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Detection and quantification of airborne inoculum of Sclerotinia sclerotiorum using quantitative PCR

机译：定量PCR检测和定量核盘菌菌空中接种物

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摘要

This paper reports the development of a new specific diagnostic technique to accurately quantify airborne inoculum of Sclerotinia sclerotiorum and discusses its potential use in disease-forecasting schemes, using examples of three contrasting epidemic seasons: 2007, when there was a severe epidemic of sclerotinia stem rot (SSR) in England and high numbers of airborne ascospores were trapped at Rothamsted, and, in contrast, 2003 and 2004, when the incidence of SSR in England was low and low numbers of airborne ascospores were trapped at Rothamsted. DNA was extracted from wax-coated plastic tapes, such as those used in Burkard (Hirst-type) spore traps and rotating-arm traps. A SYBR-green quantitative PCR (qPCR) method produced a linear relationship between ascospore numbers and S. sclerotiorum DNA (mean 0.35 pg DNA per spore) and was able to detect DNA representing as few as two ascospores. The technique was insensitive to DNA of the host plant, Brassica napus, and other plant pathogens, including Sclerotinia minor, S. trifoliorwn and Botrytis cinerea, and common airborne fungal genera such as Cladosporium and Fenicillium. There was no relationship between rainfall and numbers of airborne ascospores of S. sclerotiorum present at Rothamsted during the period of infection in the severe SSR season (2007).
机译：本文报道了一种新的特定诊断技术的发展，该技术可准确定量菌核硬化菌的空气传播接种物，并使用三个不同的流行季节的实例讨论了其在疾病预测方案中的潜在用途：2007年，当菌核菌茎腐烂严重流行时（SSR）在英国和Rothamsted捕获了大量的空中子孢子，相比之下，2003年和2004年，英格兰的SSR发病率较低，而在Rothamsted捕获了少量的空中子孢子。从涂蜡的塑料带（如Burkard（Hirst型）孢子阱和旋转臂阱中使用的胶带）提取DNA。 SYBR-green定量PCR（qPCR）方法在子囊孢子数与菌核链球菌DNA（平均每个子孢子0.35 pg DNA）之间产生线性关系，并且能够检测到代表两个子囊孢子的DNA。该技术对寄主植物，甘蓝型油菜和其他植物病原体（包括小核盘菌，三叶草和灰霉菌）以及常见的空气传播真菌属（如枝形孢子和镰孢属）的DNA不敏感。在严重的SSR季节（2007年）的感染期间，降雨与Rothamsted的硬核链球菌气生子囊孢子数量之间没有关系。

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