Toward the Integration of an Attract-and-Kill Approach with Entomopathogenic Nematodes to Control Multiple Life Stages of Plum Curculio (Coleoptera: Curculionidae).

Efforts to reduce insecticide inputs against plum curculio, Conotrachelus nenuphar, a key pest of apples in eastern North America, include perimeter-row insecticide sprays applied after the whole-orchard petal fall spray to manage dispersing adults and, more recently, insecticide sprays confined to odor-baited trap trees. Entomopathogenic nematodes (EPNs) are virulent to ground-dwelling stages of C. nenuphar, and may be applied to the ground underneath trap-tree canopies. Here, we (1) compared the efficacy of the odor-baited trap tree approach with grower-prescribed (=grower standard) sprays to manage C. nenuphar populations over a six-year period in seven commercial apple orchards in New England; and (2) assessed the performance of the EPN Steinernema riobrave at suppressing ground-dwelling stages of C. nenuphar. In addition, the performance of S. riobrave was compared against that of S. carpocapsae and S. feltiae in one year. Across the six years, percent fruit injury on trap tree plots averaged 11.3% on odor-baited trap trees and 1.4% on unbaited trees in grower standard plots, highlighting the ability of trap trees to aggregate C. nenuphar activity and subsequent injury. Mean percentage injury on fruit sampled from interior trees, the strongest measure of treatment performance, in trap tree plots did not differ significantly from that recorded on interior trees in grower standard spray plots (0.95 vs. 0.68%, respectively). Steinernema riobrave consistently reduced C. nenuphar populations as indicated by the significantly lower number of adult C. nenuphar that emerged from the soil, when compared to water control. Steinernema carpocapsae and S. riobrave performed similarly well, and both EPN species outperformed S. feltiae. Our combined findings indicate that an IPM approach that targets multiple life stages of C. nenuphar has the potential to manage this pest more sustainably in a reduced-spray environment.

Comparative genomics of Spiraeoideae-infecting Erwinia amylovora strains provides novel insight to genetic diversity and identifies the genetic basis of a low-virulence strain.

Erwinia amylovora is the causal agent of fire blight, one of the most devastating diseases of apple and pear. Erwinia amylovora is thought to have originated in North America and has now spread to at least 50 countries worldwide. An understanding of the diversity of the pathogen population and the transmission to different geographical regions is important for the future mitigation of this disease. In this research, we performed an expanded comparative genomic study of the Spiraeoideae-infecting (SI) E. amylovora population in North America and Europe. We discovered that, although still highly homogeneous, the genetic diversity of 30 E. amylovora genomes examined was about 30 times higher than previously determined. These isolates belong to four distinct clades, three of which display geographical clustering and one of which contains strains from various geographical locations ('Widely Prevalent' clade). Furthermore, we revealed that strains from the Widely Prevalent clade displayed a higher level of recombination with strains from a clade strictly from the eastern USA, which suggests that the Widely Prevalent clade probably originated from the eastern USA before it spread to other locations. Finally, we detected variations in virulence in the SI E. amylovora strains on immature pear, and identified the genetic basis of one of the low-virulence strains as being caused by a single nucleotide polymorphism in hfq, a gene encoding an important virulence regulator. Our results provide insights into the population structure, distribution and evolution of SI E. amylovora in North America and Europe.

Prevalence of Myclobutanil Resistance and Difenoconazole Insensitivity in Populations of Venturia inaequalis.

Demethylation inhibitors (DMIs) are a class of single-site fungicides with high levels of protective and curative efficacy against Venturia inaequalis, the causal agent of apple scab. To determine the prevalence of resistance to the DMI fungicide myclobutanil, 3,987 single-lesion conidial V. inaequalis isolates from 141 commercial, research, and baseline orchard populations were examined throughout New England, the mid-Atlantic, and the Midwest from 2004 to 2013. Of these orchard populations, 63% had practical resistance, 13% had reduced sensitivity, and 24% were sensitive to myclobutanil. A sensitivity baseline for the recently introduced DMI fungicide difenoconazole was established to make comparisons with myclobutanil sensitivity in orchard populations. The mean effective concentration of difenoconazole at which mycelial growth was inhibited by 50% (EC50) was determined to be 0.002 µg ml-1 for 44 baseline isolates of V. inaequalis. From 2010 to 2013, 1,012 isolates of V. inaequalis from 37 of the 141 orchard populations above were screened for sensitivity to difenoconazole. In all, 1 orchard population had reduced sensitivity to difenoconazole, while the remaining 36 orchard populations were sensitive to the fungicide. In field experiments, difenoconazole demonstrated high levels of apple scab control on mature apple fruit, despite the fact that the population of V. inaequalis had practical resistance to difenoconazole. Although our results indicate widespread resistance to myclobutanil but not difenoconazole, due to the propensity for cross-sensitivity among DMI fungicides, growers with myclobutanil resistance should be cautious when using difenoconazole for disease management.

