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韩国苹果溃疡病致病菌丁香假单胞菌丁香致病变种的鉴定与特性分析

Identification and Characterization of Pseudomonas syringae pv. syringae, a Causative Bacterium of Apple Canker in Korea.

作者信息

Lee Seunghee, Cheon Wonsu, Kwon Hyeok Tae, Lee Younmi, Kim Jungyeon, Balaraju Kotnala, Jeon Yongho

机构信息

Department of Plant Medicals, Andong National University, Andong 36729, Korea.

Agricultural Science & Technology Research Institute, Andong National University, Andong 36729, Korea.

出版信息

Plant Pathol J. 2023 Feb;39(1):88-107. doi: 10.5423/PPJ.OA.08.2022.0121. Epub 2023 Feb 1.

Abstract

In the present investigation, bacterial isolates from infected apple trees causing apple canker during winter were studied in the northern Gyeongbuk Province, Korea. The pathogen was identified as Pseudomonas syringae pv. syringae (Pss) through various physiological and biochemical characterization assays such as BIOLOG, gas chromatography of fatty acid methyl esters, and 16S rRNA. Bioassays for the production of phytotoxins were positive for syringopeptin and syringomycin against Bacillus megaterium and Geotrichum candidum, respectively. The polymerase chain reaction (PCR) method enabled the detection of toxin-producing genes, syrB1, and sypB in Pss. The differentiation of strains was performed using LOPAT and GATTa tests. Pss further exhibited ice nucleation activity (INA) at a temperature of -0.7°C, indicating an INA+ bacterium. The ice-nucleating temperature was -4.7°C for a non-treated control (sterilized distilled water), whereas it was -9.6°C for an INA- bacterium Escherichia coli TOP10. These methods detected pathogenic strains from apple orchards. Pss might exist in an apple tree during ice injury, and it secretes a toxin that makes leaves yellow and cause canker symptoms. Until now, Korea has not developed antibiotics targeting Pss. Therefore, it is necessary to develop effective disease control to combat Pss in apple orchards. Pathogenicity test on apple leaves and stems showed canker symptoms. The pathogenic bacterium was re-isolated from symptomatic plant tissue and confirmed as original isolates by 16S rRNA. Repetitive element sequence-based PCR and enterobacterial repetitive intergenic consensus PCR primers revealed different genetic profiles within P. syringae pathovars. High antibiotic susceptibility results showed the misreading of mRNA caused by streptomycin and oxytetracycline.

摘要

在本次调查中,对韩国庆尚北道北部冬季感染苹果树导致苹果溃疡病的细菌分离株进行了研究。通过多种生理生化鉴定试验,如BIOLOG、脂肪酸甲酯气相色谱分析和16S rRNA分析,将病原菌鉴定为丁香假单胞菌丁香致病变种(Pss)。针对巨大芽孢杆菌和白色念珠菌的丁香肽和丁香霉素的植物毒素产生生物测定结果均为阳性。聚合酶链反应(PCR)方法能够检测Pss中产生毒素的基因syrB1和sypB。使用LOPAT和GATTa试验对菌株进行区分。Pss在-0.7°C时进一步表现出冰核活性(INA),表明是一种INA+细菌。未处理对照(灭菌蒸馏水)的冰核形成温度为-4.7°C,而INA-细菌大肠杆菌TOP10的冰核形成温度为-9.6°C。这些方法检测到了来自苹果园的致病菌株。Pss可能在苹果遭受冻害时存在于苹果树中,并分泌一种毒素,使叶片变黄并导致溃疡症状。到目前为止,韩国尚未开发出针对Pss的抗生素。因此,有必要开发有效的病害防治措施来对抗苹果园中的Pss。对苹果叶片和茎进行的致病性试验显示出溃疡症状。从有症状的植物组织中重新分离出致病细菌,并通过16S rRNA确认为原始分离株。基于重复元件序列的PCR和肠杆菌重复基因间共有序列PCR引物揭示了丁香假单胞菌致病型内不同的遗传图谱。高抗生素敏感性结果显示链霉素和土霉素导致mRNA误读。

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