{"title":"乌克兰 Soryz 菌斑病病原体的生物特性","authors":"M.V. Reshetnikov, L. M. Butsenko, L. Pasichnyk","doi":"10.15407/microbiolj85.06.048","DOIUrl":null,"url":null,"abstract":"Soryz is a new promising agricultural crop. Sorghum leaf spots are one of the most common and harmful diseases of these crops. Improving the technology of growing agricultural crops requires the development of methods of controlling their pathogens, based on data on their distribution and properties. There is no information on the taxonomic status and properties of the causative agents of soryz bacterial spot in Ukraine. The aim of the work was to identify the causative agent of bacterial spots of a new sorghum crop — soryz in Ukraine and to study its biological properties. Methods. Identification of the causative agent of soryz bacterial spots was carried out in the Cherkasy and Kyiv regions of Ukraine in 2019-2023. Isolation of the pathogen and study of its morphological-cultural, physiological-biochemical properties were carried out by classical microbiological methods using the NEFERMtest24 (MikroLaTEST®, ErbaLachema, Czech Republic) and API 20NE (Biomerieux, France) test systems. Electron microscopy, chromatographic separation, and identification of fatty acids were conducted at the Center for Collective Research of the Zabolotny Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine (IMV). The antigenic properties of isolated bacteria were investigated by the agglutination reaction with antisera to five serological groups of Pseudomonas syringae strains. Identification of the obtained bacteria was carried out on the basis of their phenotypic properties and the results of MALDI-TOF mass spectrometry on a VITEK MS mass spectrometer. Results. Affecting by spotting was noted in 2—27% of soryz plants, and the development of symptoms of damage was 1—4 points. Bacteria with sorghum disease symptoms were isolated, from which ten virulent isolates were studied in detail. According to the phenotypic properties, 9 isolates that were similar to the characteristics of the typical strain of P. syringae UCM B-1027T, were identified as P. syringae van Hall 1902. One isolate belongs to phytopathogenic bacteria of the genus Pseudomonas according to its main properties, but its taxonomic status within the genus needs to be clarified. In the cellular lipids strains isolated from affected soryz plants, as well as in the type strain of P. syringae UCM B-1027T, such fatty acids as dodecanoic, tetradecanoic, hexadecanoic, octadecanoic, cis-9-hexadecenoic, cis-11-octadecenoic, cis-9,10-methylene hexadecanoic, cis-9,10-methylene octadecanoic acids were identified. In the cellular lipids of strains from soryz, fatty acids with an even number of carbon atoms predominate, the total content of which is more than 60% of all detected fatty acids. Isolates from soryz also contain 3-hydroxydecanoic, 2-hydroxydodecanoic, and 3-hydroxydodecanoic fatty acids, the presence of which confirms the belonging of the isolated bacteria to the species P. syringae. According to the antigenic properties, nine strains of P. syringae isolated from soryz are homogeneous and belong to serogroup I. One strain of Pseudomonas sp. does not belong to any of the studied serogroups that parasitize grain crops. Belonging to the causative agent of soryz bacterial spots to the species P. syringae was confirmed by the MALDI-TOF mass spectrometry method. Conclusions. Therefore, on the base of our results, it has been established that the main causative agent of soryz bacterial spots in Ukraine is P. syringae. According to antigenic properties, the population of the causative agent of soryz bacterial spots is homogeneous, which makes it possible to develop serological rapid tests for the detection of the causative agent. By comparing the biological properties of collection strains from sorghum, a species of Pseudomonas holci, which does not exist in modern taxonomy, with the properties of strains from soryz, the typical strain of P. syringae UCM B-1027T, we established that P. holci bacteria isolated in the Department of Phytopathogenic Bacteria of the IMV in 1968-1971 belong to the species P. syringae.","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"16 2","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2023-12-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Biological Properties of the Agent of Soryz Bacterial Spot in Ukraine\",\"authors\":\"M.V. Reshetnikov, L. M. Butsenko, L. Pasichnyk\",\"doi\":\"10.15407/microbiolj85.06.048\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"Soryz is a new promising agricultural crop. Sorghum leaf spots are one of the most common and harmful diseases of these crops. Improving the technology of growing agricultural crops requires the development of methods of controlling their pathogens, based on data on their distribution and properties. There is no information on the