The hop downy mildew pathogen Pseudoperonospora humuli.

Pseudoperonospora humuli is an obligate biotrophic oomycete that causes downy mildew, one of the most devastating diseases of cultivated hop, Humulus lupulus. Downy mildew occurs in all production areas of the crop in the Northern Hemisphere and Argentina. The pathogen overwinters in hop crowns and roots, and causes considerable crop loss. Downy mildew is managed by sanitation practices, planting of resistant cultivars, and fungicide applications. However, the scarcity of sources of host resistance and fungicide resistance in pathogen populations complicates disease management. This review summarizes the current knowledge on the symptoms of the disease, life cycle, virulence factors, and management of hop downy mildew, including various forecasting systems available in the world. Additionally, recent developments in genomics and effector discovery, and the future prospects of using such resources in successful disease management are also discussed. TAXONOMY: Class: Oomycota; Order: Peronosporales; Family: Peronosporaceae; Genus: Pseudoperonospora; Species: Pseudoperonospora humuli. DISEASE SYMPTOMS: The disease is characterized by systemically infected chlorotic shoots called "spikes". Leaf symptoms and signs include angular chlorotic lesions and profuse sporulation on the abaxial side of the leaf. Under severe disease pressure, dark brown discolouration or lesions are observed on cones. Infected crowns have brown to black streaks when cut open. Cultivars highly susceptible to crown rot may die at this phase of the disease cycle without producing shoots. However, foliar symptoms may not be present on plants with systemically infected root systems. INFECTION PROCESS: Pathogen mycelium overwinters in buds and crowns, and emerges on infected shoots in spring. Profuse sporulation occurs on infected tissues and sporangia are released and dispersed by air currents. Under favourable conditions, sporangia germinate and produce biflagellate zoospores that infect healthy tissue, thus perpetuating the infection cycle. Though oospores are produced in infected tissues, their role in the infection cycle is not defined. CONTROL: Downy mildew on hop is managed by a combination of sanitation practices and timely fungicide applications. Forecasting systems are used to time fungicide applications for successful management of the disease. USEFUL WEBSITES: https://content.ces.ncsu.edu/hop-downy-mildew (North Carolina State University disease factsheet), https://www.canr.msu.edu/resources/michigan-hop-management-guide (Michigan Hop Management Guide), http://uspest.org/risk/models (Oregon State University Integrated Plant Protection Center degree-day model for hop downy mildew), https://www.usahops.org/cabinet/data/Field-Guide.pdf (Field Guide for Integrated Pest Management in Hops).

The factors affecting a native obligate parasite, Cuscuta australis, in selecting an exotic weed, Humulus scandens, as its host.

In weed management, using native parasites to control exotic weeds is considered a better alternative than classical biological control. But the risk must be assessed because of the potential damage caused by these agents. We conducted this project to investigate the mechanism driving the choice of a native obligate parasite, Cuscuta australis, between the exotic, Humulus scandens, and native plants as its host through field and pot experiments. The results showed that C. australis preferred the exotic weed over native (naturalized) hosts and caused a notable reduction in the biomass of H. scandens in the field. In contrast, the results of the pot experimentindicated that C. australis preferred a mix of native (naturalized) hosts over the exotic weed. Both texperiments indicated that the parasitic preference of C. australis was induced more by light irradiance than plant water, carbon (C), nitrogen (N) and phosphorus (P) contents, indicating that the native parasite can only be used to control H. scandens when the exotic weed forms mono-cultures or dominates the community. Accordingly, induction and release of C. australis to control H. scandens should be conducted with great caution.

The Use of Metabolomics to Elucidate Resistance Markers against Damson-Hop Aphid.

