Fitness Attributes of Pythium aphanidermatum with Dual Resistance to Mefenoxam and Fenamidone.

Pythium aphanidermatum is the predominant species causing Pythium root rot on commercially grown poinsettias in North Carolina. Resistance to mefenoxam is common in populations of P. aphanidermatum but resistance to fenamidone and other quinone outside inhibitor fungicides has only just been reported in greenhouse floriculture crops. The in vitro sensitivity to the label rate of mefenoxam (17.6 µl active ingredient [a.i.]/ml) and fenamidone (488 µl a.i./ml) was determined for 96 isolates of P. aphanidermatum. Isolates were assigned to four fungicide phenotypes: mefenoxam-sensitive/fenamidone-sensitive (MefS, FenS), mefenoxam-sensitive/fenamidone-insensitive (MefS, FenR), mefenoxam-insensitive/fenamidone-sensitive (MefR, FenS), and mefenoxam-insensitive/fenamidone-insensitive (MefR, FenR). In all, 58% of isolates were insensitive to one (MefR, FenS = 36% and MefS, FenR = 16%) or both fungicides (MefR, FenR = 6%). A single point mutation in the cytochrome b gene (G143A) was identified in fenamidone-insensitive isolates. Mycelial growth rate at three temperatures (20, 25, and 30°C), in vitro oospore production, and aggressiveness on poinsettia were evaluated to assess relative fitness of sensitive and insensitive isolates. Isolates with dual insensitivity to mefenoxam and fenamidone had reduced radial hyphal growth at 30°C and produced fewer oospores than isolates sensitive to one or both fungicides. Isolates sensitive to both fungicides produced greater numbers of oospores. Aggressiveness on poinsettia varied by isolate but fungicide phenotype was not a good predictor of aggressiveness. These results suggest that populations of P. aphanidermatum with dual resistance to mefenoxam and fenamidone may be less fit than sensitive populations under our imposed experimental conditions but populations of P. aphanidermatum should continue to be monitored in poinsettia production systems for mefenoxam and fenamidone insensitivity.

Mefenoxam Sensitivity, Aggressiveness, and Identification of Pythium Species Causing Root Rot on Floriculture Crops in North Carolina.

Herbaceous ornamental plants exhibiting symptoms of Pythium root rot were collected from 26 greenhouses in 21 counties in North Carolina (NC) from 2010 to 2012. Plant symptoms ranged from mild stunting to severe wilting, root rot, and death. Roots were plated on selective media, and 356 isolates of Pythium were recovered from 34 host species. Selected isolates were identified by sequencing of the internal transcribed spacer (ITS) rDNA gene region. Seventeen Pythium species were identified, with P. aphanidermatum, P. irregulare, and P. myriotylum comprising 75% of the 320 isolates sequenced. Twelve of the 26 greenhouses had more than one species present. Mefenoxam sensitivity was tested in vitro by growing isolates in wells of microtiter plates containing clarified V8 agar amended with 100 µg a.i./ml mefenoxam. Colonization was scored after 24 to 48 h using a scale of 0 (no growth) to 5 (entire well colonized). Fifty-two percent of the isolates were resistant to mefenoxam (mean score ≥4). All 32 isolates of P. myriotylum were sensitive, whereas sensitivity varied among isolates of P. aphanidermatum and P. irregulare. Resistant and sensitive isolates of the same species were found within the same greenhouses. The aggressiveness of P. aphanidermatum and P. irregulare isolates was evaluated on poinsettia, Gerbera daisy, and petunia. P. aphanidermatum was more aggressive than P. irregulare on poinsettia and petunia; symptoms were mild and no differences in aggressiveness were observed on Gerbera daisy. Sensitivity to mefenoxam was not related to aggressiveness.