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(Colour online) Decay symptoms and morphological features. (a) Penicillium crustosum white mycelium and green sporulation and rot symptoms on inoculated raspberries in pathogenicity tests. (b) Penicillium crustosum conidiophores and conidia. (c) Mucor piriformis mycelia and symptoms in later stage on naturally infected cherry. (d) Sporangiophores, sporangia, collumelae and sporangiospores of Mucor piriformis. Scale bar = 10 μm.
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Cherries and raspberries are important fruit crops in Patagonia, Argentina. A high proportion (50%) of raspberry production is organic certified and sold for fresh and frozen domestic markets. Due to postharvest rot diseases, cherries and raspberries from Patagonia are kept in cold storage for a short period of time. During the storage season of 20...
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... of cherries and berries conducted on the Andean range zone of Patagonia, Argentina (Trevelin, 43°02′19S, 71°28′25W; Lago Puelo, 41°59′57S, 71°33′10W; El Bolsón, 41°56′ 42S, 71°31′13W), cherry ('Bing', 'Lapins' and 'Rainier' varieties) and raspberry ('Autumn Bliss' variety) fruits with rot symptoms were obtained from storage chambers at 0-1°C (Fig. 1a,c). Both types of fruit came from organic and conventional fields in the west area of Patagonia (Andean range zone). Small pieces (3 mm 2 ) of 50 symptomatic fruits were excised from the junction of diseased and healthy tissue, surface-sterilized in 70% ethanol for 30 s, washed in three changes of sterile distilled water, air dried, ...
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... green colonies on all media. Colonies were radially sulcate and velutinous, with clear exudate, and produced a yellow to orange reverse colour on CYA and YES. Conidiophores were terverticillate, stipes septate with rough walls, and ampulliform phialides. Conidia were smooth, spherical to subglobose, in columns, measuring 2.9-4.9 (3.6) μm (n = 30) (Fig. 1b). BLAST analysis revealed that the sequences were 100% identical to Penicillium crustosum (GenBank accession number AY674353, sequence of the type material CBS115503), and the phylogenetic analyses confirmed these relationships (Fig. ...
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... branched and unbranched, short sporangiophores branched sympodially. Sporangia were globose, black-brown at maturity, 85-212 μm in diameter. Collumelae were variable in shape, cylindrical-ellipsoidal or pyriform, 34.7-61.8 (43.1) × 23.2-60.8 (51.7) μm. Sporangiospores were ellipsoidal, subspherical and smooth 3.9-10.7 (6.2) × 4.9-12.6 μm (n = 30) (Fig. 1d). Chlamydospore-like resting structures, isogametangia and zygospores were not evident in culture. BLAST analysis of the DNA sequences identified the 10 isolates as M. piriformis sensu lato (99% identical to the ITS-5.8S rDNA sequence of M. piriformis isolate .73A, GenBank accession number JN206033, from red currant). In the ...
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... both types of fruit, temperatures and experiments; the disease incidence was 100% for both pathogens. Symptoms of decay were observed in all inoculated fruits, but not in the controls. Mucor piriformis rot was light brown, watery, soft and covered with fuzzy mycelia. Coloured sporangiophores bearing terminal sporangia protruded through the skin (Fig. 1c). Decayed areas produced by P. crustosum were light to medium brown, soft and watery, and with blue green sporulation on the surface of the lesion (Fig. 1a). Both fungi were re-isolated from infected tissues and showed the same morphological and molecular characteristics as the original ...
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... but not in the controls. Mucor piriformis rot was light brown, watery, soft and covered with fuzzy mycelia. Coloured sporangiophores bearing terminal sporangia protruded through the skin (Fig. 1c). Decayed areas produced by P. crustosum were light to medium brown, soft and watery, and with blue green sporulation on the surface of the lesion (Fig. 1a). Both fungi were re-isolated from infected tissues and showed the same morphological and molecular characteristics as the original ...
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Citations
... Most Penicillium spp. are saprophytic, but some can act as opportunistic plant pathogens and cause postharvest diseases, such as P. expansum (Louw and Korsten, 2014;Yin et al., 2017), P. crustosum (López et al., 2016), P. digitatum and P. italicum (Arras et al., 2005). More importantly, Penicillium spp. ...
