Complete genome sequence analysis of Pestalotiopsis microspora, a fungal pathogen causing kiwifruit postharvest rots.

BACKGROUND: The postharvest rot of kiwifruit is one of the most devastating diseases affecting kiwifruit quality worldwide. However, the genomic basis and pathogenicity mechanisms of kiwifruit rot pathogens are lacking. Here we report the first whole genome sequence of Pestalotiopsis microspora, one of the main pathogens causing postharvest kiwifruit rot in China. The genome of strain KFRD-2 was sequenced, de novo assembled, and analyzed. RESULTS: The genome of KFRD-2 was estimated to be approximately 50.31 Mb in size, with an overall GC content of 50.25%. Among 14,711 predicted genes, 14,423 (98.04%) exhibited significant matches to genes in the NCBI nr database. A phylogenetic analysis of 26 known pathogenic fungi, including P. microspora KFRD-2, based on conserved orthologous genes, revealed that KFRD-2's closest evolutionary relationships were to Neopestalotiopsis spp. Among KFRD-2's coding genes, 870 putative CAZy genes spanned six classes of CAZys, which play roles in degrading plant cell walls. Out of the 25 other plant pathogenic fungi, P. microspora possessed a greater number of CAZy genes than 22 and was especially enriched in GH and AA genes. A total of 845 transcription factors and 86 secondary metabolism gene clusters were predicted, representing various types. Furthermore, 28 effectors and 109 virulence-enhanced factors were identified using the PHI (pathogen host-interacting) database. CONCLUSION: This complete genome sequence analysis of the kiwifruit postharvest rot pathogen P. microspora enriches our understanding its disease pathogenesis and virulence. This study establishes a theoretical foundation for future investigations into the pathogenic mechanisms of P. microspora and the development of enhanced strategies for the efficient management of kiwifruit postharvest rots.

First Report of Fusarium fujikuroi Causing Postharvest Rot of Kiwifruit (Actinidia chinensis) in Jiangxi Province, China.

Kiwifruits (Actinidia chinensis) are among the most widely planted fruit in Jiangxi Province, China. Infected kiwifruits of the cultivars 'Hongyang' and 'Jinyan' were obtained from a commercial orchard in Fengxin county, Jiangxi Province (28°67' N; 115°42' E) from September to November 2022. The 1200 kiwifruits were collected from cold storage (cold stored for 3 months at 2°C), and moved to room temperatures (15 to 20°C), approximately 20% had symptoms of postharvest soft rot 7 days later. The infected fruits had brown or dark gray spots on the peel. Most were round or oval, with a diameter of approximately 1~3 cm. The pulp was milky white, and there was a waterlogged ring at the junction of decay. The pathogen was isolated by removing several small pieces (3×3 mm) of infected tissue from the diseased kiwifruits, which were sterilized with 75% ethanol for 30 s, dipped in 1% NaClO for 1 min, and rinsed three times with sterile distilled water. These pieces were transferred onto potato dextrose agar (PDA) and incubated for 5 days at 28°C, 75% relative humidity (RH), separated, and repurified. Eight unidentified isolates with similar morphology were obtained on PDA (D3-1 to D3-8). These isolates had abundant aerial fluffy mycelia. The colonies were white during the early stage of culture and turned light purple in the later stage. The mycelia grew 5.8 mm day-1 (n=5) on average and produced abundant conidia 10 days later. The microconidia were solitary, transparent, ovoid, with 0 to 1 septa, and 3.6 to 11.2 × 1.6 to 3.5 µm (average 6.5 × 2.9 µm, n = 50). The macroconidia were sickle-shaped, slender and slightly curved, with 3 to 5 septa, and 22.3 to 53.9 × 2.6 to 5.4 µm (average 39.5 × 4.3 µm, n = 50). Chlamydospores were absent. The morphological characteristics enabled the identification of the pathogen as Fusarium spp. (Leslie and Summerell, 2006). Isolate D3-2 was further confirmed, and the primers ITS1/ITS4 (White et al. 1990), 5F2/7CR and EF1/EF2 (O'Donnell et al. 2022) were used to amplify the internal transcribed spacer (ITS) region, RNA polymerase II largest subunit (RPB2) gene and translation elongation factor-1 alpha regions (TEF-1α). The ITS (accession no. PP077075), RPB2 (PP566653) and TEF-1α (PP566654) sequences shared 99.62 to 100% identities with ITS (ON564593.1), RPB2 (ON734380.1) and TEF-1α (ON697186.1) of F. fujikuroi from NCBI, respectively. Thus, the pathogen was identified as F. fujikuroi based on morphological and molecular characteristics. Each of the three isolates was inoculated on surface-disinfected (75% ethanol, 5 min) disease-free kiwifruits of cv. 'Jinyan' and 'Hongyang'. The six kiwifruits were pierced by a sterile inoculation needle and inoculated with 20 µl spore suspension (1×106 spores/ml), and six kiwifruits were treated with spore suspension without any wounds, four control fruits were inoculated with sterile distilled water. All the fruits were sealed in a storage box, kept at an RH of 90%-95%, and incubated at a constant temperature of 28°C for 5 days. After 3 days, the fruit rotted at the inoculation site, and after 5 days, the lesions gradually increased, and the symptoms were the same as those of the original sample. The control fruits remained disease-free. The pathogenicity tests were repeated three times. Koch's postulates were completed by reisolating the fungus from infected kiwifruits, which was identified as F. fujikuroi by sequencing. Although F. solani (Yang et al. 2018) and F. acuminatum (Wang et al. 2015) have been previously reported to rot kiwifruits in China, this is the first report of F. fujikuroi causing postharvest rot on kiwifruits in China. This discovery can alert agronomists to prevent and control this pathogen.

