Evidence for Sexual Reproduction: Identification, Frequency, and Spatial Distribution of Venturia effusa (Pecan Scab) Mating Type Idiomorphs.

Venturia effusa (syn. Fusicladium effusum), causal agent of pecan scab, is the most prevalent pathogen of pecan (Carya illinoinensis), causing severe yield losses in the southeastern United States. V. effusa is currently known only by its asexual (conidial) stage. However, the degree and distribution of genetic diversity observed within and among populations of V. effusa are typical of a sexually reproducing fungal pathogen, and comparable with other dothideomycetes with a known sexual stage, including the closely related apple scab pathogen, V. inaequalis. Using the mating type (MAT) idiomorphs from V. inaequalis, we identified a single MAT gene, MAT1-1-1, in a draft genome of V. effusa. The MAT1-1-1 locus is flanked by two conserved genes encoding a DNA lyase (APN2) and a hypothetical protein. The MAT locus spanning the flanking genes was amplified and sequenced from a subset of 14 isolates, of which 7 contained MAT1-1-1 and the remaining samples contained MAT1-2-1. A multiplex polymerase chain reaction screen was developed to amplify MAT1-1-1, MAT1-2-1, and a conserved reference gene encoding ß-tubulin, and used to screen 784 monoconidial isolates of V. effusa collected from 11 populations of pecan across the southeastern United States. A hierarchical sampling protocol representing region, orchard, and tree allowed for analysis of MAT structure at different spatial scales. Analysis of this collection revealed the frequency of the MAT idiomorphs is in a 1:1 equilibrium of MAT1-1:MAT1-2. The apparent equilibrium of the MAT idiomorphs provides impetus for a renewed effort to search for the sexual stage of V. effusa. [Formula: see text] Copyright © 2018 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license .

Population Genetic Structure of Venturia effusa, Cause of Pecan Scab, in the Southeastern United States.

Venturia effusa is the most important pathogen of pecan in the southeastern United States. Little information exists on the population biology and genetic diversity of the pathogen. A hierarchical sampling of 784 isolates from 63 trees in 11 pecan orchards in the southeastern United States were screened against a set of 30 previously characterized microsatellite markers. Populations were collected from Georgia (n = 2), Florida (n = 1), Alabama (n = 2), Mississippi (n = 1), Louisiana (n = 1), Illinois (n = 1), Oklahoma (n = 1), Texas (n = 1), and Kansas (n = 1). Clonality was low in all orchard populations (≤10.1% of isolates), and there were consistently high levels of genotypic diversity (Shannon-Weiner's index = 3.49 to 4.59) and gene diversity (Nei's measure = 0.513 to 0.713). Analysis of molecular variance showed that, although 81% of genetic diversity occurred at the scale of the individual tree, 16% occurred between orchards and only 3% between trees within orchards. All populations could be differentiated from each other (P = 0.01), and various cluster analyses indicated that some populations were more closely related compared with other pairs of populations. This is indicative of some limited population differentiation in V. effusa in the southeastern United States. Bayesian and nearest-neighbor methods suggested eight clusters, with orchards from Georgia and Florida being grouped together. A minimum spanning tree of all 784 isolates also indicated some isolate identification with source population. Linkage disequilibrium was detected in all but one population (Kansas), although 8 of the 11 populations had <20% of loci at disequilibrium. A Mantel test demonstrated a relationship between physical and genetic distance between populations (Z = 11.9, r = 0.559, P = 0.001). None of the populations were at mutation-drift equilibrium. All but 3 of the 11 populations had a deficiency of gene diversity compared with that expected at mutation-drift equilibrium (indicating population expansion); the remaining populations had an excess of gene diversity compared with that expected at mutation-drift equilibrium (indicating a recent bottleneck). These observations are consistent with the known history of pecan and pecan scab, which is that V. effusa became an issue on cultivated pecan in the last approximately 120 years (recent population expansion). Recently reported mating type genes and the sexual stage of this fungus may help explain the observed population characteristics, which bear a strong resemblance to those of other well-characterized sexually reproducing ascomycete pathogens.

