Search

Transgenic traits targeting WCR based on insecticidal proteins from Bacillus thuringiensis have proven effective in protecting maize yield in North America. High adoption has led to WCR populations that are resistant to the trait proteins.

Newly expressed proteins in genetically engineered crops are evaluated for potential cross reactivity to known allergens as part of their safety assessment. This assessment uses a weight-of-evidence approach. Two key components of this allergenicity assessment include any history of safe human exposure to the protein and/or the source organism from which it was originally derived, and bioinformatic analysis identifying amino acid sequence relatedness to known allergens. Phosphomannose-isomerase (PMI) has been expressed in commercialized genetically engineered (GE) crops as a selectable marker since 2010 with no known reports of allergy, which supports a history of safe exposure, and GE events expressing the PMI protein have been approved globally based on expert safety analysis. Bioinformatic analyses identified an eight-amino-acid contiguous match between PMI and a frog parvalbumin allergen (CAC83047.1). While short amino acid matches have been shown to be a poor predictor of allergen cross reactivity, most regulatory bodies require such matches be assessed in support of the allergenicity risk assessment. Here, this match is shown to be of negligible risk of conferring cross reactivity with known allergens.

This paper summarizes the overall results of the VIII World Engineering Education Forum, X Global Engineering Deans Council, XIV Global Student Forum, held in November 2018 in Albuquerque, NM USA, and discusses the emerging global collaboration for Peace Engineering (PEng) and Peace Engineering education. The theme of this first ever global conference on Peace Engineering was in tune with global events and developments. Participants were a confluence of practitioners, researchers, policy makers and educators from all over world, encompassing academia, multinational industry, US national labs and multilateral organizations. Results from a formal survey show that participants rated the conference as very good or excellent in all aspects and asked for more coverage of the topic of Peace Engineering and teaching material and strategies that embody Peace Engineering learning and teaching. The Conference succeeded in catalyzing conversations and idea exchanges for concrete plans for the next steps to advance a movement for Peace Engineering education, including • A Global Peace Engineering minor • Peace Engineering Certificate program • Master of Science in Peace Engineering • Database of Peace Engineering case studies • Workshops and other forums to promote discussion and planning for Peace Engineering programs and curricula • Data models, metrics, analytics and simulations: fine grain to large grain • Development of Peace Engineering as the new global engineering mindset Participants were engaged and enthusiastic about the concept and a Peace Engineering movement was born, along with the idea of creating a Peace Engineering Consortium.

The creation of the Peace Engineering Consortium is an outcome of the First Global Peace Engineering Conference, held November 2018 in Albuquerque NM, USA (www.weef-gedc2018.org). A team of conference participants are collaborating to develop the Peace Engineering Consortium. That is, to define and implement its charter, identify initiatives and preliminary activities, and continue to identify definitions of what Peace Engineering (PEng) – or Engineering for Peace – is. PEng is a call for action, in a new mindset not only for engineering, but for all existing and future disciplines to address global challenges. Current activities are: • Curricula and content – Members are collaborating to define, create and share curricula for undergraduate minors, professional certificate programs and graduate work; Creating experiential laboratories with industry and practitioner partners; Facilitating information exchange and the symmetry of information/knowledge through globally accessible digital libraries; Incorporating engineering diplomacy, diversity and cultural awareness (including the arts, music and language) into the practice for peacebuilding, peacemaking and peacekeeping. • Research and Development - The group is focused on collaborations, both domestic and international, and internal initiatives that advance the state of Peace Engineering. Key areas of focus have been developing metrics, data models, analytics, visualization and prediction. Some key initiatives include developing a modelling and visualizing conflict, Peace Data Standard, and pollution mitigation. • Entrepreneurial and Economic Development – The consortium is pursuing entrepreneurial activities in peace technology. It has been invited to produce a special edition on Peace Engineering and Innovation in the high impact Technological Forecasting and Social Change journal. • Events – In its charter year, the consortium organized a summer institute at The Hague and Peace Engineering workshops in Colombia – both models for future global workshops. • Case Studies – Members are compiling and publishing case studies and collaborating with US National Laboratories, industry, and governments to create scalable Peace Engineering case studies.

the IEEE Xplore database, Web of Science and indexed by EI Compendex and ISI Proceedings; One of the major contributions of social manufacturing is in the realm of sustainability. The apparel industry is a good example to assess contribution of social manufacturing to improve sustainability in practice. Value chains in the apparel industry are faced with various challenges regarding sustainability issues. Apparel companies pay higher attention to economic sustainability issues, and environmental and social sustainability issues of the apparel industry are often underrated. We realize that the apparel brand owners have the highest impact on improving the sustainability of the apparel industry. Thus, we design a collaborative business model empowered with social manufacturing to join the forces among the brand owners for improving sustainability of the apparel industry throughout the value chain. We chose a case study of shifting from conventional screen-printing to more environmentally sustainable digital textile printing. We suggest that this shift can be accelerated if the brand owners join theirforces together to shift from conventional printing to digital printing technology in the apparel industry.