Penicillium spp. produce a great variety of secondary metabolites, including several mycotoxins, on food sub-strates. Chestnuts represent a favorable substrate for Penicillium spp. development. In this study, the genomes of ten Penicillium species, virulent on chestnuts, were sequenced and annotated: P. bialowiezense. P. pancosmium, P. manginii, P. discolor, P. crustosum, P. palitans, P. viridicatum, P. glandicola, P. taurinense and P. terrarumae. Assembly size ranges from 27.5 to 36.8 Mb and the number of encoded genes ranges from 9,867 to 12,520. The total number of predicted biosynthetic gene clusters (BGCs) in the ten species is 551. The most represented families of BGCs are non ribosomal peptide synthase (191) and polyketide synthase (175), followed by terpene synthases (87). Genome-wide collections of gene phylogenies (phylomes) were reconstructed for each of the newly sequenced Penicillium species allowing for the prediction of orthologous relationships among our species, as well as other 20 annotated Penicillium species available in the public domain. We investigated in silico the presence of BGCs for 10 secondary metabolites, including 5 mycotoxins, whose production was validated in vivo through chemical analyses. Among the clusters present in this set of species we found andrastin A and its related cluster atlantinone A, mycophenolic acid, patulin, penitrem A and the cluster responsible for the synthesis of roquefortine C/glandicoline A/glandicoline B/meleagrin. We confirmed the presence of these clusters in several of the Penicillium species conforming our dataset and verified their capacity to synthesize them in a chestnut-based medium with chemical analysis. Interestingly, we identified mycotoxin clusters in some species for the first time, such as the andrastin A cluster in P. flavigenum and P. taurinense, and the roquefortine C cluster in P. nalgiovense and P. taurinense. Chestnuts proved to be an optimal substrate for species of Penicillium with different mycotoxigenic potential, opening the door to risks related to the occurrence of multiple mycotoxins in the same food matrix.
... Over a 3-year sampling period, we isolated 37 unique fungal genera both externally and internally from field-collected adult flies, indicating that flies both passively acquire fungal inoculum on their cuticle and feed on fungi. Many of the fungi that we isolated from are considered large, speciose taxa that contain both known fruit pathogens and fungi that colonize non-crop spaces (Bensch et al. 2015;López et al. 2016;Swett et al. 2019). For example, Cladosporium, the most commonly isolated fungal genus, has at least 170 recognized species that occupy diverse habitats including soil, indoor environments, plant detritus, and fruit (Bensch et al. 2012(Bensch et al. , 2015. ...
Interactions between microorganisms and frugivorous insects can modulate fruit rot disease epidemiology. Insect feeding and/or oviposition wounds may create opportunities for fungal infection. Passive and active dispersal of fungal inoculums by adult insects also increases disease incidence. In fall-bearing raspberries and blackberries, such vectoring interactions could increase crop damage from the invasive pestiferous vinegar fly Drosophila suzukii (spotted-wing drosophila). Periods of peak D. suzukii activity are known to overlap with several species of primary fruit rot pathogen, particularly Botrytis cinerea and Cladosporium cladosporioides, and previous work indicates that larvae co-occur with and feed on various filamentous fungi at low rates. To further our understanding of the epidemiological consequences that may emerge from these associations, we surveyed the filamentous fungal community associated with adult D. suzukii, isolating and molecularly identifying fungi externally and internally (indicating feeding) from field-collected adults over 3 years. We isolated and identified 37 unique genera of fungi in total, including known raspberry pathogens. Most fungi were detected infrequently, and flies acquired and carried fungi externally at higher richness, frequency, and density relative to internally. In a worst-case scenario laboratory vectoring assay, D. suzukii adults were able to transfer B. cinerea and C. cladosporioides to sterile media at 0, 24, 48, and 72 h after exposure to sporulating cultures in Petri dishes. These results collectively suggest an adventitious vectoring association between D. suzukii and fruit rot fungi that has the potential to alter caneberry disease dynamics.
... Raspberry is easily incurred to fruit softening and mechanical damage to tissues and higher respiratory activity, which provides a congenial environment to various fungal pathogens (Guerreiro et al., 2015). Botrytis cinerea, Rhizopus stolonifer, Colletotrichum sp., Penicillium crustosum, Mucor piriformis and yeasts have been reported to cause fruit rot in raspberries, shortening their storage life and higher postharvest losses (Guerreiro et al., 2015;López et al., 2016;Schoeneberg and Hu, 2020;Zhang et al., 2020). ...
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... Other fungal genera found in berries were Alternaria, Cladosporium, Geotrichum, Mucor, and Penicillium, which have already been reported as causative agents of postharvest disease in these fruit (Koike et al., 2003;Tournas and Katsoudas, 2005;Gordon et al., 2016;López et al., 2016;Pastrana et al., 2017;Petrasch et al., 2019). It should be noted that fungi belonging to beneficial species such as Trichoderma have also been found in berries (Santoyo et al., 2021). ...
