Although Pantoea species are widely distributed among plants, water, soils, humans, and animals, due to a lack of efficient isolation methods, the clonality of Pantoea species is poorly characterized. Therefore, we developed a new semi-selective medium designated ‘lysine-ornithine-mannitol-arginine-charcoal’ (LOMAC) to isolate these species. In an inclusive and exclusive study examining 94 bacterial strains, all Pantoea strains exhibited yellow colonies on LOMAC medium. The performance of the medium was assessed using Pantoea-spiked soils. Percent average agreement relative to the Api20E biochemical test was 97%. A total of 24 soil spot samples and 19 plant types were subjected to practical trials. Of the 91 yellow colonies selected on LOMAC medium, 81 were correctly identified as Pantoea species using the biochemical test. The sequencing of 16S rRNA (rrs) and gyrB from these isolates confirmed that Pantoea agglomerans, P. vagans, P. ananatis, and P. deleyi were present in Japanese fields. A phylogenetic analysis using rrs enabled only the limited separation of strains within each Pantoea spp., whereas an analysis using gyrB revealed higher variability and enabled the finer resolution of distinct branches. P. agglomerans isolates were divided into 3 groups, 2 of which were new clades, with the other comprising a large group including biocontrol strains. P. vagans was also in one of the new clades. The present results indicate that LOMAC medium is useful for screening Pantoea species. The use of LOMAC medium will provide new opportunities for identifying the beneficial properties of Japanese Pantoea isolates.

中文翻译：

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开发一种新的半选择性赖氨酸-鸟氨酸-甘露醇-精氨酸-木炭培养基，用于在日本从环境来源中分离泛菌物种

尽管泛菌物种广泛分布于植物、水、土壤、人类和动物中，但由于缺乏有效的分离方法，泛菌属物种的克隆性特征很差。因此，我们开发了一种新的半选择性培养基，称为“赖氨酸-鸟氨酸-甘露醇-精氨酸-木炭”（LOMAC）来分离这些物种。在一项对 94 种细菌菌株进行的全面且独家的研究中，所有泛菌菌株在 LOMAC 培养基上都显示出黄色菌落。使用 Pantoea 尖刺土壤评估培养基的性能。相对于 Api20E 生化测试的平均一致性百分比为 97%。共对24个土壤点样和19个植物类型进行了实际试验。在 LOMAC 培养基上选择的 91 个黄色菌落中，81 个使用生化测试被正确鉴定为泛菌属。来自这些分离株的 16S rRNA (rrs) 和 gyrB 的测序证实了成团泛菌、P. vagans、P. ananatis 和 P. deleyi 存在于日本田间。使用 rrs 的系统发育分析只能实现每个 Pantoea spp. 内菌株的有限分离，而使用 gyrB 的分析显示更高的可变性并实现不同分支的更精细分辨率。P. agglomerans 分离株被分为 3 组，其中 2 组是新进化枝，另一组包括一个大组，其中包括生物防治菌株。P. vagans 也是新进化枝之一。目前的结果表明 LOMAC 培养基可用于筛选泛菌属物种。LOMAC 培养基的使用将为鉴定日本泛菌分离株的有益特性提供新的机会。