Precipitation Impacts Dissemination of Three Sooty Blotch and Flyspeck Taxa on Apple Fruit.

The spatial dissemination of three prevalent taxa of sooty blotch and flyspeck (SBFS) fungi under several levels of precipitation was compared during 2015 and 2016 in an Iowa apple orchard. Overhead irrigation was used to supplement ambient precipitation in order to insure SBFS spore dissemination and colony development. There were five irrigation levels, involving 1-min-long periods of irrigation that were imposed either once or twice per hour at intervals of 3, 6, or 12 h, as well as a nonirrigated control. Preselected apple fruit were inoculated with one of the three SBFS taxa to serve as sources of inoculum. Dissemination from these inoculated apple fruit was assessed at harvest by counting SBFS colonies on water-sprayed and nontreated fruit. As a further control, additional fruit were enclosed in fruit bags throughout the fruit development period. In both 2015 and 2016, the number of colonies of the SBFS fungus Peltaster gemmifer per apple increased sharply as the duration of irrigation increased, whereas the number of colonies of Microcyclosporella mali increased to a lesser extent and Stomiopeltis sp. RS1 showed no increase. In 2015, the linear relationship between the duration of irrigation-imposed precipitation levels and the number of colonies on the water-sprayed apple fruit was similar for P. gemmifer (slope = 0.09), Stomiopeltis sp. RS1 (slope = 0.07), and Microcyclosporella mali (slope = 0.13); whereas, in 2016, the slope was higher for P. gemmifer (0.28) than for Stomiopeltis sp. RS1 (-0.09) or M. mali (0.06). The results indicated that dissemination of P. gemmifer increased sharply in response to increased irrigation-imposed precipitation, and that dissemination patterns differed considerably among the three SBFS taxa. The apparent advantage of P. gemmifer in precipitation-triggered dissemination may stem from its ability to produce spores rapidly by budding. To our knowledge, this is the first article to assess splash dispersal by SBFS fungi in the field and the first to document taxon-specific patterns of dissemination in this pathogen complex.

Stealth Pathogens: The Sooty Blotch and Flyspeck Fungal Complex.

Sooty blotch and flyspeck (SBFS) fungi produce superficial, dark-colored colonies on fruits, stems, and leaves of many plant genera. These blemishes are economically damaging on fruit, primarily apple and pear, because they reduce the sale price of fresh fruit. Fungicide spray programs can control SBFS but are costly and impair human and environmental health; thus, less chemically intensive management strategies are needed. Although the scientific study of SBFS fungi began nearly 200 years ago, recent DNA-driven studies revealed an unexpectedly diverse complex: more than 100 species in 30 genera of Ascomycota and Basidiomycota. Analysis of evolutionary phylogenetics and phylogenomics indicates that the evolution of SBFS fungi from plant-penetrating ancestors to noninvasive ectophytic parasites was accompanied by a massive contraction of pathogenicity-related genes, including plant cell wall-degrading enzymes and effectors, and an expansion of cuticle-degradation genes. This article reviews progress in understanding SBFS taxonomy and ecology and improving disease management. We also highlight recent breakthroughs in reconstructing the evolutionary origins of these unusual plant pathogens and delineating adaptations to their ectophytic niche.

Validation of a Florida Strawberry Anthracnose Fruit Rot (AFR) Warning System in Iowa.

Field validation of a disease-warning system for strawberry anthracnose fruit rot (AFR), caused by Colletotrichum acutatum sensu lato, that was originally developed for use in Florida was conducted in Iowa from 2012 to 2014. Day-neutral strawberry (cv. Tristar) was artificially inoculated with the pathogen at the start of the fruit maturation period. A factorial combination in replicated trials of two spray timing methods (the Florida warning system and prescheduled, calendar-based timing) and two fungicides (captan and pyraclostrobin) was compared, along with a nonsprayed control. The calendar-based and warning system-based treatments provided statistically equivalent control of AFR incidence compared with the nonsprayed control, and the warning system treatments required an average of 1.7 fewer fungicide sprays annually than the calendar-based treatments. Further analysis of the field data suggested that the warning system might underestimate AFR risk under high disease pressure; in these circumstances, a lower action threshold value may need to be adopted. Overall, these results indicated that the Florida warning system can be valuable for helping Midwest strawberry growers control AFR with less reliance on fungicide sprays, but it may require modification to account for periods of high inoculum pressure, subject to results of further field trials.

