Bottlebrush and Myrtle twig canker caused by Neopestalotiopsis species: an emerging canker-causing group of fungi in Italy.

Pestalotioid fungi were isolated in pure culture from symptomatic plants of Callistemonlaevis, C.viminalis, Lumaapiculata (marketed as "Myrtusluma"), Myrtuscommunissubsp.tarentina, and M.communisvar.microphylla (M.communis 'Microphylla'), showing twig canker, dieback and defoliation. The isolates were identified to species by ITS, tef1 and tub2 sequences, which revealed the presence of six species of Neopestalotiopsis (N.camelliae-oleiferae, N.hispanica, N.iberica, N.rosae, N.rosicola, and N.zakeelii) and one species of Pestalotiopsis (P.biciliata). While most species were isolated only once or twice, the majority of isolates belonged to N.rosae (13) and N.hispanica (8). Pathogenicity was investigated by pathogenicity tests on all hosts, which confirmed the pathogenicity of all Neopestalotiopsis species on at least some of the hosts tested, while P.biciliata did not cause any disease symptoms. Neopestalotiopsishispanica and N.rosae caused symptoms in all hosts of the present study, while the other Neopestalotiopsis species tested showed no symptoms on Lumaapiculata.

Fungal Species Causing Canker and Wilt of Ficus carica and Evidence of Their Association by Bark Beetles in Italy.

Field surveys conducted during 2021 and 2022 in Western Sicily, Italy, revealed the presence of common fig trees severely affected by trunk and crown root canker and bark cracking. Moreover, in conjunction with the symptomatic tissues, the same surveyed plants showed the presence of bark beetle holes and internal wood galleries. The predominant beetle Criphalus dilutus was previously reported attacking figs in Sicily. Phylogenetic analyses based on multilocus DNA data showed the presence of different fungal taxa associated with disease symptoms, including Botryosphaeria dothidea, Ceratocystis ficicola, Diaporthe foeniculina, Neocosmospora bostrycoides, N. perseae, and Neofusicoccum luteum. Pathogenicity tests conducted on potted fig plants showed that all the species were pathogenic to fig, with C. ficicola and Neocosmospora spp. as the most aggressive fungal species. Moreover, isolations conducted from the bodies of emerging adult insects recovered from disease samples confirmed the presence of C. ficicola and Neocosmospora spp., suggesting the potential involvement of C. dilutus in their dissemination.

Efficacy of Biological Control Agents and Resistance Inducer for Control of Mal Secco Disease.

Mal secco, caused by Plenodomus tracheiphilus, is an economically important fungal vascular disease in citrus-growing countries of the Mediterranean basin. Preventing fungal infections usually requires a high number of copper treatments but European legislation imposes the minimization of their accumulation in soil. In our study, biological control agents (BCAs) and a plant resistance inducer (PRI), tested in four different experiments on citrus seedlings under controlled conditions, have resulted in promising strategies to control mal secco disease. Foliar (Experiment I) and soil (Experiment II) applications of two formulations of Bacillus amyloliquefaciens strain D747 (Amylo-X® LC and Amylo-X® WG) provided similar performances in reducing the disease amount (incidence and symptoms severity) over time compared to the untreated control, whereas copper hydroxide (Kocide Opti®) used as standard was the most effective treatment over time. In the third experiment, Pythium oligandrum strain M1 (Polyversum®) and Trichoderma asperellum strain ICC012 + Trichoderma gamsii strain ICC080 (Remedier®) were able to reduce disease incidence and symptoms severity compared to the untreated control. Remedier® provided the best performances in reducing the disease amount, whereas the Polyversum® application was the least effective treatment over time. The effectiveness of the Trichoderma spp. formulation in reducing P. tracheiphilus infections did not significantly differ from the standard copper compound (Kocide Opti®). Comprehensively, in the last experiment (IV), acibenzolar-S-methyl (ASM) alone and in mixture with metalaxyl-M proved as effective as B. amyloliquefaciens strain FZB24, with no dose-response relationships observed. These findings provide important insight for the integrated management of mal secco disease.

