状态的早期诊断,本研究对田间快速检测技术进行了优化,并采集田间无症样本进行验证。结果表明,猕猴桃溃疡病病健交界处组织用金氏B培养液浸泡8 h可获得质量较高的PCR模板;在PCR反应体系中加入φ=10%的甘油能显著增强扩增效果;当检测无症带菌样本时,浸提液中加入Na2SO3,26℃浸泡8 h有利于降低PCR反应中抑制物的干扰;进一步优化PCR反应中模板DNA的添加量;无症样本检测结果表明,在猕猴桃孕蕾至开花期的新生枝条中溃疡病致病菌的检出率最高,可达91.8%。本试验优化后的PCR特异性分子检测技术,有助于及早发现无症带菌的猕猴桃溃疡病病株,对尽早有针对性实施田间防控具有重要意义。
Kiwifruit canker disease is an important bacterial disease that seriously endangers the development of the kiwifruit industry.Because pathogenic bacteria is asymptomatic during the incubation period,so the best prevention and treatment period are always missed.In order to early diagnose the asymptomatic bacteria in kiwifruit canker disease,the optimized rapid detection technology was used to detect the asymptomatic samples in the field.The results showed that margins between symptomatic and healthy tissues immersed in King’s B culture solution for 8 h were better for obtaining higher quality PCR template.Addition of 10%(V∶V)glycerol as a chemical enhancer in PCR reaction system could significantly enhance the amplification effect.During the detection of the asymptomatic infecting bacteria sample,Na2SO3 was added into the extraction template system and soaked for 8 h at 26°C,which was beneficial to reduce the interference of inhibitors in the PCR amplification reaction.The DNA template amount in the PCR reaction system was also optimized in this pape,and its results showed that the detection rate was the highest and could reached 91.8%when the new shoots of kiwifruit from the bud to the flowering stage were detected.The optimized PCR-specific molecular detection technology in this experiment is helpful for early detection of asymptomatic kiwifruit canker strains,which is of great significance for early field control.
作者
付博
王家哲
任平
张锋
李英梅
FU Bo;WANG Jiazhe;REN Ping;ZHANG Feng;LI Yingmei(Bio-Agriculture Institute of Shaanxi,Pest Control and Prevention Research Center,Xi’an 710043,China;Enzyme Engineering Research Center of Shaanxi Province,Xi’an 710600,China)
出处
《西北农业学报》
CAS
CSCD
北大核心
2020年第9期1440-1445,共6页
Acta Agriculturae Boreali-occidentalis Sinica
基金
陕西省科学院科技计划项目(2013k-16,2019k-05)
西安市科技局项目(Y18N010G18)
陕西省科技厅重点研发计划项目(2018ZDXM-NY-067)。
关键词
猕猴桃溃疡病
无症带菌
田间快速检测
Kiwi bacterial canker disease
Asymptomatic infecting bacteria
Rapid detection in the field