Severity of Scab and its Effects on Fruit Weight in Mechanically Hedge-Pruned and Topped Pecan Trees.

Scab is the most damaging disease of pecan in the southeastern United States. Pecan trees can attain 44 m in height, so managing disease in the upper canopy is a problem. Fungicide is ordinarily applied using ground-based air-blast sprayers. Although mechanical hedge-pruning and topping of pecan is done for several reasons, improved management of scab is an important reason in the humid, wet Southeast. Resulting shoot growth on cut limbs of susceptible cultivars could lead to more severe scab. In three experiments over three years, we explored the effect of hedge-pruning trees to ∼12 to 14 m compared with non-hedge-pruned trees. All trees received fungicide treatments (air-blast sprays and ≤3 aerial applications). Hedge-pruning either had no effect, or increased or decreased scab severity only slightly on leaflets, immature, or mature fruit (a -9.95 to +14.63% difference in scab severity compared with the control). However, height in the canopy invariably had a large and significant effect on scab severity, and amounted to a 0.05 to 73.77% difference in severity between the lowest and highest sample in the canopy. Fruit weight depended on sample height, with fruit most often weighing less when collected at greater sample heights. A robust relationship between fruit weight and scab severity was found at the highest sample heights where scab was also most often severe (R2 = 0.21 to 0.67, P < 0.0001). Hedge-pruning and topping pecan tree canopies to manage tree size will enable better fungicide coverage, reducing risk of a scab epidemic as more of the canopy is assured efficacious fungicide spray coverage.

Scab Susceptibility of a Provenance Collection of Pecan in Three Different Seasons in the Southeastern United States.

Pecan scab (caused by Fusicladium effusum) is the most economically destructive disease of pecan in the Southeast United States. Wet, humid conditions typical of the Southeast are known to provide conditions conducive to epidemics. A provenance collection of pecan from 19 locations representing the native range of the tree is located in Byron, Georgia, and was assessed for pecan scab severity in 1998, 2013, and 2014. There were significant differences among the 19 provenances (F = 5.6 to 62.5, P < 0.0001). Provenances from wetter locations (generally north of Texas) had the greatest proportion of scab resistant trees, while provenances from the drier southern areas (Texas and Mexico) tended to be the most susceptible to scab. The association with rainfall was borne out by correlation analysis (r = -0.625 to -0.823 [P < 0.0001 to 0.004]). Other factors consistently associated with scab severity included leaflet tilt and droop angle (r = -0.533 to -0.883 [P < 0.0001 to 0.02]). Multiple regression analysis demonstrated that leaflet droop angle was a particularly good predictor of provenance susceptibility. Leaflet characteristics vary with provenance location, and whether there is a direct relationship between scab severity and leaflet characteristics is not established. Estimates of heritability were not entirely consistent among years, but different methods were used to assess scab severity in 1998 (a 1 to 5 category scale) compared with 2013 and 2014 (the percent ratio scale). Despite using different methods, there was generally good agreement among years in regard to severity of disease on individual trees. In conclusion, trees from more northern populations (in areas with greater annual rainfall) are most likely to provide valuable and diverse sources of resistance to scab. The provenance collection contains a range of scab-resistant genotypes from diverse locations that can contribute to genetic improvement regarding scab resistance.

The Effect of Sample Height on Spray Coverage in Mature Pecan Trees.

