Use of pheromone chemistry to identify Grapholita lobarzewskii as an occasional pest of apple and plum

As shown by chemical analysis and field trapping, the sex pheromone of Grapholita lobarzewskii consists of a blend of (E)-8-dodecenyl acetate and (Z)-8-dodecenyl acetate. Maximum attractiveness was found at 80–95%. Dodecyl acetate and tetradecyl acetate, both present in the female gland, did not affect trap catch. G. lobarzewskii has recently gained importance as a pest of apple and plum, but has so far been referred to as Grapholita janthinana. The latter is attracted to the same two compounds, but in reversed portions (20% E).

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Zusammenfassung Chemische Analysen und Feldversuche zeigen, dass das Sexualpheromon des Kleinen Fruchtwicklers, G. lobarzewskii aus (E)-8-Dodecenylacetat (Z8-12:Ac) und (Z)-8-Dodecenylacetat (Z8-12:Ac) besteht. Die grösste Lockwirkung wurde mit einem Gemisch der beiden Substanzen bei einem E-Anteil von 80–95% erzielt. Dodecylacetat (12:Ac) und Tetradecylacetat (14:Ac) wurden ebenfalls in den weiblichen Pheromondrüsen nachgewiesen, hatten aber im Freiland keinen Einfluss auf die Lockwirkung. G. lobarzewskii tritt seit einiger Zeit in der Schweiz als Schädling von Apfel und Zwetschge auf und ist an ihrem charakteristischem Frassgang leicht erkennbar. Die Art wurde bisher irrtümlich als G. janthinana bezeichnet. G. janthinana lebt auf Rosaceen, und wird vom gleichen Substanzpaar angelockt, allerdings bei umgekehrtem Mischungsverhältnis (20% E).

     

Response of plum curculio (Coleoptera: Curculionidae) to odor-baited traps near woods.

Response of overwintered plum curculios, Conotrachelus nenuphar (Herbst), to odor-baited traps was evaluated from the beginning until nearly the end of emigration from overwintering sites in woods. We evaluated clear sticky Plexiglas panels and black pyramid traps placed close to woods adjacent to apple trees in an unsprayed section of an orchard. Traps were baited with aggregation pheromone (grandisoic acid) alone or in combination with one of six synthetic fruit volatiles (benzaldehvde, decyl aldehyde, E-2-hexenal, ethyl isovalerate, hexyl acetate, or limonene). Unbaited traps served as a control treatment. Plum curculio emigration from woods was divided into early-, mid-, and late-season periods based primarily on phenological stage of apple bud and fruit development (tight cluster to bloom, petal fall, and fruit set, respectively). During both early- and late season, panel and pyramid traps baited with benzaldehyde plus pheromone were significantly more attractive than any other traps (baited or unbaited), except panel traps baited with ethyl isovalerate plus pheromone in early season, which likewise captured significantly more adults than unbaited panel traps. During midseason, no lures were significantly attractive, possibly due to prevailing cool weather, unfavorable for adult activity. Over the entire season, panel or pyramid traps baited with benzaldehyde plus pheromone captured nearly six times as many plum curculios as unbaited traps of each type, whereas traps baited with pheromone alone captured about twice as many as unbaited traps of each type. We provide information on sex ratio, female maturity stage, and mating status, and several weather parameters associated with trap captures. We conclude that panel or pyramid traps, or a combination, baited with benzaldehyde plus pheromone placed at borders of plum curculio overwintering sites can be a valuable tool for monitoring the beginning, peak, and end of adult immigration into apple orchards.     

