Cost-benefits of reduced aphicide usage on dwarf hops susceptible and partially resistant to damson-hop aphid

Soil‐applied imidacloprid at full (125 g a.i. ha^?1) and half approved doses gave levels of control of damson‐hop aphid, (Phorodon humuli), similar to that provided by foliar spray(s) of tebufenpyrad on the aphid‐susceptible dwarf hop cvs First Gold and Herald. On those cultivars, aphid control was unreliable on plots treated with quarter dose imidacloprid and was generally no better than on untreated plants. Aphids were virtually eliminated from the leaves by the end of July each year in all treatments consistent with the action of natural enemies. Aphid contamination of cones reflected the numbers on foliage at flowering time, but varied widely between years. Yields and percentage α‐acids content of dried hops were unaffected by the numbers of aphids on leaves early in the season and in cones at harvest, but aphid contamination reduced the economic values of crops by as much as 80%. Few P. humuli colonised the partially aphid‐resistant breeding line 23/90/08 before their numbers were regulated and consistent with natural enemy activity. Yields, percentage α‐acids content, and commercial value of harvested cones were similar in all treatments on 23/90/08 whether or not plants were treated with aphicides. The commercial risks posed by P. humuli preclude substantial reductions in aphicide usage on aphid‐susceptible dwarf hop cultivars, but future cultivars expressing a similar level of partial resistance to aphids as 23/90/08 should not need treatment with aphicides.     

Simultaneous modification of three homoeologs of TaEDR1 by genome editing enhances powdery mildew resistance in wheat

Wheat (Triticum aestivum L.) incurs significant yield losses from powdery mildew, a major fungal disease caused by Blumeria graminis f. sp. tritici (Bgt). enhanced disease resistance1 (EDR1) plays a negative role in the defense response against powdery mildew in Arabidopsis thaliana; however, the edr1 mutant does not show constitutively activated defense responses. This makes EDR1 an ideal target for approaches using new genome‐editing tools to improve resistance to powdery mildew. We cloned TaEDR1 from hexaploid wheat and found high similarity among the three homoeologs of EDR1. Knock‐down of TaEDR1 by virus‐induced gene silencing or RNA interference enhanced resistance to powdery mildew, indicating that TaEDR1 negatively regulates powdery mildew resistance in wheat. We used CRISPR/Cas9 technology to generate Taedr1 wheat plants by simultaneous modification of the three homoeologs of wheat EDR1. No off‐target mutations were detected in the Taedr1 mutant plants. The Taedr1 plants were resistant to powdery mildew and did not show mildew‐induced cell death. Our study represents the successful generation of a potentially valuable trait using genome‐editing technology in wheat and provides germplasm for disease resistance breeding.     

Influence of hop yard ground flora on invertebrate pests of hops and their natural enemies

A 3‐year study in Washington State, USA examined the influence of hop yard ground flora both on the invertebrate pests of hops [including hop aphid (Phorodon humuli Schrank), two‐spotted spider mite (Tetranychus urticae Koch) and hop looper (Hypena humuli Harris)] and on their natural enemies. Each year half of the experimental plots were sown with a mixture of ‘insectary’ plants [including California poppy (Eschscholzia californica von Chamisso), dwarf cornflower (Centaurea cyanus L.) and buckwheat (Fagopyrum esculentum Moench)]. Season‐long average cover provided by the flowering mixture ranged from 2% to 26%, with some blooms being present for most of the season in each year. In terms of vegetation, the other main differences between cover‐cropped and control plots were in the proportions of bare soil (which was always significantly higher in control plots) and the percentage of Chenopodium album L., which was significantly higher in the cover‐cropped plots in two of the three seasons. Populations of spider mites on hop foliage were significantly lower in cover‐cropped plots than in control plots in 2 of 3 years, while mite numbers in the cones at harvest were generally low and usually did not differ between treatments. Responses of hop aphids were much more variable, with no consistent treatment effect on either leaf or cone infestations. Larval populations of hop looper tended to be higher in cover‐cropped plots in the first two generations, but usually not in the third (final) generation. Of the beneficial invertebrates, spiders and nabids consistently reached higher average densities in the ground flora of cover‐cropped plots than in control plots in all 3 years, while anthocorids, geocorids and parasitic hymenoptera reached higher mean densities in the former plots in 2 of 3 years. Adult coccinellids were higher in cover‐cropped plots only in 2006.     