Pecan scab (caused by Fusicladium effusum) is the most damaging disease of pecan in the southeastern United States. Large air-blast sprayers for orchards are used to apply fungicide to control the disease but little quantitative information exists on the spray coverage achieved in the canopy of these trees. A series of experiments using water-sensitive spray cards to record spray coverage (percent area) at different heights and locations up to 15 m in the canopy of pecan trees showed a significantly greater percentage of card area covered at the lowest sample height when compared with the highest sample height. At the lowest height (5 m), spray coverage on individual cards ranged from 4.7 to 73.5% and, at the highest sample height (15 m), spray coverage ranged from 0.02 to 9.5%. In general, there was little significant difference in spray card coverage up to at least 10 m but, at 12.5 and 15 m, there was significantly less spray coverage compared with the coverage at 5 m. Regression analysis indicated a consistent linear relationship between sample height in the tree and the percent area covered. When spray cards were positioned at different heights without possible interference from pecan limbs and foliage, similar effects of sample height on spray coverage were noted. Wind speed measurements showed that air movement declined rapidly with distance from the sprayer fan. Whereas, at 2 m from the fan, wind speeds were approximately 26 m s-1, by 10 m, speeds had declined to 2 to 4 m s-1. At distances >12 m, wind speed was approaching ambient air movement of about 1 to 3 m s-1. Although aerial application resulted in numerically greater spray coverage at sample heights >10 m, it was not significant even though a weak linear relationship (R2 = 0.21 to 0.25) suggested an effect of height. Characterizing and understanding pesticide spray coverage in pecan will allow us to discern limits imposed by existing technology, and provide the basis for improving spray application methods (or tree management) for more efficacious disease control.

Nickel affects xylem Sap RNase a and converts RNase A to a urease.

BACKGROUND: Nickel (Ni) is an essential micronutrient; however, its metabolic or physiological functions in plants and animals are largely uncharacterized. The ribonucleases (RNase, e.g., RNase A) are a large family of hydrolases found in one form or many forms facilitating nitrogen (N) cycling. It is currently unknown how either a deficiency or excess of Ni influences the functionality of ribonucleases, like RNase A. This is especially true for perennial crops possessing relatively high Ni requirements. RESULTS: We report that the 'rising' xylem sap of pecan [Carya illinoinensis (Wangenh.) K. Koch, a long-lived tree] at bud break contains a 14 kDa RNase A (aka, RNase 1), which amount has a 33% greater in Ni-deficient as in Ni-sufficient trees when exposed to Ni ions exhibits ureolytic activity. The homologous 13.4 kDa bovine pancreatic RNase A likewise exhibits ureolytic activity upon exposure to Ni ions. Ni therefore affects enzymatic function of a typically non-metalloenzyme, such as it transforms to an enzyme capable of hydrolyzing a linear amide; thus, converting an endonuclease esterase into a urease. CONCLUSIONS: We conclude that Ni potentially affects the level and activity of RNase A present in the spring xylem sap of pecan trees, and probably in other crops, it has the same influence. The catalytic property of RNase A appears to shift from a nuclease to a urease relying on Ni exposure. This is suggestive that RNase A might possess novel metabolic functionality regarding N-metabolism in perennial plants. The ability of Ni to convert the activity of plant and animal RNase A from that of a ribonuclease to a urease indicates a possible unrecognized beneficial metabolic function of Ni in organisms, while also identifying a potential detrimental effect of excessive Ni on N related metabolic activity if there is sufficient disruption of Ni homeostasis.

Pecan Scab Severity-Effects of Assessment Methods.

Pecan scab is caused by the fungus Fusicladium effusum, and is the most destructive disease of pecan in the United States. Accurate and reliable disease assessments are needed to ensure that data provide a measure of actual disease intensity. The Horsfall-Barratt (H-B) category scale and its derivatives are commonly used to assess disease. Estimates using the H-B scale were compared with nearest percent estimate (NPEs) for rating disease severity of pecan scab on valves of fruit. Both inexperienced and experienced raters were included in the experiment. Lin's concordance correlation showed that agreement using NPEs was variable (ρc = 0.57 to 0.96), whereas estimates of disease severity using the H-B scale had similar agreement among most raters (ρc = 0.59 to 0.98). Converted values of NPEs to the H-B midpoints (NPEH-B) also provided a similar range (ρc = 0.61 to 0.96). Neither experienced nor inexperienced raters were consistently better using any of the three methods. Bootstrap analysis indicated that, among experienced raters, precision (r) and agreement (ρc) were often reduced when using the H-B scale compared with NPEs. There was no consistent effect of converting NPEs to NPEH-B midpoint values compared with actual H-B values. Inter-rater reliability using the H-B scale was never better than NPEs. Bootstrap analysis indicated no difference in the length of time needed to assess disease but regression analysis suggested that raters who were inherently fast in assessing disease with NPEs were often slower when using the H-B scale; conversely, raters who were slow assessing with NPEs were often faster when using the H-B scale. Thus, there appears to be no advantage in accuracy or reliability or reduction in time when inexperienced or experienced raters used a category rating scale to assess pecan scab.

