Characterization of acetylcholinesterases of acaricide-resistant and susceptible strains of the cattle tick Boophilus microplus (Can.): I. Extraction of the critical component and comparison with enzyme from other sources

The critical acetylcholinesterase component, that is responsible for the resistance mechanism of decreased sensitivity to the inhibitor in the cattle tick Boophilus microplus (Can.), has been isolated from the organophosphorus-susceptible Yeerongpilly and resistant Biarra and Ridgelands strains. With the substrate acetylthiocholine this enzyme component gave a pH activity profile similar for all strains. Optimal substrate concentration for Ridgelands was 1.73.10^?2M compared to 2.70.10^?4M for the other strains. Computed Km values were 1.98.10^?5M, 3.65.10^?5M, and 6.43.10^?6M for Yeerongpilly, Biarra, and Ridgelands strains, respectively, while the corresponding V_max figures were 336, 135, and 21 μmoles acetylthiocholine hydrolyzed per gram of larvae per hour. These results are discussed in relation to the resistance mechanism and compared with similar parameters derived for this enzyme from bovine erythrocytes, the sheep blowfly Lucilia cuprina, and the sheep nematode Trichostrongylus colubtiformis.     

Antifungal mode of action of triforine

Rapidly growing mycelia of Aspergillus fumigatus treated with 10 μg/ml triforine (N,N′-bis-(1-formamido-2,2,2-trichloroethyl)-piperazine) showed little or no inhibition in dry weight increase prior to 2 h. By 2.5–3 h, triforine inhibited dry weight increase by 85%. The effects of triforine on protein, DNA, and RNA syntheses corresponded to the effect on dry weight increase both in time of onset and magnitude. Neither glucose nor acetate oxidation were inhibited by triforine.Ergosterol synthesis was almost completely inhibited by triforine even in the first hour after treatment. Inhibition of ergosterol synthesis was accompanied by an accumulation of the ergosterol precursors 24-methylenedihydrolanosterol, obtusifoliol, and 14α-methyl-Δ^{8, 24 (28)}-ergostadienol. Mycelia treated with 5 μg/ml of triarimol (α-(2,4-dichlorophenyl)-α-phenyl-5-pyrimidinemethanol) also accumulated the same sterols as well as a fourth sterol believed to be Δ^{5, 7}-ergostadienol.Identification of 4,4-dimethyl-Δ^{8, 24 (28)}-ergostadienol in untreated mycelia indicates that the C-14 methyl group is the first methyl group removed in the biosynthesis of ergosterol by A. fumigatus. The lack of detectable quantities of 4,4-dimethyl-Δ^{8, 24 (28)}-ergostadienol in triforine or triarimol-treated mycelia and the accumulation of C-14 methylated sterols in treated mycelia suggests that both fungicides inhibit sterol C-14 demethylation. The accumulation of Δ^{5, 7}-ergostadienol in triarimol-treated mycelia further implies that triarimol also inhibits the introduction of the sterol C-22(23) double bond.Two strains of Cladosporium cucumerinum tolerant to triforine and triarimol were also tolerant to the fungicide S-1358 (N-3-pyridyl-S-n-butyl-S′-p-t-butylbenzyl imidodithiocarbonate).     

Development of an EU protocol for the detection and diagnosis of Potato spindle tuber pospiviroid*

The work described here formed part of the EU SMT DIAGPRO project, to develop diagnostic protocols for 18 regulated pests. The Potato spindle tuber pospiviroid (PSTVd) protocol was developed primarily for testing in vitro‐ and glasshouse‐grown potato plants for the purposes of post‐entry quarantine and the production of pathogen‐tested nuclear stock. After a performance audit of methods used by 12 laboratories in Europe and America by ring testing, four methods were chosen for multilaboratory validation. For most laboratories, the detection limits were 10–20 mg of PSTVd‐infective tissue for R‐PAGE; 0.25–0.5 mg for DIG‐probe; 0.062 mg for RT‐PCR; and 0.0155 mg for TaqMan (this was the lowest weight of infective tissue tested). Some laboratories were able to extend the detection limit to 0.0155 mg for DIG‐probe and RT‐PCR. The DIG‐probe and R‐PAGE are recommended as primary detection methods, with confirmation of viroid presence by any of the four validated detection methods. Specific diagnosis requires the viroid to be sequenced. Other methods may be used for primary detection, providing that they preferably detect all PSTVd isolates and other Pospiviroids that have the potential to infect potato, and detect viroid in at least 1/10 of the tissue weight normally tested per plant.     