DOWNY MILDEW RESISTANT 6 and DMR6‐LIKE OXYGENASE 1 are partially redundant but distinct suppressors of immunity in Arabidopsis

Arabidopsis downy mildew resistant 6 (dmr6) mutants have lost their susceptibility to the downy mildew Hyaloperonospora arabidopsidis. Here we show that dmr6 is also resistant to the bacterium Pseudomonas syringae and the oomycete Phytophthora capsici. Resistance is accompanied by enhanced defense gene expression and elevated salicylic acid levels. The suppressive effect of the DMR6 oxygenase was confirmed in transgenic Arabidopsis lines overexpressing DMR6 that show enhanced susceptibility to H. arabidopsidis, P. capsici, and P. syringae. Phylogenetic analysis of the superfamily of 2‐oxoglutarate Fe(II)‐dependent oxygenases revealed a subgroup of DMR6‐LIKE OXYGENASEs (DLOs). Within Arabidopsis, DMR6 is most closely related to DLO1 and DLO2. Overexpression of DLO1 and DLO2 in the dmr6 mutant restored the susceptibility to downy mildew indicating that DLOs negatively affect defense, similar to DMR6. DLO1, but not DLO2, is co‐expressed with DMR6, showing strong activation during pathogen attack and following salicylic acid treatment. DMR6 and DLO1 differ in their spatial expression pattern in downy mildew‐infected Arabidopsis leaves; DMR6 is mostly expressed in cells that are in contact with hyphae and haustoria of H. arabidopsidis, while DLO1 is expressed mainly in the vascular tissues near infection sites. Strikingly, the dmr6‐3_dlo1 double mutant, that is completely resistant to H. arabidopsidis, showed a strong growth reduction that was associated with high levels of salicylic acid. We conclude that DMR6 and DLO1 redundantly suppress plant immunity, but also have distinct activities based on their differential localization of expression.     

Host plant recognition in monophagous weevils: Specialization ofCeutorhynchus inaffectatus to glucosinolates from its host plantHesperis matronalis

Several aphid honeydews were incorporated into sucrose solutions and presented to hop aphids, Phorodon humuli (Schrank), as artificial diets in free-choice bioassays. Small additions of honeydew collected from two species of aphid feeding on hop, Humulus lupulus L., arrested the searching behavior of the hop aphid and appeared to stimulate prolonged periods of ingestion. This effect was more dependent on the host plant honeydew source than the species of aphid that produced the honeydew. Aphid honeydews collected from plants other than hop (non-hosts to P. humuli) contained hop aphid phagostimulants that were less effective. Our results indicate that analysis of aphid honeydew could help describe chemical cues involved in the recognition of appropriate host plants by aphid species.     

Sunflower resistance to multiple downy mildew pathotypes revealed by recognition of conserved effectors of the oomycete Plasmopara halstedii

