Maize seeds infected by Stenocarpella macrospora can cause stalk and ear rot and leaf spot. Transmission of this pathogen through seeds may vary according to the cultivar, climatic conditions, and virulence of the pathogen among other factors. The aim of this study was to assess the transmission rate of S. macrospora from seeds of the maize cultivars C1-RB9308YG and C2-RB9108 using artificially infected seeds grown under two temperatures (20 ºC and 25 ºC). Seeds were inoculated by the osmotic conditioning method for 24 h (inoculum potential - P1), 48 h (P2), 72 h (P3) and 96 h (P4). After inoculation, 25 seeds were distributed individually in plastic cups with substrate, with 4 replicates per treatment. At the end of twenty-eight days of daily assessments, all plants were analyzed for the presence of the pathogen by biological methods, and some were sampled at random and analyzed Bio-PCR. The maximum percentages of dead seeds/seedlings in pre-emergence were 74.5% and 82.5% for P3 and P4, respectively. The highest total rate of transmission of the pathogen under study was 85.8% for seeds of the cultivar C1 at the highest inoculum potential (P4), grown at the temperature of 20 ºC.
-Given what is already known in regard to seed health and the availability of molecular methods for detection of the pathogens Stenocarpella maydis and Stenocarpella macrospora in maize seeds, Colletotrichum gossypii var. cephalosporioides in cotton seeds, and Corynespora cassiicola in soybean seeds, the aim of this study was to evaluate seed vigor according to different inoculum potentials. The fungus isolates were inoculated on seeds by the technique of water restriction, through which different inoculum potentials are obtained, corresponding to times of seed exposure of 0, 24, 48, and 96 hours for maize and cotton seeds, and 0, 36, 108, and 144 hours for soybean seeds. The seeds were subjected to germination, electrical conductivity, health, and qPCR tests. Results of the blotter test showed that in most pathosystems, there was a higher incidence of the fungi with an increase in inoculum potential. A decrease in germination percentage was observed in all species as inoculum potential increased, as well as further degradation of seed membranes. The qPCR test confirmed that the most damaged seeds in the tests had higher presence of the pathogens.Index terms: seed pathology, fungi, maize, soybean, cotton.Avaliação do potencial de inóculo de patógenos em sementes: sua relação com a qualidade fisiológica e quantificação do DNA por qPCR RESUMO -Diante do que já se conhece em sanidade de sementes e tendo-se em mãos métodos moleculares para a detecção dos fungos Stenocarpella maydis, Stenocarpella macrospora em sementes de milho, Colletotrichum gossypii var. cephalosporioides em algodão e Corynespora cassiicola em soja, objetivou-se neste estudo avaliar o vigor das sementes em função dos diferentes potenciais de inóculo. Os isolados dos fungos foram inoculados nas sementes por meio da técnica de restrição hídrica pela qual se obtém diferentes potenciais de inóculo, que foram representados por P0, P24, P48 e P96 e P0, P36, P108 e P144, que correspondem à exposição das sementes aos períodos de tempo de 0, 24, 48, 96 horas (milho e algodão) e nos tempos 0, 36, 108 e 144 horas (soja), respectivamente. As sementes foram submetidas aos testes de germinação, condutividade elétrica, sanidade e qPCR. Pelo blotter test, na maioria dos patossistemas, houve maior incidência do fungo com o aumento do potencial de inóculo. Foi observada uma queda na porcentagem de germinação de todas as espécies com o aumento do potencial de inóculo, assim como maior degradação das membranas das sementes. A qPCR confirmou que nas sementes mais prejudicadas havia maior quantidade de inóculo dos patógenos.Termos para indexação: patologia de sementes, fungos, milho, soja, algodão.