Impact of temperature on plum curculio (Coleoptera: Curculionidae) responses to odor-baited traps

In 2005, captures of overwintered adult plum curculios, Conotrachelus nenuphar (Herbst) (Coleoptera: Curculionidae), in standard black masonite pyramid traps deployed in apple (Malus spp.) orchards from half-inch green until fruit reached 7 mm and baited with known attractants did not result in significant captures compared with unbaited traps as they had in 2003 and 2004. These baits included the synthetic aggregation pheromone, grandisoic acid (GA) alone, a six-component synthetic host plant volatile combination (6-Tree) identified from foliar and woody tissues of a Stanley plum tree in combination with GA (6-Tree+GA), and the synthetic fruit volatile benzaldehyde (BEN) in combination with GA (BEN+GA). In 2005, the average daily temperature was below 13 degrees C, much cooler than in 2003 and 2004. We hypothesized that plum curculio could not discriminate between baited and unbaited traps because of reduced release rates of odor-bait stimuli due to their temperature-driven release system. From data collected from 2003 to 2005, we found that plum curculio captures in traps baited with GA alone, 6-Tree+GA, and BEN+GA were significantly related to temperature. We created a predictive model to determine the level of activity, i.e., trap captures in baited traps compared with unbaited traps, we would expect to observe at a particular temperature for these same odor stimuli. Our models predicts that at temperatures between approximately 11 to 13 degrees C we would expect to see no difference between captures in baited and unbaited traps. For captures in odor-baited traps to reach twice those in unbaited traps, our model predicts that temperatures must reach 19.2 degrees C for GA alone, 18.5 degrees C for 6-Tree+GA, and 15.8 degrees C for BEN+GA.     

3 using odor-baited trap trees as sentinels to monitor plum curculio (Coleoptera: Curculionidae) in apple orchards

In commercial orchards in Massachusetts in 2003, we conducted experiments aimed at developing guidelines for use of perimeter-row trap trees baited with grandisoic acid plus benzaldehyde as sentinels in a practical approach to determining need and timing of insecticide applications against overwintered plum curculios, Conotrachelus nenuphar (Herbst). Evaluations were based on percentages of sampled fruit injured by plum curculio. Trap trees baited with grandisoic acid released at approximately 1 mg/d plus benzaldehyde released at approximately 40 mg/d performed as well as or better than trap trees baited with greater or lesser amounts of these attractants in combination. The distance over which a trap tree baited with such odor was effective in aggregating damage to fruit extended to at least 31-33 m (maximum evaluated) along a perimeter row. Trap trees at corners of orchard blocks were as effective as perimeter-row trap trees midway between corner trees. Within the canopy of a trap tree, damage did not tend significantly to be localized in the vicinity of the odor source but tended to be rather evenly distributed among various sectors of the canopy. Finally, among three candidate thresholds evaluated as a trigger for insecticide application, a threshold of one freshly injured fruit proved better than thresholds of two or four freshly injured fruit out of 50 fruit sampled on a trap tree in assuring that orchard-wide damage would remain below a preset economic injury level of 1%. Our findings lead us to suggest that after a whole-orchard application of insecticide to apple trees in Massachusetts orchards shortly after petal fall, subsequent applications of insecticide against plum curculio can be confined to peripheral-row trees and be driven by a provisional threshold of one freshly injured fruit out of 50 fruit sampled on a perimeter-row trap tree baited with the above-mentioned odor.     

Susceptibility of fruit from diverse apple and crabapple germplasm to attack by plum curculio (Coleoptera: Curculionidae)

Plum curculio, Conotrachelus nenuphar (Herbst) (Coleoptera: Curculionidae), is an important apple, Malus domestica Borkh., pest that significantly hinders sustainable apple production in eastern North America. The potential for host plant resistance to plum curculio among apple germplasm has never been rigorously evaluated. Thus, studies were conducted to assess the susceptibility of a number of exotic and domestic Malus accessions housed at the USDA Plant Genetic Resources Unit (PGRU) "core" collection in Geneva, NY. Contrary to earlier published reports and promising data from a field assessment in 2005, these results suggest that there is probably little potential for genetic resistance to plum curculio among the Malus germplasm collection evaluated. More specifically, four Malus hybrid selections that have previously been released with claims of plum curculio resistance were shown to be susceptible to plum curculio attack. Because there are additional accessions housed at PGRU outside of the core collection that are currently classified as resistant, further studies are necessary to evaluate the true resistance qualities of these releases. It is also important to clarify such discrepancies in both the USDA online Germplasm Resources Information Network and in the horticultural literature. Although other Malus species exhibited some variability in fruit susceptibility, none could be classified as being truly resistant to plum curculio attack by any definition that would have relevance to commercial production and sale of apples.     

