Cross-Sample Validation Provides Enhanced Proteome Coverage in Rat Vocal Fold Mucosa

The vocal fold mucosa is a biomechanically unique tissue comprised of a densely cellular epithelium, superficial to an extracellular matrix (ECM)-rich lamina propria. Such ECM-rich tissues are challenging to analyze using proteomic assays, primarily due to extensive crosslinking and glycosylation of the majority of high Mr ECM proteins. In this study, we implemented an LC-MS/MS-based strategy to characterize the rat vocal fold mucosa proteome. Our sample preparation protocol successfully solubilized both proteins and certain high Mr glycoconjugates and resulted in the identification of hundreds of mucosal proteins. A straightforward approach to the treatment of protein identifications attributed to single peptide hits allowed the retention of potentially important low abundance identifications (validated by a cross-sample match and de novo interpretation of relevant spectra) while still eliminating potentially spurious identifications (global single peptide hits with no cross-sample match). The resulting vocal fold mucosa proteome was characterized by a wide range of cellular and extracellular proteins spanning 12 functional categories

Predicting light leaf spot (Pyrenopeziza brassicae) risk on winter oilseed rape (Brassica napus) in England and Wales, using survey, weather and crop information

Data from surveys of winter oilseed rape crops in England and Wales in growing seasons with harvests in 1987-99 were used to construct statistical models to predict, in autumn (October), the incidence of light leaf spot caused by Pyrenopeziza brassicae on winter oilseed rape crops the following spring (March/April), at both regional and individual crop scales. Regions (groups of counties) with similar seasonal patterns of incidence (percentage of plants affected) of light leaf spot were defined by using principal coordinates analysis on the survey data. At the regional scale, explanatory variables for the statistical models were regional weather (mean summer temperature and mean monthly winter rainfall) and survey data for regional light leaf spot incidence (percentage of plants with affected pods) in July of the previous season. At the crop scale, further explanatory variables were crop cultivar (light leaf spot resistance rating), sowing date (number of weeks before/after 1 September), autumn fungicide use and light leaf spot incidence in autumn. Risk of severe light leaf spot (> 25% plants affected) in a crop in spring was also predicted, and uncertainty in predictions was assessed. The models were validated using data from spring surveys of winter oilseed rape crops in England and Wales from 2000 to 2003, and reasons for uncertainty in predictions for individual crops are discussed.Peer reviewe

Effects of temperature and wetness duration on conidial infection, latent period and asexual sporulation of Pyrenopeziza brassicae on leaves of oilseed rape

Experiments in controlled environments were carried out to determine the effects of temperature and leaf wetness duration on infection of oilseed rape leaves by conidia of the light leaf spot pathogen, Pyrenopeziza brassicae. Visible spore pustules developed on leaves of cv. Bristol inoculated with P. brassicae conidia at temperatures from 4 to 20 degrees C, but not at 24 degrees C; spore pustules developed when the leaf wetness duration after inoculation was longer than or equal to approximately 6 h at 12-20 degrees C, 10 h at 8 degrees C, 16 h at 6 degrees C or 24 h at 4 degrees C. On leaves of cvs. Capricorn or Cobra, light leaf spot symptoms developed at 8 and 16 degrees C when the leaf wetness duration after inoculation was greater than 3 or 24 h, respectively. The latent period (the time period from inoculation to first spore pustules) of P. brassicae on cv. Bristol was, on average, approximately 10 days at 16 degrees C when leaf wetness duration was 24 h, and increased to approximately 12 days as temperature increased to 20 degrees C and to 26 days as temperature decreased to 4 degrees C. At 8 degrees C, an increase in leaf wetness duration from 10 to 72 h decreased the latent period from approximately 25 to 16 days; at 6 degrees C, an increase in leaf wetness duration from 16 to 72 h decreased the latent period from approximately 23 to 17 days. The numbers of conidia produced were greatest at 12-16 degrees C, and decreased as temperature decreased to 8 degrees C or increased to 20 degrees C. At temperatures from 8 to 20 degrees C, an increase in leaf wetness duration from 6 to 24 h increased the production of conidia. There were linear relationships between the number of conidia produced on a leaf and the proportion of the leaf area covered by 'lesions' (both log(10)-transformed) at different temperatures.Peer reviewe

Effects of stem canker (Leptosphaeria maculans) and light leaf spot (Pyrenopeziza brassicae) on yield of winter oilseed rape (Brassica napus) in southern England

The relationships between yield loss and incidence or severity of stem canker and light leaf spot in winter oilseed rape were analysed by correlation and regression analyses, using data from experiments at Rothamsted, England in 1992/93, 1994/95 and 1995/96. Growth stages (GS) 6,3/6,4 and 4,0/4,5 were identified as the critical points for relating percentage yield loss to stem canker and light leaf spot (on stems), respectively. Critical point (CP) and area under disease progress curve (AUDPC) models relating percentage yield loss to combined incidence or severity of stem canker and light leaf spot (stems) in each experiment were constructed by linear regression. There were no differences in the CP models for incidence between 1992/93, 1994/95 and 1995/96 experiments, or in the AUDPC models for incidence between 1992/93 and 1994/95 experiments. Therefore, a general CP model relating percentage yield loss (Delta Y) to combined incidence of stem canker (S-i) at GS 6,3/6,4 and light leaf spot (stems) (L-i) at GS 4,0/4,5 was constructed using data from the three experiments: Delta Y = 0.85 + 0.079S(i) + 0.065L(i) (R-2 = 43.7%, P < 0.001, 92 df). A general AUDPC model relating Delta Y to the AUDPC of combined incidence of stem canker (S-ia) from GS 5.7 to GS 6.5 and light leaf spot (stems) (L-ia) from GS 4.0 to GS 6.3 was constructed using data from the 1992/93 and 1994/95 experiments: Delta Y = 0.07 + 0.00096S(ia) + 0.0026L(ia) (R-2 = 43.6%, P < 0.001, 68 df). These two general yield-loss models were tested with data from Rothamsted in 1993/94 and Boxworth in 1992/93. The predictive accuracy of the CP model based on combined incidence of stem canker and light leaf spot (stems) was better than that of the AUDPC model. Yield losses predicted by summing the estimates from individual models for incidence of stem canker alone (GS 6,3/6,4) and light leaf spot alone (on leaves at GS 3,3) were greater than observed yield losses in experiments at Rothamsted in 1992/93, 1993/94, 1994/95 and 1995/96 and at Boxworth in 1992/93.Peer reviewe