Spatial patterns of the Citrus leprosis virus and its associated mite vector in systems without intervention

Leprosis is caused by the Citrus leprosis virus cytoplasmic type and is vectored by the mite Brevipalpus yothersi. Miticide applications, which cost $54 million annually, are based on inspection for the presence of mites. The aim of the present study was to characterize the spatial patterns of B. yothersi-infested trees and trees with leprosis symptoms for further improvement in sampling and disease control. The presence of mites and the occurrence of leprosis were assessed over two years in 1160 Valencia trees and 720 Natal trees in a commercial sweet orange grove in Sao Paulo State, Brazil. To assess the natural growth and dispersal of mites and leprosis, mite populations were not controlled during the experimental period. Maps of mite-infested trees and trees with leprosis symptoms were analysed at three different levels of spatial hierarchy using complementary methods, i.e. among adjacent trees within and across rows, within quadrats, and the strength and orientation of aggregation among quadrats. The study showed that the spatial patterns of virus-infected and mite-infested trees were different, with a strong aggregation pattern of trees with leprosis symptoms that increased over time. Conversely, the spatial pattern of B. yothersi showed randomness or weak aggregation at all three spatial hierarchical levels. Disease incidence increased steadily in plots of both cultivars, unlike in mite-infested trees where incidence fluctuated over time. These results have important implications for the development of better management strategies for leprosis. Sampling methods and action thresholds for mite control should consider primary disease inoculum in addition to the incidence of mites.     

Influence of temperature,humidity duration and growth stage on the infection and mycotoxin production by Fusarium langsethiae and Fusarium poae in oats

High occurrence of Fusarium poae (FP) and Fusarium langsethiae (FL) and their mycotoxins nivalenol (NIV) and T-2/HT-2 have been observed in Swiss oats. Early prediction of mycotoxin levels is important for farmers and the cereal industry to minimize the risk of contaminated food and feed. Therefore, climate chamber experiments were conducted to investigate the influence of different temperatures (10, 15, 20 °C) and durations (4, 8, 12 h) at 99% relative humidity (RH) on the infection of oats with FP and FL. In addition, to discover the most susceptible period of oats, artificial FL inoculations were conducted at different growth stages. Field experiments were performed to observe the dispersal of these fungal species within the field and to investigate the weather conditions that influence the dispersal. The climate chamber experiments revealed higher contamination with NIV and T-2/HT-2 in the 10 °C treatments and with a prolonged humidity duration of 12 h 99% RH. Inoculations of oat plants at early (DC 61) and mid (DC 65) anthesis, led to higher FL infection and T-2/HT-2 accumulation in the grains compared with treatments at earlier growth stages, which might be due to an increased susceptibility during anthesis. No indication for spore dispersal was observed in the field experiments. The results obtained, together with the cropping factors that influence infection and mycotoxin production, could be used as a first step in developing forecasting models to predict the contamination of oats with the mycotoxins NIV and T-2/HT-2.     

Multilocus sequence typing analysis of Italian Xanthomonas campestris pv. campestris strains suggests the evolution of local endemic populations of the pathogen and does not correlate with race distribution

Xanthomonas campestris pv. campestris (Xcc) is the causal agent of black rot in Brassicaceae. It is widespread in Italy and severe outbreaks occur under conditions that favour disease development. In this study a multilocus sequence typing approach (MLST) based on the partial sequence of seven loci was applied to a selection of strains representative of the main areas of cultivation and hosts. The aim was to investigate whether the long tradition of brassica crops in Italy has influenced the evolution of different Xcc populations. All loci were polymorphic; 14 allelic profiles were identified of which 13 were unique to Italian strains. Based on the seven loci, the most common genotype within the Italian Xcc strains (AP1) was also the most representative genotype found in worldwide Xcc strains. This genotype was included in a new clonal complex in addition to three other clonal complexes already identified in Xcc populations. The phylogenetic reconstruction using a concatenated dataset of four conserved protein-coding genes, dnaK, fuyA, gyrB and rpoD, showed that the Italian strains belonged to two genetic groups. Physiological races were also investigated for the first time in Italy. The race structure of Xcc was determined by inoculating eight differential Brassica lines belonging to five species and showed that, in Italy, race 4 is the most widespread, followed by races 1 and 6. No correlation was found between allelic profiles, host of isolation, geographical origin and races, although a prevalent race was identified within the same clonal complex.     

Genetic diversity and aggressiveness of Calonectria pteridis in Eucalyptus spp.