Prevalence and Stability of Qualitative QoI Resistance in Populations of Venturia inaequalis in the Northeastern United States.

Quinone-outside-inhibitor (QoI) fungicides are a safe and effective means of managing apple scab caused by Venturia inaequalis. To determine the prevalence of both quantitative (partial) and qualitative (complete) QoI resistance in V. inaequalis in the northeastern United States, we sampled single-lesion conidial isolates (n = 4,481) from 120 commercial and research orchards from 2004 to 2011 with a range of exposure to QoI fungicides from none to several applications a year. In all, 67% of these orchard populations of V. inaequalis were sensitive to QoI fungicides, 28% exhibited QoI practical resistance, and 5% were not sensitive QoI fungicides but had not become practically resistant. Isolates with qualitative QoI resistance, conferred by the G143A cytochrome b gene mutation, were found in 13 of the 34 QoI-resistant orchard populations. To evaluate the stability of the G143A mutation, 27 isolates were selected from different orchard populations to represent the scope of regional populations. These isolates were subcultured continuously in the presence or absence of the QoI fungicide trifloxystrobin. All isolates that initially possessed qualitative resistance maintained the resistant genotype (G143A) for six transfers over 6 months in both the absence and presence of trifloxystrobin. Given the observed QoI resistance in orchard populations of V. inaequalis and the stability of the G143A mutation in individual isolates, apple scab management paradigms must encompass strategies to limit selection of QoI resistance in the sensitive orchard populations remaining in the region.

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.

Variability Among Forecast Models for the Apple Sooty Blotch/Flyspeck Disease Complex.

Several disease forecast models have been developed to guide treatment of the sooty blotch and flyspeck (SBFS) disease complex of apple. Generally, these empirical models are based on the accumulation of hours of leaf wetness (leaf wetness duration [LWD]) from a biofix at or near the phenological growth stage petal fall, when apple flower petals senesce and drop. The models recommend timing of the initial fungicide application targeting SBFS. However, there are significant differences among SBFS forecast models in terms of biofix and the length of LWD thresholds. A comparison of models using a single input data set generated recommendations for the first SBFS fungicide application that differed by up to 5 weeks. In an attempt to improve consistency among models, potential sources for differences were examined. Leaf wetness (LW) is a particularly variable parameter among models, depending on whether on-site or remote weather data were used, the types of sensors and their placement for on-site monitors, and the models used to estimate LW remotely. When SBFS models are applied in the field, recommended treatment thresholds do not always match the method of data acquisition, leading to potential failures. Horticultural factors, such as tree size, canopy density, and cultivar, and orchard site factors such as the distance to potential inoculum sources can impact risk of SBFS and should also be considered in forecast models. The number of fungal species identified as contributors to the SBFS disease complex has expanded tremendously in recent years. A lack of understanding of key epidemiological factors for different fungi in the complex, and which fungi represent the most challenging management problems, are obvious issues in the development of improved SBFS models. If SBFS forecast models are to be adopted, researchers will need to address these issues.

A New View of Sooty Blotch and Flyspeck.

Sooty blotch and flyspeck (SBFS) fungi colonize the surface wax layer of the fruit of apple, pear, persimmon, banana, orange, papaya, and several other cultivated tree and vine crops. In addition to colonizing cultivated fruit crops, SBFS fungi also grow on the surfaces of stems, twigs, leaves, and fruit of a wide range of wild plants. The disease occurs worldwide in regions with moist growing seasons. SBFS is regarded as a serious disease by fruit growers and plant pathologists because it can cause substantial economic damage. The smudges and stipples of SBFS often result in downgrading of fruit from premium fresh-market grade to processing use. This review describes the major shifts that have occurred during the past decade in understanding the genetic diversity of the SBFS complex, clarifying its biogeography and environmental biology, and developing improved management strategies.