Peltaster gemmifer: A new species in the sooty blotch and flyspeck species complex from the United States.

Sooty blotch and flyspeck (SBFS) fungi infect the cuticle of fruit, including apple fruit, and produce pigmented colonies. A new member of this fungal complex in the genus Peltaster is described on the basis of molecular and morphological evidence. The SBFS complex is a diverse group of ectophytic fungi that reside primarily within the order Capnodiales. Sooty blotch and flyspeck isolates from apple orchards in the central United States were subjected to parsimony and Bayesian analyses based on the internal transcribed spacer regions of nuc rDNA, the partial translation elongation factor 1-α gene, and the partial mitochondrial small subunit rRNA gene. Phylogenetic analysis delineated a new species, Peltaster gemmifer, from P. cerophilus and P. fructicola. Peltaster gemmifer conidiophores bear primary conidia that produce secondary conidia either through budding or through microcyclic conidiation; these were not seen in cultures of P. cerophilus and P. fructicola. On cellulose membrane that was placed on water agar amended with apple juice, P. gemmifer produced brown to black pycnothyria in a superficial brownish mycelial mat, similar to the colonies produced on apple fruit. Findings from the present study add to the >80 named and putative SBFS species so far described worldwide.

Evaluating the Performance of a Relative Humidity-Based Warning System for Sooty Blotch and Flyspeck in Iowa.

A warning system for the sooty blotch and flyspeck (SBFS) fungal disease complex of apple, developed originally for use in the southeastern United States, was modified to provide more reliable assessment of SBFS risk in Iowa. Modeling results based on previous research in Iowa and Wisconsin had suggested replacing leaf wetness duration with cumulative hours of relative humidity (RH) ≥97% as the weather input to the SBFS warning system. The purpose of the present study was to evaluate the performance of a RH-based SBFS warning system, and to assess the potential economic benefits for its use in Iowa. The warning system was evaluated in two separate sets of trials-trial 1 during 2010 and 2011, and trial 2 during 2013-2015-using action thresholds based on cumulative hours of RH ≥97% and ≥90%, respectively, in conjunction with two different fungicide regimes. The warning system was compared with a traditional calendar-based system that specified spraying at predetermined intervals of 10 to 14 days. In trial 1, use of the RH ≥97% threshold caused substantial differences between two RH sensors in recording number of hours exceeding the threshold. When both RH thresholds were compared for 2013-2015, on average, RH ≥90% resulted in a 53% reduction in variation of cumulative hours between two identical RH sensors placed adjacent to each other in an apple tree canopy. Although both the SBFS warning system and the calendar-based system resulted in equivalent control of SBFS, the warning system required fewer fungicide sprays than the calendar-based system, with an average of 3.8 sprays per season (min = 2; max = 5) vs. 6.4 sprays per season (min = 5; max = 8), respectively. The two fungicide regimes provided equivalent SBFS control when used in conjunction with the warning system. A partial budget analysis showed that using the SBFS warning system with a threshold of RH ≥90% was cost effective for orchard sizes of >1 ha. The revised warning system has potential to become a valuable decision support tool for Midwest apple growers because it reduces fungicide costs while protecting apples as effectively as a calendar-based spray schedule. The next step toward implementation of the SBFS warning system in the North Central U.S. should be multiyear field testing in commercial orchards throughout the region.

Bacterial Wilt of Cucurbits: Resurrecting a Classic Pathosystem.

Bacterial wilt threatens cucurbit crop production in the Midwestern and Northeastern United States. The pathogen, Erwinia tracheiphila, is a xylem-limited bacterium that affects most commercially important cucurbit species, including muskmelon, cucumber, and squash. Bacterial wilt is transmitted and overwintered by striped and spotted cucumber beetles. Since there are few commercially available resistant cultivars, disease management usually relies on use of insecticides to suppress vector populations. Although bacterial wilt was initially described more than 100 years ago, our knowledge of disease ecology and epidemiology advanced slowly for most of the 20th century. However, a recent wave of research has begun to fill in missing pieces of the bacterial wilt puzzle. This article-the first review of research toward understanding the cucurbit bacterial wilt pathosystem-recounts early findings and updates our understanding of the disease cycle, including pathogen and vector biology. We also highlight research areas that could lead to more efficient and ecologically based management of bacterial wilt.

