Genetic modification of asexual Epichloë endophytes with the perA gene for peramine biosynthesis.

Development of grass-endophyte associations with minimal or no detrimental effects in combination with beneficial characteristics is important for pastoral agriculture. The feasibility of enhancing production of an endophyte-derived beneficial alkaloid through introduction of an additional gene copy was assessed in a proof-of-concept study. Sexual and asexual Epichloë species that form symbiotic associations with cool-season grasses of the Poaceae sub-family Pooideae produce bioactive alkaloids that confer resistance to herbivory by a number of organisms. Of these, peramine is thought to be crucial for protection of perennial ryegrass (Lolium perenne L.) from the Argentinian stem weevil, an economically important exotic pest in New Zealand, contributing significantly to pasture persistence. A single gene (perA) has been identified as solely responsible for peramine biosynthesis and is distributed widely across Epichloë taxa. In the present study, a functional copy of the perA gene was introduced into three recipient endophyte genomes by Agrobacterium tumefaciens-mediated transformation. The target strains included some that do not produce peramine, and others containing different perA gene copies. Mitotically stable transformants generated from all three endophyte strains were able to produce peramine in culture and in planta at variable levels. In summary, this study provides an insight into the potential for artificial combinations of alkaloid biosynthesis in a single endophyte strain through transgenesis, as well as the possibility of using novel genome editing techniques to edit the perA gene of non-peramine producing strains.

Overexpression of the ZmDEF1 gene increases the resistance to weevil larvae in transgenic maize seeds.

Plant defensins are divided into 18 groups and are multifunctional proteins. The Zea mays defensin 1 (ZmDEF1) gene encodes the defensin 1 protein, which can inhibit alpha-amylase in the insect gut. In this study, the ZmDEF1 gene was transferred into two maize cultivars, LC1 and LVN99, to improve weevil resistance in maize. The recombinant ZmDEF1 protein was assessed for its ability to inhibit alpha-amylase in the gut of the larvae of the maize weevil (Sitophilus zeamais Motsch.). ZmDEF1 was cloned into a pBetaPhaso-dest vector, which harbours phaseolin, a seed-specific promoter, and the Agrobacterium tumefaciens strain C58 harbouring the pBetaPhaso-ZmDEF1 vector was used to transfer the ZmDEF1 gene into two maize cultivars using immature embryos. Transformed calluses were selected on selection media containing kanamycin. The stable integration of the ZmDEF1 transgene into the transgenic maize plant genome was confirmed using Southern blotting. The recombinant ZmDEF1 protein of approximately 10 kDa was expressed in three transgenic maize lines from the LC1 cultivar (C1, C3, and C5) and two transgenic maize lines from the LVN99 cultivar (L1 and L3). The ZmDEF1 transgenic efficiency based on the results of PCR, as well as Southern and Western blotting, was 1.32% and 0.82%, respectively, which depends on the genotypes of LC1 and LVN99. The recombinant ZmDEF1 protein inhibited the alpha-amylase activity of the maize weevil larvae, and its ability to inhibit alpha-amylase is 54.52-63.09% greater than the ZmDEF1 protein extracted from non-transgenic plants.

Transgenic cotton expressing Cry10Aa toxin confers high resistance to the cotton boll weevil.