Phorodon humuli (Damson-hop aphid) is one of the major pests of hops in the northern hemisphere. It causes significant yield losses and reduces hop quality and economic value. Damson-hop aphid is currently controlled with insecticides, but the number of approved pesticides is steadily decreasing. In addition, the use of insecticides almost inevitably results in the development of resistant aphid genotypes. An integrated approach to pest management in hop cultivation is therefore badly needed in order to break this cycle and to prevent the selection of strains resistant to the few remaining registered insecticides. The backbone of such an integrated strategy is the breeding of hop cultivars that are resistant to Damson-hop aphid. However, up to date mechanisms of hops resistance towards Damson-hop aphids have not yet been unraveled. In the experiments presented here, we used metabolite profiling followed by multivariate analysis and show that metabolites responsible for hop aroma and flavor (sesquiterpenes) in the cones can also be found in the leaves, long before the hop cones develop, and may play a role in resistance against aphids. In addition, aphid feeding induced a change in the metabolome of all hop genotypes particularly an increase in a number of oxidized compounds, which suggests this may be part of a resistance mechanism.

Scanning the European corn borer (Ostrinia spp.) genome for adaptive divergence between host-affiliated sibling species.

It has recently been shown that the European corn borer, a major pest of maize crops, is actually composed of two genetically differentiated and reproductively isolated taxa, which are found in sympatry over a wide geographical range in Eurasia. Each taxon is adapted to specific host plants: Ostrinia nubilalis feeds mainly on maize, while O. scapulalis feeds mainly on hop or mugwort. Here, we present a genome scan approach as a first step towards an integrated molecular analysis of the adaptive genomic divergence between O. nubilalis and O. scapulalis. We analysed 609 AFLP marker loci in replicate samples of sympatric populations of Ostrinia spp. collected on maize, hop and mugwort, in France. Using two genome scan methods based on the analysis of population differentiation, we found a set of 28 outlier loci that departed from the neutral expectation in one or the other method (of which a subset of 14 loci were common to both methods), which showed a significantly increased differentiation between O. nubilalis and O. scapulalis, when compared to the rest of the genome. A subset of 12 outlier loci were sequenced, of which 7 were successfully re-amplified as target candidate loci. Three of these showed homology with annotated lepidopteran sequences from public nucleotide databases.

Did the introduction of maize into Europe provide enemy-free space to Ostrinia nubilalis? Parasitism differences between two sibling species of the genus Ostrinia.

We examined whether maize offers enemy-free space (EFS) to its pest Ostrinia nubilalis, and may thereby have contributed to its divergence from the sibling species, Ostrinia scapulalis, feeding mainly on mugwort, when introduced into Europe five centuries ago. We collected Ostrinia larvae on maize (70 populations, 8425 individuals) and mugwort (10 populations, 1184 individuals) and recorded parasitism using both traditional (counting emerging parasitoids) and molecular methods (detection by specific polymerase chain reaction). The main parasitoid was Macrocentrus cingulum (Braconidae). On mugwort, parasitism was twice that on maize, and parasitoid-related mortality was 8 times higher. This suggests that maize affords substantial EFS to Ostrinia feeding on it. The lower Mortality:Infestation ratio in maize suggests that O. nubilalis' immune response might be stronger than that of O. scapulalis. If so, adapting to maize and diverging from O. scapulalis would decrease the impact of parasitism on O. nubilalis at both ecological and evolutionary levels.

Effects of powdery mildew fungicide programs on twospotted spider mite (Acari: Tetranychidae), hop aphid (Hemiptera: Aphididae), and their natural enemies in hop yards.

Twospotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), and hop aphid, Phorodon humuli (Schrank) (Hemiptera: Aphididae), are the most important arthropod pests of hop (Humulus lupulus L.) in the Northern Hemisphere. A potential barrier for greater adoption of conservation biological control strategies for spider mites and hop aphid is the extensive use of fungicides for management of hop powdery mildew, Podosphaera macularis (Wallr.:Fr.) U. Braun & S. Takamatsu. Field studies conducted in experimental plots in Oregon and Washington in 2005 and 2006 quantified the effects of powdery mildew fungicide programs (i.e., sulfur, paraffinic oil, and synthetic fungicides) on arthropod pests and natural enemies on hop. Fungicide treatment significantly affected spider mite populations in all four studies. Multiple applications of sulfur fungicides applied before burr development resulted in 1.4-3.3-fold greater spider mite populations during summer. Near the cessation of the sulfur applications, or after a lag of 20-30 d, spider mite populations increased significantly faster on sulfur treated plants compared with water-treated plants in three of four experiments. The effect of paraffinic oil on spider mites was varied, leading to exacerbation of spider mites in Oregon and Washington in 2005, suppression of mites in Oregon in 2006, and no significant effect compared with water in Washington in 2006. Significant relative treatment effects for cone damage due to spider mite feeding were detected in Oregon in 2005 in plots treated with sulfur and paraffinic oil compared with water and synthetic fungicides. Mean populations of hop aphids were similar among treatments in Oregon, although sulfur treatment suppressed hop aphid populations in Washington in 2005 and 2006. Populations of individual predacious insect species and cumulative abundance of macropredators were not consistently suppressed or stimulated by treatments in all trials. However, predatory mite abundance in Washington was affected by fungicide treatments, with plots treated with sulfur consistently having 10-fold fewer phytoseiids per leaf compared with the other treatments. Based on the results of these studies, powdery mildew fungicide programs that minimize or eliminate applications of sulfur and paraffinic oil would tend to conserve predatory mites and minimize the severity of spider mite outbreaks. However, mechanisms other than direct or indirect toxicity to phytoseiid mites likely are associated with exacerbation of spider mite outbreaks on hop.

Treating hop plants with (Z)-jasmone increases colonization by Phorodon humuli (Hemiptera: Aphididae) spring migrants.

Hop plants were sprayed with (Z)-jasmone, at a rate of 50 g ha(-1), during the spring migration of the damson-hop aphid Phorodon humuli (Schrank) in 2002 and 2003. Numbers of P. humuli spring migrants colonizing hop plants, Humulus lupulus L., 2-6 and 7-11 days after applying this treatment were assessed in both years. During the first five-day period, significantly more spring migrants were found on hop plants treated with (Z)-jasmone, in comparison with control plants, in 2002. By contrast, no significant difference was evident in the second five-day period. Although the migration in 2003 was much lighter than in 2002, greater numbers of migrants were again removed from treated plants. Indeed, more spring migrants were removed from plants sprayed with (Z)-jasmone in this year during both five-day periods (11 and 44%, respectively) compared with the 23% greater numbers removed in the first five-day period in 2002. Therefore, unlike some other species of aphid, where numbers were consistently lower on plots sprayed with (Z)-jasmone, spraying the secondary host of P. humuli with this compound appears to increase colonization by spring migrants.

Genetic isolation between two sympatric host plant races of the European corn borer, Ostrinia nubilalis Hubner. II: assortative mating and host-plant preferences for oviposition.

The European corn borer, Ostrinia nubilalis Hubner, colonized maize (Zea mays L.) after its introduction into Europe about 500 years ago and is now considered one of the main pests of this crop. In northern France, two sympatric host races have been described: one feeding on maize and the other on mugwort (Artemisia vulgaris L.) and hop (Humulus lupulus L.). In a previous study, we showed that mating between the two races may be impeded by differences in the timing of moth emergence and in the composition of the sex pheromone produced by the females. In this study, we further investigated the genetic isolation of these two races using strains from the maize (Z strain) and mugwort (E strain) races selected for diagnostic alleles at two allozyme loci. In a cage containing maize and mugwort plants and located in natural conditions, mating between individuals of the same strain occurred more often than mating between males and females of the E and Z strains. In particular, we obtained no evidence for crosses between Z females and E males. We also found that females of the Z strain laid their eggs almost exclusively on maize, whereas females of the E strain laid their eggs preferentially, but not exclusively, on mugwort. These results suggest that the genetic differentiation between the two host races may also be favored by host-plant preference, one of the first steps toward sympatric speciation.

Monitoring of insecticide resistance in damson hop aphid, Phorodon humuli Schrank (Hemiptera: Aphididae) from German hop gardens.

