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.     

An isolate of Wheat streak mosaic virus from foxtail overcomes Wsm2 resistance in wheat

Wheat streak mosaic virus (WSMV) is an economically important pathogen of wheat (Triticum aestivum) causing major yield losses in regions where severe infection occurs. To detect the presence of any new virus or new WSMV isolates, green foxtail (Setaria viridis) plants exhibiting virus-like symptoms were sampled in a summer-fallowed wheat field at the Agricultural Research Center-Hays, Kansas State University, Hays, Kansas. These plants were tested serologically for four wheat viruses: WSMV, Triticum mosaic virus (TriMV), High Plains wheat mosaic virus (HPWMoV) and Foxtail mosaic virus (FoMV). Among 38 plant samples exhibiting virus-like symptoms, 29 contained WSMV as indicated by ELISA. Four isolates from samples with relatively strong reactions were transferred to healthy wheat seedlings by mechanical inoculation in a growth chamber for pathogenicity testing. Three isolates were avirulent to a wheat variety RonL, which contains Wsm2, a gene providing temperature-sensitive resistance to currently prevalent isolates of WSMV. However, one isolate, KSH294, was able to infect RonL and showed more virulence on two other varieties/lines containing Wsm2. Further sequence and phylogenetic analysis of KSH294 confirmed that this isolate displays a sequence homology with WSMV, but has sequence differences making it distinct from previously identified WSMV isolates included in the phylogenetic analysis.     

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.     

Phytophthora oleae,a new root pathogen of wild olives

Wild olive (Olea europaea subsp. europaea var. sylvestris) is an important component of Mediterranean forests and a key genetic source for olive improvement programmes. Since 2009, a severe decline caused by Phytophthora cryptogea and P. megasperma has been detected in a protected wild olive forest of high ecological value (Dehesa de Abajo, Seville, Spain). In this natural forest, sampling of roots and soil was carried out on 25 wild olives with symptoms in 2014 and 2015. Apart from the already known P. cryptogea A1 and P. megasperma, a third Phytophthora species was consistently isolated from wild olive rootlets with symptoms. These isolates conformed morphologically with the newly described species P. oleae and were confirmed by analysis of their ITS regions and cox1 sequences. Temperature–growth relationships showed a maximum growth at 19.9 °C on carrot agar medium, making it the lowest temperature Phytophthora species infecting wild olive roots. Pathogenicity was confirmed on 1-year-old healthy wild olive seedlings and was similar to the previously known pathogenic phytophthoras. As temperature requirements are quite different, the three Phytophthora species may be active against wild olive roots in different seasons. However, the prevalence of P. oleae infecting wild olives in recent years could be due to its introduction as a new invasive pathogen. The probable invasive nature of P. oleae, together with increasing rain episodes concentrated in short periods frequent in southern Spain, would allow the outbreak of infections in wild olive forests, and also put cultivated olive orchards at risk.     

Exogenous application of melatonin improves plant resistance to virus infection

In this study, melatonin (MEL)-mediated plant resistance to tobacco mosaic virus (TMV) was examined to study local infection in Nicotiana glutinosa and systemic infection in Solanum lycopersicum. Exogenous application of 100 µm MEL increased anti-virus infection activity to 37.4% in virus-infected N. glutinosa plants. The same treatment significantly reduced relative levels of virus RNA analysed by qRT-PCR and virus titres measured by dot-ELISA, and increased the relative expression levels of the PR1 and PR5 genes analysed by qRT-PCR, in virus-infected S. lycopersicum. MEL treatment induced considerable accumulations of salicylic acid (SA) and nitric oxide (NO) but did not significantly affect production of hydrogen peroxide (H₂O₂) in the virus-infected S. lycopersicum plants. Transgenic nahG N. tabacum was used to determine whether MEL-induced TMV resistance was dependent on the SA pathway. The results showed that the relative RNA level of the TMV analysed by qRT-PCR and virus titres analysed by dot-ELISA were not reduced by the MEL treatment in the nahG transgenic N. tabacum seedlings treated twice with 100 µm MEL. The increased relative expression levels of PR1 and PR5 were greatly reduced when cPTIO, an NO scavenger, was included in the MEL treatment. A working model of MEL-mediated plant resistance to TMV is proposed. MEL-mediated plant resistance to viruses provides a new avenue to control plant viral diseases.     