... Mould counts in the early stages of 'aqueous spot' were beneath the detection thresholds, with inconsistent presence of various well-known rotting species. Monilia laxa [9], A. alternata [37], P. crustosum [11], F. oxysporum [38] and B. cinerea [13] have been associated with different diseases of sweet cherries. Inoculation trials showed a high disease incidence in most cases. ...
The pre and postharvest disease named ‘aqueous spot’ is an emerging risk for sweet cherries growing in Jerte Valley (Cáceres, Spain). Early stages of the disease appear in the tree, but it is usually detected after harvesting, during the postharvest period. Symptoms include the appearance of skin discolouration and translucency in the shoulder areas. At the most advanced stages, a mycelium of white colour partially or completely covers the fruit. This manuscript provides a detailed description of the microbes involved in this disease, such as bacteria, yeasts, and moulds. Microbes of different cherry cultivars were studied during two consecutive seasons (2019 and 2020). The counts of bacteria and yeast in damaged tissues were higher (7.05 and 6.38 log10 CFU/g for total aerobic mesophilic microbes and yeasts, respectively) than sound tissues (6.08 and 5.19 log10 CFU/g, respectively). The Enterobacterales order dominated the bacteria population. Among yeasts, Yarrowia lipolytica, in 2019, and Metschnikowia pulcherrima and Metschnikowia viticola, in 2020, were consistently isolated from all samples. The presence of moulds was inconsistently detected at the early stage of this disease by plate counts. However, microscopic observations revealed the presence of hyphae in cherry flesh. Different pathogenic moulds were identified, although white mycelium, identified as Botrytis cinerea by molecular methods, was consistently isolated at later stages. Inoculation tests confirmed the involvement of white-mycelium B. cinerea in the development of this new postharvest disease in the Jerte Valley. Its combination with Enterobacterales enhanced the evolution of rotting, whereas the combination with yeasts decreased and delayed the symptoms. This work presents the first report of a consortia of microorganisms implicated in the development of ‘aqueous spot’, an emerging disease in sweet cherry cultivars in the Jerte Valley.
... Other fungal genera found in berries were Alternaria, Cladosporium, Geotrichum, Mucor, and Penicillium, which have already been reported as causative agents of postharvest disease in these fruit (Koike et al., 2003;Tournas and Katsoudas, 2005;Gordon et al., 2016;López et al., 2016;Pastrana et al., 2017;Petrasch et al., 2019). It should be noted that fungi belonging to beneficial species such as Trichoderma have also been found in berries . ...
In recent decades, various bacterial species have been characterized as biocontrol agents for plant crop diseases; however, only a few genera have been predominantly reported in literature. Therefore, identification of for new antagonists against phytopathogens is essential for boosting sustainable food production systems. In this study, we evaluated the role of strain SER3 from the recently discovered Rouxiella badensis as a biocontrol agent. SER3 was isolated from the phyllosphere of decaying strawberry fruit (Fragaria × ananassa) and showed different grades of antagonism against 20 fungal pathogens of berries, based on confrontation assays, due to the action of its diffusible and volatile compounds. These fungal pathogens were isolated from decayed strawberry, blackberry, and blueberry fruit and were characterized through internal transcribed spacer (ITS) sequencing and homology searches, exhibiting similarity with well-known postharvest pathogens such as Botrytis, Fusarium, Gotrichum, Mucor, Penicillium, Alternaria, and Botryosphaeria. Koch's postulates were confirmed for most pathogens by re-infecting berry fruit. SER3 showed good capacity to inhibit the growth of Botrytis cinerea and Fusarium brachygibbosum in strawberry fruit, affecting mycelial development. To gain better understanding of the genetic and metabolic capacities of the SER3 strain, its draft genome was determined and was found to comprise a single chromosome of 5.08 Mb, 52.8% G + C content, and 4545 protein-coding genes. Phylogenetic analysis indicated that the SER3 strain is affiliated with the R. badensis species, with an average nucleotide identity > 96% and a genome-to-genome distance > 70%. Comparison of the genomic properties of R. badensis SER3 and other close bacterial relatives showed several genes with potential functions in biocontrol activities, such as those encoding siderophores, nonribosomal peptide synthetases, and polyketide synthases. This is the first study to demonstrate a novel role of the recently discovered R. badensis species (and any other species of the genus Rouxiella) as a biocontrol agent against postharvest fungal pathogens.
... When the disease progresses growth of white mycelium and brownish to grey sporangia can be observed. Finally, the infected tissue is broken down and disintegrates in a watery rot (Kwon and Hong 2005;López et al. 2016). Phylogram generated from RAxML analysis based on combined sequences of ITS and LSU of Mucorand Backusella species. ...