Molecular and morphological analysis reveals five new species of Zygophiala associated with flyspeck signs on plant hosts from China.

Species in the genus Zygophiala are associated with sooty blotch and flyspeck disease on a wide range of hosts. In this study, 63 Zygophiala isolates collected from flyspeck colonies on a range of plants from several regions of China were used for phylogeny, host range and geographic distribution analysis. Phylogenetic trees were constructed on four genes--internal transcribed spacer (ITS), partial translation elongation factor 1-alpha (TEF), ß-tubulin (TUB2), and actin (ACT)--both individually and in combination. Isolates were grouped into 11 clades among which five new species, Z. emperorae, Z. trispora, Z. musae, Z. inaequalis and Z. longispora, were described. Species of Zygophiala differed in observed host range and geographic distribution. Z. wisconsinensis and Z. emperorae were the most prevalent throughout the sampled regions of China, whereas Z. trispora, Z. musae, Z. inaequalis and Z. longispora were collected only in southern China. The hosts of Z. wisconsinensis and Z. emperorae were mainly in the family Rosaceae whereas Z. trispora, Z. musae, Z. inaequalis and Z. longispora were found mainly on banana (Musa spp.). Cross inoculation tests provided evidence of host specificity among SBFS species.

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.

Phylogenetic placement of plant pathogenic Sclerotium species among teleomorph genera.

Phylogenetic analyses and morphological characteristics were used to assess the taxonomic placement of eight plant-pathogenic Sclerotium species. Members of this genus produce only sclerotia and no fruiting bodies or spores, so Sclerotium species have been difficult to place taxonomically. Sequences of rDNA large subunit (LSU) and internal transcribed spacer (ITS) regions were determined for isolates of Sclerotium cepivorum, S. coffeicola, S. denigrans, S. hydrophilum, Ceratorhiza oryzae-sativae, S. perniciosum, S. rhizodes, S. rolfsii and S. rolfsii var. delphinii. Parsimony analysis grouped two species previously thought to be in the Basidiomycota, S. denigrans and S. perniciosum, within the Ascomycota; these species were found to have affinities with the teleomorph genera Sclerotinia and Stromatinia and the asexual Sclerotium cepivorum, which was known earlier to be related to Sclerotinia species. The other Sclerotium species were placed in one of two basidiomycetous groups, genera Athelia or Ceratobasidium. Based on rDNA analysis and morphology the basidiomycetous Sclerotium hydrophilum and S. rhizodes were transferred to genus Ceratorhiza, the anamorph of Ceratobasidium species. Sclerotium coffeicola was found to be close to S. rolfsii var. delphinii and S. rolfsii var. rolfsii, which was shown earlier to have an Athelia teleomorph.

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.

Spatial variability of leaf wetness duration in different crop canopies.

The spatial variability of leaf wetness duration (LWD) was evaluated in four different height-structure crop canopies: apple, coffee, maize, and grape. LWD measurements were made using painted flat plate, printed-circuit wetness sensors deployed in different positions above and inside the crops, with inclination angles ranging from 30 to 45 degrees. For apple trees, the sensors were installed in 12 east-west positions: 4 at each of the top (3.3 m), middle (2.1 m), and bottom (1.1 m) levels. For young coffee plants (80 cm tall), four sensors were installed close to the leaves at heights of 20, 40, 60, and 80 cm. For the maize and grape crops, LWD sensors were installed in two positions, one just below the canopy top and another inside the canopy. Adjacent to each experiment, LWD was measured above nearby mowed turfgrass with the same kind of flat plate sensor, deployed at 30 cm and between 30 and 45 degrees. We found average LWD varied by canopy position for apple and maize (P<0.05). In these cases, LWD was longer at the top, particularly when dew was the source of wetness. For grapes, cultivated in a hedgerow system and for young coffee plants, average LWD did not differ between the top and inside the canopy. The comparison by geometric mean regression analysis between crop and turfgrass LWD measurements showed that sensors at 30 cm over turfgrass provided quite accurate estimates of LWD at the top of the crops, despite large differences in crop height and structure, but poorer estimates for wetness within leaf canopies.