Calonectria leaf blight, caused by Calonectria pteridis, is currently one of the main foliar diseases in eucalypt plantations in Brazil. In warm and high rainfall regions, the disease can be a limiting factor for eucalypt production when planting susceptible genotypes. The most effective method for controlling this disease in the field is the use of resistant genotypes, which requires knowledge of the genetic variability and aggressiveness of the pathogen population for effective deployment of plant resistance. This work evaluated the genetic diversity and aggressiveness of C. pteridis populations obtained from infected eucalypt plants in Monte Dourado (Pará state) and Imperatriz (Maranhão state), Brazil. To study the genetic diversity, 16 ISSR primers were tested, five of which amplified polymorphic, reproducible and informative bands. Thirty-one closely related genotypes were identified from 84 isolates studied, indicating that the population has a low genetic diversity. The aggressiveness of seven isolates, selected according to geographic origin and their clustering in the ISSR-based dendogram, was determined by inoculation of a hybrid Eucalyptus grandis × E. urophylla clone under controlled conditions. Disease severity was assessed by both measuring the percentage of plant defoliation and assigning a score according to a diagrammatic scale of symptoms. A high correlation between the two evaluation methods was observed, which revealed significant differences in aggressiveness among the isolates. The diagrammatic scale is recommended for disease evaluation because results are obtained much faster, before the occurrence of severe defoliation. No correlation between clustering in the ISSR-based phylogenetic analysis and aggressiveness was observed.     

Cryptic species and population genetic structure of Plasmopara viticola in São Paulo State,Brazil

Downy mildew (Plasmopara viticola) is one of the most important diseases in grape-growing areas worldwide, including Brazil. To examine pathogen population biology and structure, P. viticola was sampled during the 2015/16 growing season from 516 lesions on nine grape cultivars in 11 locations in subtropical areas of São Paulo State, Brazil. For identification of cryptic species, a subsample of 130 isolates was subjected to cleaved amplified polymorphic sequence (CAPS) analysis, and for 91 of these isolates the ITS1 region was sequenced. These analyses suggest that the population of P. viticola in São Paulo State consists of a single cryptic species, P. viticola clade aestivalis. Seven microsatellite markers were used to determine the genetic structure of all 516 P. viticola isolates, identifying 23 alleles and 55 multilocus genotypes (MLGs). Among these MLGs, 34.5% were clonal and represented 93% of the isolates sampled. Four dominant genotypes were present in at least five different locations, corresponding to 65.7% of the isolates sampled. Genotypic diversity (Ĝ = 0.21–0.89) and clonal fraction (0.58–0.96) varied among locations (populations). Most populations showed significant deviation from Hardy–Weinberg expectations; in addition, excess of heterozygosity was verified for many loci. However, principal coordinate analysis revealed no clusters among locations and no significant isolation by distance was found, suggesting high levels of migration. The results indicate that downy mildew epidemics result from multiple clonal infections caused by a few genotypes of P. viticola, and reproduction of P. viticola in São Paulo State is predominantly asexual.     

P0 protein of cotton leafroll dwarf virus-atypical isolate is a weak RNA silencing suppressor and the avirulence determinant that breaks the cotton Cbd gene-based resistance

Cotton blue disease (CBD) is the most important disease present in cotton crops in South America and cotton leafroll dwarf virus (CLRDV) is the causal agent. The disease has been controlled by sowing cotton varieties resistant to CLRDV. However, in the 2009/10 growing season, an outbreak due to an atypical CLRDV isolate (CLRDV-at) occurred in northwest Argentina. Although CLRDV and CLRDV-at genomes are very closely related, the symptoms they produce in cotton plants are quite different. P0 is the most divergent protein between the isolates and in CLRDV is a silencing suppressor protein. This work characterized the silencing suppressor activity of the P0 protein encoded by CLRDV-at (P0^CL-at) and evaluated its role in Cbd-resistance break in cotton plants. It was demonstrated that P0^CL-at, despite having a mutation in the consensus of the F-box-like motif, was able to suppress local RNA silencing, but displayed lower activity than P0^CL. P0^CL and P0^CL-at showed no differences in the interaction with Gossypium hirsutum SKP1 orthologue (GSK1) and Nicotiana benthamiana SKP1 and both P0 proteins triggered destabilization of ARGONAUTE1. However, when the ability to enhance PVX symptoms was evaluated, P0^CL-at was shown to be a weaker pathogenicity factor than P0^CL in N. benthamiana. Interestingly, trans-expressed P0^CL-at enabled CLRDV to systemically infect CBD-resistant plants, and a chimeric CLRDV-P0^CL-at infectious clone succeeded in establishing infection in CBD-resistant cotton varieties with symptoms resembling those produced by CLRDV-at. These results strongly suggest that P0^CL-at is the avirulence (Avr) determinant involved in breaking cotton Cbd gene-based resistance.     