Over the last 40 years, new sunflower downy mildew isolates (Plasmopara halstedii) have overcome major gene resistances in sunflower, requiring the identification of additional and possibly more durable broad‐spectrum resistances. Here, 354 RXLR effectors defined in silico from our new genomic data were classified in a network of 40 connected components sharing conserved protein domains. Among 205 RXLR effector genes encoding conserved proteins in 17 P. halstedii pathotypes of varying virulence, we selected 30 effectors that were expressed during plant infection as potentially essential genes to target broad‐spectrum resistance in sunflower. The transient expression of the 30 core effectors in sunflower and in Nicotiana benthamiana leaves revealed a wide diversity of targeted subcellular compartments, including organelles not so far shown to be targeted by oomycete effectors such as chloroplasts and processing bodies. More than half of the 30 core effectors were able to suppress pattern‐triggered immunity in N. benthamiana, and five of these induced hypersensitive responses (HR) in sunflower broad‐spectrum resistant lines. HR triggered by PhRXLRC01 co‐segregated with Pl22 resistance in F3 populations and both traits localized in 1.7 Mb on chromosome 13 of the sunflower genome. Pl22 resistance was physically mapped on the sunflower genome recently sequenced, unlike all the other downy mildew resistances published so far. PhRXLRC01 and Pl22 are proposed as an avirulence/resistance gene couple not previously described in sunflower. Core effector recognition is a successful strategy to accelerate broad‐spectrum resistance gene identification in complex crop genomes such as sunflower.     

Biology of the hop cyst nematode Heterodera humuli

In an intensively sampled English hop garden, cysts of Heterodera humuli were found in every sample taken. Most occurred in the top 15 cm of soil, decreasing in number by half with every 15 cm in depth. Numbers of second-stage juveniles fluctuated during the vegetative period in a pattern which indicated that more than one generation, possibly two to three, were produced. The generation time was about 1.5 months. In pot experiments under controlled conditions (20°C; 16 h light/day) the life cycle lasted 40 days. The optimal temperature was 20°C for the development of H. humuli in the roots of its host Humulus lupulus. Most H. humuli juveniles invaded hop roots at 15°C, but egg hatch was greatest at 20°C. In moist soil, second-stage juveniles survived for at least 54 days and thereafter invaded roots and reproduced.     

Survival, growth, and reproduction of two aphid species on sucrose solutions containing host or non-host honeydews

Aphids, like most phloem-feeding insects, commonly exhibit a high degree of host specificity. Plant-specific chemical compounds are likely to serve as important host selection cues for monophagous aphids and such substances could be present in aphid honeydew. Apterous virginoparae ofMyzus persicae (Sulzer) andPhorodon humuli (Schrank) were reared on a buffered sucrose solution containing various aphid honeydews or a mixture of amino acids. In two separate experiments, the host-specificP. humuli (hop aphid) could grow and reproduce only on diets containing honeydew collected from hop (Humulus lupulus L.).M. persicae (the green peach aphid, GPA) did not perform well on diets containing hop honeydew, perhaps because hop is a poor GPA host. Honeydew collected from preferred GPA host plants rape,Brassica napus L., and jimsonweed,Datura stramonium L., allowed GPA growth and reproduction. Hop aphids, however, performed poorly on rape and jimsonweed honeydew diets. Bell pepper,Capsicum annuum L., honeydew supported neither the hop aphid nor GPA. The study of aphid honeydew components may contribute towards a more complete understanding of host preference and selection phenomena in aphids.     

Secondary metabolites and plant/environment interactions: a view through Arabidopsis thaliana tinged glasses

Arabidopsis thaliana is a successful model plant for studying wide‐ranging topics including plant development, genetics and pathogen resistance. In addition, significant research has been conducted in the area of secondary metabolite biochemical genetics. The secondary metabolites in Arabidopsis include glucosinolates, terpenoids, phenylpropanoids, the alkaloid‐like camalexin, and other uncharacterized compounds. The genetic tools developed in studying secondary metabolite biochemistry are now being used to study how secondary metabolites control various biological processes. This includes compounds involved in plant/insect and plant/pathogen interactions, compounds preventing UV‐B damage, and compounds involved in hormone homeostasis. This review will describe what light Arabidopsis is shedding on the biological and ecological importance of specific secondary metabolites.     