First report of seedling stem blight of mango caused by Neofusicoccum parvum in Italy.

Seedling blight of mango (Mangifera indica L.) was observed in a commercial nursery located in Messina province (eastern Sicily, Italy) during winter of 2021. More than 30% of 3,000 seedlings, about three to six months old, of mango cv. Gomera 3 showed symptoms of basal stem blight. The symptoms started from seed, led to the decline and subsequent death of the plants. Necrotic lesions appeared at crown level two months after sowing. The stem tissues of ten symptomatic plants were cut, surface sterilized, dipped in 1.5% sodium hypochlorite for 1 min and transferred onto potato dextrose agar medium (PDA) and incubated at 25°C for four days. Approximately 60% of stem tissues developed very similar fungal colonies, resembling to Botryosphaeriaceae. A total of four representative isolates were collected through single hyphal-tip and stored at 4 °C. The internal transcriber spacer region (ITS) was amplified with primers, ITS5/ITS4 (White et al., 1990), and EF1-728F and EF1-986R (Carbone and Kohn, 1999) were used to amplify part of the translation elongation factor 1alpha gene (tef1-α), and primers Bt2a/Bt2b (Glass and Donaldson, 1995) were used for the partial ß-tubulin (tub2). The obtained sequences were deposited in GenBank with accession numbers: ON911292-95 for the ITS, ON933621-24 for tef1-α and ON933625-28 for tub2.To compare the results, 50 additional sequences were selected and inserted in the alignment according to the recent literature on the Botryosphaeriaceae (Bezerra et al., 2021; Zhang et al., 2021). Maximum parsimony analysis (MP) of concatenated dataset (ITS + tef1-α + tub2) was performed in PAUP v.4.0a. Clade support was assessed by 1,000 bootstrap replicates and Botryosphaeria dothidea was used as an outgroup. Our isolates clustered within the group of Neofusicoccum parvum (71% bootstrap value) (ex-type CMW9081). Based on these results, and morphological data (50 conidia length × width average: 18.1 × 6.6, respectively) our isolates (named MC) were identified and confirmed as Neofusicoccum parvum (Pennycook & Samuels) Crous, Slippers & A.J.L. Phillips. Pathogenicity tests were also conducted on 18 mango cv. Gomera 3 seedlings. The crown roots of each seedling were mechanically wounded and a mycelial plug of the isolate MC14 was placed onto them and covered with soil. Controls (three seedlings) were inoculated with sterile PDA only. Seedlings were maintained in a growth chamber with a 12 hrs photoperiod at 25°C ± 1°C and watered regularly. After five days, stem lesions appeared externally (1.6 cm) and one month after the inoculation, all the inoculated seedlings died. However, controls did not show any obvious symptoms. Re-isolations were conducted as described above and fulfilled Koch's postulates confirming pathogenicity. Among the diseases affecting mango plants, Botryosphaeriaceae represent a serious threat in Sicily as reported by Aiello et al., 2022. The endophytic behaviour of Botryosphaeriaceae is well known, making them latent pathogens (Slippers and Wingfield, 2007). In Italy, N. parvum was detected in mango orchards since 2013 (Ismail et al. 2013), but symptoms of seedlings stem blight have never been reported in the nursery. In Sicily, an increase of Botryosphaeriaceae infection has been observed recently, especially in nurseries, where N. parvum has been identified as a most destructive pathogen (Aiello et al., 2020; Gusella et al., 2021). To our knowledge, this is the first report worldwide of N. parvum causing mango seedling blight. The high incidence of infected seedlings detected in this study highlights the potential risk during propagation in the nursery, representing a significant source of inoculum for the field.

Neopestalotiopsis siciliana sp. nov. and N. rosae Causing Stem Lesion and Dieback on Avocado Plants in Italy.