AfIP-1A/1B is a two-component insecticidal protein identified from the soil bacterium Alcaligenes faecalis that has high activity against western corn rootworm (WCR; Diabrotica virgifera virgifera LeConte). Previous results revealed that AfIP-1A/1B is cross-resistant to the binary protein from Bacillus thuringiensis (Bt), Cry34Ab1/Cry35Ab1 (also known as Gpp34Ab1/Tpp35Ab1; Crickmore et al., 2020), which was attributed to shared binding sites in WCR gut tissue (Yalpani et al., 2017). To better understand the interaction of AfIP-1A/1B with its receptor, we have systematically evaluated the binding of these proteins with WCR brush border membrane vesicles (BBMVs). Our findings show that AfIP-1A binds directly to BBMVs, while AfIP-1B does not; AfIP-1B binding only occurred in the presence of AfIP-1A which was accompanied by the presence of stable, high molecular weight oligomers of AfIP-1B observed on denaturing protein gels. Additionally, we show that AfIP-1A/1B forms pores in artificial lipid membranes. Finally, binding of AfIP-1A/1B was found to be reduced in BBMVs from Cry34Ab1/Cry35Ab1-resistant WCR where Cry34Ab1/Cry35Ab1 binding was also reduced. The reduced binding of both proteins is consistent with recognition of a shared receptor that has been altered in the resistant strain. The coordination of AfIP-1B binding by AfIP-1A, the similar structures between AfIP-1A and Cry34Ab1, along with their shared binding sites and cross-resistance, suggest a similar role for AfIP1A and Cry34Ab1 in receptor recognition and docking site for their cognate partners, AfIP-1B and Cry35Ab1, respectively.

Western corn rootworm (WCR), Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae), is a serious insect pest in the major corn growing areas of North America and in parts of Europe. WCR populations with resistance to Bacillus thuringiensis (Bt) toxins utilized in commercial transgenic traits have been reported, raising concerns over their continued efficacy in WCR management. Understanding the modes of action of Bt toxins is important for WCR control and resistance management. Although different classes of proteins have been identified as Bt receptors for lepidopteran insects, identification of receptors in WCR has been limited with no reports of functional validation. Our results demonstrate that heterologous expression of DvABCB1 in Sf9 and HEK293 cells conferred sensitivity to the cytotoxic effects of Cry3A toxins. The result was further validated using knockdown of DvABCB1 by RNAi which rendered WCR larvae insensitive to a Cry3A toxin. However, silencing of DvABCB2 which is highly homologous to DvABCB1 at the amino acid level, did not reduce the sensitivity of WCR larvae to a Cry3A toxin. Furthermore, our functional studies corroborate different mode-of-actions for other insecticidal proteins including Cry34Ab1/35Ab1, Cry6Aa1, and IPD072Aa against WCR. Finally, reduced expression and alternatively spliced transcripts of DvABCB1 were identified in a mCry3A-resistant strain of WCR. Our results provide the first clear demonstration of a functional receptor in the molecular mechanism of Cry3A toxicity in WCR and confirmed its role in the mechanism of resistance in a mCry3A resistant strain of WCR.

Summary The coleopteran insect western corn rootworm (WCR, Diabrotica virgifera virgifera) is an economically important pest in North America and Europe. Transgenic corn plants producing Bacillus thuringiensis (Bt) insecticidal proteins have been useful against this devastating pest, but evolution of resistance has reduced their efficacy. Here, we report the discovery of a novel insecticidal protein, PIP‐47Aa, from an isolate of Pseudomonas mosselii. PIP‐47Aa sequence shows no shared motifs, domains or signatures with other known proteins. Recombinant PIP‐47Aa kills WCR, two other corn rootworm pests (Diabrotica barberi and Diabrotica undecimpunctata howardi) and two other beetle species (Diabrotica speciosa and Phyllotreta cruciferae), but it was not toxic to the spotted lady beetle (Coleomegilla maculata) or seven species of Lepidoptera and Hemiptera. Transgenic corn plants expressing PIP‐47Aa show significant protection from root damage by WCR. PIP‐47Aa kills a WCR strain resistant to mCry3A and does not share rootworm midgut binding sites with mCry3A or AfIP‐1A/1B from Alcaligenes that acts like Cry34Ab1/Cry35Ab1. Our results indicate that PIP‐47Aa is a novel insecticidal protein for controlling the corn rootworm pests.

