Life cycle of Puccinia acroptili on Rhaponticum (= Acroptilon) repens.

Russian knapweed (Rhaponticum repens) is a rangeland weed pest in the western United States. One candidate fungus for biological control of R. repens is Puccinia acroptili, which causes a rust disease. Understanding the life cycle of candidate rust fungi for weed biological control is an essential component in risk assessments and evaluations, and for P. acroptili such was unknown. For this reason greenhouse studies were undertaken to clarify the life cycle of P. acroptili under artificial conditions. Spermogonia with spermatia developed on R. repens following plant inoculation with teliospores. Artificial transfer of spermatia between spermogonia resulted in the development of aecia with uredinioid aeciospores. Inoculation with aeciospores or urediniospores resulted in uredinia containing urediniospores and occasional amphispores. Telia with teliospores and occasional mesospores developed later. Teliospores produced typical basidia with four basidiospores. These results suggest that the life cycle of P. acroptili is macrocyclic and autoecious. Inoculation with teliospores also frequently resulted in production of sori that were morphologically similar to aecia but which were not associated with spermogonia or the classical transfer of spermatia. The ontology of these sori is unknown. This is the first description of spermogonia and the first report and description of basidiospores, aecia, aeciospores, amphispores and mesospores of P. acroptili.

Effect of temperature and moisture period on infection of Rhododendron 'Cunningham's White' by Phytophthora ramorum.

We investigated the temperature and moisture conditions that allow Phytophthora ramorum to infect Rhododendron 'Cunningham's White'. Most experiments were performed with a single P. ramorum isolate from the NA1 clonal lineage. For whole plants incubated in dew chambers at 10 to 31 degrees C, the greatest proportion of diseased leaves, 77.5%, occurred at the optimum temperature of 20.5 degrees C. Disease occurred over the entire range of temperatures tested, although amounts of disease were minor at the temperature extremes. For whole plants exposed to varying dew periods at 20 degrees C and then incubated at 20 degrees C for 7 days, a dew period as short as 1 h resulted in a small amount of disease; however, at least 4 h of dew were required for >10% of the leaves to become diseased. Moisture periods of 24 and 48 h resulted in the greatest number of diseased leaves. In detached-leaf, temperature-gradient-plate experiments, incubation at 22 degrees C resulted in the greatest disease severity, followed by 18 degrees C and then 14 degrees C. In detached-leaf, moisture-tent experiments, a 1-h moisture period was sufficient to cause disease on 67 to 73% of leaves incubated for 7 days at 20 degrees C. A statistical model for disease development that combined the effects of temperature and moisture period was generated using nonlinear regression. Our results define temperature and moisture conditions which allow infection by P. ramorum on Cunningham's White rhododendron, and show that P. ramorum is able to infect this host over a wide range of temperatures and moisture levels. The results indicate that P. ramorum has the potential to become established in parts of the United States that are outside its current range.

Survival of Phytophthora ramorum hyphae after exposure to temperature extremes and various humidities.

We examined the effect of short-term exposure to high and low temperatures and a range of relative humidity (RH) on survival of Phytophthora ramorum hyphae. Spore-free hyphal colonies were grown on dialysis squares atop V8 medium. Colonies were transferred to water agar plates positioned at 27.5-50 C on a thermal gradient plate and incubated 2.5-480 min. For low temperature trials colonies were transferred to vials of distilled water and incubated in a water bath at -5 to -25 C for 1-24 h. In the relative humidity trials hyphal colonies were transferred to sealed humidity chambers containing various concentrations of glycerin for 1-8 h. Relative humidity was 41-93% at 20 C and 43-86% at 28 C. Survival in all trials was characterized by growth from dialysis squares into V8 medium. Temperatures of 37.5-40 C were lethal to P. ramorum hyphae within several hours, and temperatures of 42.5-50 C were lethal within minutes. Exposure to 32.5 and 35 C resulted in reduced survival over 8 h, while 30 C had no effect on three of four isolates. Hyphal colonies demonstrated considerable tolerance to cold, with all isolates surviving a 24 h exposure to -5 C. Survival diminished over time at lower temperatures, however a few colonies survived 24 h exposure to -25 C. Temperature also affected the ability of hyphal colonies to withstand reduced humidity. A RH of 41-43% was lethal in 2 h at 28 C compared to 8 h at 20 C. Three of four isolates were unaffected by an 8 h exposure to 81 and 95% RH at 20 C, and 73 and 86% RH at 28 C. Isolate differences were apparent in tolerance to freezing temperatures and reduced humidity. From these results it is apparent that the cold temperatures found in the northeastern USA are not likely to prevent the establishment of P. ramorum. There is also the potential for hyphae, and presumably spores, to survive periods of high humidity on the leaf surface in the absence of free water.

Recovery of Phytophthora ramorum Following Exposure to Temperature Extremes.

We examined the impact of exposure to high and low temperature extremes on recovery of Phytophthora ramorum both as free chlamydospores and within infected rhododendron tissue over a 7-day period. Chlamydospores held in moistened sand were incubated at 30, 35, 40, 0, -10, and -20°C for up to 7 days. Infected Rhododendron 'Cunningham's White' leaf disks held in sandy loam, loam, or sand at two different soil moisture levels also were subjected to these temperatures for up to 7 days, and to a variable temperature regimen for 12 weeks. Recovery was characterized by growth of P. ramorum on selective agar medium following exposures to temperature treatments. Chlamydospores held in moistened sand showed a high rate of recovery at 30°C, steadily declining recovery at 35°C, and no recovery at 40°C over the 7-day period. Chlamydospores were recovered from 0°C after 7 days, with little or no recovery observed at -10 or -20°C. In infected rhododendron tissue, P. ramorum was recovered at 20 and 30°C after 7 days but, at 35°C, the pathogen showed a decline within 2 days and no recovery by 4 days. A 40°C treatment allowed no recovery of P. ramorum from infected tissue after 2 days. For cold treatments, P. ramorum was recovered from infected leaf disks at 0 and -10°C after 7 days. At -20°C, recovery declined rapidly after 1 to 3 days and no recovery was obtained after 4 days. P. ramorum showed nearly 100% recovery from leaf disks subjected to a 12-week variable temperature treatment based on ambient summer temperatures in Lewisburg, TN. The results suggest that P. ramorum is capable of surviving some highly adverse temperature conditions for at least 7 days both as free chlamydospores in sand and within infected host tissue. Thus, P. ramorum present as free chlamydospores or within tissue of infected plants shipped to the eastern United States has the potential to survive some of the adverse conditions encountered in summer and winter in many eastern states.

In vitro germination of Striga hermonthica and Striga aspera seeds by 1-aminocyclopropane-1-carboxylic acid.

Treatment of conditioned seeds of four isolates of Striga hermonthica and one isolate of Striga aspera with various concentrations of the ethylene precursor, 1-aminocyclopropane-1-carboxylic acid (ACC), caused complex stimulation of germination patterns. GR 24, the strigol analogue served as a positive control and its stimulatory activity was comparable to that of ACC. When conditioned Striga seeds were treated with negative control that did not contain ACC, the stimulatory effect was lost. Overall, the germination data suggested a hormonal mode of action by ACC, which involves indirect stimulation of biosynthesis of ethylene that then triggers seed germination. The various mechanisms that have been proposed for the chemical and biological oxidation of ACC to generate ethylene are discussed.