Postharvest Pathogens Research Articles

Selection and identification of potential bacteria against postharvest fungal pathogens on strawberry fruit

Strawberries are economically important owing to their unique flavour, health benefits, and nutritional aspects. Decay after harvest is a major issue in the development of strawberry industry. Currently, the most effective method for controlling decay after harvest is storage of strawberries at low temperature combined with usage of chemical fungicides. However, long-term usage of chemical fungicides not only causes pathogen resistance but also is harmful for human health and environment. Biocontrol method for the management of disease after strawberry harvest has great practical significance. In this study, totally 200 bacterial strains isolated from the surface of healthy strawberry fruit were tested for antagonistic activity against five fungal pathogens. Among them, VK199 strain was potent against all five postharvest spoilage fungi on strawberry fruit and then utilized for further study. This strain was identified as Bacillus siamensis based on sequence analysis of 16S rDNA. The extracellular lytic enzymes, including cellulase, amylase, protease and chitinase released by B. siamensis VK199 were detected. Furthemore, in vivo, the results of biological control efficacy test showed that B. siamensis VK199 suppressed the occurrence of diseases caused by Botrytis cinerea, Colletotrichum gloeosporioides, Mucor nidicola and Rhizopus delemar of strawberry during storage.

The pH signalling transcription factor PacC modulate growth, development, stress response and pathogenicity of Trichothecium roseum.

pH is one of the important environmental factors that affect the growth, development and pathogenicity of postharvest pathogen. The transcription factor PacC dominates the pH signal pathway. PacC in Trichothecium roseum showed three typical conserved zinc finger domains and closest homology to Fusarium graminearum. T. roseum increased the environmental pH both in vitro and in vivo. Expression patterns of TrpacC under different pH showed that at increasing pH from 3 to 5, the wild-type (WT) strain induced the expression of TrPacC in parallel to increased fungal growth; however, TrPacC expression decline at higer pH than 5, while fungal growth continued to increase. Development of a ΔTrPacC mutant down-regulated the expression of TrbrlA, TrabaA and TrwetA, reduced sporulation and delayed spore germination, resulting in smaller spores and sparse hyphae. ΔTrPacC mutant was sensitive to ionic stress, oxidative stress and cell wall integrity stress compared to the WT strain, especially the ionic stress. In addition, ∆TrPacC mutant showed reduced pathogenicity to muskmelon and tomato fruits. Taken together, T. roseum is an alkalinizing fungus, and the acidic environment could induce TrPacC expression. TrPacC positively regulates fungal growth and development as well as pathogenicity showing effect on fungal response to different stresses.

Exploring the transcriptome signature associated with tolerance to Penicillium expansum in apple through feature selection algorithms and differential gene expression analysis

ABSTRACT Penicillium expansum is one of the most destructive post-harvest pathogens that causes fruit spoilage in apple. This study explored the molecular response of apple to this pathogen via microarray data mining based on attribute weighting algorithms (AWAs), differential expression and pathway analyses, and multivariate statistical analysis. A total of 124 probe sets were identified by at least one AWA and these yielded 29 genes with significantly differential expression. Leave-one-out cross-validation showed that the 29-gene signature could differentiate resistant (R) and susceptible (S) cultivars with 98.65% accuracy, with most of the genes showing significant expression changes in these cultivars at 1 week post-inoculation (wpi). Many genes encoding hydrolases, reductases and proteins with zinc-finger domains showed an increased expression in the R cultivars. In particular, disease resistance protein CC-NBS-LRR, resistance-related protein E3-ubiquitin and cuticle wax biosynthesis genes displayed increased expression, especially in R cultivars at 1 and 6 wpi. The secondary metabolite biosynthesis pathway, metabolic pathway and amino acid biosynthesis pathway were commonly upregulated in both R and S cultivars, but with a greater intensity in R cultivars, while the flavonoid biosynthesis pathway was downregulated in the S cultivars at 1 h post-inoculation. This study identifies candidate targets for developing resistant cultivars.