Avocado (Persea americana) represents an important emerging tropical crop in Italy, especially in the southern regions. In this study, young plants of avocado showing symptoms of stem and wood lesion, and dieback, were investigated. Isolations from symptomatic tissues consistently yielded colonies of Neopestalotiopsis-like species. The characterization of representative isolates was based on the observation of morphological characters, the effect of temperature on mycelial growth rate, and on the sequencing of three different gene regions, specifically ITS, TEF1, and TUB2. Phylogenetic analyses were conducted based on maximum parsimony and maximum likelihood approaches. The results showed the presence of two species, viz. Neopestalotiopsis rosae and N. siciliana, the latter of which is here described as a new species. Pathogenicity tests were conducted using the mycelial plug technique on young potted avocado trees for both Neopestalotiopsis species. The results showed that both species were pathogenic to avocado. This study represents the first report of these two species affecting avocado and results in the description of a new species within the genus Neopestalotiopsis. Based on phylogeny, Pestalotiopsis coffeae-arabicae is combined in Neopestalotiopsis.

A New Disease for Europe of Ficus microcarpa Caused by Botryosphaeriaceae Species.

The Indian laurel-leaf fig (Ficus microcarpa) is an important ornamental tree widely distributed in the urban areas of Italy. Surveys conducted in 2019 and 2020 on several tree-lined streets, squares, and public parks in Catania and Siracusa provinces (Sicily, southern Italy) revealed the presence of a new disease on mature trees. About 9% of approximately 450 mature plants showed extensive branch cankers and dieback. Isolations from woody tissues obtained from ten symptomatic plants consistently yielded species belonging to the Botryosphaeriaceae family. The identification of the recovered fungal isolates was based on a multi-loci phylogenetic (maximum parsimony and maximum likelihood) approach of the ITS, tef1-α, and tub2 gene regions. The results of the analyses confirmed the presence of three species: Botryosphaeria dothidea, Neofusicoccum mediterraneum, and N. parvum. Pathogenicity tests were conducted on potted, healthy, 4-year-old trees using the mycelial plug technique. The inoculation experiments revealed that all the Botryosphaeriaceae species identified in this study were pathogenic to this host. Previous studies conducted in California showed similar disease caused by Botryosphaeriaceae spp., and the pathogenic role of these fungi was demonstrated. To our knowledge, this is the first report of Botryosphaeriaceae affecting Ficus microcarpa in Europe.

A New Strategy to Improve Management of Citrus Mal Secco Disease Using Bioformulates Based on Bacillusamyloliquefaciens Strains.

The effectiveness of biological commercial products based on Bacillus amyloliquefaciens strains was evaluated through in vitro and in vivo experiments against Plenodomus tracheiphilus. The activity of bacterial cells, volatile organic compounds (VOCs), and culture filtrates of bacteria were tested in vitro against different isolates of P. tracheiphilus. Afterwards, the virulence of these isolates was evaluated on Citrus volkameriana plants to select the most virulent isolate to use in the in vivo experiments. To evaluate the effectiveness of products, C. volkameriana seedlings were pre-treated, twice with biological products and once with standard fungicides, before pathogen inoculation. Moreover, in order to determine the endophytic ability of the bacteria, the population density within the treated citrus stem was determined. Comprehensively, bacterial cells, filtrates, and VOCs were able to significantly reduce the average mycelial diameter of P. tracheiphilus, with some variability according to pathogen isolate. In planta experiments showed that the biological products on average were less effective than fungicides, although all formulates were able to significantly reduce disease incidence and symptom severity, except B. amyloliquefaciens strain D747 (Amylo-X) for symptom severity (SS) 20 days after inoculation. Bacteria were re-isolated from the internal woody tissue of treated plants, showing strong endophytic ability. This work is important as commercial biological products based on B. amyloliquefaciens strains could represent a promising and sustainable alternative for the integrated management of mal secco disease.

Impact of Calonectria Diseases on Ornamental Horticulture: Diagnosis and Control Strategies.