-Stenocarpella maydis is one of the main fungi associated with maize seeds, being a causative agent of stalk and ear rot, a disease which causes considerable losses for crop-producing regions in Brazil. The organism is considered to be a pest, subject to sanitary standardisation in current programs of seed certification in the country. The aim of this study was to evaluate the transmission rate of the fungus from infected maize seeds. Seeds were inoculated with two isolates using a method of physiological conditioning, in which the seeds are kept in contact with colonies of the fungus for 24 (P1), 48 (P2), 72 (P3) and 96 (P4) hours. Two cultivars were used, one susceptible (C1) and one moderately resistant (C2), and the trial carried out at two temperatures (20 °C and 25 °C). The inoculated seeds were distributed individually into plastic cups containing substrate. The plants were evaluated daily for stand and the appearance of post-emergent symptoms. Based on the number of dead seeds, transmission rates reached a maximum of 90.5% at the P4 inoculum potential, this rate being greater than transmission rates achieved for symptomatic and asymptomatic infection in emerged plants. For the total transmission rate, transmission of the pathogen was seen at all inoculum potentials; these values varying from 25% for cultivar C2 at potential P1 and a temperature of 20 °C, to 93% for cultivar C2 at potential P3 and a temperature of 25 °C. RESUMO -Stenocarpella maydis é um dos principais fungos que se associam às sementes de milho, sendo um dos agentes causais da podridão do colmo e da espiga, que ocasiona perdas consideráveis em regiões produtoras desta cultura no Brasil. O referido organismo tem sido enquadrado como uma praga sujeita a padronização sanitária em programas de certificação de sementes vigentes no país. O objetivo nesse trabalho foi avaliar a taxa de transmissão do referido fungo a partir de sementes de milho infectadas. As sementes foram inoculadas com dois isolados pelo método de condicionamento fisiológico no qual as sementes são mantidas em contato com colônias do fungo por 24 (P1), 48 (P2), 72 (P3) e 96 (P4) horas. Foram utilizadas duas cultivares, uma suscetível (C1) e outra moderadamente resistente (C2) e o ensaio conduzido em duas temperaturas (20 e 25 °C). As sementes inoculadas foram distribuídas individualmente em copos plásticos contendo substrato. As plantas foram avaliadas diariamente quanto ao estande e aparecimento de sintomas em pós-emergência. Com base em sementes mortas, as taxas de transmissão alcançaram o percentual máximo de 90,5% para o potencial de inóculo P4, sendo esta taxa mais alta do que as alcançadas pelas taxas de transmissão para infecção sintomática e assintomática em plantas emergidas. Em relação à taxa de transmissão total, observou-se a transmissão do patógeno em todos os potenciais de inóculo, variando estes valores entre 25% para C2 no potencial P1 sob temperatura de 20 °C e 93% para C2 no potencial P3 sob temperatura de 25 °C.Palavras-chave: Fitopatologia. Patol...
-The association of the fungus Stenocarpella maydis with corn seed may cause a reduction of seed germination and vigor of the emerged seedlings. This work was carried out in order to evaluate the effects of S. maydis on corn seed quality as well as on its early development. To evaluate such effects, seeds of cultivars RB9308YG (C1) and RB9108 (C2) were inoculated by the osmotic conditioner technique with two S. maydis isolates for 24(P1), 48(P2), 72(P3) and 96 hours (P4). Plants were grown in a room chamber at 20 °C and 25 °C and daily assessed until 28 days after emergence. Seed germination, incidence of S maydis, electrical conductivity, speed of emergence index (SEI), initial and final seedling population and dry weight of emerged plants, were assessed. The longer the exposition times of the seeds to the fungal colony, the more severe negative effects of the pathogens on seed vigor were observed. S. maydis caused reduced seed vigor in the speed of seedling emergence in the final stand and early development of corn seedlings.
-The fungus Corynespora cassiicola, causal agent of target spot in soybeans, can be transmitted by soybean seeds and as of that point cause severe damage. This disease may be diagnosed at an early stage by seed testing, but knowledge in this area is insufficient. Because of that and increased attack by the disease in soybean areas in Brazil, further studies are required. The aim of this study was to evaluate the use of conventional PCR in detecting C. cassiicola in soybean seeds. The GA4-F/GA4-R primers described in the literature were tested for their specificity and sensitivity for detection of C. cassiicola in pure culture and in soybean seeds. Uninoculated and inoculated seed samples were used with different incidence levels -100%, 10%, 1%, 0.5%, 0.25%, and 0% of preestablished inoculum potentials, P0, P1, P2, and P3. Detection of C. cassiicola in P1 inoculum potential was observed in samples with incidence levels of 10% to 100%. In the P3 potential, detection of the pathogen was successful in samples at the low level of 0.25%.Index terms: seed pathology, target spot, electrophoresis.Uso de PCR convencional para a detecção de Corynespora cassiicola em sementes de soja RESUMO -O fungo Corynespora cassiicola, agente causal da mancha alvo em soja, pode ser transmitido por sementes de soja e a partir daí causar danos severos. A diagnose precoce desta doença pode ser realizada por meio de testes de sementes, cujo conhecimento é insuficiente. Por esta razão e pelo aumento da área de soja atacada pela referida doença no Brasil, estudos mais aprofundados são necessários. Neste trabalho, o objetivo foi avaliar o uso da técnica PCR convencional na detecção de C. cassiicola em amostras de sementes de soja. Os primers GA4-F/GA4-R, relatados na literatura, foram testados para verificar a sua especificidade e sensibilidade para detecção de C. cassiicola em cultura pura e em sementes de soja. Foram utilizadas amostras de sementes, inoculadas e não inoculadas, com diferentes níveis de incidência, de 100%, 10%, 1%, 0,5%, 0,25% e 0%, por potenciais de inóculo preestabelecidos, P0, P1, P2 e P3. A detecção de C. cassiicola no potencial de inóculo P1, foi observada em amostras com níveis de incidência de 10% a 100%. No potencial P3, a detecção do patógeno foi bem-sucedida em amostras com nível mínimo de 0,25%.Termos para indexação: patologia de sementes, mancha alvo, eletroforese.
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