Genetically modified (GM) cotton plants that effectively control cotton boll weevil (CBW), which is the most destructive cotton insect pest in South America, are reported here for the first time. This work presents the successful development of a new GM cotton with high resistance to CBW conferred by Cry10Aa toxin, a protein encoded by entomopathogenic Bacillus thuringiensis (Bt) gene. The plant transformation vector harbouring cry10Aa gene driven by the cotton ubiquitination-related promoter uceA1.7 was introduced into a Brazilian cotton cultivar by biolistic transformation. Quantitative PCR (qPCR) assays revealed high transcription levels of cry10Aa in both T0 GM cotton leaf and flower bud tissues. Southern blot and qPCR-based 2-ΔΔCt analyses revealed that T0 GM plants had either one or two transgene copies. Quantitative and qualitative analyses of Cry10Aa protein expression showed variable protein expression levels in both flower buds and leaves tissues of T0 GM cotton plants, ranging from approximately 3.0 to 14.0 µg g-1 fresh tissue. CBW susceptibility bioassays, performed by feeding adults and larvae with T0 GM cotton leaves and flower buds, respectively, demonstrated a significant entomotoxic effect and a high level of CBW mortality (up to 100%). Molecular analysis revealed that transgene stability and entomotoxic effect to CBW were maintained in T1 generation as the Cry10Aa toxin expression levels remained high in both tissues, ranging from 4.05 to 19.57 µg g-1 fresh tissue, and the CBW mortality rate remained around 100%. In conclusion, these Cry10Aa GM cotton plants represent a great advance in the control of the devastating CBW insect pest and can substantially impact cotton agribusiness.

Plasticity and evolution of (+)-3-carene synthase and (-)-sabinene synthase functions of a sitka spruce monoterpene synthase gene family associated with weevil resistance.

The monoterpene (+)-3-carene is associated with resistance of Sitka spruce against white pine weevil, a major North American forest insect pest of pine and spruce. High and low levels of (+)-3-carene in, respectively, resistant and susceptible Sitka spruce genotypes are due to variation of (+)-3-carene synthase gene copy number, transcript and protein expression levels, enzyme product profiles, and enzyme catalytic efficiency. A family of multiproduct (+)-3-carene synthase-like genes of Sitka spruce include the three (+)-3-carene synthases, PsTPS-3car1, PsTPS-3car2, PsTPS-3car3, and the (-)-sabinene synthase PsTPS-sab. Of these, PsTPS-3car2 is responsible for the relatively higher levels of (+)-3-carene in weevil-resistant trees. Here, we identified features of the PsTPS-3car1, PsTPS-3car2, PsTPS-3car3, and PsTPS-sab proteins that determine different product profiles. A series of domain swap and site-directed mutations, supported by structural comparisons, identified the amino acid in position 596 as critical for product profiles dominated by (+)-3-carene in PsTPS-3car1, PsTPS-3car2, and PsTPS-3car3, or (-)-sabinene in PsTPS-sab. A leucine in this position promotes formation of (+)-3-carene, whereas phenylalanine promotes (-)-sabinene. Homology modeling predicts that position 596 directs product profiles through differential stabilization of the reaction intermediate. Kinetic analysis revealed position 596 also plays a role in catalytic efficiency. Mutations of position 596 with different side chain properties resulted in a series of enzymes with different product profiles, further highlighting the inherent plasticity and potential for evolution of alternative product profiles of these monoterpene synthases of conifer defense against insects.

A Model for Mountain Pine Beetle Outbreaks in an Age-Structured Forest: Predicting Severity and Outbreak-Recovery Cycle Period.

The mountain pine beetle (MPB, Dendroctonus ponderosae), a tree-killing bark beetle, has historically been part of the normal disturbance regime in lodgepole pine (Pinus contorta) forests. In recent years, warm winters and summers have allowed MPB populations to achieve synchronous emergence and successful attacks, resulting in widespread population outbreaks and resultant tree mortality across western North America. We develop an age-structured forest demographic model that incorporates temperature-dependent MPB infestations. Stability of fixed points is analyzed as a function of (thermally controlled) MPB population growth rates and indicates the existence of periodic outbreaks that intensify as growth rates increase. We devise analytical methods to predict outbreak severity and duration as well as outbreak return time. After incorporating a spatial aspect and controlling initial stand demographic variation, the model predicts cycle periods that fall within observed outbreak return time ranges. To assess future MPB impact on forests, we use climate model projected temperatures with our model-based approximation methods to predict potential severity of future outbreaks that reflect the effects of changing climate.

