Essential oils for managing anthracnose in mango (Mangifera indica): laboratory results do not translate into field efficacy.

Essential oil-based products with broad plant disease control claims are commercially available and may be a practical alternative to copper fungicides for crop protection in organic mango orchards. We evaluated the disease control efficacy and crop safety of thyme oil, savory oil, and tree tea oil through replicated in vitro, in vivo (detached leaf and potted trees), and field assays. Three Colletotrichum species associated with mango anthracnose were tested in vitro, whereas only C. siamense was used for in vivo assays. Within the range of concentrations tested in vitro (62.5 to 2,000 µl a.i./liter), thyme and savory oil displayed fungicidal activity, whereas no fungistatic or fungicidal activity was observed with tea tree oil. In the in vivo assays, none of the treatments based on a preventive application rate of thyme (1,150 µl a.i./liter), savory (2,000 µl a.i./liter), or tea tree oil (342 µl a.i./liter) were effective in preventing the development of anthracnose on wounded and artificially inoculated leaves. Although field applications of thyme or tea tree oil did not result in phytotoxicity or negative impacts on fruit yield, they were ineffective in reducing the incidence and severity of naturally occurring anthracnose. Applications of copper hydroxide approved for organic agriculture were effective in controlling anthracnose in the field, and no added benefits were found by premixing this compound with thyme oil. Results indicate that essential oil products based on thyme or tea tree oil are inefficient at controlling anthracnose in mangoes.

Genome Sequence Resource of the Avocado Scab Pathogen Elsinoe perseae.

We present a draft genome sequence of Elsinoe perseae, an economically important plant pathogen of commercially grown avocados. The 23.5-Mb assembled genome consists of 169 contigs. This report represents an important genomic resource to guide future research aimed at understanding the genetic interactions of E. perseae with its host.

First report of Diaporthe ueckerae causing stem canker on soybean (Glycine max L.) in Colombia.

In October 2018, soybean plants displaying elongated black to reddish-brown lesions on stems were observed in a field planted to the cv. BRS Serena in the locality of Puerto López (Meta, Colombia), with 20% incidence of diseased plants. Symptomatic stems were collected from five plants, and small pieces (∼5 mm2) were surface sterilized, plated on potato dextrose agar (PDA) and incubated for 2 weeks at 25°C in darkness. Three fungal isolates with similar morphology were obtained, i.e., by subculturing single hyphal tips, and their colonies on PDA were grayish-white, fluffy, with aerial mycelium, dark colored substrate mycelium, and produced circular black stroma. Pycnidia were globose, black, occurred as clusters, embedded in tissue, erumpent at maturity, with an elongated neck, and often had yellowish conidial cirrus extruding from the ostiole. Alpha conidia were observed for all isolates after 30 days growth on sterile soybean stem pieces (5 cm) on water agar, under 25ºC and 12 h light/12h darkness photoperiod. Alpha conidia (n = 50) measured 6.0 - 7.0 µm (6.4 ± 0.4 µm) × 2.0 - 3.0 µm (2.5± 0.4 µm), were aseptate, hyaline, smooth, ellipsoidal, often biguttulate, with subtruncate base. Beta conidia were not observed. Observed morphological characteristics of these isolates were similar to those reported in Diaporthe spp. by Udayanga et al. (2015). DNA from each fungal isolate was used to sequence the internal transcribed spacer region (ITS), and the translation elongation factor 1-α (TEF1) gene, using the primer pairs ITS5/ITS4 (White et al. 1990) and EF1-728F/EF1- 986R (Carbone & Kohn, 1999), respectively. Results from an NCBI-BLASTn, revealed that the ITS sequences of the three isolates (GenBank accessions MW566593 to MW566595) had 98% (581/584 bp) identity with D. miriciae strain BRIP 54736j (NR_147535.1), whereas the TEF1 sequences (GenBank accessions MW597410 to MW597412) had 97 to 100% (330-339/339 bp) identity with D. ueckerae strain FAU656 (KJ590747). The species Diaporthe miriciae R.G. Shivas, S.M. Thomps. & Y.P. Tan, and Diaporthe ueckerae Udayanga & Castl. are synonymous, with the latter taking the nomenclature priority (Gao et al. 2016). According to a multilocus phylogenetic analysis, by maximum likelihood, the three isolates clustered together in a clade with reference type strains of D. ueckerae (Udayanga et al. 2015). Soybean plants cv. BRS Serena (growth stages V3 to V4) were used to verify the pathogenicity of each isolate using a toothpick inoculation method (Mena et al. 2020). A single toothpick colonized by D. ueckerae was inserted directly into the stem of each plant (10 plants per isolate) approximately 1 cm below the first trifoliate node. Noncolonized sterile toothpicks, inserted in 10 soybean plants served as the non-inoculated control. Plants were arbitrarily distributed inside a glasshouse, and incubated at high relative humidity (>90% HR). After 15 days, inoculated plants showed elongated reddish-brown necrosis at the inoculated sites, that were similar to symptoms observed in the field. Non-inoculated control plants were asymptomatic. Fungal cultures recovered from symptomatic stems were morphologically identical to the original isolates. This is the first report of soybean stem canker caused by D. ueckerae in Colombia. Due to the economic importance of this disease elsewhere (Backman et al. 1985; Mena et al. 2020), further research on disease management strategies to mitigate potential crop losses is warranted.