Hop stunt viroid (HSVd) is an economically important pathogen that reduces growth and yield of hops. Visual symptoms of infected hop are highly dependent on cultivar. A study was conducted using six cultivars of hop to determine the impact on yield. Average dry cone yields of infected ‘Glacier’, ‘Cascade’, and ‘Willamette’ were reduced by 62, 14, and 34%, respectively, relative to noninoculated healthy plants. No significant yield reduction was observed for ‘Nugget’, ‘Columbus’, and ‘Galena’. The α-acid and β-acid contents showed a parallel pattern. Horticultural parameters of Willamette and Nugget were measured in the final year of the study. Internode length, shoot length, and side-arm length were reduced by 29, 26, and 73%, respectively, for infected Willamette bines relative to noninfected bines; no effects were observed resulting from infection of Nugget. To understand the current potential impact of HSVd, a survey was conducted to determine its distribution in central Washington. The survey revealed that 17% of hop plants tested are infected. Hop yield and hop plant longevity will be significantly affected by this level of infection. [Formula: see text] Copyright © 2017 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

The incidences of Hop latent virus (HpLV), Hop mosaic virus (HpMV), and American hop latent virus (AHLV), members of the genus Carlavirus, and Prunus necrotic ringspot virus and Apple mosaic virus, members of the genus Ilarvirus, were assessed for two hop cultivars, Horizon and Nugget, in Washington State. The spatial distribution of plants infected by the carlaviruses was assessed in two Horizon gardens in 2000 and one Nugget garden in 1993, 1994, and 1995. In the first Horizon garden (garden 1) and the Nugget garden, plants were separated by 2.1 m within and between rows. In these gardens, cultivation and the wide plant spacing discouraged contact between plants in either direction. In the second Horizon garden (garden 2), plants were separated by 4.3 m between rows and 1.0 m within rows. In all gardens, mechanical operations operated predominantly along rows; however, the closer plant spacing within rows in garden 2 permitted contact between adjacent plants within rows. In both Horizon gardens, the distribution of plants infected with HpMV was aggregated within rows. However, the distribution of plants infected with HpLV and AHLV was strongly influenced by contact between plants. In the Nugget garden, the distribution of plants infected by all three carlaviruses was autocorrelated within rows by 1995.

A novel Bacillus thuringiensis Cry protein, Cry8Hb, active against Diabrotica virgifera virgifera (Western corn rootworm, WCRW) was discovered. Unexpectedly, the anti-rootworm activity of the Cry8Hb toxin was enhanced significantly by fusing Escherichia coli maltose binding protein (MBP) to this Cry toxin. While the exact mechanism of the activity enhancement remains indefinite, it is probable that the enhancement is a result of increased solubility of the MBP-Cry8Hb fusion in the rootworm midgut. This hypothesis was examined using a synthetic Cry3 protein called IP3-1, which was not soluble at a neutral pH like Cry8Hb and marginally active to WCRW. When IP3-1 was fused to MBP, its anti-WCRW activity was enhanced 13-fold. To further test the hypothesis, DNA shuffling was performed on IP3-1 to increase the solubility without MBP. Screening of shuffled libraries found six new IP3 variants showing very high anti-WCRW activity without MBP. Sequence and 3D structure analysis of those highly active, shuffled IP3 variants revealed several charge-altering mutations such as Lys to Glu on the putative MBP-attaching side of the IP3 molecule. It is likely that those mutations make the protein acidic to substitute the functions of MBP including enhancing the solubility of IP3 at a neutral pH.

Pseudoperonospora humuli populations from Oregon and Washington were analyzed for genetic variation using random amplified polymorphic DNA (RAPD) and DNA amplification fingerprinting (DAF) markers. The genetic structure of the Oregon and Washington populations differed considerably. There was little genetic diversity in Washington, with only five RAPD and six DAF groups detected among 40 isolates tested. One genotype was predominant in Washing-ton. In contrast, 18 RAPD and 34 DAF groups were found among the 40 isolates tested from Oregon. No unique band profile associated with host cultivar was observed. It is suggested that the distinct difference in population structure between the two geographic regions might be due to climatic differences resulting in a higher frequency of sexual reproduction of P. humuli in Oregon than in Washington.