An integrated genomic, proteomic and biochemical analysis of (+)-3-carene biosynthesis in Sitka spruce (Picea sitchensis) genotypes that are resistant or susceptible to white pine weevil.

Conifers are extremely long-lived plants that have evolved complex chemical defenses in the form of oleoresin terpenoids to resist attack from pathogens and herbivores. In these species, terpenoid diversity is determined by the size and composition of the terpene synthase (TPS) gene family and the single- and multi-product profiles of these enzymes. The monoterpene (+)-3-carene is associated with resistance of Sitka spruce (Picea sitchensis) to white pine weevil (Pissodes strobi). We used a combined genomic, proteomic and biochemical approach to analyze the (+)-3-carene phenotype in two contrasting Sitka spruce genotypes. Resistant trees produced significantly higher levels of (+)-3-carene than susceptible trees, in which only trace amounts were detected. Biosynthesis of (+)-3-carene is controlled, at the genome level, by a small family of closely related (+)-3-carene synthase (PsTPS-3car) genes (82-95% amino acid sequence identity). Transcript profiling identified one PsTPS-3car gene (PsTPS-3car1) that is expressed in both genotypes, one gene (PsTPS-3car2) that is expressed only in resistant trees, and one gene (PsTPS-3car3) that is expressed only in susceptible trees. The PsTPS-3car2 gene was not detected in genomic DNA of susceptible trees. Target-specific selected reaction monitoring confirmed this pattern of differential expression of members of the PsTPS-3car family at the proteome level. Kinetic characterization of the recombinant PsTPS-3car enzymes identified differences in the activities of PsTPS-3car2 and PsTPS-3car3 as a factor contributing to the different (+)-3-carene profiles of resistant and susceptible trees. In conclusion, variation of the (+)-3-carene phenotype is controlled by copy number variation of PsTPS-3car genes, variation of gene and protein expression, and variation in catalytic efficiencies.

A model for long-distance dispersal of boll weevils (Coleoptera: Curculionidae).

The boll weevil, Anthonomus grandis (Boheman), has been a major insect pest of cotton production in the US, accounting for yield losses and control costs on the order of several billion US dollars since the introduction of the pest in 1892. Boll weevil eradication programs have eliminated reproducing populations in nearly 94%, and progressed toward eradication within the remaining 6%, of cotton production areas. However, the ability of weevils to disperse and reinfest eradicated zones threatens to undermine the previous investment toward eradication of this pest. In this study, the HYSPLIT atmospheric dispersion model was used to simulate daily wind-aided dispersal of weevils from the Lower Rio Grande Valley (LRGV) of southern Texas and northeastern Mexico. Simulated weevil dispersal was compared with weekly capture of weevils in pheromone traps along highway trap lines between the LRGV and the South Texas/Winter Garden zone of the Texas Boll Weevil Eradication Program. A logistic regression model was fit to the probability of capturing at least one weevil in individual pheromone traps relative to specific values of simulated weevil dispersal, which resulted in 60.4% concordance, 21.3% discordance, and 18.3% ties in estimating captures and non-captures. During the first full year of active eradication with widespread insecticide applications in 2006, the dispersal model accurately estimated 71.8%, erroneously estimated 12.5%, and tied 15.7% of capture and non-capture events. Model simulations provide a temporal risk assessment over large areas of weevil reinfestation resulting from dispersal by prevailing winds. Eradication program managers can use the model risk assessment information to effectively schedule and target enhanced trapping, crop scouting, and insecticide applications.

Conifer bark beetles.

Fettig and Audley introduce the bark beetles-a large and diverse group of insects of which some are commonly recognized as important disturbance agents in conifer forests.

Cross-scale interaction of host tree size and climatic water deficit governs bark beetle-induced tree mortality.

