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中国油料作物学报 ›› 2019, Vol. 41 ›› Issue (3): 409-.doi: 10.7505/j.issn.1007-9084.2019.03.013

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Molecular detection and diagnosis of clubroot caused by Plasmodiophora brassicae in oilseed rape

  

  1. 1.College of Plant Protection, Shenyang Agricultural University, Shenyang 110866, China;
     2. College of Horticulture, Shenyang Agricultural University, Shenyang 110866, China
  • Online:2019-06-28 Published:2019-09-02

Abstract:

Clubroot has recently become one of the main diseases of oilseed rape and cruciferous crops in China, which seriously affected the yield of cruciferous crops. Therefore, an efficient method with a high efficiency for molecular detection of P. brassicae was developed for accurate diagnosis in various samples using a pair of specific primers (PbITS1) designed on the basis of ITS (internal transcribed spacer) sequence, and the molecular detection of P. brassicae and disease assessment were carried out by PCR procedure. The results indicated that PCR system could specifically detect P. brassicae without any amplification product from representative soil-borne pathogenic fungi, bacteria, nematodes and endophytes. Sensitivity assay showed that the minimum concentration of template DNA was 1×10-3 ng/μL, the minimum number of spores in the contaminated soil was 1×103 spores/g soil, and the minimum level of spore with seeds was 1×105 spores/g seeds. Moreover, rape and other cruciferous plants (e.g., tissues and seeds) and soils could be applied in this detection system, meanwhile roots and surrounding soils collected from the different growth stages could be used for disease assessment. The benefit of this detection system was convenient, sensitive and specific with a wide range of detectable samples, which provided insights on detection of P. brassicae as well as the diagnosis, monitoring epidemic pattern, and integrated management of clubroot.

Key words: Plasmodiophora brassicae, Molecular detection, Clubroot