Ge B B, Cheng Y, Liu Y, Liu B H, Zhang K C
State Key Laboratory of Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Haidian District, Beijing, China.
Lett Appl Microbiol. 2015 Dec;61(6):596-602. doi: 10.1111/lam.12500. Epub 2015 Nov 13.
We developed a real-time PCR assay to specifically detect and quantify the efficacy of a biological fungicide from Streptomyces ahygroscopicus var. wuyiensis on tomato leaves. This fungicide, the natural secondary metabolite wuyiencin, is an antifungal agent against Botrytis cinerea. Specific primers were designed based on the β-actin gene sequences, which were used to detect a 303 bp fragment from B. cinerea isolates. Our assay is highly sensitive and can be used to reliably detect and quantify as little as 1·75 pg of B. cinerea DNA. We used this detection method to monitor the progression of B. cinerea infection in inoculated plant material under preventive (wuyiencin) and nonpreventive treatment. After 5 days, plants under preventive treatment exhibited a sharp decrease in fungal biomass and no symptoms, whereas plants under nonpreventive treatment displayed severe disease symptoms. The results demonstrate that wuyiencin has significant effects on B. cinerea in tomato plants and that real-time PCR is a reliable method for evaluating the effects of Streptomyces wuyiensis CK-15 on B. cinerea.
Botrytis cinerea commonly produces latent or nonsymptomatic infection on and within plant tissues, which can develop into symptomatic infection when triggered by changes in environmental conditions or host plant physiology. In this study, we develop a specific, sensitive real-time PCR assay for detecting and quantifying B. cinerea on tomato leaves to determine the control efficacy of Streptomyces ahygroscopicus var. wuyiensis as a biological fungicide. Our findings demonstrate that wuyiencin has significant effects on B. cinerea in tomato plants and that real-time PCR is a reliable method for evaluating the effects of biological fungicides on plant pathogens.
我们开发了一种实时荧光定量PCR检测方法，用于特异性检测和量化来自武夷菌素吸水链霉菌变种对番茄叶片的生物杀菌剂的功效。这种杀菌剂，即天然次生代谢产物武夷菌素，是一种抗灰葡萄孢菌的抗真菌剂。基于β-肌动蛋白基因序列设计了特异性引物，用于检测灰葡萄孢菌分离株的303 bp片段。我们的检测方法高度灵敏，可用于可靠地检测和定量低至1.75 pg的灰葡萄孢菌DNA。我们使用这种检测方法监测在预防性（武夷菌素）和非预防性处理下接种植物材料中灰葡萄孢菌感染的进展。5天后，预防性处理的植物真菌生物量急剧下降且无病症，而非预防性处理的植物则表现出严重的病害症状。结果表明，武夷菌素对番茄植株中的灰葡萄孢菌有显著影响，实时荧光定量PCR是评估武夷菌素吸水链霉菌CK-15对灰葡萄孢菌作用效果的可靠方法。
灰葡萄孢菌通常在植物组织上和组织内产生潜伏性或无症状感染，当环境条件或寄主植物生理发生变化触发时，可发展为有症状感染。在本研究中，我们开发了一种特异性、灵敏的实时荧光定量PCR检测方法，用于检测和量化番茄叶片上的灰葡萄孢菌，以确定武夷菌素吸水链霉菌变种作为生物杀菌剂的防治效果。我们的研究结果表明，武夷菌素对番茄植株中的灰葡萄孢菌有显著影响，实时荧光定量PCR是评估生物杀菌剂对植物病原菌作用效果的可靠方法。