taxonomic status and properties of the causative agents of soryz bacterial spot in Ukraine. The aim of the work was to identify the causative agent of bacterial spots of a new sorghum crop — soryz in Ukraine and to study its biological properties. Methods. Identification of the causative agent of soryz bacterial spots was carried out in the Cherkasy and Kyiv regions of Ukraine in 2019-2023. Isolation of the pathogen and study of its morphological-cultural, physiological-biochemical properties were carried out by classical microbiological methods using the NEFERMtest24 (MikroLaTEST®, ErbaLachema, Czech Republic) and API 20NE (Biomerieux, France) test systems. Electron microscopy, chromatographic separation, and identification of fatty acids were conducted at the Center for Collective Research of the Zabolotny Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine (IMV). The antigenic properties of isolated bacteria were investigated by the agglutination reaction with antisera to five serological groups of Pseudomonas syringae strains. Identification of the obtained bacteria was carried out on the basis of their phenotypic properties and the results of MALDI-TOF mass spectrometry on a VITEK MS mass spectrometer. Results. Affecting by spotting was noted in 2—27% of soryz plants, and the development of symptoms of damage was 1—4 points. Bacteria with sorghum disease symptoms were isolated, from which ten virulent isolates were studied in detail. According to the phenotypic properties, 9 isolates that were similar to the characteristics of the typical strain of P. syringae UCM B-1027T, were identified as P. syringae van Hall 1902. One isolate belongs to phytopathogenic bacteria of the genus Pseudomonas according to its main properties, but its taxonomic status within the genus needs to be clarified. In the cellular lipids strains isolated from affected soryz plants, as well as in the type strain of P. syringae UCM B-1027T, such fatty acids as dodecanoic, tetradecanoic, hexadecanoic, octadecanoic, cis-9-hexadecenoic, cis-11-octadecenoic, cis-9,10-methylene hexadecanoic, cis-9,10-methylene octadecanoic acids were identified. In the cellular lipids of strains from soryz, fatty acids with an even number of carbon atoms predominate, the total content of which is more than 60% of all detected fatty acids. Isolates from soryz also contain 3-hydroxydecanoic, 2-hydroxydodecanoic, and 3-hydroxydodecanoic fatty acids, the presence of which confirms the belonging of the isolated bacteria to the species P. syringae. According to the antigenic properties, nine strains of P. syringae isolated from soryz are homogeneous and belong to serogroup I. One strain of Pseudomonas sp. does not belong to any of the studied serogroups that parasitize grain crops. Belonging to the causative agent of soryz bacterial spots to the species P. syringae was confirmed by the MALDI-TOF mass spectrometry method. Conclusions. Therefore, on the base of our results, it has been established that the main causative agent of soryz bacterial spots in Ukraine is P. syringae. According to antigenic properties, the population of the causative agent of soryz bacterial spots is homogeneous, which makes it possible to develop serological rapid tests for the detection of the causative agent. By comparing the biological properties of collection strains from sorghum, a species of Pseudomonas holci, which does not exist in modern taxonomy, with the properties of strains from soryz, the typical strain of P. syringae UCM B-1027T, we established that P. holci bacteria isolated in the Department of Phytopathogenic Bacteria of the IMV in 1968-1971 belong to the species P. syringae.\",\"PeriodicalId\":18628,\"journal\":{\"name\":\"Mikrobiolohichnyi zhurnal\",\"volume\":\"16 2\",\"pages\":\"\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2023-12-21\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Mikrobiolohichnyi zhurnal\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.15407/microbiolj85.06.048\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q4\",\"JCRName\":\"Biochemistry, Genetics and Molecular Biology\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Mikrobiolohichnyi zhurnal","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.15407/microbiolj85.06.048","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"Biochemistry, Genetics and Molecular Biology","Score":null,"Total":0}
引用次数: 0
摘要
