RESUMO

Verticillium wilt of olive (VWO) caused by Verticillium dahliae is considered a major olive (Olea europaea) disease in Mediterranean-type climate regions. The lack of effective chemical products against VWO makes it necessary to search for alternatives such as biological control. The main goal of this study was to evaluate the effect of six Streptomyces spp. strains as biological control agents (BCAs) against VWO. All of them were molecularly characterized by sequencing 16S or 23S rRNA genes and via phylogenetic analysis. Their effect was evaluated in vitro on the mycelial growth of V. dahliae (isolates V004 and V323) and on microsclerotia (MS) viability using naturally infested soils. Bioassays in olive plants inoculated with V. dahliae were also conducted to evaluate their effect against disease progress. In all the experiments, the reference BCAs Fusarium oxysporum FO12 and Aureobasidium pullulans AP08 were included for comparative purposes. The six strains were identified as Streptomyces spp., and they were considered as potential new species. All the BCAs, including Streptomyces strains, showed a significant effect on mycelial growth inhibition for both V. dahliae isolates compared to the positive control, with FO12 being the most effective, followed by AP08, while the Streptomyces spp. strains showed an intermediate effect. All the BCAs tested also showed a significant effect on the inhibition of germination of V. dahliae MS compared to the untreated control, with FO12 being the most effective treatment. Irrigation treatments with Streptomyces strain CBQ-EBa-21 or FO12 were significantly more effective in reducing disease severity and disease progress in olive plants inoculated with V. dahliae compared to the remaining treatments. This study represents the first approach to elucidating the potential effect of Streptomyces strains against VWO.

RESUMO

Eggplant (Solanum melongena L.) is an important solanaceous crop that is produced mainly in tropical and subtropical regions and is widely consumed worldwide. In 2018, eggplant production in Mexico was approximately 80000 t, and Sinaloa State contributed 96% of this production; however, this crop suffers significant losses from plant pathogens. In December 2019, fruits from commercial orchards (geographical coordinates: 24°45'39.39''N, 107°26'57.30''O) with visible brown soft rot and profuse white mycelia were analysed. On V8 medium, pieces of tissue obtained from the border of lesions were plated and incubated between 25°C and 36°C. After five days, a dense cottony mycelium with a slightly petaloid pattern was observed at 25°C and did not grow at 36°C. Isolates of that pathogen were heterothallic, and microscopic preparations showed development of coenocytic mycelium and spheroid sporangia that were noncaducous and papillate, measuring 35.6 ± 5.8 x 27.1 ± 4.4. Based on morphological characteristics, the eggplant soft rot causal agent was identified as Phytophthora nicotianae Breda de Haan (Erwin and Ribeiro, 1996). From a representative isolate denominated PhySm01, two DNA regions (internal transcribed spacer (ITS) and the large subunit ribosomal (28S)) were amplified and sequenced with ITS1-ITS4 and NL1-LR3 primers, respectively. The obtained ITS sequence (GenBank accession number MT508842) showed 100% identity with several P. nicotianae sequences (Access MT065840, MH290435 and MG570057) with 100% query coverage and 740 matching nucleotides. For the 28S sequence (accession number MT508843), the identity with strains N° Access EU080889 and EU080508 of P. nicotianae was 99.86%, with 100 query coverage and 729 matching nucleotides. Further, phylogenetic analysis from P. nicotianae strain PhySm01 and GenBank reference sequences was carried out by Maximum Likelihood method with Mega 7 software based on the ITS sequences, which verified the species identification. To fulfill Koch's postulates, a suspension containing 1 x 104 zoospores/mL of the oomycete isolated from the original diseased eggplant fruit was used to inoculate ten healthy and disinfested fruits. Sterile water was used as a control. Three wounds per fruit were made with a sterile needle, and 20 µL of the zoospore suspension (or water) was placed on each. All fruits were placed into plastic bags with moistened paper and incubated at 25°C for three days. Thisest was repeated twice with similar results. Initial symptoms developed 24 hours after inoculation with brown soft tissue forming around the inoculated area, and profuse soft rot accompanied by white mycelium was observed two days after inoculation. No symptoms developed on the control fruits during this time. Pieces of necrotic tissue were plated on V8 medium and incubated as described previously. The reisolated pathogen was compared to the original isolate and had the same morphological characteristics. Phytophthora nicotianae has a worldwide distribution and can infect multiple solanaceous crops, including tobacco (Gallup et al., 2018). In addition, it has been reported on other hosts outside of the Solanaceae family, causing economically important losses in citrus and strawberry. In India, it causes necrosis of citrus fruits and roots, leading to tree decline (Das et al., 2016), and in the United States, it causes crown rot of strawberry (Marin et al., 2018). In Mexico, P. nicotianae has been reported on vinca (Alvarez-Rodriguez et al., 2013); however, this is the first report of P. nicotianae causing fruit soft rot of eggplant, and this pathogen represents a new threat when the environment is favourable for disease outbreaks. References Alvarez-Rodriguez, B. et al. 2013. Plant Dis. 97: 1257. https://doi.org/10.1094/PDIS-04-13-0400-PDN Das, A. K. et al. 2016. J. Plant Pathol. 98: 55. DOI: 10.4454/JPP.V98I1.038 Erwin, D. C. and Ribeiro, O. K. 1996. Phytophthora Diseases Worldwide. The American Phytopathological Society, St. Paul, MN. 562 Pp. Gallup, C. A. et al. 2018. Plan Dis. 102: 1108. https://doi.org/10.1094/PDIS-02-17-0295-RE Marin, M. V. et al. 2018. Plant Dis. 102:1463. https://doi.org/10.1094/PDIS-08-17-1333-PDN.

