First Report of Insensitivity to Cyazofamid Among Isolates of Phytophthora capsici from the Southeastern United States.

Abstract

Phytophthora capsici is rapidly becoming an important limiting factor in vegetable production in the southeastern United States, particularly on cucurbits as fruit rots. One of the strategies used to manage diseases caused by P. capsici is the regular application of fungicides. Recently the new fungicide cyazofamid (trade name Ranman, FRAC Group 21, FMC Corporation, EPA Reg. No. 71512-3-279) was registered for management of P. capsici on cucurbits. Cyazofamid has been reported to be very effective against P. capsici on peppers (1). In a recent evaluation, we observed that cyazofamid was not very effective on fruit rot of watermelon in a field artificially infested with P. capsici (3). Hence, we evaluated our collection of isolates for sensitivity to cyazofamid. We confirmed our isolates as P. capsici based on morphology of colonies and sporangia and amplification of internal transcribed spacer regions using specific PCR primers (4). Mycelial growth of 28 isolates from the southeastern United States including North (NC) and South Carolina (SC), Georgia (GA), and Florida (FL) was evaluated on Ranman amended (0, 25, 100, 310, 518, and 1,000 mg/liter of the active ingredient cyazofamid) V8 juice agar using similar techniques as described before (2). The EC50 (50% effective concentration) values ranged from 3.8 to 535 mg/liter. Thirteen isolates (8 GA, 3 SC, 1 NC, and 1 FL) had EC50 >100 mg/liter. Similar results were obtained when technical grade cyazofamid was used. The same 28 isolates were evaluated on media amended with technical grade cyazofamid (0, 1, 10, and 100 mg/liter) and 100 mg/liter of salicylhydroxaymic acid, which was added to inhibit the alternative oxidase enzyme. The EC50 values ranged from <1 to >100 mg/liter. Six isolates (5 GA and 1 NC) had EC50 >100 mg/liter. Three isolates, one sensitive and two insensitive, were used to inoculate cucumber (Cucumis sativus) fruits treated with commercial Ranman at 0, 10, 100, 300, and 1,000 mg/liter of cyazofamid plus the surfactant Silwett L-77 (0.52 ml/liter). Mycelial plugs (7-mm diameter) were placed on nonwounded fruits. Fruits were kept under high humidity at 25 ± 1°C in an incubator for 3 days. Two measurements of each lesion at right angles were averaged to get the lesion diameter. The EC50 value for lesion diameter on fruits varied from 13 mg/liter for the sensitive isolate to >233 mg/liter for the insensitive isolates. EC50 values for diameter of the lesion with sporulation ranged from 3 to 107 mg/liter. Relative lesion diameters of the insensitive isolates at 100 mg/liter treatment compared with nonsprayed check were 70 to 93%, and at 300 mg/liter, it was 38 to 80%. Similarly in another experiment, watermelon (Citrullus lanatus var. lanatus) fruits were sprayed with a recommended field rate of Ranman (284 mg of cyazofamid/liter) plus Silwett L-77 (0.52 ml/liter) till runoff and inoculated with four isolates. The relative lesion diameter for insensitive isolates on Ranman treated watermelon fruits were 76 to 100% of nonsprayed fruits. To our knowledge, these insensitive isolates were collected from fields that were never sprayed with Ranman. Because of the existence of cyazofamid insensitive P. capsici isolates, it should be rotated with fungicides from other chemical classes to prevent extensive selection of insensitive isolates.