A rapid method of DNA detection of the beet cyst nematode, Heterodera schachtii, in infested field soil

Abstract

Infestation of vegetable fields by the beet cyst nematode, Heterodera schachtii, was first reported in Japan in 2017. After pesticide treatments, fields should be monitored for the possible re-establishment of the nematode population. This paper describes a rapid method of DNA detection by direct extraction of DNA from soil using saturated phosphate buffer, followed by real-time PCR using H. schachtii-specific primers. Accurate handling of many soil samples collected from large fields is possible; however, the method has a limited success rate of one second-stage juvenile (J2) or ten cysts per 10 g of soil, which was determined by adding J2s or cysts in vitro. Using this method on 141 soil samples collected from an infested field, a clear correlation between DNA level and the number of hatched J2s was found. This method enables rapid detection and semi-quantitative measurement of cyst nematodes in the soil.