Genetic Control of Resistance to the Piperidine Fungicide Piperalin in Ustilago maydis

Mutants of Ustilago maydis resistant to the piperidine fungicide piperalin were isolated in a mutation frequency of 2.4 × 10^–5 after UV-irradiation and selection on media containing 50gml^–1 piperalin. Genetic analysis with 15 such mutant isolates resulted in the identifications of two unlinked chromosomal loci, the U/ppl-1 locus with two allelic genes (U/ppl-1A and U/ppl-1B) and the U/ppl-2 locus. The U/ppl-2 and U/ppl-1A mutations are responsible for two levels of moderate and high resistance to piperalin (resistance factor, Rf: 54 and 135, respectively, based on effective concentration causing a 50% reduction in the growth rate, EC₅₀), while the U/ppl-1B mutation gives only a small reduction (approximately 8-fold) in piperalin sensitivity. Cross-resistance studies with other SBIs shows that the major gene (U/ppl-2 and U/ppl-1A) mutants are resistant to fenpropidin (Rf: 43 and 68), fenpropimorph (Rf: 261 and 283) and tridemorph (Rf: 9 and 10), but not to the inhibitors of C-14 demethylase (DMIs) and squalene epoxidase. The minor gene mutation U/ppl-1B codes a low-level of resistance (approximately 5—12-fold) to the above morpholine-type fungicides, but in contrast with the major gene mutations it increases 2–10 times the sensitivity to triazoles: triadimefon, triadimenol, propiconazole and flusilazole. Crosses between mutants carrying the U/ppl-genes with compatible isolates carrying the U/fpd, U/fpm or U/tdm mutations, which have been identified in previous genetic works for resistance to morpholine-type fungicides, yielded, with the exception of U/ppl-2 ×U/fpm-2 cross, a large number of recombinants with wild-type sensitivity, indicating that the mutant genes involved in these crosses, were not allelic. An additive gene effect was observed only between nonallelic minor genes U/ppl-1B and U/fpm-1B or U/tdm-1,2. Studies of the fitness of piperalin-resistant isolates showed that the reduced sensitivity of major gene mutants was not associated with changes on the phytopathogenic fitness determining characteristics, such as growth in liquid culture and pathogenicity on young corn plants. Conversely, the minor gene mutation U/ppl-1B appeared to be pleiotropic, having significantly adverse effects on the phytopathogenic fitness.     

Inhibitory activities of venom alkaloids of Red Imported Fire Ant against Clavibacter michiganensis subsp. michiganensis in vitro and the application of piperidine alkaloids to manage symptom development of bacterial canker on tomato in the greenhouse

Tomato bacterial canker caused by Clavibacter michiganensis subsp. michiganensis (CMM) is a highly destructive disease that has caused major economic losses in tomato production worldwide. In seeking disease management alternatives, the inhibitory activity of alkaloids extracted from the Red Imported Fire Ant was studied in the laboratory and the greenhouse. Piperidine and piperideine alkaloids each significantly inhibited CMM growth on nutrient agar plates. The inhibitory activity of piperidine alkaloids was stable at 4 ^° C and 22 ^° C for 12 weeks and at 54 ^° C for 4 weeks. The growth of CMM was negatively correlated with the concentration of piperidine alkaloids in nutrient broth. In the greenhouse, piperidine alkaloids also significantly reduced the symptom development on two tomato cultivars, Better Boy and DRK7018F1. This is the first demonstration that piperidine and piperideine alkaloids from the Red Imported Fire Ant are highly inhibitory against a plant-pathogenic bacterium, viz. CMM. Piperidine alkaloids could provide satisfactory management of CMM bacterial canker on tomato seedlings in the greenhouse. Our findings may lead to the development of a new group of bactericides.     

Effects of piperidine and piperideine alkaloids from the venom of red imported fire ants,Solenopsis invicta Buren,on Pythium ultimum Trow growth in vitro and the application of piperideine alkaloids to control cucumber damping‐off in the greenhouse

BACKGROUND: Pythium ultimum is a plant pathogen that causes significant yield losses on many economically important crops. Chemical treatment has been used for disease control. In searching for alternatives, venom piperidine and piperideine alkaloids from red imported fire ants were tested against P. ultimum in vitro, and piperideines were employed to control cucumber damping‐off in the greenhouse as drench treatments. Results Piperidine and piperideine alkaloids of the red imported fire ant significantly inhibited mycelium growth of P. ultimum. Piperidine alkaloids were stable at both room and elevated temperatures. The inhibitory activity positively correlated with the concentrations of piperidine alkaloids in the medium, and the EC₅₀ = 17.0 µg ml^?1. Germination of sporangia of P. ultimum was negatively correlated with the concentrations of piperidine alkaloids in the medium, and the EC₅₀ = 12.3 µg ml^?1. The piperideine alkaloid drenching treatment significantly improved seedling emergence and seedling height of cucumber. CONCLUSION: This is the first report describing the use of venom alkaloids from the red imported fire ant to inhibit P. ultimum in the laboratory and the application of piperideine alkaloids to control damping‐off disease caused by P. ultimum in the greenhouse. These findings may lead to the development of a new group of fungicides. Copyright © 2012 Society of Chemical Industry