The recent Californian hot drought (2012-2016) precipitated unprecedented ponderosa pine (Pinus ponderosa) mortality, largely attributable to the western pine beetle (Dendroctonus brevicomis; WPB). Broad-scale climate conditions can directly shape tree mortality patterns, but mortality rates respond non-linearly to climate when local-scale forest characteristics influence the behavior of tree-killing bark beetles (e.g., WPB). To test for these cross-scale interactions, we conduct aerial drone surveys at 32 sites along a gradient of climatic water deficit (CWD) spanning 350 km of latitude and 1000 m of elevation in WPB-impacted Sierra Nevada forests. We map, measure, and classify over 450,000 trees within 9 km2, validating measurements with coincident field plots. We find greater size, proportion, and density of ponderosa pine (the WPB host) increase host mortality rates, as does greater CWD. Critically, we find a CWD/host size interaction such that larger trees amplify host mortality rates in hot/dry sites. Management strategies for climate change adaptation should consider how bark beetle disturbances can depend on cross-scale interactions, which challenge our ability to predict and understand patterns of tree mortality.

A one-dimensional map to study multi-seasonal coffee infestation by the coffee berry borer.

The coffee berry borer (CBB, Hypothenemus hampei) is the most serious insect pest of coffee worldwide; understanding the dynamics of its reproduction is essential for pest management. The female CBB penetrates the coffee berry, eats the seed, and reproduces inside it. A mathematical model of the infestation progress of the coffee berry by the CBB during several coffee seasons is formulated. The model represents the interaction among five populations: uninfested, slightly infested, and severely infested coffee berries, and free and encapsulated CBBs. Coffee harvesting is also included in the model. A one-dimensional map is derived for tracking the population dynamics subject to certain coffee harvesting percentages over several seasons. Stability analysis of the map's fixed points shows that CBB infestation could be eliminated or controlled to a specific level over multiple seasons of coffee harvesting. However, the percent of coffee harvesting required is determined by the level of CBB infestation at the beginning of the first season and in some cases it is impossible to achieve that percentage.

Joint Spatial Modeling of Recurrent Infection and Growth with Processes under Intermittent Observation.

In this article, we present a new statistical methodology for longitudinal studies in forestry, where trees are subject to recurrent infection, and the hazard of infection depends on tree growth over time. Understanding the nature of this dependence has important implications for reforestation and breeding programs. Challenges arise for statistical analysis in this setting with sampling schemes leading to panel data, exhibiting dynamic spatial variability, and incomplete covariate histories for hazard regression. In addition, data are collected at a large number of locations, which poses computational difficulties for spatiotemporal modeling. A joint model for infection and growth is developed wherein a mixed nonhomogeneous Poisson process, governing recurring infection, is linked with a spatially dynamic nonlinear model representing the underlying height growth trajectories. These trajectories are based on the von Bertalanffy growth model and a spatially varying parameterization is employed. Spatial variability in growth parameters is modeled through a multivariate spatial process derived through kernel convolution. Inference is conducted in a Bayesian framework with implementation based on hybrid Monte Carlo. Our methodology is applied for analysis in an 11-year study of recurrent weevil infestation of white spruce in British Columbia.

Genetic Basis of Maize Resistance to Multiple Insect Pests: Integrated Genome-Wide Comparative Mapping and Candidate Gene Prioritization.

Several species of herbivores feed on maize in field and storage setups, making the development of multiple insect resistance a critical breeding target. In this study, an association mapping panel of 341 tropical maize lines was evaluated in three field environments for resistance to fall armyworm (FAW), whilst bulked grains were subjected to a maize weevil (MW) bioassay and genotyped with Diversity Array Technology's single nucleotide polymorphisms (SNPs) markers. A multi-locus genome-wide association study (GWAS) revealed 62 quantitative trait nucleotides (QTNs) associated with FAW and MW resistance traits on all 10 maize chromosomes, of which, 47 and 31 were discovered at stringent Bonferroni genome-wide significance levels of 0.05 and 0.01, respectively, and located within or close to multiple insect resistance genomic regions (MIRGRs) concerning FAW, SB, and MW. Sixteen QTNs influenced multiple traits, of which, six were associated with resistance to both FAW and MW, suggesting a pleiotropic genetic control. Functional prioritization of candidate genes (CGs) located within 10-30 kb of the QTNs revealed 64 putative GWAS-based CGs (GbCGs) showing evidence of involvement in plant defense mechanisms. Only one GbCG was associated with each of the five of the six combined resistance QTNs, thus reinforcing the pleiotropy hypothesis. In addition, through in silico co-functional network inferences, an additional 107 network-based CGs (NbCGs), biologically connected to the 64 GbCGs, and differentially expressed under biotic or abiotic stress, were revealed within MIRGRs. The provided multiple insect resistance physical map should contribute to the development of combined insect resistance in maize.