RESUMO

Diseases incited by soilborne fungi are responsible for reducing the yield and cause significant impact to almost all crops. Among then, Rhizoctonia solani AG-4 is considered the most important in the cotton, common bean and soybean crops in Brazil. The use of diagrammatic scale or rating scales, as a tool to help in the quantification of the severity of a particular disease, is more common for foliar diseases. Considering the lack of a standardized, illustrated and easy-to-apply methodology for assessing the severity of R. solani lesions in cotton, soybean and common bean seedlings, a simple and precise rating scale was developed with the objective to fill this gap. The proposed scale shows four levels of disease severity, with the descriptions and illustrations for each type of lesion observed in the cotton, soybean and common bean seedlings. This developed scale was validated in many experiments and proved to be adequate for severity assessments of R. solani lesions on cotton, soybean and common bean seedlings.

Doenças causadas por fungos de solo reduzem a produtividade e impactam de forma significativa quase todas as culturas. Dentre elas, Rhizoctonia solani AG-4 é considerada a mais importante nas culturas de algodão, feijão e soja no Brasil. A utilização de escalas diagramáticas ou escalas de notas, como ferramenta para auxiliar na quantificação da severidade de uma determinada doença, é mais comum quando se trata de doenças foliares. Considerando a inexistência, até então, de metodologia padronizada, ilustrada e de fácil aplicação para a avaliação da severidade de lesões de R. solani em plântulas de algodão, soja e feijão, desenvolveu-se uma escala de notas, simples e precisa, com o objetivo de suprir essa lacuna. A escala proposta apresenta quatro níveis de severidade de doença, com descrições e ilustrações para cada tipo de lesão observada nas plântulas de algodão, soja e feijão. Essa escala foi validade em inúmeros experimentos, provando ser adequada para a avaliação da severidade de lesões de R. solani nas plântulas de algodão, soja e feijão

Assuntos

RESUMO

Studies were conducted to determine the effect of osmotic and matric stress on germination and growth of two Fusarium solani strains, the etiological agent responsible of peanut brown root rot. Both strains had similar osmotic and matric potential ranges that allowed growth, being the latter one narrower. F. solani showed the ability to grow down to -14 MPa at 25 °C in non-ionic modified osmotic medium, while under matric stress this was limited to -8.4 MPa at 25 °C. However, both strains were seen to respond differently to decreasing osmotic and matric potentials, during early stages of germination. One strain (RC 338) showed to be more sensitive to matric than osmotic (non ionic) and the other one (RC 386) showed to be more sensitive to osmotic than matric imposed water stress. After 24 h of incubation, both isolates behaved similarly. The minimum water potential for germination was -8.4 MPa on glycerol amended media and -5.6 MPa for NaCl and PEG amended media, respectively. The knowledge of the water potential range which allow mycelia growth and spore germination of F. solani provides an inside to the likely behaviour of this devastating soilborne plant pathogen in nature and has important practical implications.

