During 2004 to 2005, an unreported disease of maize (Zea mays) was observed on commercial fields in the Northwest and Mpumalanga Provinces of South Africa. Infected plants were stunted, with a vertical crack at the first internode. Inside the stem, a dark-brown, narrow lesion was present along the crack. Internal browning inside the stem extended upward, reaching the top internode in some plants. Seed cobs were underdeveloped. Diseased plants were scattered in the fields and 10 to 70% of the crop was affected. Gram-negative, facultatively anaerobic bacteria were consistently isolated from diseased tissues. Pathogenicity tests established that representative strains induced disease symptoms similar to those observed on maize plants in the field. Physiological and biochemical characterization using the API 20E and API 50CHE systems and 16S rRNA gene sequence analyses showed that the strains belonged to the genus Pantoea. The results of these tests also separated the strains into two groups. The first group, giving a positive reaction in the indole test, was similar to Pantoea ananatis. The second group of strains was indole negative and resembled P. agglomerans. The fluorescent amplified fragment length polymorphism (F-AFLP) genomic fingerprints generated by the indole-positive strains and P. ananatis reference strains were similar and clustered together in the dendrogram, confirming that the indole-positive bacteria causing brown stalk rot on maize were P. ananatis. The F-AFLP fingerprints produced by the indole-negative strains were distinctly different from those generated by P. ananatis, P. agglomerans, P. dispersa, P. citrea, P. stewartii subsp. stewartii, and P. stewartii subsp. indologenes. The results indicated that indole-negative bacteria causing brown stalk rot on maize might belong to a previously undescribed species of the genus Pantoea. This is the first report of a new disease on maize, brown stalk rot, caused by two bacterial species, P. ananatis and an undescribed Pantoea sp.
Eight yellow-pigmented, Gram-negative, rod-shaped, oxidase-negative, motile, facultatively anaerobic bacteria were isolated from onion seed in South Africa and from an onion plant exhibiting centre rot symptoms in the USA. The isolates were assigned to the genus Pantoea on the basis of phenotypic and biochemical tests. 16S rRNA gene sequence analysis and multilocus sequence analysis (MLSA), based on gyrB, rpoB, infB and atpD sequences, confirmed the allocation of the isolates to the genus Pantoea. MLSA further indicated that the isolates represented a novel species, which was phylogenetically most closely related to Pantoea ananatis and Pantoea stewartii. Amplified fragment length polymorphism analysis also placed the isolates into a cluster separate from P. ananatis and P. stewartii. Compared with type strains of species of the genus Pantoea that showed .97 % 16S rRNA gene sequence similarity with strain BD 390 T , the isolates exhibited 11-55 % whole-genome DNA-DNA relatedness, which confirmed the classification of the isolates in a novel species. The most useful phenotypic characteristics for the differentiation of the isolates from their closest phylogenetic neighbours are production of acid from amygdalin and utilization of adonitol and sorbitol. A novel species, Pantoea allii sp. nov., is proposed, with type strain BD 390 T (5LMG 24248 T ).
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