Seasonal changes in population structure of the ambrosia beetle Xylosandrus compactus and its associated fungi in a southern Mediterranean environment.

Exotic ambrosia beetles are increasing in Europe due to global trade and global warming. Among these xylomycetophagous insects, Xylosandrus compactus (Eichhoff) (Coleoptera: Curculionidae) is a serious threat for several Mediterranean host plants. Carob trees growing in Sicily (Italy) have been extensively attacked by beetles leading to rapid tree decline. Although X. compactus has been found in Europe for several years, most aspects of its ecology are still unknown. We thus studied the population structure and dynamics of X. compactus, together with its twig size preference during a sampling of infested carob trees in south east Sicily. In addition, fungi associated with insects or galleries were isolated and characterized. The results showed that, in this newly-colonized environment and host plant, adult X. compactus overwinters inside twigs and starts to fly and reproduce in mid spring, completing five generations before overwintering in late fall. The mean diameter of carob twigs infested by the beetle varied significantly over the seasons, with the insect tending to infest larger twigs as season progresses. The mean number of adults/gallery was 19.21, ranging from 6 to 28. The minimum temperature significantly affected the overwintering adult mortality. Ambrosiella xylebori and Fusarium solani were the main symbionts associated with the pest in this study. Acremonium sp. was instead recorded for the first time in Europe inside X. compactus galleries. Several other fungi species were also found for the first time in association with X. compactus. Our findings provide useful insights into the sustainable management of this noxious pest.

Reconsiderations on evaluating methodology of repellent effects: validation of indices and statistical analyses.

Many pest repellents are in household use, and a variety of procedures, indices, and statistical tests have been used to examine their effectiveness. However, the effect of action mechanism on these indices has not been adequately discussed. In addition, the interreplication variance of data have not been taken into account in some cases. This study considered the validity of tests used to evaluate repellents and proposed methods to statistically evaluate them. First, various repellents with different mechanisms of action were virtually assumed, and their repelling processes were formulated as differential equations. Numerical and analytical analyses of the formulae revealed that, in some action mechanisms, the indices could be altered by parameters other than the repellent effect and that the indices should be used only for repellents of a specific mechanism. Furthermore, the effect of an actual repellent, by using two different experimental designs and the maize weevil, Sitophilus zeamais Motschulsky (Coleoptera: Curculionidae), were evaluated. This demonstrated that the evaluated effectiveness could vary with experimental design and suggested that the design should reflect the situation in which repellents are actually used. Therefore, statistical methods to quantify the effectiveness of repellents are proposed herein and applied to the results of the above-mentioned experiments. Particular emphasis was placed on the importance of incorporating the interreplication variance in statistical models and estimating confidence intervals. Finally, the validity and limitations of evaluation using indices of repellents' effects, and the identification of important criteria that should be considered when assessing the effects of repellents also are discussed.

Curative activity of insecticides against plum curculio (Coleoptera: Curculionidae) in tart cherries.