Integrated Genomic and Metabolomic Analysis Illuminates Key Secreted Metabolites Produced by the Novel Endophyte Bacillus halotolerans Cal.l.30 Involved in Diverse Biological Control Activities.

The endophytic strain Cal.l.30, isolated from the medicinal plant Calendula officinalis, was selected among seven Bacillus strains with plant growth promoting activity and strong biological potential against the postharvest fungal pathogen Botrytis cinerea. Treatment by inoculating Cal.l.30 bacterial cell culture or cell free supernatant on harvested grapes and cherry tomato fruits, significantly reduced gray mold disease severity index and disease incidence. Based on 16S rRNA sequence analysis and whole genome phylogeny, Cal.l.30 was identified as Bacillus halotolerans. Genome mining revealed that B. halotolerans Cal.l.30 is endowed with a diverse arsenal of secondary metabolite biosynthetic gene clusters (SM-BGCs) responsible for metabolite production with antimicrobial properties. A sub-set of the identified SM-BGCs (mojavensin A, ‘bacillunoic acid’) appears to be the result of recent horizontal gene transfer events. Its genome was also mined for CAZymes associated with antifungal activity. Further UHPLC-HRMS analysis indicated that Cal.l.30 synthesizes and secretes secondary metabolites with antimicrobial activity, including the lipopeptides, fengycin, surfactin and mojavensin A, bacillaene isoforms, L-dihydroanticapsin and bacillibactin. Other compounds with known antimicrobial activity were also detected, such as azelaic acid, 15- hydroxypentadecanoid acid and 2-hydroxyphenylacetic acid. The genomic and metabolomic features of the B. halotolerans Cal.l.30 provided new perspectives on the exploitation of novel Bacillus sp. as a biocontrol agent.

Wood Waste from Fruit Trees: Biomolecules and Their Applications in Agri-Food Industry.

In the European Union (EU), a total of 11,301,345 hectares are dedicated to the cultivation of fruit trees, mainly olive orchards, grapevines, nut trees (almond, walnut, chestnut, hazelnut, and pistachio), apple and pear trees, stone fruit trees (peach, nectarine, apricot, cherry, and plum), and citrus fruit trees (orange, clementine, satsuma, mandarin, lemon, grapefruit, and pomelo). Pruning these trees, together with plantation removal to a lesser extent, produces a huge amount of wood waste. A theoretical calculation of the wood waste in the European Union estimates approximately 2 and 25 million tons from wood plantation removal and pruning, respectively, per year. This wood waste is usually destroyed by in-field burning or crushing into the soil, which result in no direct economic benefits. However, wood from tree pruning, which is enriched in high added-value molecules, offers a wide spectrum of possibilities for its valorization. This review focuses on the contribution of wood waste to both sustainability and the circular economy, considering its use not only as biomass but also as a potential source of bioactive compounds. The main bioactive compounds found in wood are polyphenols, terpenes, polysaccharides, organic compounds, fatty acids, and alkaloids. Polyphenols are the most ubiquitous compounds in wood. Large amounts of hydroxytyrosol (up to 25 g/kg dw), resveratrol (up to 66 g/kg dw), protocatechuic acid (up to 16.4 g/kg), and proanthocyanins (8.5 g/kg dw) have been found in the wood from olive trees, grapevines, almond trees and plum trees, respectively. The bioactivity of these compounds has been demonstrated at lower concentrations, mainly in vitro studies. Bioactive compounds present antioxidant, antimicrobial, antifungal, biostimulant, anti-inflammatory, cardioprotective, and anticarcinogenic properties, among others. Therefore, wood extracts might have several applications in agriculture, medicine, and the food, pharmaceutical, nutraceutical, and cosmetics industries. For example, olive tree wood extract reduced thrombin-induced platelet aggregation in vitro; grapevine tree wood extract acts a preservative in wine, replacing SO2; chestnut tree wood extract has antifungal properties on postharvest pathogens in vitro; and stone tree wood extracts are used for aging both wines and brandies. Moreover, the use of wood waste contributes to the move towards both a more sustainable development and a circular economy.