Overwintering of Sphaerotheca humuli, the cause of hop powdery mildew

The ability of Sphaerotheca humuli to overwinter as cleistocarps in infected hop cones and leaves and in aerial buds on rootstocks was examined during the winters of 1970-1, 1971-2 and 1972-3. Periodical examination of cleistocarps, collected in October and overwintered in Terylene bags on the soil of a hop garden, consistently revealed two periods of maturation ending in November and in March, when over 50% contained eight, well-defined ascospores. In laboratory tests cleistocarps, kept either in the hop garden or dry at 4, 8 or 18^oC during the winter, could not be encouraged to dehisce earlier than April when naturally dehisced cleistocarps were first detected in the field. More ascospores were discharged from cleistocarps, and germination of ascospores in laboratory tests was greater, at 18 than at 4, 8 or 24^oC. Colonies of S. humuli arose on leaves of potted plants exposed to overwintered cleistocarps in the hop garden and were observed microscopically to originate from ascospores. However, a Burkard spore trap, operated amidst the cleistocarps in this garden in 1972 and 1973, failed to detect ascospores. Ascospores, discharged onto susceptible leaves in the laboratory, germinated but failed to produce colonies. It was demonstrated that S. humuli can perennate in aerial, dormant buds on hop rootstocks. Examination of buds in autumn revealed mycelium external to and between the bud scales. At budburst the mycelium was still present internally. Cleistocarps were occasionally associated with hibernating mycelium. Primarily infected shoots arose from plants bearing infected buds in conditions which precluded chance infection. Some evidence was obtained that conditions during the winter determine the success of survival in buds. The fungus appeared to be incapable of infecting a selection of weeds common to hop gardens and their vicinity.     

Susceptibility of hop to Phorodon humuli

A portion of the USDA World Hop (Humulus lupulus L.) Germ Plasm Collection was screened for antibiosis to the hop aphid, Phorodon humuli (Shrank). Hop aphid development and reproduction on the retained accessions 58016, 21090M, 21211, 21240, and 60038 were then compared to hop aphids reared on the commercial hop cultivars (cvs) L-8, Perle, Chinook, Galena, Eroica, Willamette, and Cascade. In general, hop aphids matured faster and reproduced more on the commercial cvs than on 21090M, 58016, 21211, or 21240. Accession 60038, of note because it supported unusually high numbers of aphids during screening, was found to be the most susceptible of any hop tested. Mortality and natality schedules were then combined to construct matrix models of aphid population growth on 60038, 58016, Cascade, and Perle (the most and least susceptible noncommercial and commercial hops, respectively). Beginning with one one-day-old nymph per leaf, the models predicted that P. humuli would require 24 days to surpass an action threshold of 100 aphids per leaf on 58016. This was seven days longer than on the susceptible Cascade and five days longer than on Perle. A commercial hop with the antibiosis of 58016 would probably require fewer insecticide applications during the growing season, thus retarding the development of insecticide resistance in P. humuli and enhancing the effectiveness of integrated management programs by protecting beneficial insects.

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Zusammenfassung Ein Teil der Genbank des USDA von Hopfen (Humulus lupulus L.) wurde auf Resistenz gegen Phorodon humuli (Shrank) getestet. Dabei wurden die besonders widerstands fähigen Klone 58016, 21090M, 21211, 21240 sowie Klon 60038, der ungewöhnlich viele Blattläuse ertrug, ausgewählt. Anschließend wurde die Entwicklung und Vermehrung der Blattläuse auf den ausgewählten Klonen und auf den Handelssorten (cvs) L-8, Perle, Chinook, Galena, Eroica, Willamette, Cascade untersucht. Im Allgemeinen entwickelten sich die Blattläuse auf den Handelssorten reascher und vermehrten sich stärker als auf den resistenten Klonen. Klon 60038 war dagegen die anfälligste Sorte. Mortalitäts- und Natlitätsdaten wurden kombiniert um Matrixmodelle des Populationswachstums der Blattläuse auf 60038, 58016, Cascade und Perle (die am meisten und am wenigsten anfälligen kommerziellen und nichtkommerziellen Sorten) zu konstruieren. Die Modelle begannen bei 1 Larve pro Blatt und sagten voraus, dass die Aktionsgrenze von 100 Tieren pro Blatt auf 58016 nach 24 Tagen erreicht wird. Das ist 7 Tage später als auf den anfälligen Sorten Cascade und 5 Tage später als auf Perle. Eine kommerzielle Sorte mit der Anfälligkeit von 58016 würde vermutlich während der Vegetationszeit weniger Insektizidspritzungen benötigen. Damit würde die Resistenzentwicklung bei Phorodon humuli verlangsamt und die Wirkung von IPM Programmen durch Schutz der natürlichen Feinde verbessert.