Functional modes and residue flexibility control the anisotropic response of guanylate kinase to mechanical stress

The coupling between the mechanical properties of enzymes and their biological activity is a well-established feature that has been the object of numerous experimental and theoretical works. In particular, recent experiments show that enzymatic function can be modulated anisotropically by mechanical stress. We study such phenomena using a method for investigating local flexibility on the residue scale that combines a reduced protein representation with Brownian dynamics simulations. We performed calculations on the enzyme guanylate kinase to study its mechanical response when submitted to anisotropic deformations. The resulting modifications of the protein's rigidity profile can be related to the changes in substrate binding affinity observed experimentally. Further analysis of the principal components of motion of the trajectories shows how the application of a mechanical constraint on the protein can disrupt its dynamics, thus leading to a decrease of the enzyme's catalytic rate. Eventually, a systematic probe of the protein surface led to the prediction of potential hotspots where the application of an external constraint would produce a large functional response both from the mechanical and dynamical points of view. Such enzyme-engineering approaches open the possibility to tune catalytic function by varying selected external forces.     

Expression profile analysis to identify circular RNA expression signatures in hair follicle of Hu sheep lambskin

CircRNAs are involved in the regulation of various cellular and biological processes, but none of the studies have focused on hair follicle in sheep. In this study, the expression profile of circRNAs between small waves (SM) and straight wool (ST) groups was investigated using RNA-Seq. The results showed that a total of 5,527 circRNAs were identified and 114 of them were differentially expressed between two groups. Enrichment analysis revealed that the host genes with DE circRNAs were mainly enriched in TGF-beta pathway, Notch pathway. Miranda software was used to found that 129 miRNAs might be binding to 114 DE circRNAs, including miR-10a, miR-143, miR-let-7a, miR-199a-3p, miR-200a, which also had important influence on hair follicle morphogenesis. Furthermore, the coding potential of circRNAs was predicted, and 11 circRNAs were simultaneously identified with coding potential. In summary, circRNAs have important effects on hair follicle growth and development, and these results will provide a basis for molecular mechanism of pattern formation.     

Use of selection with recurrent backcrossing and QTL mapping to identify loci contributing to southern leaf blight resistance in a highly resistant maize line

B73 is a historically important maize line with excellent yield potential but high susceptibility to the foliar disease southern leaf blight (SLB). NC292 and NC330 are B73 near-isogenic lines (NILs) that are highly resistant to SLB. They were derived by repeated backcrossing of an elite source of SLB resistance (NC250P) to B73, with selection for SLB resistance among and within backcross families. The goal of this paper was to characterize the loci responsible for the increased SLB resistance of NC292 and NC330 and to determine how many of the SLB disease resistance quantitative trait loci (dQTL) were selected for in the development of NC292 and NC330. Genomic regions that differentiated NC292 and NC330 from B73 and which may contribute to NC292 and NC330s enhanced SLB resistance were identified. Ten NC250P-derived introgressions were identified in both the NC292 and NC330 genomes of which eight were shared between genomes. dQTL were mapped in two F_2:3 populations derived from lines very closely related to the original parents of NC292 and NC330—(B73rhm1 × NC250A and NC250A × B73). Nine SLB dQTL were mapped in the combined populations using combined SLB disease data over all locations (SLB AllLocs). Of these, four dQTL precisely colocalized with NC250P introgressions in bins 2.05–2.06, 3.03, 6.01, and 9.02 and three were identified near NC250P introgressions in bins 1.09, 5.05–5.06, and 10.03. Therefore the breeding program used to develop NC292 and NC330 was highly effective in selecting for multiple SLB resistance alleles. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Use of antibiotic resistance analysis to identify nonpoint sources of fecal pollution.