Phylogenetic Diversity and Phenotypic Characterization of Phlyctema vagabunda (syn. Neofabraea alba) and Neofabraea kienholzii Causing Postharvest Bull's Eye Rot of Apple in Northern Italy.

Bull's eye rot, caused by Phlyctema vagabunda and Neofabraea species, is one of the most important postharvest diseases of apple. South Tyrol (northern Italy) is the largest continuous apple-producing area in Europe, with approximately 1 million tons being produced yearly and conserved in technologically advanced storage facilities for several months. Still, studies on the pathogen species causing postharvest bull's eye rot of apple, as well as their diversity and biology, are lacking for this region. Therefore, the main purpose of the present work was to identify and characterize fungal isolates obtained from decayed apple fruit with symptoms of bull's eye rot that were collected in 2018 and 2019 in different packinghouses in South Tyrol. Among more than 1,000 fungal isolates that were obtained, 419 could be assigned to the genera Phlyctema and/or Neofabraea based on rot symptoms on apple fruit and colony morphology on potato dextrose agar. A smaller subset of 101 representative isolates was further analyzed by DNA sequencing of the internal transcribed spacer region. Furthermore, partial segments of the β-tubulin gene, the translation elongation factor 1α gene, and the 16S mitochondrial ribosomal RNA gene were studied. The phylogenetic analyses, including sequences of reference species, showed that P. vagabunda is the dominant species associated with bull's eye rot of apple in the study area, whereas Neofabraea kienholzii was found only on a small number of apple fruit samples. The combination of multilocus sequence data revealed 11 unique genotypes that belonged to P. vagabunda and four to N. kienholzii. To the best of our knowledge, this study is the first to report N. kienholzii as a postharvest pathogen of apple in Italy. Finally, a pathogenicity test demonstrated different degrees of virulence among selected isolates of P. vagabunda and N. kienholzii on the cultivar Golden Delicious. The present study emphasizes the importance of accurate species identification, because different species may vary in their biological and pathogenic characteristics, and consequently require distinct disease management strategies, both in the field and during the postharvest stages.[Formula: see text] Copyright © 2022 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Wide distribution of resistance to the fungicides fludioxonil and iprodione in Penicillium species.

Fludioxonil and iprodione are effective fungicides widely used for crop protection and are essential for controlling plant pathogenic fungi. The emergence of fungicide-resistant strains of targeted pathogens is regularly monitored, and several cases have been reported. Non-targeted fungi may also be exposed to the fungicide residues in agricultural fields. However, there are no comprehensive reports on fungicide-resistant strains of non-targeted fungi. Here, we surveyed 99 strains, representing 12 Penicillium species, that were isolated from a variety of environments, including foods, dead bodies, and clinical samples. Among the Penicillium strains, including non-pathogenic P. chrysogenum and P. camembertii, as well as postharvest pathogens P. expansum and P. digitatum, 14 and 20 showed resistance to fludioxonil and iprodione, respectively, and 6 showed multi-drug resistance to the fungicides. Sequence analyses revealed that some strains of P. chrysogenum and Penicillium oxalicum had mutations in NikA, a group III histidine kinase of the high-osmolarity glycerol pathway, which is the mode of action for fludioxonil and iprodione. The single nucleotide polymorphisms of G693D and T1318P in P. chrysogenum and T960S in P. oxalicum were only present in the fludioxonil- or iprodione-resistant strains. These strains also exhibited resistance to pyrrolnitrin, which is the lead compound in fludioxonil and is naturally produced by some Pseudomonas species. This study demonstrated that non-targeted Penicillium strains distributed throughout the environment possess fungicide resistance.