Diseases caused by fungi in the genus Calonectria pose a significant threat to the ornamental horticulture industries in Europe and the United States. Calonectria spp. are particularly challenging pathogens to manage in ornamental production systems and the urban landscape for multiple reasons. A high level of species diversity and poorly resolved taxonomy in the genus makes proper pathogen identification and disease diagnosis a challenge, though recent molecular phylogenetic studies have made significant advances in species delimitation. From a disease management perspective, Calonectria spp. produce long-lived survival structures (microsclerotia) that contaminate nursery production systems and can survive multiple years in the absence of a susceptible plant host. Latent infection of plant material is poorly understood but likely contributes to long-distance dissemination of these fungal pathogens, including the clonal Calonectria spp. responsible for the global emergence of boxwood blight. Breeding for disease resistance represents a sustainable strategy for managing Calonectria diseases but is challenging due to the perennial nature of many ornamental plants and high levels of susceptibility in commercial cultivars. Ultimately, long-term sustainable management of Calonectria diseases will require an improved understanding of pathogen biology as well as integration of multiple disease management strategies.

Etiology of Botryosphaeria Panicle and Shoot Blight of Pistachio (Pistacia vera) Caused by Botryosphaeriaceae in Italy.

Pistachio (Pistacia vera) is an important crop in Italy, traditionally cultivated in Sicily (southern Italy) for several decades now. In recent years, new orchards have been planted in new areas of the island. Field surveys conducted in 2019 revealed the presence of symptomatic trees showing shoot dieback, cankers, fruit spots, and leaf lesions. Isolations from symptomatic samples consistently yielded fungal species in the Botryosphaeriaceae family. Identification of collected isolates was conducted using morphological and molecular analyses. Morphological characterization was based on conidia measurements of representative isolates and also effects of temperatures on mycelial growth was evaluated. DNA data derived from sequencing the ITS, tef1-α, and tub2 gene regions were analyzed via phylogenetic analyses (maximum parsimony and maximum likelihood). Results of the analyses confirmed the identity of Botryosphaeria dothidea, Neofusicoccum hellenicum, and N. mediterraneum. Pathogenicity tests were conducted on detached twigs and in the fields both on shoots as well as on fruit clusters using the mycelial plug technique. The inoculation experiments revealed that among the Botryosphaeriaceae species identified in this study N. hellenicum (occasionally detected) and N. mediterraneum were the most aggressive based on lesion length on shoots and fruits. N. mediterraneum was the most widespread among the orchards while B. dothidea can be considered a minor pathogen involved in this complex disease of pistachio. Moreover, to our knowledge, this is the first report of N. hellenicum in Italy.

Update of pistachio leaf spot caused by Septoria pistaciarum in light of new taxonomic advances in Italy.

Septoria leaf spot is one the most widespread disease affecting pistachio (Pistacia vera) in countries of the Mediterranean region. Traditionally, three species have been associated with pistachio, including Septoria pistaciae, Septoria pistaciarum and S. pistacina. However, recent taxonomic studies have reordered and clarified the status of Septoria and septoria-like pathogens affecting pistachio. In our study, field surveys conducted in the traditional Sicilian pistachio production area of Bronte revealed the presence of trees showing characteristic septoria-like leaf spot. Collected isolates were morphologically and molecularly characterized. Morphological characterization was based on conidia measurements and evaluation of mycelial growth on different artificial media. Tested media included CMA, MEA, OA, PDA, and SNA. Phylogenetic analysis was conducted on a multi-locus approach (ITS + tef1 + tub2) based on Maximum Parsimony and Maximum Likelihood. Results showed that our isolates clustered with S. pistaciarum. Pathogenicity test was conducted in the field using conidia suspensions in order to fulfill Koch's postulates. Presence of characteristic rounded spots and pycnidia was evaluated on the inoculated leaves 9 and 23 days after inoculation. This study represents the first update on S. pistaciarum in Italy since its first identification in 1934.