Tart cherry, Prunus cerasus L. variety Montmorency, fruit were infested with plum curculio, Conotrachelus nenuphar (Herbst) (Coleoptera: Curculionidae), and treated with insecticides to target late instars, neonates, and eggs. The organophosphates azinphos-methyl and phosmet and the neonicotinoid thiamethoxam reduced larval emergence rates by >90% for all life stage targets; after >30 d, few surviving larvae were found inside fruit. Acetamiprid and thiacloprid also had curative activity and yielded >75% reductions in emergence and few surviving larvae in the fruit after >30 d. The juvenile hormone analog pyriproxyfen reduced larval emergence, but 66% of fruit that was treated to target late-instars still had live larvae inside of them after >30 d. Novaluron, chlorantraniliprole, and esfenvalerate had no curative activity. Indoxacarb had limited curative activity, and all targeted life stages had larval emergence. Internal and external residues were analyzed and are discussed in relation to their penetration and curative potential. The curative activity of azinphos-methyl has played an important role in meeting federal standards for infestation-free tart cherries at processing. Regulatory changes are eliminating the use of this compound, and new integrated pest management programs for plum curculio will need to address the loss of azinphos-methyl's curative activity.

Fine-tuned intruder discrimination favors ant parasitoidism.

A diversity of arthropods (myrmecophiles) thrives within ant nests, many of them unmolested though some, such as the specialized Eucharitidae parasitoids, may cause direct damage to their hosts. Ants are known to discriminate between nestmates and non-nestmates, but whether they recognize the strength of a threat and their capacity to adjust their behavior accordingly have not been fully explored. We aimed to determine whether Ectatomma tuberculatum ants exhibited specific behavioral responses to potential or actual intruders posing different threats to the host colony and to contribute to an understanding of complex ant-eucharitid interactions. Behavioral responses differed significantly according to intruder type. Ants evicted intruders that represented a threat to the colony's health (dead ants) or were not suitable as prey items (filter paper, eucharitid parasitoid wasps, non myrmecophilous adult weevils), but killed potential prey (weevil larvae, termites). The timing of detection was in accordance with the nature and size of the intruder: corpses (a potential source of contamination) were detected faster than any other intruder and transported to the refuse piles within 15 min. The structure and complexity of behavioral sequences differed among those intruders that were discarded. Workers not only recognized and discriminated between several distinct intruders but also adjusted their behavior to the type of intruder encountered. Our results confirm the previously documented recognition capabilities of E. tuberculatum workers and reveal a very fine-tuned intruder discrimination response. Colony-level prophylactic and hygienic behavioral responses through effective removal of inedible intruders appears to be the most general and flexible form of defense in ants against a diverse array of intruders. However, this generalized response to both potentially lethal and harmless intruders might have driven the evolution of ant-eucharitid interactions, opening a window for parasitoid attack and allowing adult parasitoid wasps to quickly leave the natal nest unharmed.

Organic mulches reduce crop attack by sweetpotato weevil (Cylas formicarius).

Mulching with organic materials is a management practice with long history for weed suppression, soil water conservation and erosion control. Its potential impact on crop pests is less well explored. Here we report its utility for reducing crop damage by the serious pest, sweetpotato weevil (Cylas formicarius). Laboratory bioassays measured the response of adult female weevils to sweetpotato storage roots beneath mulches of fresh or dried plant materials. Weevils were significant repelled by fresh basil, catnip, basil lime and dry eucalyptus, cypress, lucerne and sugarcane. A subsequent field study found that mulches of dry cypress, eucalyptus and lucerne reduced movement of weevils from a release point to reach sweetpotato plants and lowered level of damage to storage roots. Results demonstrate that mulching with organic materials merits further testing as part of the integrated management of sweetpotato weevil, particularly to protect developing storage roots during dry periods when soil cracking can facilitate access by pests.

Construction of a high density linkage map and genome dissection of bruchid resistance in zombi pea (Vigna vexillata (L.) A. Rich).