Melatonin enhanced the disease resistance by regulating reactive oxygen species metabolism in postharvest jujube fruit

Melatonin (MT), as a putative pleiotropic signal molecule, could participate in inducing postharvest disease resistance and inhibiting postharvest pathogen infection in fruit. In this study, we investigate the mechanism of induced resistance by MT treatment against Alternaria rot of jujube. The results showed that 2 and 20 μmol/L MT did not have direct antimicrobial activity, 200 μmol/L MT had little influence on antifungal activity against Alternaria alternata in vitro. MT significantly reduced the lesion area of fruit inoculated with A. alternata. MT decreased the relative electrolytic leakage, malondialdehyde content, hydrogen peroxide, and the rate of superoxide anion production, while enhanced the ascorbic acid and reduced glutathione contents. Furthermore, MT observably increased the defensive enzyme activities of superoxide dismutase, the catalase, ascorbate peroxidase, peroxidase. Accordingly, these results indicated that MT treatment increased the disease resistance of jujube fruit by regulating reactive oxygen species metabolism. Novelty impact statement The results showed that MT treatment could inhibit the expansion of lesion diameter, reduce the decay rate of winter jujube, and reduce the accumulation of reactive oxygen species by activating the oxidase system, thus enhancing the resistance of winter Jujube fruit to Alternaria alternata infection by adjusting the ROS mechanism.

Green synthesized nanomaterials as an alternative to restricted chemicals in postharvest preservation of horticultural crops: a review

Synthetic chemicals have been successfully used to control economic postharvest pathogens such as Botrytis cinerea, Penicillium spp., Rhizopus spp., etc., thus reducing postharvest losses. However, their active ingredient and co-formulants have resulted in their use being restricted internationally due to health concerns and consumers’ requests for safe and natural alternatives. As an alternative to chemicals, green synthesized nanomaterials incorporated with edible coatings in a polymer matrix have been successfully applied on horticultural crops to improve their shelf-life Due to their known antimicrobial activities, plant extracts and nanoparticles such as silver and zinc oxide have been successfully used in food processing applications. Green sources are used as stabilizer and reducing agents to synthesize shape and sizecontrolled metal and metal oxide nanoparticles. Thus, eco-friendly nanoparticles are produced without using toxic materials and are cost-effective. Edible coatings form a thin layer that prevents oxidation, maintains the product quality, can be safely eaten as part of the product, and does not add unfavourable properties. Studies showed that edible coatings with or without green nanomaterials have been successfully used to control postharvest losses and improve the shelf-life of fruit crops such as plums, raspberries, etc.

Effect of Low Pressure and Low Oxygen Treatments on Fruit Quality and the In Vivo Growth of Penicillium digitatum and Penicillium italicum in Oranges

Penicillium digitatum and P. italicum are the major postharvest pathogens in citrus. To reduce postharvest decay, the use of low-oxygen (0.9 kPa O2) (LO) or low-pressure (6.6 kPa) (LP) treatments were evaluated during the storage of navel oranges for four or eight days. The results showed that exposure to both LO and LP treatments reduced in vivo pathogen growth compared to the untreated (UTC) oranges, with LO being the most effective. The effects of LO and LP on fruit metabolism and quality were further assessed, and it was found that there was no effect on fruit ethylene production, respiration rate, TSS (total soluble solids), TA (titratable acidity) or fruit firmness. However, both LO and LP treatments did have an effect on juice ethanol concentration and fruit weight-loss. The effect of adding exogenous ethylene at either LP (1 µL/L) or atmospheric pressure (AP) (at either 0.1, 1 µL/L) was also evaluated, and results showed that the addition of ethylene at these concentrations had no effect on mould diameter at LP or AP. Therefore, both LO of 0.9 kPa O2 and LP of 6.6 kPa at 20 °C are potential non-chemical postharvest treatments to reduce mould development during storage with minimal effects on fruit quality.

Antifungal and Antitoxigenic Effects of Selected Essential Oils in Vapors on Green Coffee Beans with Impact on Consumer Acceptability.