Ray blight of pyrethrum in Australia: a review of the current status and future opportunities

Ray blight caused by Stagonosporopsis tanaceti is one of the most important diseases of pyrethrum (Tanacetum cinerariifolium), a perennial herbaceous plant cultivated for the extraction of insecticidal pyrethrins in Australia. The disease is responsible for complete yield loss in severe outbreaks. Infected seed is considered as the principal source of S. tanaceti. Infection hyphae remain only in the seed coat and not in the embryo, resulting in pre- and post-emergence death of seedlings and latent infection. Therefore, quantification of the level of infection by S. tanaceti within seed using a qPCR assay is important for efficient management of the disease. Stagonosporopsis tanaceti completes its life cycle within 12 days after leaf infection through production of pycnidia and can infect every tissue of the pyrethrum plant except the vascular and root tissues. Ray blight epidemics occur in pyrethrum fields through splash dispersal of pycnidiospores between adjacent plants. Besides steam sterilization, thiabendazole/thiram and fludioxonil are effective seed-treating chemicals in controlling S. tanaceti before planting begins. Ray blight is currently managed in the field through the foliar application of strobilurin fungicides in the first 1–2 years of crop establishment. Later on, difenoconazole and multisite specific fungicides in the next 2–3 years during early spring successfully reduce ray blight infestation. Avoiding development of resistance to fungicides will require more sustainable management of ray blight including the development and deployment of resistant cultivars.     

High vegetative compatibility diversity of Cryphonectria parasitica infecting sweet chestnut (Castanea sativa) in Britain indicates multiple pathogen introductions

Chestnut blight, caused by Cryphonectria parasitica, was identified in Devon, UK, in December 2016. Intensive surveys detected the disease at further sites in Devon (seven), Berkshire (one), Dorset (one), Derbyshire (four) and a cluster of eight sites in southeast London. Over 570 survey samples were tested, and 227 were positive for C. parasitica by isolation and real-time PCR. A total of 227 isolates were tested for mating type, and 197 screened for vegetative compatibility group (VCG) and compared with VCGs known from mainland Europe. The same isolates were also screened for the presence of Cryphonectria hypovirus 1 (CHV-1). Eleven VCGs were identified within the UK population. Five corresponded to already known European VCGs but six were unique. The European VCGs mainly came from the Devon, Dorset, Berkshire and Derbyshire disease outbreaks, whilst unique VCGs were almost exclusively from the southeast London cluster. Both mating types were detected, but only one mating type was present at each site, with the exception of a single Devon site. Perithecia of C. parasitica were never observed at any site. CHV-1 was found in seven isolates from three different locations and was always subtype-I, which has limited hypovirulence. Therefore, although CHV-1 is associated with C. parasitica at some outbreaks, it probably has limited impact on virulence. The diversity of VCGs and their distribution at outbreak sites, together with findings of CHV-1, suggests C. parasitica has been introduced to the UK multiple times over at least two decades through international plant trade.     

Synchytrium endobioticum – risk from biogas plants?

Synchytrium endobioticum is one of the most important pathogens of potato and is known for its persistent propagation structures. Under favourable conditions, infection of highly susceptible potato cultivars leads to clearly visible cauliflower‐like tissue warts, the typical symptom of potato wart disease. However, unfavourable infection conditions or low infection pressure may result in symptoms being overlooked. Thus, the introduction of pathogen structures into stages of the processing industry cannot be ruled out. As the amounts of processed potato products continue to rise, phytosanitary risks from processing discarded potatoes and potato waste in biogas plants have to be considered. Hence, the resilience of resting spores against mesophilic anaerobic digestion was analysed in stirred tank reactors. Laboratory‐scale results show that S. endobioticum not only withstands mesophilic anaerobic digestion but also subsequent storage of the digestate for at least 4 weeks. Large numbers of viable resting spores were detectable by microscopic assessment in all samples. Viability was proved and verified additionally by bioassay. Consequently, potatoes, potato waste and processing water from potato processing industries used in biogas plants pose a phytosanitary risk if the accruing digestates are returned as fertilizer to arable land.     

Selection,optimization and characterization of molecular tests for the detection of Tobacco ringspot virus (TRSV)

The aim of the study was to select, optimize and characterize RT‐PCR tests for the detection of Tobacco ringspot virus (TRSV) in post‐entry quarantine. Among five different tests, the Poojari et al. (Journal of Virological Methods 235, 112–118) and Jossey & Babadoost (Plant Disease 90, 1361) conventionnal RT‐PCR tests were chosen and optimized for this purpose. Ten TRSV isolates, 17 healthy plants from the genera Vitis, Prunus and Malus, four other Nepoviruses or Secoviridae isolates and serial dilutions of three TRSV isolates were used for the complete characterization of the optimized tests. In the tested conditions, the Poojari and Jossey & Babadoost tests respectively showed 100% and 95% inclusivity, 100% and 100% exclusivity, 100% and 96.4% analytical specificity, 100% and 100% diagnostic specificity, [10^?7; 10^?1] and [10^?8; 10^?2] limit of detection dilution factors (analytical sensitivity), 91.1% and 90.0% repeatability, 90.3% and 91.1% reproducibility and 100% and 100% selectivity. Due to its higher inclusivity, the optimized Poojari test is recommended for the detection of TRSV isolates for post‐entry quarantine purposes, but can also be used for global surveys. The optimized Jossey & Babadoost test showed the best analytical sensitivity, suggesting that combining both tests can further enhance detection.