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.     

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.     

Assessing the efficacy and impact of management of an invasive species in a protected area: Poa annua on sub‐Antarctic Macquarie Island

Plant eradication is difficult, particularly in remote, protected areas. The Southern Ocean Islands are very isolated and highly protected, but the flora contains many alien plants. Small restricted populations have been eradicated, but eradication of established species has proven difficult. A better understanding of the efficacy of control methods at sub‐Antarctic temperatures and their off‐target impacts may increase eradication success. With interest in controlling non‐native Poa annua in the region, we aimed to determine if physical and chemical methods can control P. annua (the sub‐Antarctic biotype) in sub‐Antarctic conditions and examined their impact on native plants. We quantified the effectiveness of physical control methods on P. annua in situ on sub‐Antarctic Macquarie Island through field‐based experiments and assessed their selectivity on P. annua compared with native grasses. We also quantified the effectiveness of several herbicides on P. annua at sub‐Antarctic temperatures and assessed their selectivity on native grasses. Of the four physical disturbance methods tested, none effectively reduced P. annua cover as one‐off treatments. Of the herbicide treatments, glyphosate and trifloxysulfuron sodium were effective and were less damaging to native grass species, indicating potential selectivity. Physical control was of limited effectiveness, but did not affect native species richness. An integrated weed management programme utilising the strategic use of selective herbicides with follow‐up chemical and physical intervention may balance control and biodiversity outcomes. This research highlights the importance of site‐specific testing of control methods and understanding off‐target impacts of control when managing alien plant species in protected areas.     

Evaluating diverse systems of tobacco genetic resistance to Phytophthora nicotianae in Yunnan,China

Black shank, caused by the soilborne pathogen Phytophthora nicotianae, is one of the most devastating diseases affecting tobacco production in China. The most effective strategy for reducing economic loss from this pathogen is development and use of resistant tobacco varieties. Multiple sources and systems of resistance have been developed in the Western Hemisphere; however, populations of P. nicotianae are variable around the world, including the predominance of different races. Different P. nicotianae isolates may react differently on tobacco plants with different systems of resistance, a possibility that could complicate the breeding of cultivars with resistance that is effective in different tobacco production regions worldwide. The objective of this research was to evaluate an array of tobacco germplasm possessing different systems of genetic resistance to black shank disease in tobacco-growing regions of Yunnan, China. Resistance types included simply inherited resistance mechanisms introgressed from wild Nicotiana relatives and polygenic partial resistance systems of N. tabacum origin. The loci of Wz exhibited high level resistance to black shank in the five diverse disease environments in Yunnan, China. K326 Php/−Wz/− genotype and Beinhart 1000 exhibited the greatest levels of resistance in both 2015 and 2016. Field observed results for 13 tobacco genotypes were highly correlated with those tested in growth chamber evaluation. These findings suggest that both Wz− and Beinhart 1000-mediated resistance have important commercial value in flue-cured tobacco breeding programmes in China. Cultivars developed for black shank resistance in China may also have utility in other tobacco-growing areas.     

Tomato brown rugose fruit disease: current distribution,knowledge and future prospects

Tomato is the most economically important fruit/vegetable crop grown worldwide. However, viral diseases remain an important factor limiting its productivity, with estimated quantitative and qualitative yield losses in tomato crops often reaching up to 100%. Many viruses infecting tomato have been reported, while new viral diseases have also emerged. The climatic changes the world is experiencing can be a contributing factor to the successful spread of newly emerging viruses, as well as the establishment of disease in areas that were previously either unfavourable or where the disease was absent. Because antiviral products are not available, strategies to mitigate viral diseases rely on genetic resistance/tolerance to infection, control of vectors, improvement in crop hygiene, roguing of infected plants and seed certification. Tomato brown rugose fruit virus (ToBRFV) is an emerging viral threat to tomato productivity and is currently spreading into new areas, which is of great concern to the growing global production in the absence of mitigation measures. This review presents the current knowledge about ToBRFV and future prospects for an improved understanding of the virus, which will be needed to support effective control and mitigation of the impact it is likely to cause.     

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.