Historical range of variability in eastern Cascades forests,Washington, USA

The historical range of variability (HRV) has been suggested as a coarse filter approach to maintain ecosystem sustainability and resiliency. The historical range of variability in forest age structure for the central eastern Cascade Range in Washington State, USA was developed from historical fire return intervals and the manner in which fire acted as both cyclic and stochastic processes. The proportions of seven forest structural stages calculated through these processes were applied to the area of each forest series within the central eastern Cascades landscape. Early successional forest stages were more common in high elevation forest than low elevation forest. The historical proportion of old growth and late successional forest varied from 38 to 63 percent of the forested landscape. These process-based estimates are consistent with those developed from forest structural information. HRV is a valuable planning tool for ecosystem conservation purposes, but must be applied to real landscapes with consideration of both temporal and spatial scale. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effects of Latent Infection of Stock Plants and Abundance of Thrips on the Occurrence of <Emphasis Type="Italic">Tomato spotted wilt virus </Emphasis>in Chrysanthemum Fields

DAS-ELISA proved to be reliable enough to detect a latent infection by Tomato spotted wilt virus (TSWV) in asymptomatic stock plants of chrysanthemum. A high density of Frankliniella occidentalis, the predominant vector, in the presence of latently infected stock plants resulted in a high incidence of disease in the chrysanthemum production field. The incidence of disease was low when the vector thrips were not abundant in spite of the presence of latently infected stock plants. These results suggest that an infestation of the vector thrips causes severe secondary spread of TSWV originating from latently infected stock plants in chrysanthemum production fields. Received 27 July 2001/ Accepted in revised form 27 November 2001     

Indole-3-acetic acid biosynthesis in <Emphasis Type="Italic">Aciculosporium take</Emphasis>, a causal agent of witches' broom of bamboo

 Aciculosporium take (Ascomycota; Clavicipitaceae) is a causal agent of witches' broom of bamboo plants. The symptoms of this disease are believed to be induced by plant hormones, particularly auxins. Indole-3-acetic acid (IAA) was identified in cultures of this fungus in an l-tryptophan-supplemented liquid medium. IAA production was confirmed on 30 isolates of A. take from various hosts and locations at levels up to 1 mg/l. The biosynthetic pathway of IAA in A. take culture was examined by analyzing intermediate products and by feeding experiments. The results showed that the indole-3-pyruvic acid pathway (l-tryptophan → indole-3-pyruvic acid → indole acetaldehyde → IAA) was the dominant pathway in A. take. Received: June 3, 2002 / Accepted: July 25, 2002     

Inconspicuous restraint of rice seedling growth by root-infecting fungi in soil of a rice paddy field

 Rice seedling growth, estimated by plant height and root development and discoloration, was better in pasteurized soil than in unpasteurized soil obtained from a flooded rice field. Rice seedlings also grew better in sterilized soil modified by adding roots harvested from the pasteurized soil than in soil modified by adding roots harvested from the unpasteurized soil. The results demonstrate that seedling growth in the rice field soil was inhibited by soil microorganisms, even though no typical symptoms such as seedling blight or damping-off appeared. Pythium aristosporum is suggested to be involved in the inhibition. Thus, it appears that inconspicuous restraint of rice seedling growth could occur in soils of rice paddy fields. Received: May 20, 2002 / Accepted: October 16, 2002 Acknowledgments The authors thank Dr. T. Ichitani, former professor at Osaka Prefectural University, for providing an isolate of Pythium aristosporum for comparison, and Mr. Mitsuaki Sato of Akita Prefectural College of Agriculture for technical assistance.     

Functional heterogeneity in resources within landscapes and herbivore population dynamics

Large mammalian herbivores are notorious for their propensity towards population irruptions and crashes, yet many herbivore populations remain relatively stable. I explore how resource heterogeneity within landscapes dampens population instability, using a metaphysiological modelling approach considering patch state distributions. Resource heterogeneity is functionally stabilizing through spreading consumption away from preferred resources before these become critically depleted. Lower-quality resources act as a buffer against starvation during critical periods of the seasonal cycle. Enriching resource quality is destabilizing, even if patch diversity is maintained, because food quantity then becomes the limitation. The potential consequences of landscape fragmentation are explored using the Serengeti ecosystem, characterised by broadscale resource gradients, as a hypothetical example. Further insights provided by the model are illustrated with specific examples concerning the effects of patch scales and waterpoint distribution. A metaphysiological modelling approach enables the basic consequences of landscape heterogeneity to be distinguished from further effects that may arise from specific patch scales and configurations, without the distracting detail of spatially explicit models.     

Characterizing historical and modern fire regimes in Michigan (USA): A landscape ecosystem approach

We studied the relationships of landscape ecosystems to historical and contemporary fire regimes across 4.3 million hectares in northern lower Michigan (USA). Changes in fire regimes were documented by comparing historical fire rotations in different landscape ecosystems to those occurring between 1985 and 2000. Previously published data and a synthesis of the literature were used to identify six forest-replacement fire regime categories with fire rotations ranging from very short (<100 years) to very long (>1,000 years). We derived spatially-explicit estimates of the susceptibility of landscape ecosystems to fire disturbance using Landtype Association maps as initial units of investigation. Each Landtype Association polygon was assigned to a fire regime category based on associations of ecological factors known to influence fire regimes. Spatial statistics were used to interpolate fire points recorded by the General Land Office. Historical fire rotations were determined by calculating the area burned for each category of fire regime and dividing this area by fifteen (years) to estimate area burned per annum. Modern fire rotations were estimated using data on fire location and size obtained from federal and state agencies. Landtype Associations networked into fire regime categories exhibited differences in both historical and modern fire rotations. Historical rotations varied by 23-fold across all fire rotation categories, and modern forest fire rotations by 13-fold. Modern fire rotations were an order of magnitude longer than historical rotations. The magnitude of these changes has important implications for forest health and understanding of ecological processes in most of the fire rotation categories that we identified.This revised version was published online in May 2005 with corrections to the Cover Date.