高粱是一种新的有前途的农作物。高粱叶斑病是这些作物最常见、最有害的病害之一。要提高农作物的种植技术,就必须根据病原体的分布和特性数据,开发控制病原体的方法。目前还没有关于乌克兰 soryz 细菌性斑点病病原体的分类状况和特性的信息。这项工作的目的是确定乌克兰一种新的高粱作物--soryz--的细菌性斑点病原菌,并研究其生物学特性。研究方法2019-2023 年在乌克兰的切尔卡瑟州和基辅州进行了高粱细菌性斑点病原菌的鉴定工作。通过经典微生物学方法,使用 NEFERMtest24(MikroLaTEST®,ErbaLachema,捷克共和国)和 API 20NE (Biomerieux,法国)测试系统分离病原体并研究其形态-文化、生理-生化特性。电子显微镜、色谱分离和脂肪酸鉴定在乌克兰国家科学院扎博洛特尼微生物学和病毒学研究所集体研究中心(IMV)进行。分离出的细菌的抗原特性是通过与五组血清学意义上的丁香假单胞菌菌株抗血清的凝集反应来研究的。根据细菌的表型特性和在 VITEK MS 质谱仪上进行的 MALDI-TOF 质谱分析结果,对获得的细菌进行了鉴定。结果。2-27%的高粱植株受到斑点病的影响,损害症状发展为 1-4 点。分离出了具有高粱病害症状的细菌,并对其中的 10 个毒力分离物进行了详细研究。根据表型特征,9 个分离物与典型的 P. syringae UCM B-1027T 菌株特征相似,被鉴定为 P. syringae van Hall 1902。根据其主要特性,其中一个分离物属于假单胞菌属植物病原菌,但其在假单胞菌属中的分类地位有待明确。在从受影响的 soryz 植物中分离出的细胞脂质菌株以及 P. syringae UCM B-1027T 型菌株中,发现了十二酸、十四酸、十六酸、十八酸、顺式-9-十六烯酸、顺式-11-十八烯酸、顺式-9,10-亚甲基十六烷酸、顺式-9,10-亚甲基十八烷酸等脂肪酸。在索里兹菌株的细胞脂质中,碳原子数为偶数的脂肪酸占主导地位,其总含量占所有检测到的脂肪酸的 60% 以上。从 soryz 分离出来的菌株还含有 3-羟基癸酸、2-羟基十二酸和 3-羟基十二酸脂肪酸,这些脂肪酸的存在证实了分离出来的细菌属于 P. syringae 菌种。根据抗原特性,从 soryz 中分离出的 9 株 P. syringae 具有同质性,属于血清组 I。通过 MALDI-TOF 质谱分析方法证实,沙雷氏菌病的病原菌属于 P. syringae。得出结论。因此,根据我们的研究结果,可以确定乌克兰 Soryz 菌斑病的主要病原体是 P. syringae。根据抗原特性,浆液性细菌病斑的致病菌群是同质的,这使得开发用于检测致病菌的血清学快速测试成为可能。通过比较从高粱中采集的菌株(现代分类学中不存在的一种全缘假单胞菌)的生物学特性与从沙里兹中采集的菌株(典型的 P. syringae 菌株 UCM B-1027T)的特性,我们确定 1968-1971 年在 IMV 植物病原菌部分离的全缘假单胞菌属于 P. syringae 菌种。
Biological Properties of the Agent of Soryz Bacterial Spot in Ukraine
Soryz is a new promising agricultural crop. Sorghum leaf spots are one of the most common and harmful diseases of these crops. Improving the technology of growing agricultural crops requires the development of methods of controlling their pathogens, based on data on their distribution and properties. There is no information on the taxonomic status and properties of the causative agents of soryz bacterial spot in Ukraine. The aim of the work was to identify the causative agent of bacterial spots of a new sorghum crop — soryz in Ukraine and to study its biological properties. Methods. Identification of the causative agent of soryz bacterial spots was carried out in the Cherkasy and Kyiv regions of Ukraine in 2019-2023. Isolation of the pathogen and study of its morphological-cultural, physiological-biochemical properties were carried out by classical microbiological methods using the NEFERMtest24 (MikroLaTEST®, ErbaLachema, Czech Republic) and API 20NE (Biomerieux, France) test systems. Electron microscopy, chromatographic separation, and identification of fatty acids were conducted at the Center for Collective Research of the Zabolotny Institute of Microbiology and Virology of the National Academy of Sciences of Ukraine (IMV). The antigenic properties of isolated bacteria were investigated by the agglutination reaction with antisera to five serological groups of Pseudomonas syringae strains. Identification of the obtained bacteria was carried out on the basis of their phenotypic properties and the results of MALDI-TOF mass spectrometry on a VITEK MS mass spectrometer. Results. Affecting by spotting was noted in 2—27% of soryz plants, and the development of symptoms of damage was 1—4 points. Bacteria with sorghum disease symptoms were isolated, from which ten virulent isolates were studied in detail. According to the phenotypic properties, 9 isolates that were similar to the characteristics of the typical strain of P. syringae UCM B-1027T, were identified as P. syringae van Hall 1902. One isolate belongs to phytopathogenic bacteria of the genus Pseudomonas according to its main properties, but its taxonomic status within the genus needs to be clarified. In the cellular lipids strains isolated from affected soryz plants, as well as in the type strain of P. syringae UCM B-1027T, such fatty acids as dodecanoic, tetradecanoic, hexadecanoic, octadecanoic, cis-9-hexadecenoic, cis-11-octadecenoic, cis-9,10-methylene hexadecanoic, cis-9,10-methylene octadecanoic acids were identified. In the cellular lipids of strains from soryz, fatty acids with an even number of carbon atoms predominate, the total content of which is more than 60% of all detected fatty acids. Isolates from soryz also contain 3-hydroxydecanoic, 2-hydroxydodecanoic, and 3-hydroxydodecanoic fatty acids, the presence of which confirms the belonging of the isolated bacteria to the species P. syringae. According to the antigenic properties, nine strains of P. syringae isolated from soryz are homogeneous and belong to serogroup I. One strain of Pseudomonas sp. does not belong to any of the studied serogroups that parasitize grain crops. Belonging to the causative agent of soryz bacterial spots to the species P. syringae was confirmed by the MALDI-TOF mass spectrometry method. Conclusions. Therefore, on the base of our results, it has been established that the main causative agent of soryz bacterial spots in Ukraine is P. syringae. According to antigenic properties, the population of the causative agent of soryz bacterial spots is homogeneous, which makes it possible to develop serological rapid tests for the detection of the causative agent. By comparing the biological properties of collection strains from sorghum, a species of Pseudomonas holci, which does not exist in modern taxonomy, with the properties of strains from soryz, the typical strain of P. syringae UCM B-1027T, we established that P. holci bacteria isolated in the Department of Phytopathogenic Bacteria of the IMV in 1968-1971 belong to the species P. syringae.