Zombi pea (Vigna vexillata) is a legume crop that is resistant to several biotic and abiotic stresses. Callosobruchus maculatus and Callosobruchus chinensis are serious stored-insect pests of legume crops. We constructed a high-density linkage map and performed quantitative trait loci (QTLs) mapping for resistance to these insect species in zombi pea. An F2 population of 198 individuals from a cross between 'TVNu 240' (resistant) and 'TVNu 1623' (susceptible) varieties was used to construct a linkage map of 6,529 single nucleotide polymorphism markers generated from sequencing amplified fragments of specific loci. The map comprised 11 linkage groups, spanning 1,740.9 cM, with an average of 593.5 markers per linkage group and an average distance of 0.27 cM between markers. High levels of micro-synteny between V. vexillata and cowpea (Vigna unguiculata), mungbean (Vigna radiata), azuki bean (Vigna angularis) and common bean (Phaseolus vulgaris) were found. One major and three minor QTLs for C. chinensis resistance and one major and one minor QTLs for C. maculatus resistance were identified. The major QTLs for resistance to C. chinensis and C. maculatus appeared to be the same locus. The linkage map developed in this study will facilitate the identification of useful genes/QTLs in zombi pea.

Development of an SNP-based high-density linkage map and QTL analysis for bruchid (Callosobruchus maculatus F.) resistance in black gram (Vigna mungo (L.) Hepper).

Black gram (Vigna mungo var. mungo) is an important pulse crop in Asia. The cowpea weevil (Callosobruchus maculatus) is a stored-seed insect pest (seed weevil/bruchid) that causes serious postharvest losses in pulse crops, including black gram. In this study, we constructed a high-density linkage map for black gram and identified quantitative trait loci (QTLs) for C. maculatus resistance. A recombinant inbred line (RIL) population of 150 lines from a cross between BC48 [cultivated black gram (var. mungo); bruchid-susceptible] and TC2210 [wild black gram (var. silvestris); bruchid-resistant] were used to construct a linkage map of 3,675 SNP markers from specific-locus amplified fragment sequencing. The map comprised 11 linkage groups spanning 1,588.7 cM with an average distance between adjacent markers of 0.57 cM. Seeds of the RIL population grown in 2016 and 2017 were evaluated for C. maculatus resistance through two traits; the percentage of damaged seeds (PDS) and infestation severity progress (AUDPS). Inclusive composite interval mapping identified three QTLs each for PDS and AUDPS. Two QTLs, qVmunBr6.1 and qVmunBr6.2, mapped about 10 cM apart on linkage group 6 were common between PDS and AUDPS. Comparative genome analysis revealed that qVmunBr6.1 and qVmunBr6.2 are new loci for C. maculatus resistance in Vigna species and that genes encoding a lectin receptor kinase and chitinase are candidates for qVmunBr6.2. The high-density linkage map constructed and QTLs for bruchid resistance identified in this study will be useful for molecular breeding of black gram.

Screening hybrid poplar clones for susceptibility to Cryptorhynchus lapathi (Coleoptera: Curculionidae).

The poplar-and-willow borer, Cryptorhynchus lapathi (L.) (Coleoptera: Curculionidae), is a wood-boring pest of economic importance in irrigated hybrid poplar (Populus spp.) farms in eastern Washington and Oregon. There is no practical insecticide control tactic against either the larval or adult stage of C. lapathi. To assess variability in C. lapathi toward clone preference, we initiated a no-choice study on 180 caged trees that consisted of five clones in a randomized complete block design. C. lapathi was significantly more successful at establishing a population in two clones with Populus trichocarpa X P. deltoides (TxD) parentage (female x male) than in either of two clones with P. deltoides x P. nigra (DxN) parentage (female x male), or a single clone with P. deltoides x P. maximowiczii (DxM) parentage (female X male). There were no significant differences in the rate of weevil development among infested clones, with the exception of DxM trees. Larvae in DxM clones developed on average to the fourth size grouping and those in the two TxD clones developed on average to the fifth size grouping, and this difference was significant. These results corroborate our general damage surveys conducted in the field. Our findings provide growers with the option to choose less susceptible varieties when replanting.