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Specific and Sensitive Primers Developed by Comparative Genomics to Detect Bacterial Pathogens in Grains.
The Plant Pathology Journal ( IF 2.3 ) Pub Date : 2018-04-01 , DOI: 10.5423/ppj.oa.11.2017.0250 Kwang Yeol Baek 1 , Hyun-Hee Lee 2 , Geun Ju Son 1 , Pyeong An Lee 1 , Nazish Roy 1 , Young-Su Seo 2 , Seon-Woo Lee 1
The Plant Pathology Journal ( IF 2.3 ) Pub Date : 2018-04-01 , DOI: 10.5423/ppj.oa.11.2017.0250 Kwang Yeol Baek 1 , Hyun-Hee Lee 2 , Geun Ju Son 1 , Pyeong An Lee 1 , Nazish Roy 1 , Young-Su Seo 2 , Seon-Woo Lee 1
Affiliation
Accurate and rapid detection of bacterial plant pathogen is the first step toward disease management and prevention of pathogen spread. Bacterial plant pathogens Clavibacter michiganensis subsp. nebraskensis (Cmn), Pantoea stewartii subsp. stewartii (Pss), and Rathayibacter tritici (Rt) cause Goss's bacterial wilt and blight of maize, Stewart's wilt of maize and spike blight of wheat and barley, respectively. The bacterial diseases are not globally distributed and not present in Korea. This study adopted comparative genomics approach and aimed to develop specific primer pairs to detect these three bacterial pathogens. Genome comparison among target pathogens and their closely related bacterial species generated 15-20 candidate primer pairs per bacterial pathogen. The primer pairs were assessed by a conventional PCR for specificity against 33 species of Clavibacter, Pantoea, Rathayibacter, Pectobacterium, Curtobacterium. The investigation for specificity and sensitivity of the primer pairs allowed final selection of one or two primer pairs per bacterial pathogens. In our assay condition, a detection limit of Pss and Cmn was 2 pg/μl of genomic DNA per PCR reaction, while the detection limit for Rt primers was higher. The selected primers could also detect bacterial cells up to 8.8 × 103 cfu to 7.84 × 104 cfu per gram of grain seeds artificially infected with corresponding bacterial pathogens. The primer pairs and PCR assay developed in this study provide an accurate and rapid detection method for three bacterial pathogens of grains, which can be used to investigate bacteria contamination in grain seeds and to ultimately prevent pathogen dissemination over countries.
中文翻译:
比较基因组学开发的特异性和灵敏引物,用于检测谷物中的细菌病原体。
准确,快速地检测细菌性植物病原体是迈向疾病管理和预防病原体传播的第一步。细菌植物病原体密歇根杆菌亚种。nebraskensis(Cmn),Pantoea stewartii亚种。stewartii(Pss)和Rathayibacter tritici(Rt)分别引起戈斯的玉米枯萎病,斯图尔特的玉米枯萎病和小麦和大麦的穗枯病。细菌性疾病不在全球范围内分布,并且在韩国不存在。这项研究采用比较基因组学方法,旨在开发特定的引物对以检测这三种细菌病原体。目标病原体及其密切相关的细菌种类之间的基因组比较,每个细菌病原体产生15-20个候选引物对。引物对通过常规PCR的特异性评估针对33种的棍状杆菌,泛菌属,Rathayibacter,果胶杆菌属,Curtobacterium属。对引物对的特异性和敏感性的研究允许每个细菌病原体最终选择一个或两个引物对。在我们的测定条件下,每个PCR反应的Pss和Cmn的检出限为2 pg /μl基因组DNA,而Rt引物的检出限更高。选择的引物还可以检测高达8.8×10 3 cfu至7.84×10 4的细菌细胞每克谷物种子每英亩cfu人工感染了相应的细菌病原体。本研究开发的引物对和PCR分析方法为谷物的三种细菌病原体提供了一种准确,快速的检测方法,可用于调查谷物种子中的细菌污染并最终防止病原体在各国传播。
更新日期:2020-08-21
中文翻译:
比较基因组学开发的特异性和灵敏引物,用于检测谷物中的细菌病原体。
准确,快速地检测细菌性植物病原体是迈向疾病管理和预防病原体传播的第一步。细菌植物病原体密歇根杆菌亚种。nebraskensis(Cmn),Pantoea stewartii亚种。stewartii(Pss)和Rathayibacter tritici(Rt)分别引起戈斯的玉米枯萎病,斯图尔特的玉米枯萎病和小麦和大麦的穗枯病。细菌性疾病不在全球范围内分布,并且在韩国不存在。这项研究采用比较基因组学方法,旨在开发特定的引物对以检测这三种细菌病原体。目标病原体及其密切相关的细菌种类之间的基因组比较,每个细菌病原体产生15-20个候选引物对。引物对通过常规PCR的特异性评估针对33种的棍状杆菌,泛菌属,Rathayibacter,果胶杆菌属,Curtobacterium属。对引物对的特异性和敏感性的研究允许每个细菌病原体最终选择一个或两个引物对。在我们的测定条件下,每个PCR反应的Pss和Cmn的检出限为2 pg /μl基因组DNA,而Rt引物的检出限更高。选择的引物还可以检测高达8.8×10 3 cfu至7.84×10 4的细菌细胞每克谷物种子每英亩cfu人工感染了相应的细菌病原体。本研究开发的引物对和PCR分析方法为谷物的三种细菌病原体提供了一种准确,快速的检测方法,可用于调查谷物种子中的细菌污染并最终防止病原体在各国传播。