Gent, D. H., Nelson, M. E., and Grove, G. G. 2008. Persistence of phenylamide insensitivity in Pseudoperonospora humuli. Plant Dis. 92:463-468. Downy mildew, caused by Pseudoperonospora humuli, is an important disease of hop in most production regions in the northern hemisphere. Insensitivity to phenylamide fungicides was detected in isolates of P. humuli in production regions in Oregon and Idaho in 1992, and these fungicides since have been used on a limited basis. In this study, the prevalence of phenylamide insensitivity among isolates of P. humuli collected from 2005 to 2007 in the northwestern United States was quantified using a leaf disk assay with a discriminating dose (25 µg/ml) of either metalaxyl or mefenoxam with inoculum derived from 201 systemically infected diseased shoots (basal spikes) collected from 6, 10, and 11 hop yards in Idaho, Washington, and Oregon, respectively. A subset of 47 basal spike isolates and 42 monosporic isolates collected from two yards in Idaho and nine yards in Oregon during 2006 and 2007 were assayed using a dilution series of metalaxyl to determine the effective dose that inhibited 50% of the incidence of sporulation (ED50). Insensitivity to mefenoxam was detected in 31 of 74 (41.9%) basal spikes collected from 13 hop yards. Insensitivity to the related compound metalaxyl was detected in 52 of 80 (65%) spikes collected from nine hop yards, including four hop yards in Washington. Log ED50 values ranged from –2.25 to 2.67 for basal spike isolates and –2.27 to 2.98 for monosporic isolates and had a similar distribution. Log ED50 values for monosporic isolates and entire basal spike isolates were significantly associated. However, the slope of the regression line was less than 1, indicating that the log ED50 values obtained from entire basal spike isolates were greater than the corresponding log ED50 values obtained from a monosporic isolate obtained from that spike. This research suggests that insensitivity to phenylamide fungicides is a stable phenotype in P. humuli. Management of downy mildew should rely on measures other than these fungicides in yards or regions where phenylamide insensitivity is prevalent, which is now known to include certain hop yards in Idaho, Oregon, and Washington.

Gent, D. H., Nelson, M. E., Grove, G. G., Mahaffee, W. F., Turechek, W. W., and Woods, J. L. 2012. Association of spring pruning practices with severity of powdery mildew and downy mildew on hop. Plant Dis. 96:1343-1351. Downy mildew (caused by Pseudoperonospora humuli) and powdery mildew (caused by Podosphaera macularis) are important diseases of hop in the Pacific Northwest United States, and cultural practices may affect the severity of both diseases. The association of spring pruning quality and timing with severity of downy mildew and powdery mildew was assessed through analysis of survey data collected from commercial hop yards in Oregon and Washington. Among 149 hop yards surveyed, the most common pruning method was chemical desiccation (48% of yards), mechanical pruning (23%), or a combination of these practices (15%). The quality of pruning was assessed using a threecategory ordinal scale (“excellent”, “moderate”, or “poor”) based on the amount of foliage remaining on plants following pruning. Excellent pruning quality was attained more often in yards pruned twice (74.6 to 82.1% of yards) versus once (33.8% of yards), independent of pruning method. Seasonal severity of downy mildew in Oregon increased approximately twofold with reduction in pruning quality from excellent to moderate to poor. Pruning quality was not significantly related to levels of powdery mildew on leaves or cones in Oregon. Under more severe disease pressure in Washington, however, seasonal severity of powdery mildew on leaves and the incidence of cones with powdery mildew were significantly greater in yards that had poor pruning compared with excellent pruning. Moreover, yards that had excellent pruning quality received, on average, 1.1 to 1.5 fewer fungicide applications per season for downy mildew or powdery mildew compared with yards that had moderate or poor pruning quality. This savings was associated with delayed initiation of the first application by 7.5 to 14.2 days in yards with excellent pruning quality. Replicated experiments in commercial yards in Oregon quantified the effect of delaying pruning timing 5 to 21 days compared with growers’ standard practices on the diseases and yield. Downy mildew suppression by delayed pruning was dependent on cultivar and year of sampling, being significantly reduced fivefold only in ‘Willamette’ in 2007. Severity of powdery mildew and cone yield was similar between plots that received the delayed or standard pruning timing treatments. Collectively, these studies emphasize that early spring sanitation measures are associated with reduced primary inoculum and are critically important for managing both downy mildew and powdery mildew. A savings of at least one fungicide application per year appears achievable when spring pruning is conducted thoroughly and slightly delayed compared with growers’ current practices. Cultural disease management practices are actions that involve the manipulation of the plant, its components, crop refuse, and the environment to reduce or prevent disease (5,26,31,34). In hop (Humulus lupulus L.), cultural practices are an important component of integrated management of the two most important foliar diseases of