High Genetic Diversity in Predominantly Clonal Populations of the Powdery Mildew Fungus Podosphaera leucotricha from U.S. Apple Orchards.

Apple powdery mildew (APM), caused by Podosphaera leucotricha, is a constant threat to apple production worldwide. Very little is known about the biology and population structure of this pathogen in the United States and other growing regions, which affects APM management. A total of 253 P. leucotricha isolates, sampled from 10 apple orchards in Washington, New York, and Virginia, were genetically characterized with novel single sequence repeat and mating type markers. Eighty-three multilocus genotypes (MLGs) were identified, most of which were unique to a given orchard. Each isolate carried either a MAT1-1 or a MAT1-2 idiomorph at the mating type locus, indicating that P. leucotricha is heterothallic. Virulence tests on detached apple leaves showed that the 10 most frequent P. leucotricha MLGs were avirulent on a line containing a major resistance gene. Analysis of molecular variance showed significant differentiation (P < 0.001) among populations, a result supported by principal coordinate analysis revealing three genetic groups, each represented by nonoverlapping MLGs from Washington, New York, and Virginia. A Bayesian cluster analysis showed genetic heterogeneity between Washington populations, and a relative migration analysis indicated substantial gene flow among neighboring orchards. Random mating tests indicated that APM epidemics during the active cycle were dominated by clonal reproduction. However, the presence of sexual structures in orchards, the likelihood that five repeated MLGs resulted from sexual reproduction, and high genotypic diversity observed in some populations suggest that sexual spores play some role in APM epidemics. IMPORTANCE Understanding the population biology and epidemiology of plant pathogens is essential to develop effective strategies for controlling plant diseases. Herein, we gathered insights into the population biology of P. leucotricha populations from conventional and organic apple orchards in the United States. We showed genetic heterogeneity between P. leucotricha populations in Washington and structure between populations from different U.S. regions, suggesting that short-distance spore dispersal plays an important role in the disease's epidemiology. We presented evidence that P. leucotricha is heterothallic and that populations likely result from a mixed (i.e., sexual and asexual) reproductive system, revealing that the sexual stage contributes to apple powdery mildew epidemics. We showed that the major resistance gene Pl-1 is valuable for apple breeding because virulent isolates have most likely not emerged yet in U.S. commercial orchards. These results will be important to achieve sustainability of disease management strategies and maintenance of plant health in apple orchards.

No Evidence of Resistance to Trifloxystrobin, Triflumizole, and Boscalid in Podosphaera leucotricha Isolates From U.S. Commercial Apple Orchards.

Apple powdery mildew, caused by Podosphaera leucotricha, continues to be a challenge in commercial apple orchards in the U.S. Pacific Northwest and worldwide. In this study, P. leucotricha isolates were collected in 2018 and 2019 from two organic (baseline) and eight conventional (exposed) apple orchards in Washington, New York, and Virginia, and assessed for their sensitivity to trifloxystrobin (TRI, n = 232), triflumizole (TFZ, n = 217), and boscalid (BOS, n = 240) using a detached leaf assay. Effective concentrations inhibiting 50% growth (EC50) were not significantly different between baseline and exposed isolates, and ranged from 0.001 to 0.105, 0.09 to 6.31, and 0.05 to 2.18 µg/ml, for TRI, TFZ, and BOS, respectively. Reduction in sensitivity by factors of 105, 63, and 22 to TRI, TFZ, and BOS, respectively, were observed in some isolates, but all isolates were controlled by the commercial label rates of the three fungicides on detached leaves. Sequencing of the cytochrome b (cytb), cytochrome P450 sterol 14α-demethylase (CYP51), and the iron-sulfur protein subunit (SdhB) genes in isolates with high EC50 revealed no mutation previously reported to confer resistance to these fungicides in other fungi, and presence of a group I intron after codon 143 in the cytb gene. Significant (P < 0.001) moderate positive correlations (r = 0.38) observed between sensitivity to TRI and TFZ warrant continuous rotations of fungicides with different modes of action in conventional orchards. The established baseline sensitivities and the molecular markers will help in selecting discriminatory doses and bypassing the challenging in vivo testing for future sensitivity monitoring in P. leucotricha.