Vertical Distribution of Scab in Large Pecan Trees.

Pecan scab (Fusicladium effusum) is a destructive disease of pecan in the southeastern United States. This study was conducted to investigate the vertical distribution of scab in tall pecan trees (14 to 16 m tall) in three experiments in 2010 and 2011. Although 2010 had average rainfall, a factor that drives scab epidemics, 2011 was a very dry year with a consequently low scab severity. A total of eight trees were included in each experiment, four were nontreated controls, and four were sprayed using a ground-based air-blast sprayer. Trees were assessed for foliar and fruit scab at 0-5.0, 5.0+-7.5, 7.5+-10.0, 10.0+-12.5, and 12.5+-15.0 m. Mixed model analysis showed main effects of height, fungicide treatment, and height*treatment interactions in all three experiments, although on foliage the effects were less consistent (P value = 0.003-0.8), perhaps due to delayed fungicide applications early in the season. However, fruit of nontreated trees had more severe scab low in the canopy compared to fungicide-treated trees, with a consistent height*treatment interaction (P value = <0.0001-0.04). Most often the severity of scab in the upper canopy was similar in trees on fungicide-treated and nontreated trees, suggesting that fungicide had less impact at heights ≥10.0 m compared to <10.0 m in the canopy. There was a consistent reduction in scab severity on foliage and on immature fruit in August due to fungicide treatment at heights ≤10.0 m. Above 10.0 m, the effect was inconsistent, but late in the season (October), the fungicide-treated trees showed lower scab severity throughout the canopy. A metallic tracer study using cerium (Ce) showed an exponential relationship between quantities of Ce recovered and sample height in the canopy, with the quantity of Ce at ≥10 m being statistically equal to background levels. The relationship between scab severity on fruit and sample height in the canopy of nontreated trees was most often described by a negative linear function, but there was no discernible relationship on fungicide-treated trees, as the severity of scab in the lower canopy was most often similar to that in the upper canopy. Gradients in fungicide coverage and scab severity have ramifications for scab management options and potentially for the development of fungicide resistance in F. effusum.

The Effect of Horsfall-Barratt Category Size on the Accuracy and Reliability of Estimates of Pecan Scab Severity.

Pecan scab (Fusicladium effusum) is a destructive pecan disease. Disease assessments may be made using interval-scale-based methods or estimates of severity to the nearest percent area diseased. To explore the effects of rating method-Horsfall-Barratt (H-B) scale estimates versus nearest percent estimates (NPEs)-on the accuracy and reliability of severity estimates over different actual pecan scab severity ranges on fruit valves, raters assessed two cohorts of images with actual area (0 to 6, 6+ to 25%, and 25+ to 75%) diseased. Mean estimated disease within each actual disease severity range varied substantially. Means estimated by NPE within each actual disease severity range were not necessarily good predictors of the H-B scale estimate at <25% severity. H-B estimates by raters most often placed severity in the wrong category compared with actual disease. Measures of bias, accuracy, precision, and agreement using Lin's concordance correlation depended on the range of actual severity, with improvements increasing with actual disease severity category (from 0 to 6 through 25+ to 75%); however, the improvement was unaffected by the H-B assessments. Bootstrap analysis indicated that NPEs provided either equally good or more accurate and precise estimate of disease compared with the H-B scale at severities of 25+ to 75%. Inter-rater reliability using NPEs was greater at 25+ to 75% actual disease severity compared with using the H-B scale. Using NPEs compared with the H-B scale will more often result in more precise and accurate estimates of pecan scab severity, particularly when estimating actual disease severities of 25+ to 75%.