Resistant cultivars of hop (Humulus lupulus) have been grown, with the aim of helping to manage powdery mildew in the Pacific Northwest since the first report of the disease in the field in 1997 (4). A major objective of many breeding programs is development of resistance to powdery mildew, and this has generally been achieved by single resistance genes (qualitative resistance). One such gene, R6 (3), has been utilized extensively in new cultivars and has prevented epidemics of the disease in those cultivars across the Pacific Northwestern United States for approximately 15 years. In 2011, a grower in Washington State reported outbreaks of powdery mildew on cv. Apollo, which is thought to possess powdery mildew resistance derived from R6. Fungicides and cultural control measures were applied, and the grower reported no substantial crop damage from the disease. During the winter of 2012, the same grower planted rhizomes of cv. Apollo in a greenhouse in the Yakima Valley of Washington State and later found the plants to be affected by powdery mildew. Affected leaves from plants of cvs. Apollo, Newport, and Nugget (all reported [3] or assumed to possess R6 based on pedigree) grown in the same greenhouse were later provided to the authors. Conidia obtained from each affected plants were transferred to plants of the highly susceptible cv. Symphony, which is not known to contain any resistance genes. After 10 to 14 days of incubation, resultant conidia from each cultivar above (total of three isolates) were transferred to greenhouse grown plants of cvs. Nugget and Symphony and incubated at 18°C. Within 7 days, all three isolates produced powdery mildew colonies characteristic of P. macularis (2) on both cultivars. Cleistothecia did not develop in any colonies. In addition, Nugget and Symphony plants were inoculated with a field population of P. macularis originating from cultivars lacking R6 in Oregon. These inoculations on Nugget did not develop powdery mildew whereas Symphony plants did. Non-inoculated controls remained free of powdery mildew. Results were identical in two additional experiments. The sequence of the mating type idiomorph, MAT1-1, was obtained to confirm identity of the pathogen as P. macularis as described previously (1). The sequences were identical among the three isolates obtained from the greenhouse in Washington and isolates of P. macularis obtained previously from Oregon and Washington. MAT1-2 idiomorph was not detected in the isolates collected. While R6-virulent strains have been detected previously in race characterization experiments, these strains have not caused widespread epidemics of powdery mildew. The increasing prevalence of virulent strains of P. macularis and outbreaks of powdery mildew on formerly resistant cultivars necessitates changes in breeding strategies and disease management efforts to minimize damage resulting from the disease. The distribution of virulent strains of the pathogen and susceptibility of formerly resistance cultivars to powdery mildew are currently under investigation. References: (1) B. Asalfet et al. Phytopathology 103:717, 2013. (2) R. Bélanger et al. The Powdery Mildews: a Comprehensive Treatise. APS Press, St. Paul, MN, 2002. (3) P. Darby. Brew Hist. 121:94, 2005. (4) C. Ocamb et al. Plant Dis. 83:1072, 1999.

Host resistance, both quantitative and qualitative, is the preferred long-term approach for disease management in many pathosystems, including powdery mildew of hop (Podosphaera macularis). In 2012, an epidemic of powdery mildew occurred in Washington and Idaho on previously resistant cultivars whose resistance was putatively based on the gene designated R6. In 2013, isolates capable of causing severe disease on cultivars with R6-based resistance were confirmed in Oregon and became widespread during 2014. Surveys of commercial hop yards during 2012 to 2014 documented that powdery mildew is now widespread on cultivars possessing R6 resistance in Washington and Oregon, and the incidence of disease is progressively increasing. Pathogenic fitness, race, and mating type of R6-virulent isolates were compared with isolates of P. macularis lacking R6 virulence. All isolates were positive for the mating type idiomorph MAT1-1 and were able to overcome resistance genes Rb, R3, and R5 but not R1 or R2. In addition, R6-virulent isolates were shown to infect differential cultivars reported to possess the R6 gene and also the R4 gene, although R4 has not yet been broadly deployed in the United States. R6-virulent isolates were not detected from the eastern United States during 2012 to 2015. In growth chamber studies, R6-virulent isolates of P. macularis had a significantly longer latent period and produced fewer lesions on plants with R6 as compared with plants lacking R6, indicating a fitness cost to the fungus. R6-virulent isolates also produced fewer conidia when compared with isolates lacking R6 virulence, independent of whether the isolates were grown on a plant with or without R6. Thus, it is possible that the fitness cost of R6 virulence occurs regardless of host genotype. In field studies, powdery mildew was suppressed by at least 50% on plants possessing R6 as compared with those without R6 when coinoculated with R6-virulent and avirulent isolates. R6 virulence in P. macularis appears to be race specific and, at this time, imposes a measurable fitness penalty on the fungus. Resistance genes R1 and R2 appear to remain effective against R6-virulent isolates of P. macularis in the U.S. Pacific Northwest.

Several Bt maize events expressing various insecticidal Cry protein genes have been commercialized for management of western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae). We used high efficacy (>99.7%) experimental maize events that express mCry3A for selections under laboratory conditions to develop a western corn rootworm colony resistant to mCry3A at higher levels than published results. The resistance ratio (RR) to mCry3A was >97-fold based on LC50 values in diet-based bioassays after six generations of selections when compared to that of an unselected Control colony. Using a sublethal seedling assay (SSA) method, we confirmed that the colony had no cross-resistance to maize event DAS-59122-7, which expresses Cry34/35Ab. Reciprocal crosses between the mCry3A-resistant colony and the susceptible colony were performed to test the inheritance of resistance. Larval survival and development evaluated by the SSA method indicated that resistance to mCry3A was inherited autosomally and was incompletely recessive (h = 0.23–0.25). Specific binding of mCry3A to brush border membrane vesicles of midgut tissue revealed reduced binding in the resistant colony when compared to binding in the susceptible colony. This is the first report where resistance in western corn rootworm has been shown to involve reduced binding of a Cry3-class protein in midgut tissue.

