Melanin biosynthesis in Pyricularia oryzae: Site of tricyclazole inhibition and pathogenicity of melanin-deficient mutants

Tricyclazole (5-methyl-1,2,4-triazolo[3,4-b]benzothiazole) inhibits melanin synthesis in Pyricularia oryzae at concentrations less than 0.01 μg/ml. The primary site of inhibition in the biosynthetic pathway occurs between scytalone and vermelone. Accumulation of several metabolites derived from melanin precursors along branch pathways is associated with inhibition of melanin biosynthesis. At low tricyclazole concentrations (0.01–1 μg/ml), predominant accumulation of 2-hydroxyjuglone and 3,4-dihydro-3,4,8-trihydroxy-1-(2H)-naphthalenone (3,4,8-DTN) occurs as a result of the primary block between 1,3,8-trihydroxynaphthalene and vermelone. As the concentration of tricyclazole is increased from 1 to 10 μg/ml, flaviolin accumulation is markedly enhanced, whereas that of 3,4,8-DTN and 3,4-dihydro-4,8-dihydroxy-1-(2H)-naphthalenone is depressed, indicating possible secondary sites of inhibition in the main and branch pathways. Five melanin-deficient mutants of P. oryzae that phenotypically resemble the tricyclazole-treated wild-type strain were nonpathogenic or rarely infected two rice varieties. Three of the mutants studied were genetically defective in the melanin biosynthetic pathway at the site blocked by tricyclazole in the wild type. The wild-type strain converted both scytalone and vermelone to melanin; whereas the three mutants and the tricyclazole-treated wild type converted only vermelone to melanin. The data suggest a relationship between melanin biosynthesis and pathogenicity in P. oryzae.     

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).     

A coat protein transgene from a Scottish isolate of potato mop-top virus mediates strong resistance against Scandinavian isolates which have similar coat protein genes

Resistance tests were made on seedlings of transformed lines of Nicotiana benthamiana which contain a transgene encoding the coat protein (CP) gene of a Scottish isolate of potato mop-top virus (PMTV). This transgene has been reported to confer strong resistance to the PMTV isolate from which the transgene sequence was derived and also to a second Scottish isolate. Plants of lines of the transgenic N. benthamiana were as resistant to two Swedish and two Danish PMTV isolates as to a Scottish isolate, and of five lines tested, greater than 93.5% of transgenic plants were immune. The coat protein gene sequences of these four Scandinavian isolates were very similar to those of the two Scottish isolates. The greatest divergence between the isolates was three amino acid changes and there was less than 2% change in CP gene nucleotide sequence. It is concluded that the PMTV CP transgene used in these experiments could confer resistance against isolates from different geographical areas because it is becoming apparent that the CP genes of PMTV isolates are highly conserved.     

Can additional N fertiliser ameliorate the elevated CO2‐induced depression in grain and tissue N concentrations of wheat on a high soil N background?

Elevated CO₂ stimulates crop yields but leads to lower tissue and grain nitrogen concentrations [N], raising concerns about grain quality in cereals. To test whether N fertiliser application above optimum growth requirements can alleviate the decline in tissue [N], wheat was grown in a Free Air CO₂ Enrichment facility in a low‐rainfall cropping system on high soil N. Crops were grown with and without addition of 50–60 kg N/ha in 12 growing environments created by supplemental irrigation and two sowing dates over 3 years. Elevated CO₂ increased yield and biomass (on average by 25%) and decreased biomass [N] (3%–9%) and grain [N] (5%). Nitrogen uptake was greater (20%) in crops grown under elevated CO₂. Additional N supply had no effect on yield and biomass, confirming high soil N. Small increases in [N] with N addition were insufficient to offset declines in grain [N] under elevated CO₂. Instead, N application increased the [N] in straw and decreased N harvest index. The results suggest that conventional addition of N does not mitigate grain [N] depression under elevated CO₂, and lend support to hypotheses that link decreases in crop [N] with biochemical limitations rather than N supply.     

Level and Timing of Nitrogen Fertilizer Application to Early and Semi-early Potatoes (Solanum tuberosum L.) Grown with Irrigation on Light Soils in Norway

Trials were performed with early and semi-early potatoes to test the effects of nitrogen (N) fertilizer level (0-160 kg N ha^-1) and timing (all at planting versus half then and half either soon after emergence or 3 weeks later). All seven trials with earlies were irrigated as required, whilst different irrigation regimes (moderate versus intensive) were compared in two trials with semi-earlies. No benefit was derived from splitting the N application. Haulm growth and N uptake increased in all cases almost linearly up to the highest N level, but tuber yield did not respond in the same way. The optimum N level was 80 kg N ha^-1 for a yield of 15 Mg ha^-1, rising to 120 kg N ha^-1 for a yield of 40 Mg ha^-1. Tuber quality was lowered by the use of excess N fertilizer, particularly in the case of earlies. The quantity of mineralised N present in the soil after harvest rose sharply with above optimum fertilizer use, and the amount of N present in crop residues also increased. The likely leaching after early potatoes was estimated to be up to 80 kg N ha^-1. The proportion of fertilizer N which was not accounted for in either tuber yield, crop residues or mineral N in soil was 26% in earlies and 38% in semi-earlies.     

North American veterinary pathology residency training programs: an overview of where they are today and where they were five years ago, with an analysis of trends

An e-mail/telephone survey of all active North American residency training programs in veterinary pathology was conducted in September 2005. The purpose of this survey was to determine current numbers of trainees, their program length and type, and salaries; to compare current numbers to five years earlier; and, finally, to gauge interest in expanding current programs. All 41 training institutions contacted responded to the survey. Briefly, the survey found that there are currently 235 veterinary pathology residents, for a mean of 5.7 residents per training program. The number of residents currently in training programs and the number of applicants for these programs has increased compared to five years earlier. There is widespread interest in further expanding capacity in these programs, and the coalition of the American College of Veterinary Pathologists and the Society of Toxicologic Pathology is a well-known source of possible funding for additional residents. This survey report further documents the numbers of combined residency/PhD programs, average starting salaries for new residents, outside sponsorship effects on pathology training programs, and some of the common concerns regarding veterinary pathology training programs voiced by the respondents. While residency training capacity has expanded in the last five years, and there is widespread desire to further expand these training programs, a shortage of veterinary pathologists for future market needs will need to be addressed by increased funding from as yet unspecified sources.     

Modelling optimal timing and frequency of insecticide sprays for knockdown in preparation for other control measures

A modelling investigation was conducted into optimizing the number of sprays and inter-spray interval to reduce an insect population to a low level, for example, prior to pheromone trapping or the release of sterile males. The model population was age-structured and density-dependent. If spray mortality is 100% for each spray, then the ideal spraying schedule is easily determined from the durations of the various life stages. For spray mortality of less than 100%, a simulation was used to determine optimal spraying schedules. Relative length of the larval period, fertility rate and age to first oviposition were found to be the most important biotic parameters for this determination. Their importance is magnified as spray mortality decreases. The stage targeted by sprays and the percent mortality caused by each spray are also important in determining the required number of sprays. Using medfly (Ceratitis capitata Wiedmann) biotic parameters as an example when the spray targets adults, it appeared that neither the stage at which density-dependent mortality takes effect, nor the form of the adult survivorship curve are important in determining the optimal spray schedule.