A study was conducted to determine the reliability and repeatability of antibiotic resistance analysis as a method of identifying the sources of fecal pollution in surface water and groundwater. Four large sets of isolates of fecal streptococci (from 2,635 to 5,990 isolates per set) were obtained from 236 samples of human sewage and septage, cattle and poultry feces, and pristine waters. The patterns of resistance of the isolates to each of four concentrations of up to nine antibiotics were analyzed by discriminant analysis. When isolates were classified individually, the average rate of correct classification (ARCC) into four possible types (human, cattle, poultry, and wild) ranged from 64 to 78%. When the resistance patterns of all isolates from each sample were averaged and the resulting sample-level resistance patterns were classified, the ARCCs were much higher (96 to 100%). These data confirm that there are measurable and consistent differences in the antibiotic resistance patterns of fecal streptococci isolated from various sources of fecal pollution and that antibiotic resistance analysis can be used to classify and identify these sources.     

Effectiveness of odor-baited trap trees for plum curculio (Coleoptera: Curculionidae) monitoring in commercial apple orchards in the northeast

The plum curculio, Conotrachelus nenuphar (Herbst), is a key pest of pome and stone fruit in eastern and central North America. For effective management of this insect pest in commercial apple (Malus spp.) orchards in the northeastern United States and Canada, one of the greatest challenges has been to determine the need for and timing of insecticide applications that will protect apple fruit from injury by adults. In a 2004-2005 study, we assessed the efficacy and economic viability of a reduced-risk integrated pest management strategy involving an odor-baited trap tree approach to determine need for and timing of insecticide use against plum curculio based on appearance of fresh egg-laying scars. Evaluations took place in commercial apple orchards in seven northeastern U.S. states. More specifically, we compared the trap-tree approach with three calendar-driven whole-block sprays and with heat-unit accumulation models that predict how long insecticide should be applied to orchard trees to prevent injury by plum curculio late in the season. Trap tree plots received a whole-plot insecticide spray by the time of petal fall, and succeeding sprays (if needed) were applied to peripheral-row trees only, depending on a threshold of one fresh plum curculio egg-laying scar out of 25 fruit sampled from a single trap tree. In both years, level of plum curculio injury to fruit sampled from perimeter-row, the most interior-row trees and whole-plot injury in trap tree plots did not differ significantly from that recorded in plots subject to conventional management or in plots managed using the heat-unit accumulation approach. The amount of insecticide used in trap tree plots was reduced at least by 43% compared with plots managed with the conventional approach. Advantages and potential pitfalls of the bio-based trap tree approach to plum curculio monitoring in apple orchards are discussed.     

Use of SSCP analysis to identify germline mutations in HNPCC families fulfilling the Amsterdam criteria

Hereditary non-polyposis colorectal cancer (HNPCC) is a clinical syndrome characterised by an inherited predisposition to early onset colorectal and uterine cancers and an increased incidence of other cancers. It is caused by germline defects in the human mismatch repair genes. Defects in two of the known mismatch repair genes (namely hMSH2 and hMLH1) account for over 90% of mutations found in HNPCC families. In this study we have identified 14 families that fulfilled the clinical criteria for HNPCC and screened the hMSH2 and hMLH1 genes for germline mutations using single-strand conformational polymorphism (SSCP) analysis and DNA sequencing. Seven mutations were identified. Of these, there were five frameshifts, one missense mutation and a further novel mutation that involved separate transition and transversion changes in successive amino acid residues. Three of the mutations were in hMSH2 and four in hMLH1. The identification of germ-line mutations in an HNPCC family enables targeted surveillance and the possibility of early curative intervention. SSCP is a simple and effective method for identifying most mutations in the human mismatch repair genes using DNA from fresh, frozen or archival material. Received: 24 July 1996 / Revised: 26 September 1996     

