摘要
近年来芸薹根肿菌引起的根肿病已成为我国油菜的主要病害,严重影响油菜及其它十字花科作物的产量及品质。为建立一套在土壤、植物组织和种子中精准检测根肿菌的高效分子方法,利用根肿菌ITS(internal transcribed spacer)序列,设计一对特异性引物(Pb ITS1),通过PCR反应优化体系进行根肿菌的分子检测及病害评估。结果表明,该引物及方法能够特异性检测根肿菌DNA,不受土传病原真菌、细菌、线虫及寄主植物内生菌的DNA干扰。灵敏性检测表明,模板DNA最低浓度达1pg·μL^-1,土壤带菌量最低为1×10^3个孢子/g土,种子带菌量最低1×10^5个孢子/g种子。此外,该体系能够用于检测油菜及其它十字花科寄主植物(如组织、种子)和土壤类型。同时,可用于田间油菜不同生长阶段油菜根部及周围土壤的根肿菌检测。本研究建立的检测体系操作简便、适用范围广、灵敏度高、特异性强,可检测样品种类丰富,可为油菜及其它十字花科根肿病的早期诊断、流行监测与预警、综合防控等提供科学依据。
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 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 1×10^3 spores/g soil,and the minimum level of spore with seeds was 1 × 10^5 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.
作者
关格格
邢曼竹
庞文星
夏子豪
杨新宇
朴钟云
吴元华
梁月
GUAN Ge-ge;XING Man-zhu;PANG Wen-xing;XIA Zi-hao;YANG Xin-yu;PIAO Zhong-yun;WU Yuan-hua;LIANG Yue(College of Plant Protection, Shenyang Agricultural University, Shenyang 110866, China;College of Horticulture, Shenyang Agricultural University, Shenyang 110866, China)
出处
《中国油料作物学报》
CAS
CSCD
北大核心
2019年第3期409-414,共6页
Chinese Journal of Oil Crop Sciences
基金
国家油菜产业技术体系(CARS-12)
辽宁省“兴辽英才计划”项目(XLYC1807242)
沈阳农业大学引进人才科研启动费项目(20153040)
关键词
芸薹根肿菌
分子检测
根肿病
Plasmodiophora brassicae
molecular detection
clubroot