The main objective of this study is to evaluate the effect of selected essential oils thyme chemotype linalool (Thymus zygis L.), thyme chemotype tymol (Thymus vulgaris L.), eucalyptus (Eucalyptus globulus Labill.), lavender (Lavandula angustifolia Mill.), mint (Mentha piperita L.), almond (Prunbus dulcis Mill.), cinnamon bark (Cinnamomum zeylanicum Nees), litsea (Litsea cubeba Lour. Pers), lemongrass (Cympogon citrati L. Stapf), and ginger (Zingiber officinalis Rosc.) in the vapor phase on growth, sporulation, and mycotoxins production of two Aspergillus strains (Aspergillus parasiticus CGC34 and Aspergillus ochraceus CGC87), important postharvest pathogens of green and roasted coffee beans. Moreover, the effect of the essential oils (EOs) on the sensory profile of the coffee samples treated with EOs was evaluated. The major components of tested EOs were determined by gas chromatography and mass spectrometry (GC–MS) and gas chromatography with flame ionization detector (GC-FID). The results showed that almond, cinnamon bark, lemongrass, and litsea EOs are able to significantly inhibit the growth, sporulation, and mycotoxins production by toxigenic fungi. Sensory evaluation of coffee beans treated with EOs before and after roasting showed that some EOs (except lemongrass and litsea) do not adversely affect the taste and aroma of coffee beverages. Thus, application of the vapors of almond and cinnamon EOs appears to be an effective way that could serve to protect coffee during its transport and storage from toxigenic fungi.

Incidence, Speciation, and Morpho-Genetic Diversity of Penicillium spp. Causing Blue Mold of Stored Pome Fruits in Serbia.

Blue mold, caused by Penicillium spp., is one of the most economically important postharvest diseases of pome fruits, globally. Pome fruits, in particular apple, is the most widely grown pome fruit in Serbia, and the distribution of Penicillium spp. responsible for postharvest decay is unknown. A two-year survey was conducted in 2014 and 2015, where four pome fruits (apple, pear, quince, and medlar) with blue mold symptoms were collected from 20 storage locations throughout Serbia. Detailed morphological characterization, analysis of virulence in three apple cultivars, and multilocus phylogeny revealed three main Penicillium spp. in order of abundance: P. expansum, P. crustosum, and P. solitum. Interestingly, P. expansum split into two distinct clades with strong statistical support that coincided with several morphological observations. Findings from this study are significant and showed previously undocumented diversity in blue mold fungi responsible for postharvest decay including the first finding of P. crustosum, and P. solitum as postharvest pathogens of quince and P. crustosum of medlar fruit in the world, and P. expansum of quince in Serbia. Data from this study provide timely information regarding phenotypic, morphological and genotypic plasticity in P. expansum that will impact the design of species-specific detection tools and guide the development of blue mold management strategies.

Organic acid, a virulence factor for pathogenic fungi, causing postharvest decay in fruits.

Decay due to fungal infection is a major cause of postharvest losses in fruits. Acidic fungi may enhance their virulence by locally reducing the pH of the host. Several devastating postharvest fungi, such as Penicillium spp., Botrytis cinerea, and Sclerotinia sclerotiorum, can secrete gluconic acid, oxalic acid, or citric acid. Emerging evidence suggests that organic acids secreted by acidic fungi are important virulence factors. In this review, we summarized the research progress on the biosynthesis of organic acids, the role of the pH signalling transcription factor PacC in regulating organic acid, and the action mechanism of the main organic acid secreted via postharvest pathogenic fungi during infection of host tissues. This paper systematically demonstrates the relationships between tissue acidification and postharvest fungal pathogenicity, which will motivate the study of host–pathogen interactions and provide a better understanding of virulence mechanisms of the pathogens so as to design new technical strategies to prevent postharvest diseases.

Antifungal activities of fluoroindoles against the postharvest pathogen Botrytis cinerea: In vitro and in silico approaches