Lipoxygenase activity in apples in relation to storage and physiological disorders

Different methods for the preparation of active lipoxygenase (LOX) extracts from apples were compared. Highest activities were obtained using a 0.25 M phosphate buffer (pH 7) containing 1% Triton X-100 and 10^?2 M metabisulphite as extraction solvent. LOX activity during storage was investigated in the core, flesh, and peel. Activity was always highest in the core and peel. On storage, activity was increased in each part of the fruit but especially in the core and peel. Increase in LOX preceded the browning of the core. LOX may be responsible for the browning and may be concerned in the induction of superficial scald.     

Use of granulated fertilizers with Actellic EC 50 insecticide in the control of mosquito larvae

Granulated fertilizers [ammonium nitrate, synthetic fertilizer] were used as carriers to prepare an insecticide granulate suitable for the control of Aedes cantans and Aedes vexans mosquito larvae under field conditions. The insecticide granulate was obtained by mixing 1 volume unit of Actellic EC 50 with 50 volume units of fertilizer. The optimal dose required to ensure and effective control of Aedes mosquito larvae was 2--5 g of granulate per 1 m3 of water [i.e. 2--5 kg/ha per each 1- cm depth of water] in hatching areas with a relatively clean water or during periods of slow larval growth and development, and 10 g of granulate per 1 m3 of water for hatching places polluted with organic substances or for periods of accelerated larval growth during summer months. The described Actellic EC 50 application practice ensures a complete survival of numerous non-target water organisms, including Tubifex species, Rhynchelmis species, molluscs, flatworms [Turbellaria], Asellus aquaticus, water mites [Hydrachna sp.] water striders (Gerris lacustris] and Dixella species larvae].     

Use of representational difference analysis to identify Escherichia coli O157-specific DNA sequences

Whereas several important virulence factors in Escherichia coli O157 have been identified, studies suggest they are not always essential and are probably insufficient to account for the severe clinical manifestation of E. coli O157 infection. Identification of putative virulence determinants is crucial to the understanding of bacterial pathogenesis and genomic comparison analysis may aid the characterisation of unidentified virulence attributes. In this study, representational difference analysis (RDA) was used for genomic comparison of E. coli O157 with the proposed ancestral strain, E. coli O55. Unique E. coli O157 gene sequences were isolated and one, termed RDA-1, taken forward for further analysis. Southern blotting with labelled RDA-1 as a probe showed it to be present in 77% of E. coli O157 isolates and absent in all non-E. coli O157 screened. Sequence flanking RDA-1 was obtained from a genomic clone identified by hybridisation, and contained an open reading frame predicted to encode a novel iron-regulated outer membrane protein.     

Use of a gut content ELISA to detect whitefly predator feeding activity after field exposure to different insecticide treatments

A 2-year commercial-scale study was conducted to qualitatively evaluate the effect of different insecticide treatment regimes on the predator complex attacking Bemisia tabaci (Gennadius) in cotton. In 1996 three insecticide regimes were compared: a rotation of conventional broad-spectrum insecticides or one of two different regimes based on the initial use of the insect growth regulators (IGRs), buprofezin and pyriproxyfen. In 1997 the same three regimes plus an untreated control were compared; split-plots were sprayed once for Lygus hesperus Knight control using a broad-spectrum insecticide. Relative feeding activity for each predator species was compared between treatment regimes by analyzing the gut contents of predators for the presence of whitefly remains using a whitefly-specific enzyme-linked immunosorbent assay (ELISA). The ELISA results were combined with predator density data to obtain a qualitative pesticide impact index for each predator group. In total, we analyzed the gut contents of 32 262 field-collected predators, representing nine different taxa. Of these, Pseudatomocelis seriatus (Reuter), Spanagonicus albofasciatus (Reuter), and spiders consisting primarily of Misumenops celer (Hentz) are shown to be whitefly predators for the first time. Predator populations were usually reduced in plots that received applications of broad-spectrum insecticides for B. tabaci and L. hesperus control, but there were few treatment differences in the proportions of predators containing whitefly remains in their guts. However, the feeding activity of certain predator species in fields sprayed with broad spectrum insecticides was significantly reduced compared with those in IGR-based and control treatments. Overall, insecticide regimes using IGRs were less lethal to the whitefly predator complex than regimes consisting of only conventional, broad-spectrum insecticides, but differences in predator feeding activity on whitefly between the various insecticide treatment regimes were minimal.     