Neofusicoccum actinidiae and Neofusicoccum guttata, Two New Species Causing Kiwifruit Rot in China.

Kiwi is a popular fruit consumed worldwide. A number of fungal pathogens have been reported to cause postharvest rot of kiwifruit, and Botryosphaeriaceae species are the major causal agents of the disease. In this study, 18 isolates belonging to the genus Neofusicoccum (family Botryosphaeriaceae) were isolated from 247 symptomatic kiwifruits of the cultivars Jinyan, Jintao, and Jinkui collected from orchards in Hubei and Jiangxi provinces, China. Among the isolates, three grouped with various known Neofusicoccum parvum isolates, whereas the remaining 15 formed two independent clades. On the basis of further phylogenetic analyses with concatenated sequences of ITS and three genes encoding translation elongation factor 1-alpha (TEF), ß-tubulin (TUB), and DNA-dependent RNA polymerase II subunit (RPB2), as well as morphological characteristics, two new species, N. actinidiae and N. guttata, were proposed. Their pathogenicity to kiwi, apple, and citrus fruits was also confirmed.

Utilization of the antagonistic yeast, Wickerhamomyces anomalus, combined with UV-C to manage postharvest rot of potato tubers caused by Alternaria tenuissima.

Postharvest rot of potato tubers caused by fungal pathogens is the main cause of significant economic losses, while also raising potential food safety issues. Integrated disease management, utilizing bio-safe and eco-friendly methods, represents a sustainable strategy for reducing postharvest losses in crops, including potato. In the current study, the application of the antagonistic yeast, Wickerhamomyces anomalus, combined with a UV-C treatment was evaluated for the management of postharvest Alternaria rot of potato tubers, caused by Alternaria tenuissima. Both W. anomalus and UV-C as individual treatments reduced the size of A. tenuissima infections on potato tubers, relative to the control, while the combined treatment of W. anomalus and UV-C exhibited the highest level of inhibition. W. anomalus or UV-C alone, and especially when used in combination, induced the expression of defense-related genes, including polyphenol oxidase, peroxidase, and ß-1,3-glucanase, and also increased the level of flavonoids and lignin in potato tubers. Our findings indicate that the mechanism of action by which UV-C enhances the biocontrol effect of W. anomalus against postharvest Alternaria rot includes the activation of defense-related response in potato tubers. The integration of biocontrol agents and physical treatments (e.g., UV-C) represents an effective, eco-friendly hurdle technology for managing postharvest rot in potato.

Pre- and postharvest measures used to control decay and mycotoxigenic fungi in potato (Solanum tuberosum L.) during storage.

Potato (Solanum tuberosum L.), a worldwide, staple food crop, is susceptible to postharvest rots caused by a variety of fungal pathogens, including Fusarium spp., Alternaria spp., Phytophthora infestans, Helminthosporium solani, Rhizoctonia solani, and Colletotrichum coccodes. Rots resulting from infections by these pathogens cause a significant reduction in potato quality and marketable yield. Importantly, some of these decay fungi also produce mycotoxins that represent a potential risk to human health. In the present review, an overview and discussion are provided on the epidemiology and pathogenesis of decay fungi, especially Fusarium spp., that include recent data derived from genomic and phylogenetic analyses. The biosynthesis and functional role of fungitoxic metabolites such as trichothecene mycotoxins and fusaric acid, produced in rotted potatoes are also reviewed. Advances in pre- and postharvest measures for rot management, especially eco-friendly methods including physical control, biological control, the use of natural compounds, and other agricultural management practices are also reviewed. Lastly, novel approaches to control potato dry rot such as the use of mycoviruses and CRISPR technology are highlighted.