Diversity and biogeography of sooty blotch and flyspeck fungi on apple in the eastern and midwestern United States.

Sooty blotch and flyspeck (SBFS) fungi on apple fruit were sampled from nine orchards in four midwestern U.S. states during 2000 and 30 orchards in 10 eastern U.S. states during 2005 in order to estimate taxonomic diversity and discern patterns of geographic distribution. Forty apple fruit per orchard were arbitrarily sampled and colonies of each mycelial phenotype were counted on each apple. Representative colonies were isolated, cultures were purified, and DNA was extracted. For representative isolates, the internal transcribed spacer (ITS) and large subunit (LSU) regions of ribosomal DNA were amplified and sequenced. In total, 60 SBFS putative species were identified based on ITS sequences and morphological characteristics; 30 of these were discovered in the 2005 survey. Modified Koch's postulates were fulfilled for all 60 species in an Iowa orchard; colonies resulting from inoculation of apple fruit were matched to the original isolates on the basis of mycelial type and ITS sequence. Parsimony analysis for LSU sequences from both surveys revealed that 58 putative SBFS species were members of the Dothideomycetes, 52 were members of the Capnodiales, and 36 were members of the Mycosphaerellaceae. The number of SBFS species per orchard varied from 2 to 15. Number of SBFS species and values of the Margalef and Shannon indexes were significantly (P < 0.05) lower in 21 orchards that had received conventional fungicide sprays during the fruit maturation period than in 14 unsprayed orchards. Several SBFS species, including Schizothyrium pomi, Peltaster fructicola, and Pseudocercosporella sp. RH1, were nearly ubiquitous, whereas other species, such as Stomiopeltis sp. RS5.2, Phialophora sessilis, and Geastrumia polystigmatis, were found only within restricted geographic regions. The results document that the SBFS complex is far more taxonomically diverse than previously recognized and provide strong evidence that SBFS species differ in geographic distribution. To achieve more efficient management of SBFS, it may be necessary to understand the environmental biology of key SBFS species in each geographic region.

Maturation of Thyriothecia of Schizothyrium pomi on the Reservoir Host Rubus allegheniensis.

Flyspeck disease caused by the Ascomycete fungus Schizothyrium pomi is a major component of a summer disease complex that has become an increasing problem on apple in the northeastern United States. This study describes the development of thyriothecia of S. pomi on one of its major reservoir hosts, wild blackberry, and suggests that temperature drives thyriothecium maturation. An empirical model relating thyriothecium maturity to degree-days from apple leaf budbreak in Massachusetts had a high coefficient of determination, R2 = 0.90. Ninety percent of thyriothecia matured between 540 and 1,625 degree-days from the budbreak biofix. S. pomi produced one generation of asci per year on blackberry. This discrete maturation period occurred well before signs of disease developed on apple fruit and before production of conidia. Considering that fungicide applications for scab management in commercial apple orchards also will protect fruit from flyspeck during early fruit development, it is unlikely that ascospores produced on reservoir hosts directly infect fruit. Instead, they probably initiate infections in reservoir hosts adjacent to commercial orchards, which produce conidia that are the inoculum which infects apple fruit in commercial production.

Summer Pruning as a Method for Reducing Flyspeck Disease on Apple Fruit.

Summer pruning of apples, as opposed to the conventional commercial practice of dormant pruning, consistently reduced the incidence of flyspeck on apple fruit by approximately 50% in each of 2 years in trees where no fungicides were applied. In commercial orchard blocks using fungicides, summer pruning also produced a slight but significant decrease in disease severity. There appear to be at least two mechanisms contributing to decreased flyspeck incidence and severity in summer-pruned apple trees. Summer pruning resulted in a small change in the apple canopy microclimate, decreasing the hours of relative humidity >95% in the canopy by 63% and increasing the evaporative potential. Summer pruning also resulted in improved spray deposition in the upper two-thirds of the tree canopy when applications were made with an airblast sprayer.