Combining permanent aerobiological networks and molecular analyses for large-scale surveillance of forest fungal pathogens: A proof-of-concept

Forest disease management relies principally on a preventive approach in which epidemiological surveillance plays a crucial role. However, efficient and cost-effective surveillance methods are not currently available for large spatial scales. Nevertheless, aerobiological networks have been set up for several decades in many countries to monitor pollen dispersal and provide real-time assessments of allergenic risk. Here, we suggest that the same approach could be used for the surveillance of forest pathogens. Using molecular methods, we analysed samples from 12 sites of the French aerobiological network, at different dates. Both metabarcoding by high-throughput sequencing (using two markers and two different bioinformatics approaches) and real-time PCR targeting eight important forest pathogens were conducted. To validate the approach, temporal and spatial trends of spore detection were compared with field disease data. The metabarcoding approach demonstrated that many fungal plant pathogens could be found in aerobiological samples. Moreover, five of the eight targeted forest pathogens were detected by real-time PCR, with temporal and spatial trends of spore capture consistent with field data. In particular, Hymenoscyphus fraxineus was detected at high frequency in aerobiological samples in the areas where ash dieback has been present for the longest period of time, and at lower frequency in areas with more recent invasion. Spore detection of seasonal pathogens showed a temporal pattern similar to that of disease reports. Overall, our study provides a proof of concept that permanent aerobiological networks combined with molecular methods may provide a useful tool for large-scale surveillance of forest pathogens.     

Genetic Diversity of Pathogenic and Nonpathogenic Populations of Fusarium oxysporum Isolated from Carnation Fields in Argentina

ABSTRACT In order to elucidate the origin of Fusarium oxysporum f. sp. dianthi in Argentina, the genetic diversity among pathogenic isolates together with co-occurring nonpathogenic isolates on carnation was investigated. In all, 151 isolates of F. oxysporum were obtained from soils and carnation plants from several horticultural farms in Argentina. The isolates were characterized using vegetative compatibility group (VCG), intergenic spacer (IGS) typing, and pathogenicity tests on carnation. Seven reference strains of F. oxysporum f. sp. dianthi also were analyzed and assigned to six different IGS types and six VCGs. Twenty-two Argentinean isolates were pathogenic on carnation, had the same IGS type (50), and belonged to a single VCG (0021). The 129 remaining isolates were nonpathogenic on carnation and sorted into 23 IGS types and 97 VCGs. The same VCG never occurred in different IGS types. Our results suggest that the pathogen did not originate in the local populations of F. oxysporum but, rather, that it was introduced into Argentina. Given the genetic homogeneity within Argentinean isolates of F. oxysporum f. sp. dianthi, either IGS type or VCG can be used for the identification of the forma specialis dianthi currently in Argentina.     

Volatile flavor constituents of fruits from Southern Africa: Mobola plum (Parinari curatellifolia)

The volatile flavor components of Mobola plum (Parinari curatellifolia), a native fruit of Southern Africa, have been isolated by a vacuum headspace concentration method. The concentrate was analyzed by hyphenated gas chromatographic techniques, including gas chromatography (GC)/mass spectrometry (MS) and GC/Fourier transform infrared (FTIR), together with other GC-coupled detection devices for the specific and selective detection of nitrogen- and sulfur-containing compounds. A total of 88 components were identified. Of these compounds, 12 contain nitrogen, including 2-aminobenzaldehyde and phenylacetaldoxime, which are detected for the first time in an edible fruit. In addition, two unusual nitrated compounds have been identified, including optically active (2-nitrobutyl)benzene, which is a new natural product. Quantitative and sensory data of the new compounds are provided.     

Effect of vitamin E supplementation in milk replacer and Shiga toxoid vaccination on serum α‐tocopherol,performance, haematology and blood chemistry in male Holstein calves

Vitamin E (vit E), an essential antioxidant for maintaining the stability of biological membranes and the function of the immune system, is considered to support adaptive immune responses and performance in cattle. The principal virulence factor of Shiga toxin (Stx)‐producing Escherichia coli (STEC), the eponymous Stx, modulates cellular immune responses in cattle, the primary STEC reservoir. Active and passive immunization of calves with Shiga toxoids (rStx_MUT) was recently shown to reduce the STEC shedding. Here, we examined the influence of vit E on calves' serum α‐tocopherol, performance, haematology, blood chemistry and its interaction with rStx_MUT immunization. Data from calves having received passive (colostrum from immunized cows) and active (intramuscularly at 5th and 8th weeks of life) vaccination with rStx_MUT (n = 24) were compared to unvaccinated controls (n = 24; fed with low anti‐Stx colostrum, placebo injected). For each vaccination group, data were analysed according to the level of vit E supplementation offered by milk replacer (188 IU all‐rac‐α‐tocopheryl acetate daily [VitE_M] vs. 354 IU [VitE_H]). An increase by 79% in daily vit E supplementation led to slightly higher serum α‐tocopherol level and earlier concentrate intake at the beginning of the experiment without significant differences in live weight gain, haematology, blood chemistry parameters and peripheral CD4⁺ and CD8⁺ T‐cell subpopulations. rStx_MUT vaccination modulated the CD4⁺/CD8⁺ ratio irrespective of vit E supplementation but decreased concentrate intake in VitE_H in a time‐dependent manner. Results of our study indicate that an increase in daily vit E supplementation vastly fails to exert effects on laboratory parameters and growth performance. However, observed interactive effects of vit E supply and vaccination on the regulation of feed intake deserves further attention.     

Expansion of industrial logging in Central Africa

Industrial logging has become the most extensive land use in Central Africa, with more than 600,000 square kilometers (30%) of forest currently under concession. With use of a time series of satellite imagery for the period from 1976 to 2003, we measured 51,916 kilometers of new logging roads. The density of roads across the forested region was 0.03 kilometer per square kilometer, but areas of Gabon and Equatorial Guinea had values over 0.09 kilometer per square kilometer. A new frontier of logging expansion was identified within the Democratic Republic of Congo, which contains 63% of the remaining forest of the region. Tree felling and skid trails increased disturbance in selectively logged areas.     

Resistance to <Emphasis Type="Italic">Meloidogyne chitwoodi</Emphasis> Identified in Wild Potato Species

Meloidogyne chitwoodi (Columbia root-knot nematode, CRKN) can cause serious damage in potato production systems, decreasing tuber value in the fresh market and processing industries. Genetic resistance to CRKN was first identified from the wild diploid potato species Solanum bulbocastanum accession SB22 and was successfully introgressed into tetraploid potato breeding material. To expand the base of genetic resistance, 40 plant accessions representing nine wild potato species were screened for their resistance to M. chitwoodi. Greenhouse screening identified fifteen clones from S. hougasii, one clone from S. bulbocastanum, and one clone from S. stenophyllidium with moderate to high levels of resistance against three isolates of M. chitwoodi. Geographical mapping showed that the resistance sources identified in this and previous studies primarily originated in the states of Jalisco and Michoacán in west-central Mexico. These new sources of resistance will be introgressed into elite potato populations to facilitate the development of potato cultivars with durable resistance to M. chitwoodi.