Control of key pecan insect pests using biorational pesticides.

Key pecan insect pests include the black pecan aphid, Melanocallis caryaefoliae (Davis), pecan weevil, Curculio caryae (Horn), and stink bugs (Hemiptera: Pentatomidae). Alternative control tactics are needed for management of these pests in organic and conventional systems. Our objective was to evaluate the potential utility of several alternative insecticides including three plant extract formulations, eucalyptus extract, citrus extract-8.92%, and citrus extract-19.4%, and two microbial insecticides, Chromobacterium subtsugae (Martin et al.) and Isaria fumosorosea (Wize). In the laboratory, eucalyptus extract, citrus extract-8.92%, citrus extract-19.4%, and C. subtsugae caused M. caryaefoliae mortality (mortality was reached approximately 78, 83, and 96%, respectively). In field tests, combined applications of I. fumosorosea with eucalyptus extract were synergistic and caused up to 82% mortality in M. caryaefoliae. In laboratory assays focusing on C. caryae suppression, C. subtsugae reduced feeding and oviposition damage, eucalyptus extract and citrus extract-19.4% were ineffective, and antagonism was observed when citrus extract-19.4% was combined with the entomopathogenic nematode, Steinernema carpocapsae (Weiser). In field tests, C. subtsugae reduced C. caryae damage by 55% within the first 3d, and caused 74.5% corrected mortality within 7 d posttreatment. In the laboratory, C. subtsugae and eucalyptus extract did not cause mortality in the brown stink bug, Euschistus servus (Say). Applications of C. subtsugae for suppression of C. caryae, and eucalyptus extract plus I. fumosorosea for control of M. caryaefoliae show promise as alternative insecticides and should be evaluated further.

Effects of combining microbial and chemical insecticides on mortality of the Pecan Weevil (Coleoptera: Curculionidae).

The pecan weevil, Curculio caryae (Horn), is a key pest of pecan [Carya illinoinensis (Wangenh.) K. Koch]. Current control recommendations are based on chemical insecticide applications. Microbial control agents such as the entomopathogenic nematode, Steinernema carpocapsae (Weiser) and the fungus Beauveria bassiana (Balsamo) Vuillemin occur naturally in southeastern U.S. pecan orchards and have shown promise as alternative control agents for C. caryjae. Conceivably, the chemical and microbial agents occur simultaneously within pecan orchards or might be applied concurrently. The objective of this study was to determine the interactions between two chemical insecticides that are used in commercial C. caryae control (i.e., carbaryl and cypermethrin applied below field rates) and the microbial agents B. bassiana and S. carpocapsae. In laboratory experiments, pecan weevil larval or adult mortality was assessed after application of microbial or chemical treatments applied singly or in combination (microbial + chemical agent). The nature of interactions (antagonism, additivity, or synergy) in terms of weevil mortality was evaluated over 9 d (larvae) or 5 d (adults). Results for B. bassiana indicated synergistic activity with carbaryl and antagonism with cypermethrin in C. caryae larvae and adults. For S. carpocapsae, synergy was detected with both chemicals in C. caryae larvae, but only additive effects were detected in adult weevils. Our results indicate that the chemical-microbial combinations tested are compatible with the exception of B. bassiana and cypermethrin. In addition, combinations that exhibited synergistic interactions may provide enhanced C. caryae control in commercial field applications; thus, their potential merits further exploration.

Gibberellic acid decreases Melanocallis caryaefoliae (Hemiptera: Aphididae) population density and chlorotic feeding injury to foliage in pecan orchards.