     

Insecticide resistance in damson-hop aphid, Phorodon humuli in commercial hop gardens in Kent

From 1966 to 1976, samples of Phorodon humuli were collected annually from five commerical hop gardens in Kent and from other hop gardens where problems in control occurred. A susceptible stock was obtained from wild hop growing in northern England in 1969. The samples were cultured in isolation on potted hops and bioassayed against insecticides in common use. The level of resistance to demeton-S-methyl was c. 10X in 1966 after 10 yr use, and more than doubled from 1968–1974 apparently due to the spread of a more resistant type; there was a further increase to c. 50X in 1975–1976. There was also resistance of C. 20–30X to omethoate, 2–7X to methidathion and 4–8X to methomyl. Assays and field results show an increased resistance to methidathion and less certainly to methomyl after 5 yr use. There was no clear change in response to endosulfan. The LC₅₀'s estimated from a single dose and a mean probit slope were found to agree satisfactorily with the LC₅₀'s calculated from serial doses and so should be adequate for monitoring trends in resistance.     

Behavior and Population Development of <Emphasis Type="Italic">Phorodon humuli</Emphasis> (Schrank) (Homoptera: Aphididae) on Two Hop Cultivars of Different Susceptibility

The two hop cultivars “Hallertauer Magnum” (HM) and “Spalter Select” (SE) are regarded by growers as extremely different in their susceptibility to the damson-hop aphid Phorodon humuli (Schrank). To investigate these anecdotal observations, spring migration and initial population development of P. humuli were monitored on the two cultivars in 1998 and 1999 in an experimental hop garden. Numbers of migrant aphids on SE were significantly lower, comprising 18.8 and 30.2% as compared to HM in 1998 and 1999, respectively. Population development of apterous aphids on these two cultivars differed significantly. At the end of the monitoring period numbers of aphids on SE were 7.5 and 14.2% as compared to HM in 1998 and 1999, respectively. In behavioral studies of P. humuli alates released on glasshouse plants, those on SE spent significantly more time in motile behavior patterns than aphids on HM. In the glasshouse, population development also differed significantly and the number of aphids developing on SE was 12.9% of that on HM after 28 days. It is concluded that SE exhibits a certain repellent effect on P. humuli and, compared to HM, is possibly nutritionally less suitable to the aphid.     

Identification and characterization of a fusarium head blight resistance gene TaACT in wheat QTL‐2DL

Fusarium head blight (FHB) resistance in wheat is considered to be polygenic in nature. Cell wall fortification is one of the best resistance mechanisms in wheat against Fusarium graminearum which causes FHB. Metabolomics approach in our study led to the identification of a wide array of resistance‐related (RR) metabolites, among which hydroxycinnamic acid amides (HCAAs), such as coumaroylagmatine and coumaroylputrescine, were the highest fold change RR metabolites in the rachis of a resistant near‐isogenic line (NIL‐R) upon F. graminearum infection. Placement of these metabolites in the secondary metabolic pathway led to the identification of a gene encoding agmatine coumaroyl transferase, herein referred to as TaACT, as a candidate gene. Based on wheat survey sequence, TaACT was located within a FHB quantitative trait loci on chromosome 2DL (FHB QTL‐2DL) between the flanking markers WMC245 and GWM608. Phylogenetic analysis suggested that TaACT shared closest phylogenetic relationship with an ACT ortholog in barley. Sequence analysis of TaACT in resistant and susceptible NILs, with contrasting levels of resistance to FHB, led to the identification of several single nucleotide polymorphisms (SNPs) and two inversions that may be important for gene function. Further, a role for TaACT in FHB resistance was functionally validated by virus‐induced gene silencing (VIGS) in wheat NIL‐R and based on complementation studies in Arabidopsis with act mutant background. The disease severity, fungal biomass and RR metabolite analysis confirmed TaACT as an important gene in wheat FHB QTL‐2DL, conferring resistance to F. graminearum.     