Botrytis cinerea is a common necrotrophic fungal pathogen, leading cause of gray mold diseases in plants and fruit. Several benzimidazoles are used for controlling B. cinerea-associated infection in fruit and vegetables, but benzimidazoles resistance restricts its further uses. Therefore, it is a need for alternative drugs that control B. cinerea. Indoles are multi-faceted compounds and their structural similarities with antifungal benzimidazoles make them a choice for further investigation. Thus, the main objective of the study was to investigate the antifungal potencies of indoles against B. cinerea and to decipher the molecular mechanism involved. We conducted in vitro antifungal assays, fruit assays, and computational studies of interactions between indoles and fungal microtubule polymerase. Of the 16 halogenated indoles examined, 4-fluoroindole, 5-fluoroindole, and 7-fluoroindole (MIC range 2–5 mg/L) were found to be more potent than the fungicides fluconazole and natamycin. Fluoroindoles inhibited or eradicated B. cinerea infections in tangerines and strawberries. Molecular dynamic simulation and density functional theory showed that these fluoroindoles stably interacted with microtubule polymerase. Quantitative structure-activity relationship analyses of halogenated indoles revealed that the presence of a fluoro group in the indole moiety is essential for anti-Botrytis activity. The plausibility of the underlying antifungal mechanism was confirmed by in vitro tubulin polymerization. Collective outcomes of this study indicates that fluoroindoles could be used as alternative fungicidal agents against B. cinerea.

Double-stranded RNA targeting fungal ergosterol biosynthesis pathway controls Botrytis cinerea and postharvest grey mould.

SummaryPathogenic fungi cause major postharvest losses. During storage and ripening, fruit becomes highly susceptible to fungi that cause postharvest disease. Fungicides are effective treatments to limit disease. However, due to increased public concern for their possible side effects, there is a need to develop new strategies to control postharvest fungal pathogens. Botrytis cinerea, a common postharvest pathogen, was shown to uptake small double‐stranded RNA (dsRNA) molecules from the host plant. Such dsRNA can regulate gene expression through the RNA interference system. This work aimed to develop a synthetic dsRNA simultaneously targeting three essential transcripts active in the fungal ergosterol biosynthesis pathway (dsRNA‐ERG). Our results show initial uptake of dsRNA in the emergence zone of the germination tube that spreads throughout the fungus and results in down‐regulation of all three targeted transcripts. Application of dsRNA‐ERG decreased B. cinerea germination and growth in in vitro conditions and various fruits, leading to reduce grey‐mould decay. The inhibition of growth or decay was reversed by the addition of ergosterol. While dual treatment with dsRNA‐ERG and ergosterol‐inhibitor fungicide reduced by 100‐fold the required amount of fungicide to achieve the same protection rate. The application of dsRNA‐ERG induced systemic protection as shown by decreased decay development at inoculation points distant from the treatment point in tomato and pepper fruits. Overall, this study suggests that dsRNA‐ERG can effectively control B. cinerea growth and grey‐mould development suggesting its efficacy as a future method for postharvest control of fungal pathogens.

Inhibitory Activity of Shrimp Waste Extracts on Fungal and Oomycete Plant Pathogens

(1) Background: This study was aimed at determining the in vitro inhibitory effect of new natural substances obtained by minimal processing from shrimp wastes on fungi and oomycetes in the genera Alternaria, Colletotrichum, Fusarium, Penicillium, Plenodomus and Phytophthora; the effectiveness of the substance with the highest in vitro activity in preventing citrus and apple fruit rot incited by P. digitatum and P. expansum, respectively, was also evaluated. (2) Methods: The four tested substances, water-extract, EtOAc-extract, MetOH-extract and nitric-extract, were analyzed by HPLC-ESI-MS-TOF; in vitro preliminary tests were carried out to determine the minimal inhibitory/fungicidal concentrations (MIC and MFC, respectively) of the raw dry powder, EtOAc-extract, MetOH-extract and nitric-extract for each pathogen. (3) Results: in the agar-diffusion-assay, nitric-extract showed an inhibitory effect on all pathogens, at all concentrations tested (100, 75, 50 and 25%); the maximum activity was on Plenodomus tracheiphilus, C. gloeosporioides and Ph. nicotianae; the diameters of inhibition halos were directly proportional to the extract concentration; values of MIC and MFC of this extract for all pathogens ranged from 2 to 3.5%; the highest concentrations (50 to 100%) tested in vivo were effective in preventing citrus and apple fruit molds. (4) Conclusions: This study contributes to the search for natural and ecofriendly substances for the control of pre- and post-harvest plant pathogens.

