SdhC-I86F Mutation in Phakopsora pachyrhizi Is Stable and Can Be Related to Fitness Penalties.

Succinate dehydrogenase inhibitors (SDHIs) fungicides are used to control Asian soybean rust (Phakopsora pachyrhizi), and the SdhC-I86F mutation is related to pathogen resistance. The objective of this study was to determine whether fitness penalties are associated with SDHI resistance (SdhC-I86F mutation) in P. pachyrhizi populations. Moreover, the study investigated whether the SdhC-I86F mutation remained stable after the fungus propagation both in the absence and presence of fungicide. The populations used in this study presented mutations for all genes analyzed (Cyp51, Cytb, and SdhC), except for a wild-type population (WTSdhC) found with no SdhC-I86F mutation. The frequencies of the SdhC-I86F mutant populations were stable after 36 generations in the absence of fungicide. However, in the case of the WTSdhC population, the SdhC-I86F mutation was further detected after one generation of the fungus in the presence of the SDHI fungicide, according to the results of a detached leaf assay. Three tests were performed to evaluate fitness components and sensitivity to fungicides (half maximal effective concentration). SdhC-I86F mutant populations were more sensitive to osmotic and oxidative stress than the WTSdhC population; however, the sensitivity to ultraviolet radiation was similar for both populations. All mutated populations were less sensitive than the WTSdhC when using SDHI (azoxystrobin + benzovindiflupyr), but more sensitive to mancozeb. The presence of fitness penalties, the mutation stability, and the sensitivity to mancozeb presented by the SdhC-I86F mutant populations can be relevant to the management of the disease in the field.

Sensitivity of the Colletotrichum acutatum Species Complex From Apple Trees in Brazil to Dithiocarbamates, Methyl Benzimidazole Carbamates, and Quinone Outside Inhibitor Fungicides.

Glomerella leaf spot (GLS) and bitter rot (BR) on apples are often caused by Colletotrichum acutatum in Paraná State, Brazil. GLS control is difficult because of its rapid development, with an incubation period of only 2 days under favorable conditions. Therefore, producers use successive fungicide applications every season; however, failure to control GLS has been commonly reported. The objectives of this study were to determine the sensitivity of isolates of the C. acutatum species complex obtained from apple orchards in Brazil to mancozeb, thiophanate-methyl, and azoxystrobin fungicides. Isolates from the different parts of the plant (leaves, flowers, buds, and twigs) and cultivars (Gala and Eva) showed different levels of sensitivity to mancozeb, thiophanate-methyl, and azoxystrobin. For mancozeb, the frequencies of isolates were 25% highly resistant, 50% low-resistance, and 25% sensitive. For thiophanate-methyl, the frequencies of isolates were 72.2% highly resistant, 11.1% resistant, and 16.7% moderately resistant. For azoxystrobin, the frequencies of isolates were 11.1% highly resistant, 5.6% resistant, and 83.3% sensitive. Interestingly, no mutations in the ß-tubulin and cytochrome b genes were observed in any of the isolates resistant to thiophanate-methyl and azoxystrobin fungicides.

Competitive Fitness of Phakopsora pachyrhizi Isolates with Mutations in the CYP51 and CYTB Genes.

Soybean rust (Phakopsora pachyrhizi) in Brazil is mainly controlled with applications of fungicides, including demethylation inhibitors (DMI) and quinone outside inhibitors (QoI). Isolates with less sensitivity to DMI and QoI have been reported, and these have been found to have mutations in the CYP51 and CYTB genes, respectively. There have been no reports of fitness costs in isolates with mutations in CYP51 and CYTB, and the aim of this work was to compare the competitive ability of isolates with lower DMI or QoI sensitivities with that of sensitive (wild-type) isolates. Urediniospores of sensitive wild-type isolates and isolates with different CYP51 or CYTB alleles were mixed and inoculated on detached soybean leaves. After 3 weeks, urediniospores were harvested and used as inoculum for the next disease cycle. Frequencies of relevant target site mutations were monitored using the pyrosequencing method over four disease cycles. Isolates with lower DMI sensitivity and different CYP51 alleles had competitive disadvantages compared with a DMI-sensitive, wild-type CYP51 isolate. In contrast, the isolate with the F129L mutation in the CYTB gene competed equally well with a QoI-sensitive, wild-type CYTB isolate under the conditions of this experiment. The CYP51 and CYTB alleles were stable in all isolates over four disease cycles when cultivated alone.