Assuntos

RESUMO

Studies were conducted to determine the effect of osmotic and matric stress on germination and growth of two Fusarium solani strains, the etiological agent responsible of peanut brown root rot. Both strains had similar osmotic and matric potential ranges that allowed growth, being the latter one narrower. F. solani showed the ability to grow down to -14 MPa at 25 °C in non-ionic modified osmotic medium, while under matric stress this was limited to -8.4 MPa at 25 °C. However, both strains were seen to respond differently to decreasing osmotic and matric potentials, during early stages of germination. One strain (RC 338) showed to be more sensitive to matric than osmotic (non ionic) and the other one (RC 386) showed to be more sensitive to osmotic than matric imposed water stress. After 24 h of incubation, both isolates behaved similarly. The minimum water potential for germination was -8.4 MPa on glycerol amended media and -5.6 MPa for NaCl and PEG amended media, respectively. The knowledge of the water potential range which allow mycelia growth and spore germination of F. solani provides an inside to the likely behaviour of this devastating soilborne plant pathogen in nature and has important practical implications.

Assuntos

RESUMO

Studies were conducted to determine the effect of osmotic and matric stress on germination and growth of two Fusarium solani strains, the etiological agent responsible of peanut brown root rot. Both strains had similar osmotic and matric potential ranges that allowed growth, being the latter one narrower. F. solani showed the ability to grow down to -14 MPa at 25 °C in non-ionic modified osmotic medium, while under matric stress this was limited to -8.4 MPa at 25 °C. However, both strains were seen to respond differently to decreasing osmotic and matric potentials, during early stages of germination. One strain (RC 338) showed to be more sensitive to matric than osmotic (non ionic) and the other one (RC 386) showed to be more sensitive to osmotic than matric imposed water stress. After 24 h of incubation, both isolates behaved similarly. The minimum water potential for germination was -8.4 MPa on glycerol amended media and -5.6 MPa for NaCl and PEG amended media, respectively. The knowledge of the water potential range which allow mycelia growth and spore germination of F. solani provides an inside to the likely behaviour of this devastating soilborne plant pathogen in nature and has important practical implications.

Assuntos

RESUMO

Crop management may modify soil characteristics, and as a consequence, alter incidence of diseases caused by soilborne pathogens. This study evaluated the suppressiveness to R. solani in 59 soil samples from a microbasin. Soil sampling areas included undisturbed forest, pasture and fallow ground areas, annual crops, perennial crops, and ploughed soil. The soil samples were characterized according to abiotic variables (pH; electrical conductivity; organic matter content; N total; P; K; Ca; Mg; Al; H; S; Na; Fe; Mn; Cu; Zn; B; cation exchange capacity; sum of bases and base saturation) and biotic variables (total microbial activity evaluated by the CO2 evolution and fluorescein diacetate hydrolysis; culturable bacterial, fungal, actinomycetes, protozoa, fluorescent Pseudomonas and Fusarium spp. communities). The contribution and relationships of these variables to suppression to R. solani were assessed by path analysis. When all samples were analyzed together, only abiotic variables correlated with suppression of R. solani, but the entire set of variables explained only 51% of the total variation. However, when samples were grouped and analyzed by vegetation cover, the set of evaluated variables in all cases accounted for more than 90% of the variation in suppression of the pathogen. In highly suppressive soils of forest and pasture/fallow ground areas, several abiotic variables and fluorescein diacetate hydrolysis correlated with suppression of R. solani and the set of variables explained more than 98% of suppressiveness.

