Maize white spot lesions caused by Pantoea ananatis has contributed substantially to yield reduction of maize crops in many countries, including Brazil. The initial symptoms of the disease include watersoaked lesions on the leaves, which later become necrotic and straw-colored. Basic knowledge regarding the biology and the infection mechanisms of this pathogen is lacking. In this study, 15 P. ananatis isolates obtained from maize white spot lesions were examined for their ice nucleation activity (INA). The INAs of individual bacterial isolates was determined by tube nucleation tests. Bacterial isolates were grown on tryptic soy broth medium and an aliquot of 0.1 mL of culture was added to test tubes containing 1 mL of sterile distilled water. The tubes were packed in an ice bath, which had a temperature below -10°C, for approximately 2 min. Instantaneous formation of ice in the tube revealed a positive INA phenotype of the isolate. Only 9 of the 15 studied isolates showed the INA + phenotype. Pathogenicity tests were performed using whole plants and detached leaves. Symptoms were reproduced in both tests, but only for the inoculations using INA + isolates. Electron microscopy allowed visualization of protein vesicles under outer cell wall of isolates characterized as INA + . Key words: INA, vesicle protein, transmission electron microscopy ResumoLesões de mancha branca do milho causadas por Pantoea ananatis contribuiram substancialmente para a redução da produtividade nos cultivos de milho, em muitos países, incluindo o Brasil. Os sintomas iniciais da doença incluem lesões anasarcas nas folhas, que mais tarde tornam-se necróticas e de cor palha. O conhecimento básico sobre a biologia e os mecanismos de infecção deste patógeno são escassos. Neste estudo, 15 isolados de P. ananatis obtidos a partir de lesões da mancha branca do milho foram examinados quanto à sua atividade de nucleação de gelo (INA). A INA de isolados bacterianos individuais foi determinada por testes de nucleação em tubos. Isolados bacterianos foram cultivados em meio de caldo de soja tríptico e uma alíquota de 0,1 mL de cultura foi adicionada à tubos de ensaio contendo 1 mL de água destilada esterelizada. Os tubos foram colados em banho de gelo, o qual continha temperatura inferior a -10 ºC, por aproximadamente 2 min. A formação de gelo instantânea no tubo revelou um fenótipo INA positivo do isolado. Apenas 9 dos 15 isolados estudados apresentaram o fenótipo INA+. Testes de patogenicidade foram realizados com plantas inteiras e folhas destacadas.
ResumoO objetivo deste trabalho foi avaliar a eficiência de diferentes produtos no controle da mancha branca do milho, doença foliar causada pela bactéria Pantoea ananatis. Para o experimento foram utilizados seis produtos pertencentes a diferentes grupos químicos, testados no híbrido suscetível HS200. Avaliaramse as variáveis severidade, área abaixo da curva de progresso da doença (AACPD) e a produtividade de grãos. O tratamento com oxitetraciclina resultou maior produtividade de grãos da cultura do milho. Produtos naturais testados no experimento, como Rocksil e Ácido Pirolenhoso, utilizados como suplementos nutricionais para as plantas, não foram eficientes no controle da mancha branca do milho. Palavras-chave: Zea mays L., controle químico, Pantoea ananatis, antibiótico AbstractThe aim of this work was the evaluation of different products used to control maize white spot, a leaf disease caused by the bacterium Pantoea ananatis. Six products from different chemical groups were used for this experiment, and they were tested on the susceptible hybrid HS200. Parameters, such as severity (%), area under disease progress curve (AUDPC), and productivity, have been estimated. The treatment with oxytetracycline resulted higher grain productivity for maize. Natural products, such as Rocksil and Pyroligneous acid, even though they are used as supplement for plants, were not effective in controlling the maize white spot.
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