Podosphaera macularis, the causal agent of hop powdery mildew, is a recurrent threat to hops in the Pacific Northwest because of the potential to reduce cone yield and quality. Early-season pruning is a common practice in hop production for horticultural reasons. Studies were conducted over a 3-year period in a commercial hop yard to quantify the effect of pruning method and timing on disease development, yield, and cone quality factors. A 4-week delay in pruning reduced the incidence of leaves with powdery mildew from 46 to 10% and cones from 9 to 1%, with the specific effect being season dependent. Pruning using chemical desiccants rather than by mechanical means had similar effects on disease levels on leaves. On cones, though, chemical pruning had a small but significant reduction in the incidence of powdery mildew compared with mechanical pruning. Cone yield, levels of bittering-acids, and color were not negatively affected in any individual year or cumulatively over three seasons when pruning treatments were applied repeatedly to the same plots during the study period. Delayed pruning may offer a low-cost means of reducing both the incidence of powdery mildew and early-season fungicide inputs in certain cultivars.

Canopy management is an important aspect of control of powdery mildew diseases and may influence the intensity of fungicide applications required to suppress disease. In hop, powdery mildew (caused by Podosphaera macularis) is most damaging to cones when infection occurs during bloom and the juvenile stages of cone development. Experiments were conducted over 3 years to evaluate whether fungicide applications could be ceased after the most susceptible stages of cone development (late July) without unduly affecting crop yield and quality when disease pressure was moderated with varying levels of basal foliage removal. In experimental plots of ‘Galena’ hop, the incidence of leaves with powdery mildew was similar whether fungicides were ceased in late July or made in late August. Disease levels on leaves were unaffected by the intensity of basal foliage removal, whereas the intensity of basal foliage removal interacted with the duration of fungicide applications to affect disease levels on cones. Similar experiments conducted in large plots of ‘Tomahawk’ hop in a commercial hop yard similarly found no significant impact on disease levels on leaves from either the duration of fungicide applications or intensity of basal foliage removal. In contrast, on cones, application of fungicides into August had a modest, suppressive effect on powdery mildew. There was also some evidence that the level of powdery mildew on cones associated with fungicide treatment was influenced by the intensity of basal foliage removal. When fungicide applications ceased in late July, there was a progressive decrease in the incidence of cones with powdery mildew with increasing intensity of basal foliage removal. Removing basal foliage two to three times allowed fungicide applications to be terminated in late July rather than late August without diminishing disease control on cones, yield, or cone quality factors. Thus, this study further establishes that fungicide applications made during the early stages of hop cone development have the strongest effect on suppression of powdery mildew on cones. The additive effect of fungicide applications targeted to the periods of greatest cone susceptibility and canopy management to reduce disease favorability may obviate the need for fungicide applications later in the season. This appears to be a viable strategy in mature hop yards of certain cultivars when disease pressure is not excessively high.

The western corn rootworm (WCR, Diabrotica virgifera virgifera) gene, dvssj1, is a putative homolog of the Drosophila melanogaster gene, snakeskin (ssk). This gene encodes a membrane protein associated with the smooth septate junction (SSJ) which is required for the proper barrier function of the epithelial lining of insect intestines. Disruption of DVSSJ integrity by RNAi technique has been shown previously to be an effective approach for corn rootworm control, by apparent suppression of production of DVSSJ1 protein leading to growth inhibition and mortality. To understand the mechanism that leads to the death of WCR larvae by dvssj1 double-stranded RNA, we examined the molecular characteristics associated with SSJ functions during larval development. Dvssj1 dsRNA diet feeding results in dose-dependent suppression of mRNA and protein; this impairs SSJ formation and barrier function of the midgut and results in larval mortality. These findings suggest that the malfunctioning of the SSJ complex in midgut triggered by dvssj1 silencing is the principal cause of WCR death. This study also illustrates that dvssj1 is a midgut-specific gene in WCR and its functions are consistent with biological functions described for ssk.