As atividades agrícolas podem modificar as características do solo e, como conseqüência, alterar a incidência de patógenos veiculados pelo solo. Este trabalho avaliou a supressividade a R. solani em 59 amostras de solos de uma microbacia. As áreas amostradas foram selecionadas quanto à vegetação, incluindo mata, pasto/pousio, culturas anuais, culturas perenes e solo arado. As amostras de solo foram caracterizadas quanto às propriedades abióticas (pH, condutividade elétrica, teor de matéria orgânica, N total, P, K, Ca, Mg, Al, H, S, Na, Fe, Mn, Cu, Zn, B, capacidade de troca catiônica, soma de bases e saturação de bases) e bióticas (atividade microbiana do solo, avaliada pelo desprendimento de CO2 e hidrólise de diacetato de fluoresceína; comunidades de bactérias, fungos, actinomicetos, protozoários, Pseudomonas fluorescentes e Fusarium spp.). A contribuição e relação dessas variáveis para a supressividade a R. solani foram quantificadas por análise de coeficientes de trilha. Quando se avaliaram todas as amostras em conjunto, somente variáveis abióticas apresentaram correlação com a supressão a R. solani, mas o conjunto das variáveis explicou somente 51% da variação total. Entretanto, quando as amostras foram agrupadas e analisadas considerando o tipo de cobertura vegetal, o conjunto de variáveis explicou mais de 90% da variação da supressividade. Para as áreas de floresta e pasto/pousio, as quais foram classificadas como as mais supressivas, algumas variáveis abióticas e a hidrólise de diacetato de fluoresceína apresentaram correlação com a supressão de R. solani e o conjunto de variáveis explicou mais de 98% da supressividade nesses solos.

RESUMO

Crop management may modify soil characteristics, and as a consequence, alter incidence of diseases caused by soilborne pathogens. This study evaluated the suppressiveness to R. solani in 59 soil samples from a microbasin. Soil sampling areas included undisturbed forest, pasture and fallow ground areas, annual crops, perennial crops, and ploughed soil. The soil samples were characterized according to abiotic variables (pH; electrical conductivity; organic matter content; N total; P; K; Ca; Mg; Al; H; S; Na; Fe; Mn; Cu; Zn; B; cation exchange capacity; sum of bases and base saturation) and biotic variables (total microbial activity evaluated by the CO2 evolution and fluorescein diacetate hydrolysis; culturable bacterial, fungal, actinomycetes, protozoa, fluorescent Pseudomonas and Fusarium spp. communities). The contribution and relationships of these variables to suppression to R. solani were assessed by path analysis. When all samples were analyzed together, only abiotic variables correlated with suppression of R. solani, but the entire set of variables explained only 51% of the total variation. However, when samples were grouped and analyzed by vegetation cover, the set of evaluated variables in all cases accounted for more than 90% of the variation in suppression of the pathogen. In highly suppressive soils of forest and pasture/fallow ground areas, several abiotic variables and fluorescein diacetate hydrolysis correlated with suppression of R. solani and the set of variables explained more than 98% of suppressiveness.

As atividades agrícolas podem modificar as características do solo e, como conseqüência, alterar a incidência de patógenos veiculados pelo solo. Este trabalho avaliou a supressividade a R. solani em 59 amostras de solos de uma microbacia. As áreas amostradas foram selecionadas quanto à vegetação, incluindo mata, pasto/pousio, culturas anuais, culturas perenes e solo arado. As amostras de solo foram caracterizadas quanto às propriedades abióticas (pH, condutividade elétrica, teor de matéria orgânica, N total, P, K, Ca, Mg, Al, H, S, Na, Fe, Mn, Cu, Zn, B, capacidade de troca catiônica, soma de bases e saturação de bases) e bióticas (atividade microbiana do solo, avaliada pelo desprendimento de CO2 e hidrólise de diacetato de fluoresceína; comunidades de bactérias, fungos, actinomicetos, protozoários, Pseudomonas fluorescentes e Fusarium spp.). A contribuição e relação dessas variáveis para a supressividade a R. solani foram quantificadas por análise de coeficientes de trilha. Quando se avaliaram todas as amostras em conjunto, somente variáveis abióticas apresentaram correlação com a supressão a R. solani, mas o conjunto das variáveis explicou somente 51% da variação total. Entretanto, quando as amostras foram agrupadas e analisadas considerando o tipo de cobertura vegetal, o conjunto de variáveis explicou mais de 90% da variação da supressividade. Para as áreas de floresta e pasto/pousio, as quais foram classificadas como as mais supressivas, algumas variáveis abióticas e a hidrólise de diacetato de fluoresceína apresentaram correlação com a supressão de R. solani e o conjunto de variáveis explicou mais de 98% da supressividade nesses solos.