The baseline sensitivities for mycelial growth of foliar fungal pathogens of citrus, Colletotrichum acutatum, Alternaria alternata, Elsinoe fawcettii, Diaporthe citri, and Mycosphaerella citri, the causal agents of postbloom fruit drop, brown spot of tangerine, citrus scab, melanose, and greasy spot, respectively, were determined in vitro for azoxystrobin, pyraclostrobin, and fenbuconazole. The effective dose to reduce growth by 50% (ED50 values) was determined for each pathogen-fungicide combination using five isolates from different citrus areas of Florida and eight fungicide concentrations. A discriminatory dose for each combination was selected near the ED50, and the range of sensitivity of 50 to 62 isolates of each fungal species was determined. The effect of salicylhydroxamic acid (SHAM) on the sensitivity of the five fungal species to azoxystrobin and pyraclostrobin was determined. Since mycelial growth of A. alternata was insensitive to azoxystrobin, the effect of that fungicide with and without SHAM on spore germination was assessed. The ED50 values for most fungal pathogens of citrus were relatively high compared with foliar pathogens of other tree crops. Values for azoxystrobin ranged from a low of 0.06 μg/ml with E. fawcettii to a high of >100 μg/ml with A. alternata. With pyraclostrobin, the values ranged from a low of 0.019 μg/ml with D. citri to a high of 0.87 μg/ml with A. alternata. With fenbuconazole, the lowest ED50 value was 0.21 μg/ml with M. citri and the highest was 1.01 μg/ml with C. acutatum, but A. alternata and D. citri were not tested. SHAM was inhibitory to all species and reduced growth of D. citri greatly. Inclusion of SHAM in the medium did not greatly affect the sensitivity of mycelial growth of these fungi to azoxystrobin or pyraclostrobin, nor did it affect the ED50 values for conidial germination of A. alternata. The coefficients of variation for the sensitivity of 50 to 62 isolates of each species to these fungi ranged from 7.3% with the pyraclostrobin-C. acutatum combination to a high of 55.0% with the fenbuconazole- M. citri combination. Discriminatory doses have been established for these pathogen- fungicide combinations that should be useful for detecting major shifts in fungicide sensitivity.
Phylogenetic analysis revealed that isolates of Alternaria alternata causing black rot of citrus were associated with six well-supported evolutionary lineages. Isolates recovered from brown spot lesions on Minneola tangelo, leaf spot lesions on rough lemon, and healthy citrus tissue and noncitrus hosts were related closely to isolates from black-rotted fruit. Phylogenies estimated independently from DNA sequence data from an endopolygalacturonase gene (endoPG) and two anonymous regions of the genome (OPA1-3 and OPA2-1) had similar topologies, and phylogenetic analysis was performed on the combined data set. In the combined phylogeny, isolates from diverse ecological niches on citrus and noncitrus hosts were distributed in eight clades. Isolates from all lineages, regardless of ecological or host association, caused black rot in fruit inoculation assays, demonstrating that small-spored Alternaria isolates associated with different ecological niches on citrus and other plant hosts are potential black rot pathogens. These data also indicated that the fungi associated with black-rotted fruit do not form a natural evolutionary group distinct from other Alternaria pathogens and saprophytes associated with citrus. The use of the name A. citri to describe fungi associated with citrus black rot is not justified and it is proposed that citrus black rot fungi be referred to as A. alternata.
Alternaria brown spot, caused by Alternaria alternata, results in serious yield losses of tangerines and their hybrids in Florida. The Alter-Rater model predicts the need for fungicide applications based on daily cumulative points that are assigned on the basis of rainfall, leaf wetness, and temperature. Previously, Alter-Rater threshold or trigger values of 50, 75, 100, and 150 points for application of copper fungicides were suggested for groves with different cultivars and disease histories. In this study, we evaluated thresholds of 50, 100, and 150 points in four Minneola tangelo and Murcott tangor groves in 2000 and 2001. For comparison, copper fungicides were applied according to the DISC Copper Model in 2000 and according to calendar sprays in 2001. Use of the Alter-Rater model resulted in fewer sprays in three of the four groves in 2000 and better fruit quality in the other grove than the Copper Model. Compared to a calendar spray schedule in 2001, use of the Alter-Rater model resulted in fewer sprays in two of the four groves but more sprays in one grove. The results confirmed that the Alter-Rater is a valuable tool for timing fungicide applications and that its use results in better disease control or fewer sprays.
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