Use of systemic insecticide to measure antixenosis to aphids in plant choice experiments

Antixenosis in three wheat varieties (Timmo, Moghan 2 and Ommid) toRhopalosiphum padi (L.) was measured by pairing plants of any two varieties with one plant of each pair treated with a soil drench of the systemic insecticide pirimicarb. Aphids were then allowed access to equivalent leaf areas from the two plants in a leaf cage. The results were compared with similar choice experiments without insecticide. By either technique, the two Iranian varieties (Moghan 2 and Ommid) showed antixenosis in comparison with Timmo. The statistical analysis of the data, using a heterogeneity ² test, is described.

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Zusammenfassung Die Antixenose gegenüberRhopalosiphum padi (L.) in den drei Weizensorten Timmo, Moghan 2 und Ommid, von denen die zwei letzteren Sorten aus dem Iran stammen, wurde unter Verwendung des systemischen Insektizids Pirimicarb gemessen. Jeweils zwei Pflanzen aus verschiedenen Sorten wurden als Paar verwendet (in allen Kombinationen der drei Sorten), pro Paar ein Topf mit Pirimicarb gegossen, und zwanzig Blattlaüse über Blattflächen von beiden Sorten gekäfigt. Ähnliche Versuche wurden auch ohne Insektizid ausgeführt. Beide Methoden zeigten deutliche Antixenose von Moghan 2 und Ommid in Vergleich mit Timmo. Die Insektizidmethode zeigte auch eine Bevorzugung von Moghan 2 gegenüber Ommid, ein Resultat, das mit der schon bekannten relativen Antibiose dieser zwei Sorten übereinstimmte, während sich ohne Insektizid kein solcher Unterschied erwies. Die statistische Analyse solcher Versuche mit einem Insektizid, die eine Modifikation der Berechnung des erwarteten Werts mit dem ² Test enthält, ist beschrieben.

     

Use of a genome-wide approach to identify new genes that control resistance of Saccharomyces cerevisiae to ionizing radiation

We have used the recently completed set of all homozygous diploid deletion mutants in budding yeast, S. cerevisiae, to screen for new mutants conferring sensitivity to ionizing radiation. In each strain a different open reading frame (ORF) has been replaced with a cassette containing unique 20-mer sequences that allow the relative abundance of each strain in a pool to be determined by hybridization to a high-density oligonucleotide array. Putative radiation-sensitive mutants were identified as having a reduced abundance in the pool of 4,627 individual deletion strains after irradiation. Of the top 33 strains most sensitive to radiation in this assay, 14 contained genes known to be involved in DNA repair. Eight of the remaining deletion mutants were studied. Only one, which deleted for the ORF YDR014W (which we name RAD61), conferred reproducible radiation sensitivity in both the haploid and diploid deletions and had no problem with spore viability when the haploid was backcrossed to wild-type. The rest showed only marginal sensitivity as haploids, and many had problems with spore viability when backcrossed, suggesting the presence of gross aneuploidy or polyploidy in strains initially presumed haploid. Our results emphasize that secondary mutations or deviations from euploidy can be a problem in screening this resource for sensitivity to ionizing radiation.