The western corn rootworm (WCR) Diabrotica virgifera virgifera causes substantial damage in corn. Genetically modified (GM) plants expressing some Bacillus thuringiensis (Bt) insecticidal Cry proteins efficiently controlled this pest. However, changes in WCR susceptibility to these Bt traits have evolved and identification of insecticidal proteins with different modes of action against WCR is necessary. We show here for the first time that Cyt1Aa from Bt exhibits toxicity against WCR besides to the dipteran Aedes aegypti larvae. Cyt1Aa is a pore-forming toxin that shows no cross-resistance with mosquitocidal Cry toxins. We characterized different mutations in helix α-A from Cyt1Aa. Two mutants (A61C and A59C) exhibited reduced or absent hemolytic activity but retained toxicity to A. aegypti larvae, suggesting that insecticidal and hemolytic activities of Cyt1Aa are independent activities. These mutants were still able to form oligomers in synthetic lipid vesicles and to synergize Cry11Aa toxicity. Remarkably, mutant A61C showed a five-fold increase insecticidal activity against mosquito and almost 11-fold higher activity against WCR. Cyt1Aa A61C mutant was as potent in killing WCR that were selected for resistance to mCry3A as it was against unselected WCR indicating that this toxin could be a useful resistance management option in the control of WCR.

Bacillus thuringiensis Cry1Ab and Cry1Fa toxins are environmentally safe insecticides that control important insect pests. Spodoptera frugiperda is an important maize pest that shows low susceptibility to Cry1A toxins, in contrast to Cry1Fa, which is highly active against this pest and is used in transgenic maize for S. frugiperda control. The β16 region from domain III of Cry1Ab has been shown to be involved in interactions with receptors such as alkaline phosphatase (ALP) or aminopeptidase (APN) in different lepidopteran insects. Alanine-scanning mutagenesis of amino acids of Cry1Ab β16 (509STLRVN514) revealed that certain β16 mutations, such as N514A, resulted in increased toxicity of Cry1Ab for S. frugiperda without affecting the toxicity for other lepidopteran larvae, such as Manduca sexta larvae. Exhaustive mutagenesis of N514 was performed, showing that the Cry1Ab N514F, N514H, N514K, N514L, N514Q, and N514S mutations increased the toxicity toward S. frugiperda. A corresponding mutation was constructed in Cry1Fa (N507A). Toxicity assays of wild-type and mutant toxins (Cry1Ab, Cry1AbN514A, Cry1AbN514F, Cry1Fa, and Cry1FaN507A) against four S. frugiperda populations from Mexico and one from Brazil revealed that Cry1AbN514A and Cry1FaN507A consistently showed 3- to 18-fold increased toxicity against four of five S. frugiperda populations. In contrast, Cry1AbN514F showed increased toxicity in only two of the S. frugiperda populations analyzed. The mutants Cry1AbN514A and Cry1AbN514F showed greater stability to midgut protease treatment. In addition, binding analysis of the Cry1Ab mutants showed that the increased toxicity correlated with increased binding to brush border membrane vesicles and increased binding affinity for S. frugiperda ALP, APN, and cadherin receptors. IMPORTANCESpodoptera frugiperda is the main maize pest in South and North America and also is an invasive pest in different African countries. However, it is poorly controlled by Bacillus thuringiensis Cry1A toxins expressed in transgenic crops, which effectively control other lepidopteran pests. In contrast, maize expressing Cry1Fa is effective in the control of S. frugiperda, although its effectiveness is being lost due to resistance evolution. Some of the Cry1Ab domain III mutants characterized here show enhanced toxicity for S. frugiperda without loss of toxicity to Manduca sexta. Thus, these Cry1Ab mutants could provide useful engineered toxins that, along with other Cry toxins, would be useful for developing transgenic maize expressing stacked proteins for the effective control of S. frugiperda and other lepidopteran pests in the field.

The current paper presents a theoretical framework for standardizing Peace Data as a means of understanding the conditions under which people’s technology use results in positive engagement and peace. Thus, the main point of our paper is that Big Data can be conceptualized in terms of its value to peace. We define peace as a set of positive, prosocial behaviors that maximize mutually beneficial positive outcomes resulting from interactions with others. To accomplish this goal, we present hypothetical and real-world, data driven examples that illustrate our thinking in this domain and present guidelines for how to identify, collect, utilize, and evaluate Peace Data generated during mediated interactions and further suggest that Peace Data has four primary components: group identity information, behavior data, longitudinal data, and metadata. This paper concludes with a call for participation in a Peace Data association and suggested for guidelines for how scholars and practitioners can identify Peace Data in their own domains. Ethical considerations and suggestions for future research are also discussed.