Fusarium nirenbergiae (Fusarium oxysporum Species Complex) Causing the Wilting of Passion Fruit in Italy.

Passion fruit (Passiflora edulis Sims.) is an ever-increasing interest crop in Italy because it is mainly cultivated for its edible fruit and, secondly, as an ornamental evergreen climber. During the summer of 2020, two-year-old plants of purple passion fruit in one of the most important expanding production areas of Sicily (southern Italy) showed symptoms of yellowing, wilting, and vascular discoloration. Fusarium-like fungal colonies were consistently yielded from symptomatic crown and stem tissues. Five representative isolates were characterized by a morphological and molecular analysis based on a multilocus phylogeny using RNA polymerase's second largest subunit (RPB2) and translation elongation factor 1-alpha (EF-1α) genes, as Fusarium nirenbergiae (Fusarium oxysporum species complex). Pathogenicity tests conducted on healthy 1-year-old passion fruit cuttings revealed symptoms similar to those observed in the field. To our knowledge, this is the first report of Fusarium wilt on passion fruit caused by Fusarium nirenbergiae. This report focuses on the phytopathological implications of this fungal pathogen, which may represent a future significant threat for the expanding passion fruit production in Italy and Europe.

Unusual Stylar-End Breakdown and Sour Rot on Key Lime (Citrus aurantiifolia) in Pre-Harvest Condition in Italy.

Key lime (Citrus aurantiifolia) is an emerging crop in Italy, especially in the Southern regions, where the environmental conditions are suitable for its cultivation. A field survey in Sicily in a commercial orchard of Key lime revealed the widespread presence of water-soaked spots and sunken/dry lesions at the stylar-end, mainly in pre-harvest condition. Water-soaked spots were attributed to Geotrichum citri-aurantii, an agent of sour rot on Citrus spp., whereas the sunken/dry lesion was attributed to the physiological disorder known as stylar-end breakdown. Sour rot and stylar-end breakdown are usually considered post-harvest diseases and rarely found in the field on fruit still attached to the tree. Although Geotrichum citri-aurantii is not responsible for the stylar-end breakdown, its association with this alteration reveals the importance of the environmental conditions and the agronomic practices in diseases/disorders development. In addition, to our knowledge, this is the first report of Geotrichum citri-aurantii on Key lime in Europe.

Genetic Diversity and Pathogenicity of Botryosphaeriaceae Species Associated with Symptomatic Citrus Plants in Europe.

This study represents the first survey studying the occurrence, genetic diversity, and pathogenicity of Botryosphaeriaceae species associated with symptomatic citrus species in citrus-production areas in five European countries. Based on morphological features and phylogenetic analyses of internal transcribed spacer (ITS) of nuclear ribosomal DNA (nrDNA), translation elongation factor 1-alpha (TEF1) and ß-tubulin (TUB2) genes, nine species were identified as belonging to the genera Diplodia, Dothiorella, Lasiodiplodia, and Neofusicoccum. Isolates of Neofusicoccum parvum and Diplodia pseudoseriata were the most frequently detected, while Dothiorella viticola had the widest distribution, occurring in four of the five countries sampled. Representative isolates of the nine Botryosphaeriaceae species used in the pathogenicity tests caused similar symptoms to those observed in nature. Isolates assayed were all re-isolated, thereby fulfilling Koch's postulates. Isolates of Diplodia pseudoseriata and Diplodia olivarum are recorded for the first time on citrus and all species found in our study, except N. parvum, are reported for the first time on citrus in Europe.

Can Biological Control Agents Reduce Multiple Fungal Infections Causing Decline of Milkwort in Ornamental Nursery?