This is a continuation of a series focused on providing a stable platform for the taxonomy of phytopathogenic fungi and fungus-like organisms. This paper focuses on one family: Erysiphaceae and 24 phytopathogenic genera: Armillaria, Barriopsis, Cercospora, Cladosporium, Clinoconidium, Colletotrichum, Cylindrocladiella, Dothidotthia,, Fomitopsis, Ganoderma, Golovinomyces, Heterobasidium, Meliola, Mucor, Neoerysiphe, Nothophoma, Phellinus, Phytophthora, Pseudoseptoria, Pythium, Rhizopus, Stemphylium, Thyrostroma and Wojnowiciella. Each genus is provided with a taxonomic background, distribution, hosts, disease symptoms, and updated backbone trees. Species confirmed with pathogenicity studies are denoted when data are available. Six of the genera are updated from previous entries as many new species have been described.
... The full ITS region of rDNA (ITS1, ITS2 and the intervening 5.8S RNA gene) was amplified. Filamentous yeasts were amplified using primer pairs ITS1/ITS4 (White et al., 1990), setting thermocycling conditions according to López et al. (2016). 'Zygomycota' isolates were amplified using ITS1/ITS4 universal primer pairs (White et al., 1990) setting PCR conditions according to Walther et al. (2013). ...
Temperate forests ecosystems are threatened by declines and diseases worldwide. Causes have been investigated during the last decades allowing, in some cases, to relate such deterioration to climate change or pathogens but, in many cases, causes have not yet been elucidated. In recent decades, a phenomenon of grouped mortality, whose etiology remains unknown, has been observed in the two most distributed Nothofagus species of North Patagonian forests. This study aimed to assess sapwood-inhabiting fungal diversity of N. dombeyi and N. pumilio trees in healthy and affected stands in order to determine whether health condition shape these fungal communities, to characterize such patterns and, to seek for the likely pathogens associated with tree damage. Seven sites in Los Alerces National Park (Chubut, Argentina), were sampled seasonally for two years. Eighty-eight fungal taxa were recovered and identified. Spatial heterogeneity across plant compartments was found, involving community composition and structure, with stem harbouring greater diversity than root. It was found that the sapwood in both Nothofagus species was inhabited by different fungal species, in high richness and showing different patterns according to health condition and plant compartment. Health condition was a stronger driver of N. dombeyi sapwood-inhabiting fungal community than it was for N. pumilio; the latter evidenced plant compartment as a stronger driver of such community. Nothofagus pumilio presented higher frequency of decomposition agents in both affected and healthy trees than it was in N. dombeyi, indicating a greater wood deterioration of its stands. More decomposition agents and potential pathogens have been found in symptomatic stands of N. dombeyi than in healthy stands, although their frequency patterns do not allow the inference of conclusions about the primary fungal agent causing tree mortality. Our findings suggest a secondary role of living-wood-inhabiting mycobiota in tree damage processes as an expression of tree stress caused by climatic factors.
Cherry trees (Cerasus pseudocerasus (Lindl.) Loudon) in Bai Lu Yuan, Shaanxi Province, China, are susceptible to a serious disease that causes substantial economic losses. The pathogens causing this disease, designated black spot disease, are currently unknown. To identify the relationship between microbial community structure and the status of plant health, we obtained samples from the rhizosphere and tissues of both healthy cherry trees and those with symptoms of black spot disease. We used Illumina sequencing of 16S rRNA gene and internal transcribed spacer (ITS) amplicons to examine the bacterial and fungal communities in these samples. An alpha diversity analysis showed that there were no significant differences between diseased and healthy soils, while fungal diversity in diseased tissue was significantly higher than in healthy tissue (P < 0.01), suggesting that an imbalance in fungal flora may be related to black spot disease. Taxonomic classification and linear discriminant analysis effect size (LEfSe) indicated an increase in the relative abundance of some bacterial genera (Pseudomonas, Burkholderia, Sphingobium, Erwinia, Sphingomonas and Bradyrhizobium) and fungal genera (Phoma, Penicillium, Fusarium, Cystobasidium, Aureobasidium and Nakazawaea) in the diseased tissues (p<0.05). Among these, the increased relative abundance of the bacteria Pseudomonas and Erwinia and fungi Penicillium, Phoma and Fusarium may be associated with blackspot of cherry tree. The bacterial genera Burkholderia, Sphingobium, Sphingomomas and Bradyrhizobium are probably antagonists of the pathogen(s) of black spot. These findings are an initial step in the identification of potential candidates for the biological control of the disease.