Western corn rootworm (Diabrotica virgifera virgifera) is a serious agricultural pest known for its high adaptability to various management strategies, giving rise to a continual need for new control options. Transgenic maize expressing insecticidal RNAs represents a novel mode of action for rootworm management that is dependent on the RNA interference (RNAi) pathways of the insect for efficacy. Preliminary evidence suggests that western corn rootworm could develop broad resistance to all insecticidal RNAs through changes in RNAi pathway genes; however, the likelihood of field-evolved resistance occurring through this mechanism remains unclear. In the current study, eight key genes involved in facilitating interference in the microRNA and small interfering RNA pathways were targeted for knockdown in order to evaluate impact on fitness of western corn rootworm. These genes include drosha, dicer-1, dicer-2, pasha, loquacious, r2d2, argonaute 1, and argonaute 2. Depletion of targeted transcripts in rootworm larvae led to changes in microRNA expression, decreased ability to pupate, reduced adult beetle emergence, and diminished reproductive capacity. The observed effects do not support evolution of resistance through changes in expression of these eight genes due to reduced insect fitness.

Results of 28 field trials conducted over a 12-year period investigating management of hop powdery mildew caused by Podosphaera macularis were quantitatively summarized by meta-analysis to compare product efficacy and use patterns by mode of action as defined by Fungicide Resistance Action Committee (FRAC) groups. Availability of original observations enabled individual participant data meta-analysis. Differences in control of powdery mildew on leaves and cones were apparent among fungicide FRAC groups when individual products were evaluated over the course of a growing season. FRAC groups 13, 3, and U13 provided the most efficacious control of powdery mildew on leaves. Percent disease control on cones was influenced by midseason foliar disease and fungicide mode-of-action. FRAC 13 provided significantly better disease control on cones than all other groups except U13, 3, and premixes of 7 with 11. Disease control on leaves was similar when a rotational program of fungicides was used, independent of the modes of action, but improved on cones if FRAC groups 13 and 3 were both included compared with programs consisting of FRAC groups 11 and 3, 11 and 5, or 3 and 5. Disease control on cones was improved from 32 to 52%, on average, when the fungicide quinoxyfen (FRAC 13) was applied at least once during the early stages of cone development, defined in this analysis as 20 July to 10 August, as compared with all other treatments. Efficacy of disease control on cones by quinoxyfen was moderated by and interacted with the incidence of leaves with powdery mildew. Disease control on cones was further improved if two applications of quinoxyfen were made during this period. Collectively, these findings suggest that disease control during juvenile stages of cone development largely influences the success of fungicide programs and point to the critical importance of focusing management efforts during this stage of development, independent of what actual management strategy is employed.

Viroids are the smallest known plant pathogens that exploit host systems for their replication and cause diseases in many hosts. In this study, the host response of hop plants to Hop stunt viroid (HSVd) infection was studied through transcriptome analysis. RNA sequence analysis of hop leaves infected with HSVd revealed dynamic changes in hop gene expression. Defense-related genes and genes involved in lipid and terpenoid metabolism are the major categories that showed differential expression due to HSVd infection. Additionally, the effect of HSVd on development of hop powdery mildew (Podospheara macularis) (HPM) was studied. Transcriptome analysis followed by quantitative reverse transcription-polymerase chain reaction analysis showed that transcript levels of pathogenesis-related (PR) genes such as PR protein 1, chitinase, and thaumatin-like protein genes are induced in leaves infected with HPM alone. The response in these genes to HPM is significantly down-regulated in leaves with HSVd-HPM mixed infection. These results confirm that HSVd alters host metabolism, physiology, and plant defense responses. Nevertheless, in detached leaf assays, HPM consistently expanded faster on HSVd-negative leaves relative to HSVd-positive leaves. Although HSVd infection suppresses elements associated with the host immunity response, infection by HSVd is antagonistic to HPM infection of hops.

Crops expressing Bacillus thuringiensis (Bt)-derived insecticidal protein genes have been commercially available for over 15 years and are providing significant value to growers. However, there remains the need for alternative insecticidal actives due to emerging insect resistance to certain Bt proteins. A screen of bacterial strains led to the discovery of a two-component insecticidal protein named AfIP-1A/1B from an Alcaligenes faecalis strain. This protein shows selectivity against coleopteran insects including western corn rootworm (WCR). Transgenic maize plants expressing AfIP-1A/1B demonstrate strong protection from rootworm injury. Surprisingly, although little sequence similarity exists to known insecticidal proteins, efficacy tests using WCR populations resistant to two different Cry proteins show that AfIP-1A/1B and mCry3A differ in their mode of action while AfIP-1A/1B and the binary Cry34Ab1/Cry35Ab1 protein share a similar mode. These findings are supported by results of competitive binding assays and the similarity of the x-ray structure of AfIP-1A to Cry34Ab1. Our work indicates that insecticidal proteins obtained from a non-Bt bacterial source can be useful for developing genetically modified crops and can function similarly to familiar proteins from Bt.

This is the publisher’s final pdf. The article was published by John Wiley & Sons Ltd. and is in the public domain. The published article can be found at: http://onlinelibrary.wiley.com/journal/10.1111/%28ISSN%291365-3059.