Hormetic UV-C seed treatments for the control of tomato diseases

Hormesis is a dose response phenomenon in which low, non-damaging doses of a stressor bring about a positive response in the organism undergoing treatment. Evidence is provided here that hormetic UV-C treatments of tomato seed can control disease caused by Botrytis cinerea, Fusarium oxysporum f. sp. lycopersici (FOL) and f. sp. radicis-lycopersici (FORL) on tomato (Solanum lycopersicum). Treating seeds with a 4 kJ m⁻² dose of UV-C significantly reduced both the disease incidence and progression of B. cinerea, with approximately 10% reductions in both on cv. Shirley. Disease severity assays for FOL and FORL on cv. Moneymaker showed dose-dependent responses: UV-C treatments of 4 and 6 kJ m⁻² significantly reduced the disease severity scores of FOL, whilst only the 6 kJ m⁻² showed significant reductions for FORL. To determine the effects of treatment on germination and seedling growth, UV-C doses of 4, 8 and 12 kJ m⁻² were performed on cv. Shirley. No negative impacts on germination or seedling growth were observed for any of the treatments. However, the 8 kJ m⁻² treatment showed significant biostimulation, with increases in seedling, root and hypocotyl dry weight of 11.4%, 23.1% and 12.0%, respectively, when compared to the control. Furthermore, significant increases in the root-mass fraction (10.6%) and root:shoot ratio (13.1%) along with a decrease in shoot-mass fraction (2.0%) indicates that the 8 kJ m⁻² treatment stimulated root growth to the greatest extent. There was no effect on hypocotyl and primary root length or the number of lateral roots, indicating no adverse effects to basic root architecture or seedling growth.     

Rhynchosporium leaf scald disease incidence: seed source and spatial pattern

A programme of field trials for the study of the winter barley–Rhynchosporium commune pathosystem is reported. The associated seedborne disease rhynchosporium leaf scald is regarded as having an important impact on barley yields. The analysis in this study relates to the impact of the seed source (commercial or farm-saved seed) on disease incidence and to the spatial pattern of rhynchosporium leaf scald disease incidence. Disease incidence data were calculated from field data recorded as disease severity. Mean disease incidence was higher in the crops grown from farm-saved seed than in those grown from commercial seed, although great agronomic significance cannot be attached to this result. The spatial pattern of rhynchosporium leaf scald disease incidence was characterized in terms of the binary power law (BPL) and was indicative of an aggregated pattern. Programme-wide BPL results were described using a novel phytopathological application of a random coefficients model. These results have application in field sampling for rhynchosporium leaf scald disease.     

Effect of apple cultivars and storage periods on the virulence of Neofabraea spp.

Bull’s eye rot is a typical quiescent postharvest apple disease in major fruit-growing areas. The susceptibility of different apple cultivars to Neofabraea spp. (N. vagabunda and N. malicorticis) was assessed, with Granny Smith showing the most resistance and Cripps Pink the most susceptibility. To assess the factors involved in conidial germination, Neofabraea spp. were grown on crude protein extracts (CPEs) collected from apple fruits at different storage periods. Fungal germ tube growth rate and pathogenic enzyme (cellulase and xylanase) activity were assessed. Results showed that CPEs collected after 2 and 4 months of storage progressively stimulated conidial germination and germ tube elongation, while a lesser effect was observed from CPEs after 1 month of storage. Xylanase proved to be the main degrading enzyme secreted by all the isolates, while cellulase was produced only by N. vagabunda isolates. Overall, the isolate ID02 was the most virulent, based on more rapid germ tube elongation and greater activity of the lytic enzymes.     

New Pseudococcidae species on Cupressus macrocarpa in Tunisia: first report of Planococcus vovae

In May 2017 and 2018, Planococcus vovae (Nasonov) (Hemiptera: Pseudococcidae) was collected in Hammam Sousse (Tunisia) on the ornamental plant Cupressus macrocarpa (Cupressaceae). The infestation was observed on needles which showed symptoms of desiccation and chlorosis. This is the first record of P. vovae in Tunisia and in North Africa.