Control of Penicillium expansum by an Epiphytic Basidiomycetous Yeast

Postharvest biocontrol agents are considered a viable alternative to the use of synthetic chemicals as demonstrated by extensive research conducted by scientists and companies worldwide. In the present investigation, the biocontrol potential of a carotenoid-producing basidiomycetous yeast isolated from table grape flowers was analyzed. The strain RY1 proved to be Sporobolomyces roseus. In vitro and in vivo tests were conducted to assess its efficacy against Penicillium expansum, one of the most important postharvest pathogens and producer of the mycotoxin patulin. The yeast proved to control both fungal growth and patulin production, and, in addition, to greatly affect disease incidence and severity on apples. Its mode of action is presumably related both to the competition for nutrients and the production of antifungal volatiles. As such, although further large-scale trials are needed, our S. roseus strain represents a potential interesting biocontrol agent to be applied after harvest.

An Analysis of Postharvest Fungal Pathogens Reveals Temporal-Spatial and Host-Pathogen Associations with Fungicide Resistance-Related Mutations.

Fungicides are the primary tools to control a wide range of postharvest fungal pathogens. Fungicide resistance is a widespread problem that has reduced the efficacy of fungicides. Resistance to FRAC-1 (Fungicide Resistance Action Committee-1) chemistries is associated with mutations in amino acid position 198 in the β-tubulin gene. In our study, we conducted a meta-analysis of β-tubulin sequences to infer temporal, spatial, plant host, and pathogen genus patterns of fungicide resistance in postharvest fungal pathogens. In total, data were acquired from 2,647 specimens from 12 genera of fungal phytopathogens residing in 53 countries on >200 hosts collected between 1926 and 2020. The specimens containing a position 198 mutation were globally distributed in a variety of pathosystems. Analyses showed that there are associations among the mutation and the year an isolate was collected, the pathogen genus, the pathogen host, and the collection region. Interestingly, fungicide-resistant β-tubulin genotypes have been in a decline since their peak between 2005 and 2009. FRAC-1 fungicide usage data followed a similar pattern in that applications have been in a decline since their peak between 1997 and 2003. The data show that, with the reduction of selection pressure, FRAC-1 fungicide resistance in fungal populations will decline within 5 to 10 years. Based on this line of evidence, we contend that a β-tubulin position 198 mutation has uncharacterized fitness cost(s) on fungi in nature. The compiled dataset can inform end users on the regions and hosts that are most prone to contain resistant pathogens and assist decisions concerning fungicide resistance management strategies.

First Report of Postharvest Blue Mold decay caused by Penicillium expansum on Lemon (Citrus limon) fruit in China.