The damson hop aphid, Phorodon humuli (Schrank) (Hemiptera: Aphididae), is one of the most important sucking pests of many hop-growing areas world-wide. In this study we determined the efficacy of several insecticides against strains collected throughout the year 2001. All strains were collected in different hop gardens in the Hallertau (Bavaria), Germany, the largest hop-growing area of the world. First of all we established a leaf dip bioassay, carried out using six-well tissue culture plates and appropriate for monitoring susceptibility against imidacloprid, oxydemeton-methyl, cyfluthrin, amitraz, pymetrozine and pirimicarb. Four of these compounds, imidacloprid, cyfluthrin, pymetrozine and amitraz, are currently registered for the control of sucking pests in German hop gardens and are useful against P. humuli. The leaf-dip bioassay system turned out to be very reliable and robust. Ten P. humuli strains were collected in May 2001 and maintained in the laboratory to assess their resistance to the different insecticides in comparison with two laboratory reference strains (H2 and H5). Using diagnostic concentrations, resistance monitoring for imidacloprid and cyfluthrin was investigated during July and August 2001 on 53 populations from 30 sites around the Hallertau, an area of ca 2500 km2. Resistance to diagnostic concentrations (LC95 for reference strains) of imidacloprid, amitraz and pymetrozine was not detected in any strain received in 2001, but late-season (August) populations seemed to respond more heterogeneously than those collected mid-season (July). Overall composite mean mortalities to diagnostic concentrations of imidacloprid (13 mg litre(-1)) in collections from May, July and August were 95 (+/-2.5), 98 (+/-2.3) and 87 (+/-5.9)%, respectively. Moderate resistance to pyrethroids was observed in all strains collected in May and August using a diagnostic concentrations of 10 mg litre(-1) cyfluthrin (LC95 of the susceptible reference strain H5). Slight to moderate resistance to diagnostic concentrations of oxydemeton-methyl and pirimicarb was observed in some, but not all, strains collected early season. The results are discussed in terms of the implemention of hop aphid resistance management strategies in German hop-cultivation areas.

Host-plant-associated genetic differentiation in Northern French populations of the European corn borer.

The phytophagous insects that damage crops are often polyphagous, feeding on several types of crop and on weeds. The refuges constituted by noncrop host plants may be useful in managing the evolution in pest species of resistance to the Bacillus thuringiensis toxins produced by transgenic crops. However, the benefits of these refuges may be limited because host-plant diversity may drive genetic divergence and possibly even host-plant-mediated sympatric speciation. The European corn borer, Ostrinia nubilalis Hübner (Lepidoptera: Crambidae), is the main pest of maize in Europe and North America, where it was introduced early in the 20th century. It has a wide host range but feeds principally on mugwort (Artemisia vulgaris L.) and maize (Zea mays L.). O. nubilalis is found on mugwort only in the northern part of France, whereas it is found on maize throughout France. The extent of genetic variation at allozyme markers was investigated in populations collected from the two host plants over the entire geographical distribution of the European corn borer on mugwort in France. Allelic differentiation between pairs of populations and hierarchical analyses of pools of samples from each host plant indicate that the group of populations feeding on maize differed from the group of populations feeding on mugwort. Our results suggest (1) host-plant-related divergent selection at the genomic region surrounding the Mpi locus and (2) limited gene flow between the populations feeding on mugwort and those infesting maize fields. These data indicate that adults emerging from mugwort would not be useful for managing the evolution of resistance to the B. thuringiensis toxins in European corn borer populations.

Toxicity of imidacloprid to Galendromus occidentalis, Neoseiulus fallacis and Amblyseius andersoni (Acari: Phytoseiidae) from hops in Washington State, USA.

The toxicity of systemic and spray formulations of imidacloprid to Galendromus occidentalis Nesbitt, Neoseiulus fallacis Garman and Amblyseius andersoni (Chant) from hop yards in Washington State was evaluated in laboratory bioassays. The field rate of imidacloprid for hop aphids (0.13 g a.i. l) was highly toxic (100% mortality) to G. occidentalis and N. fallacis but less so (35.6% mortality) to A. andersoni. Half and quarter rates were also highly toxic to G. occidentalis and N. fallacis (79.5-100% mortality) but again had lower toxicity to A. andersoni (8.2-31.3% mortality). Systemic toxicity (via consumption of spider mite motiles feeding on leaf discs cut from imidacloprid-treated (0.13 g a.i. l) dwarf bean plants) was also high for G. occidentalis (98.3% mortality), as was toxicity from dried residues (93-98% mortality). Residual toxicity to N. fallacis was also high (89% mortality). The significance of these results for biological control of spider mites in hops and other crops is discussed.