BACKGROUND: Melanocallis caryaefoliae (Davis), Monellia caryella (Fitch), and Monelliopsis pecanis Bissell (Hemiptera: Aphididae) attack pecan foliage (Carya illinoinensis [Wangenh.] K. Koch). Unlike M. caryella and M. pecanis, feeding by M. caryaefoliae triggers a physiological change within foliage mimicking natural leaf senescence; it can lead to defoliation. Pretreatment of pecan foliage with gibberellic acid (GA3 ) mitigates M. caryaefoliae-elicited physiological disturbances. GA3 application to pecan was evaluated for efficacy regarding effects on M. caryaefoliae populations and possible negative side-effects on two natural enemy species and on return bloom of pecan. RESULTS: All GA3 treatment rate schedules significantly reduced M. caryaefoliae nymphs but not adults or adults and nymphs of M. caryella or M. pecanis. Percentage leaf chlorosis elicited by M. caryaefoliae was significantly reduced by GA3 (i.e., 39.5 to 197.7 g a.i./ha). No negative side-effects of GA3 treatment were detected regarding certain key natural enemy species or on return bloom of pecan. CONCULUSION: Application of GA3 to the orchard canopy protects foliage from senescence-like physiological responses triggered by M. caryaefoliae. This reduces detrimental leaflet chlorosis, both senescence and abscission processes and horticulturally significant feeding injury. Additionally, the absence of apparent negative side-effects on key natural enemies and return bloom is suggestive of a practical means for efficacious non-insecticidal control of M. caryaefoliae populations in orchards. This novel protective effect of GA3 against aphid-elicited, senescence-like physiological responses may merit investigation as an IPM tool to manage aphid species eliciting similar senescence-like damage to other crop species. Published 2020. This article is a U.S. Government work and is in the public domain in the USA.

Chlorotic feeding injury by the black pecan aphid (hemiptera: aphididae) to pecan foliage promotes aphid settling and nymphal development.

The nature of the interaction between the black pecan aphid, Melanocallis caryaefoliae (Davis) (Hemiptera: Aphididae), and the chlorosis it causes to foliage of its pecan [Carya illinoinensis (Wangenh.) K. Koch)] host is poorly understood. Laboratory experiments were conducted on the settling behavior of the black pecan aphid, when provided chlorotic pecan leaf discs resulting from previous black pecan aphid feeding and nonchlorotic leaf discs, under a normal photoperiod and constant dark. Additionally, aphid development from the first instar to the adult stage was examined when nymphs were either allowed to feed on the same leaf disc or moved daily to a new, nondamaged, same age leaf disc. After 24 h, a significantly higher percentage of black pecan aphids settled on chlorotic than on nonchlorotic leaf discs, regardless of photoperiod. When starting from the first instar, nymphs that were prevented from inducing leaf chlorosis by moving daily to new, same-age leaf discs took approximately 5 d longer to complete development, had a shorter body length, and had higher mortality than when aphids remained on the same leaf disc. These results show that black pecan aphid-induced leaf chlorosis plays an important role in the interaction of the black pecan aphid with its pecan host.

Changes of oxidase and hydrolase activities in pecan leaves elicited by black pecan aphid (Hemiptera: Aphididae) feeding.