Control of nectarine fruits postharvest fungal rots caused by Botrytis Cinerea and Rhizopus Stolonifer via some essential oils.

Nectarines (Prunus persica L. Bath) are very sensitive fruit to fungal infection. Today, the control of postharvest fruit diseases with essential oils (EOs) has been significantly noticed as a novel trend in biological preservation. In this study, volatile compounds of Cinnamon zeylanicum (CEO), Zataria multiflora (ZEO), and Satureja khuzestanica (SEO) were analyzed by Gas Chromatography-Mass spectroscopy. Also, the in vitro antifungal activities of EOs against Botrytis cinerea and Rhizopus stolonifer were evaluated at different concentrations. The in vivo antifungal activity of these EOs on artificially infected nectarine fruits was also considered. The major components were Thymol (32.68%) and Carvacrol (30.57%) for ZEO, cinnamaldehyde (80.82%) for CEO, and carvacrol (38.43%) for SEO. The application of different concentrations showed a decreasing trend in the fungus radial growth in all EOs. In the in vitro experiments, ZEO and CEO exhibited more significant mycelial inhibition results and reduction of the IC50, MIC and MFC values against Botrytis cinerea and Rhizopus stolonifer, respectively. However, in the in vitro experiments, none of the treatments were capable of completely inhibiting the growth of the fungi. According to the results of this study, ZEO and CEO could reduce the damage caused by these fungi.

Detection and epidemiological progress of quiescent avocado diseases / Detecção e progresso epidemiológico de doenças quiescentes do abacate

ABSTRACT: One of the major problems in the commercialization of avocados is the incidence of postharvest diseases, especially anthracnose (Colletotrichum spp.) and stem-end rot (Lasiodiplodia theobromae, Fusicoccum aesculi and Neofusicoccum spp.). As there is a lack of epidemiological information on these pathosystems, the objective of this study was to establish a method to detect quiescent infections and characterize their temporal progression and spatial pattern in a commercial orchard. Detection of quiescent infections was evaluated in flowers and fruits that were immature and in commercial harvest stage, treated with paraquat, ethrel or water. Treatment of flowers and immature fruits with paraquat led to rapid detection of Colletotrichum spp. In two seasons of a 'Hass' avocado orchard, the incidence of diseases was evaluated from open flowers to fruit harvest, totaling 11 evaluations at biweekly intervals. When fruits reached the harvest stage, the spatial distribution of diseased fruits in the trees was evaluated by means of dispersion index and modified Taylor's law. Considering the evaluation of temporal disease progression, anthracnose was the most important disease, presenting a high initial incidence of 60 and 86% diseased flowers in the two seasons, respectively, while fruits showed an average disease incidence of 70 and 87%, respectively. Stem-end rot was observed only in fruits since the beginning of their development and presented low incidence (<8% fruits), significantly inferior to that of anthracnose. The diseases showed random dispersion within the trees, indicating that their initial inoculum is evenly distributed in the plants.

RESUMO: Um dos grandes problemas na comercialização de abacates é a incidência de doenças pós-colheita, principalmente a antracnose (Colletotrichum spp.) e as podridões pedunculares (Lasiodiplodia theobromae, Fusicoccum aesculi e Neofusicoccum spp.). Em função da carência de informações epidemiológicas sobre esses patossistemas, objetivou-se estabelecer um método para detectar infecções quiescentes e caracterizar o progresso temporal e o padrão espacial dessas doenças em pomar comercial. A detecção de infecções quiescentes foi avaliada em flores e em frutos imaturos e no ponto de colheita comercial, tratados com paraquat, ethrel ou água. O tratamento de flores e frutos imaturos com paraquat propiciou uma rápida detecção de Colletotrichum spp. Em duas safras de um pomar de abacate 'Hass', avaliou-se a incidência das doenças a partir das flores abertas até a colheita dos frutos, totalizando 11 avaliações em intervalos quinzenais. Quando os frutos alcançaram o ponto de colheita, avaliou-se a distribuição espacial de frutos doentes nas árvores por meio do índice de dispersão e da lei de Taylor modificada. Na avaliação do progresso temporal das doenças, a antracnose foi a mais importante, apresentando elevada incidência inicial, com 60 e 86% das flores doentes nas duas safras, enquanto que os frutos apresentaram médias de 70 e 87% de incidência da doença, respectivamente. As podridões pedunculares foram constatadas em frutos desde o início de seu desenvolvimento e apresentaram baixas incidências (<8% de frutos), significativamente inferior à de antracnose. As doenças apresentaram dispersão ao acaso dentro das árvores, indicando que o inóculo inicial das doenças está distribuído de maneira uniforme nas plantas.