The Lr34 adult plant rust resistance gene provides seedling resistance in durum wheat without senescence

The hexaploid wheat (Triticum aestivum) adult plant resistance gene, Lr34/Yr18/Sr57/Pm38/Ltn1, provides broad‐spectrum resistance to wheat leaf rust (Lr34), stripe rust (Yr18), stem rust (Sr57) and powdery mildew (Pm38) pathogens, and has remained effective in wheat crops for many decades. The partial resistance provided by this gene is only apparent in adult plants and not effective in field‐grown seedlings. Lr34 also causes leaf tip necrosis (Ltn1) in mature adult plant leaves when grown under field conditions. This D genome‐encoded bread wheat gene was transferred to tetraploid durum wheat (T. turgidum) cultivar Stewart by transformation. Transgenic durum lines were produced with elevated gene expression levels when compared with the endogenous hexaploid gene. Unlike nontransgenic hexaploid and durum control lines, these transgenic plants showed robust seedling resistance to pathogens causing wheat leaf rust, stripe rust and powdery mildew disease. The effectiveness of seedling resistance against each pathogen correlated with the level of transgene expression. No evidence of accelerated leaf necrosis or up‐regulation of senescence gene markers was apparent in these seedlings, suggesting senescence is not required for Lr34 resistance, although leaf tip necrosis occurred in mature plant flag leaves. Several abiotic stress‐response genes were up‐regulated in these seedlings in the absence of rust infection as previously observed in adult plant flag leaves of hexaploid wheat. Increasing day length significantly increased Lr34 seedling resistance. These data demonstrate that expression of a highly durable, broad‐spectrum adult plant resistance gene can be modified to provide seedling resistance in durum wheat.     

Tryptophan‐derived secondary metabolites in Arabidopsis thaliana confer non‐host resistance to necrotrophic Plectosphaerella cucumerina fungi

A defence pathway contributing to non‐host resistance to biotrophic fungi in Arabidopsis involves the synthesis and targeted delivery of the tryptophan (trp)‐derived metabolites indol glucosinolates (IGs) and camalexin at pathogen contact sites. We have examined whether these metabolites are also rate‐limiting for colonization by necrotrophic fungi. Inoculation of Arabidopsis with adapted or non‐adapted isolates of the ascomycete Plectosphaerella cucumerina triggers the accumulation of trp‐derived metabolites. We found that their depletion in cyp79B2 cyp79B3 mutants renders Arabidopsis fully susceptible to each of three tested non‐adapted P. cucumerina isolates, and super‐susceptible to an adapted P. cucumerina isolate. This assigns a key role to trp‐derived secondary metabolites in limiting the growth of both non‐adapted and adapted necrotrophic fungi. However, 4‐methoxy‐indol‐3‐ylmethylglucosinolate, which is generated by the P450 monooxygenase CYP81F2, and hydrolyzed by PEN2 myrosinase, together with the antimicrobial camalexin play a minor role in restricting the growth of the non‐adapted necrotrophs. This contrasts with a major role of these two trp‐derived phytochemicals in limiting invasive growth of non‐adapted biotrophic powdery mildew fungi, thereby implying the existence of other unknown trp‐derived metabolites in resistance responses to non‐adapted necrotrophic P. cucumerina. Impaired defence to non‐adapted P. cucumerina, but not to the non‐adapted biotrophic fungus Erysiphe pisi, on cyp79B2 cyp79B3 plants is largely restored in the irx1 background, which shows a constitutive accumulation of antimicrobial peptides. Our findings imply differential contributions of antimicrobials in non‐host resistance to necrotrophic and biotrophic pathogens.