The black pecan aphid, Melanocallis caryaefoliae (Davis) (Hemiptera: Aphididae), is a foliar feeder of pecan, Carya illinoinensis (Wangenh.) K. Koch (Juglandaceae). The pest causes chlorosis of leaflet lamina, physiological damage to foliage and trees, and commonly limits the profitability of commercial pecan orchard enterprises. However, key aspects of this host-pest interaction are poorly understood. We report here the effects of M. caryaefoliae feeding on the foliar activity of oxidative (i.e., catalase, lipoxygenase [LOX]-1 and 3, and peroxidase) and hydrolytic (i.e., esterase) enzymes in relation to the degree of aphid resistance among pecan varieties. The 2-yr study showed that M. caryaefoliae-infested foliage exhibited elevated peroxidase activity only in susceptible ('Desirable', 'Sumner', and 'Schley'), but not in resistant ('Cape Fear', 'Gloria Grande', and 'Money Maker') genotypes. Susceptible genotypes also exhibited more severe leaf chlorosis in response to M. caryaefoliae feeding than the resistant genotypes; however, the aphid feeding did not influence catalase or esterase activity in all varieties, except the increase of esterase activity in Desirable and Gloria Grande. Melanocallis caryaefoliae feeding also influences activity of two lipoxygenase isozymes, with LOX3 being more frequently induced than LOX1. Foliar LOX3 activity was more frequently induced by M. caryaefoliae feeding in the moderately resistant 'Oconee' and highly resistant Money Maker and Cape Fear than in the susceptible genotypes. Therefore, the elevation of peroxidase is likely to be associated with aphid susceptibility and contributed to the severe leaf chlorosis, whereas the increase of LOX3 activity might be associated with aphid resistance in pecan. These findings contribute to our understanding of the etiology of M. caryaefoliae-elicited leaf chlorosis on pecan foliage. Such information may also be used to develop enzyme markers for identifying black pecan aphid resistance and/or susceptibility in pecan germplasm.

Comparison of application methods for suppressing the pecan weevil (Coleoptera: Curculionidae) with Beauveria bassiana under field conditions.

The pecan weevil, Curculio caryae (Horn), is a key pest of pecans. The entomopathogenic fungus Beauveria bassiana (Balsamo) Vuillemin is pathogenic to C. caryae. One approach to managing C. caryae may be application of B. bassiana directed toward adult weevils as they emerge from the soil to attack nuts in the tree canopy. Our objective was to compare different application methods for suppression of C. caryae adults. Treatments included direct application of B. bassiana (GHA strain) to soil under the tree canopy, soil application followed by cultivation, soil application in conjunction with a cover crop (Sudan grass), direct application to the tree trunk, and application to the trunk with an UV radiation-protecting adjuvant. The study was conducted in a pecan orchard in Byron, GA, in 2005 and 2006. Naturally emerging C. caryae adults, caught after crawling to the trunk, were transported to the laboratory to determine percentage mortality and signs of mycosis. When averaged over the 15-d sampling period, weevil mortality and signs of mycosis were greater in all treatments than in the nontreated control in 2005 and 2006; >75% average mortality was observed with the trunk application both years and in the trunk application with UV protection in 2005. Results indicated trunk applications can produce superior efficacy relative to ground application, particularly if the ground application is followed by cultivation. Efficacy in the cover crop treatment, however, did not differ from other application approaches. Future research should focus on elucidating the causes for treatment differences we observed and the extent to which B. bassiana-induced C. caryae mortality reduces crop damage.

Virulence of Hypocreales fungi to pecan aphids (Hemiptera: Aphididae) in the laboratory.

There is need for efficacious biocontrol agents for aphids in commercial orchards. As a preliminary step to this end we determined the virulence of several Hypocreales fungi to pecan aphids. In the first experiment we tested the virulence of Isaria fumosorosea (ARSEF 3581) blastospores to three pecan aphids Monellia caryella, Melanocallis caryaefoliae, and Monelliopsis pecanis under laboratory conditions. Rates of 1x10(7) or 1x10(8) spores per ml were applied in 2 ml via a spray tower to 90 mm Petri dishes containing 10 aphids each. Mortality and mycosis were determined after 24, 48 and 72 h. Treatment effects were observed by 48 h post-application, and by 72 h the higher application rate caused >90% mortality and mycosis in M. caryella and M. caryaefoliae, whereas <70% was observed in M. pecanis. We conducted two subsequent experiments (Experiments 2 and 3), using the same methodology, to compare the virulence of several Hypocreales species and strains against the aphid of primary economic concern to most pecan growers, M. caryaefoliae. In Experiment 2, we compared blastospores and conidia of two I. fumosorosea strains (ARSEF 3581 and ATCC 20874 [= strain 97]). The blastospores of ARSEF 3581 and conidia of ATCC 20874 showed higher virulence than other treatments and thus were included in Experiment 3, which also compared the virulence of conidia of Beauveria bassiana (GHA strain) and Metarhizium anisopliae (F52 strain). Results in Experiment 3 indicated the highest virulence in I. fumosorosea 3581 blastospores and M. anisopliae (F52) followed by I. fumosorosea (20874) conidia. The detection of pathogenicity to pecan aphids establishes the potential for commercial usage and additional study. Results reported here will narrow treatments to test in future greenhouse and field trials.