Lemon (Citrus limon) is one of the most important commercial (both dried and fresh) citrus fruits in China. In the spring of 2019, postharvest blue mold decay was observed at an incidence of 3-5% on lemon fruit at the local markets in Beijing, China. Fruit lesions were circular, brown, soft, and watery, and rapidly expanded at 25°C. To isolate the causal organism, small pieces (2 mm3) were cut from the lesions, surface-sterilized for 1 min in 1.5% NaOCl, rinsed three times with sterilized water, dried with sterile filter paper, placed onto potato dextrose agar (PDA) medium, and incubated at 25°C for 6 days. Eight morphologically similar single-colony fungal isolates were recovered from six lemon fruit. Colony surfaces were bluish-green on the upper surface and cream to yellow-brown one the reverse. Hyphae on colony margins were entirely subsurface and cream in color. Mycelium was highly branched, septate, and colorless, and conidiophores were 250 to 450 × 3.0 to 4.0 µm in size. Stipe of conidiophores were smooth-walled, bearing terminal penicilli, typically terverticillate or less commonly birverticillate, rami occurring singly, 16 to 23 × 3.0 to 4.0 µm, metulae in 3 to 6, measuring 12 to 15 × 3.0 to 4.0 µm. Phialides were ampulliform to almost cylindrical, in verticils of 5 to 8, measuring 8 to 11 × 2.5 to 3.2 µm with collula. Conidia were smooth-walled, ellipsoidal, measuring 3.0 to 3.5 × 2.5 to 3.0 µm. According to morphological characteristics, the fungus was identified as Penicillium expansum (Visagie et al. 2014). For molecular identification, genomic DNA of eight fungal isolates was extracted, regions of the beta-tubulin (TUB), and calmodulin (CAL) genes and ITS region, were amplified using Bt2a/Bt2b, CAL-228F/ CAL-737, and ITS1/ITS4 primers respectively. Obtained sequences of all isolates were identical to sequences of the representative isolate YC-IK12, which was submitted in the GenBank. BLAST results of YC-IK12 sequences (ITS; MT856700: TUB; MT856958: CAL; MT856959) showed 98 to 100% similarity with P. expansum accessions (NR-077154, LN896428, JX141581). For pathogenicity tests, 10 μl of conidial suspension (10 × 105 conidia/ml) from seven-day-old YC-IK12 culture was inoculated using a sterilized needle into the surface of each five asymptomatic disinfected lemons. As a control, three lemons were inoculated using sterile distilled water. All inoculated lemons were placed in plastic containers and incubated at 25°C for 7 days. Decay lesions, identical to the original observations, developed on all inoculated lemons, while control lemons remained asymptomatic. Fungus re-isolated from the inoculated lemon was identified as P. expansum on the basis morphology and Bt2a/Bt2b, CAL-228F/ CAL-737, and ITS1/ITS4 sequences. Previously, Penicillium spp. including P. expansum have been reported as post-harvest pathogens on various Citrus spp. (Louw & Korsten 2015). However, P. digitatum has been reported on lemons and P. expansum has been reported on stored Kiwifruit (Actinidia arguta), Malus, and Pyrus species in China (Tai, 1979; Wang et al. 2015). To our knowledge, this is the first report of blue mold caused by P. expansum on lemons in China. References Louw, J. P., Korsten, L. 2015. Plant Dis. 99:21-30. Tai, F.L. 1979. Sylloge Fungorum Sinicorum. Sci. Press, Acad. Sin., Peking, 1527 pages. 8097 Visagie, C.M. et al. 2014. Studies. Mycol.78: 343. Wang, C. W. et al. 2015. Plant Dis. 99:1037.

Antifungal Activity of Essential Oils from Three Artemisia Species against Colletotrichum gloeosporioides of Mango.

Post-harvest diseases of mango reduce fruit quality and cause severe yield losses with completely unmarketable fruits. The most common diseases of mangos are anthracnose (Colletotrichum gloeosporioides). In this study, the antibacterial activities of essential oils from Artemisia scoparia, Artemisia lavandulaefolia, and Artemisia annua against C. gloeosporioides were tested. The results showed that the essential oil of A. scoparia was more effective by the agar diffusion method; the EC50 value was 9.32 µL/mL. The inhibition rate was 100%, at a concentration of 10 μL/mL, through the spore germination method. The morphological changes of the mycelium were observed by scanning electron microscopy (SEM), the mycelia treated with essential oils showed shrinking, deformity, fracture, and dryness through SEM. A. scoparia essential oil was inoculated in vivo and subjected to paroxysm testing under natural conditions. A. scoparia had significantly inhibitory activity, and the inhibition rate was 66.23% in vivo inoculation tests after 10 days. The inhibition rate was 92.06% in the paroxysm test under natural conditions after 15 days. Finally, A. acoparia essential oil was analyzed by gas chromatography-mass spectrometry. The main compounds were 2-ethenyl-Naphthalene (23.5%), 2,4-pentadiynyl-Benzene (11.8%), 1,2-dimethoxy-4-(2-propenyl)-Benzene (10.0%), β-Pinene (8.0%), and 1-methyl-4-(1-methylethyl)-1,4-Cyclohexadiene (6.3%). The results have revealed the potential use of A. scoparia essential oil against post-harvest fungal pathogens C. gloeosporioides.