Draft genome sequence of Venturia carpophila, the causal agent of peach scab.

Venturia carpophila causes peach scab, a disease that renders peach (Prunus persica) fruit unmarketable. We report a high-quality draft genome sequence (36.9 Mb) of V. carpophila from an isolate collected from a peach tree in central Georgia in the United States. The genome annotation is described and a phylogenetic analysis of the pathogen is presented. The genome sequence will be a useful resource for various studies on the pathogen, including the biology and ecology, taxonomy and phylogeny, host interaction and coevolution, isolation and characterization of genes of interest, and development of molecular markers for genotyping and mapping.

Taxonomic and Biological Characterization of Steinernema rarum Found in the Southeastern United States.

Two Steinernema isolates found in Louisiana and Mississippi were later identified as isolates of S. rarum. DNA sequences of ITS regions of the United States isolates are identical with sequences of Argentinean S. rarum strains Samiento and Noetinger and differ by two bases from the Arroyo Cabral isolate from Córdoba, Argentina. SEM observations revealed several new structures in the isolates from the US: female face views have a hexagonal-star perioral disc and eye-shaped lips; some females do not have cephalic papillae; lateral fields of infective juveniles are variable; there are two openings observed close to the posterior edge of the cloaca. Virulence of the US isolates to Anthonomus grandis, Diaprepes abbreviatus, Solenopsis invicta, Coptotermes formosanus, Agrotis ipsilon, Spodoptera frugiperda, and Trichoplusia ni and reproductive potential were evaluated in comparison with other heterorhabditid and steinernematid nematodes. Results such as particularly high virulence to S. frugiperda indicate that the biocontrol potential of the new S. rarum strains merits further study.

Effect of Soil Moisture and a Surfactant on Entomopathogenic Nematode Suppression of the Pecan Weevil, Curculio caryae.

Our overall goal was to investigate several aspects of pecan weevil, Curculio caryae, suppression with entomopathogenic nematodes. Specifically, our objectives were to: 1) determine optimum moisture levels for larval suppression, 2) determine suppression of adult C. caryae under field conditions, and 3) measure the effects of a surfactant on nematode efficacy. In the laboratory, virulence of Heterorhabditis megidis (UK211) and Steinernema carpocapsae (All) were tested in a loamy sand at gravimetric water contents of negative 0.01, 0.06, 0.3, 1.0, and 15 bars. Curculio caryae larval survival decreased as moisture levels increased. The nematode effect was most pronounced at -0.06 bars. At -0.01 bars, larval survival was

Draft genome sequence of Fusicladium effusum, cause of pecan scab.

Pecan scab, caused by the plant pathogenic fungus Fusicladium effusum, is the most destructive disease of pecan, an important specialty crop cultivated in several regions of the world. Only a few members of the family Venturiaceae (in which the pathogen resides) have been reported sequenced. We report the first draft genome sequence (40.6 Mb) of an isolate F. effusum collected from a pecan tree (cv. Desirable) in central Georgia, in the US. The genome sequence described will be a useful resource for research of the biology and ecology of the pathogen, coevolution with the pecan host, characterization of genes of interest, and development of markers for studies of genetic diversity, genotyping and phylogenetic analysis. The annotation of the genome is described and a phylogenetic analysis is presented.