This research evaluates biological control agents (BCAs) and fungicide alone and in combination for the management of decline caused by multiple fungi on milkwort (Polygala myrtifolia). Four experiments were performed in a greenhouse within a nursery located in Catania province (southern Italy). The activity of fungicides and biological control agents was evaluated by calculating the plant mortality (%) and recovery frequency (%) of different fungi associated with symptomatic tissue. Comprehensively, boscalid + pyraclostrobin and fosetyl-Al showed the best results in managing disease complex on milkwort. Biological control agents provided, on average, the lowest performances; nevertheless, in most cases, they were able to significantly reduce multiple infections and sometimes when combined with fungicide enhanced the effectiveness. The molecular analysis of 86 isolates obtained from symptomatic tissue allowed to identify the fungi involved in the disease as Calonectriapauciramosa, C. pseudomexicana, Fusariumoxysporum, Neocosmospora solani (syn. F. solani) and binucleate Rhizoctonia AG-R. Calonectriapseudomexicana never reported on milkwort and in Europe was inoculated on P. myrtifolia potted healthy cuttings and produced crown and root rot after 40 days. Our findings represent the first worldwide report about disease complex of milkwort caused by several fungi (Calonectria spp., Fusarium spp. and binucleate Rhizoctonia) and on the effects of integrated control strategies to manage this disease in the nursery.

Seasonal changes in population structure of the ambrosia beetle Xylosandrus compactus and its associated fungi in a southern Mediterranean environment.

Exotic ambrosia beetles are increasing in Europe due to global trade and global warming. Among these xylomycetophagous insects, Xylosandrus compactus (Eichhoff) (Coleoptera: Curculionidae) is a serious threat for several Mediterranean host plants. Carob trees growing in Sicily (Italy) have been extensively attacked by beetles leading to rapid tree decline. Although X. compactus has been found in Europe for several years, most aspects of its ecology are still unknown. We thus studied the population structure and dynamics of X. compactus, together with its twig size preference during a sampling of infested carob trees in south east Sicily. In addition, fungi associated with insects or galleries were isolated and characterized. The results showed that, in this newly-colonized environment and host plant, adult X. compactus overwinters inside twigs and starts to fly and reproduce in mid spring, completing five generations before overwintering in late fall. The mean diameter of carob twigs infested by the beetle varied significantly over the seasons, with the insect tending to infest larger twigs as season progresses. The mean number of adults/gallery was 19.21, ranging from 6 to 28. The minimum temperature significantly affected the overwintering adult mortality. Ambrosiella xylebori and Fusarium solani were the main symbionts associated with the pest in this study. Acremonium sp. was instead recorded for the first time in Europe inside X. compactus galleries. Several other fungi species were also found for the first time in association with X. compactus. Our findings provide useful insights into the sustainable management of this noxious pest.

Ochraceocephala foeniculi gen. et sp. nov., a new pathogen causing crown rot of fennel in Italy.

A new disease of fennel is described from Sicily (southern Italy). Surveys of the disease and sampling were conducted during spring 2017 and 2018 in Adrano and Bronte municipalities (Catania province) where this crop is widely cultivated. Isolations from the margin of symptomatic tissues resulted in fungal colonies with the same morphology. Pathogenicity tests with one isolate of the fungus on 6-month-old plants of fennel reproduced similar symptoms to those observed in nature. Inoculation experiments to assess the susceptibility of six different fennel cultivars to infection by the pathogen showed that the cultivars 'Narciso', 'Apollo', and 'Pompeo' were more susceptible than 'Aurelio', 'Archimede', and 'Pegaso'. Phylogenetic analyses based on a matrix of the internal transcribed spacer (ITS), the large subunit (LSU), and the small subunit (SSU) rDNA regions revealed that the isolates represent a new genus and species within the Leptosphaeriaceae, which is here described as Ochraceocephala foeniculi gen. et sp. nov. This study improves the understanding of this new fennel disease, but further studies are needed for planning effective disease management strategies. According to the results of the phylogenetic analyses, Subplenodomus iridicola is transferred to the genus Alloleptosphaeria and Acicuseptoria rumicis to Paraleptosphaeria.

Liberomycespistaciae sp. nov., the causal agent of pistachio cankers and decline in Italy.

A new canker and decline disease of pistachio (Pistaciavera) is described from Sicily (Italy). Observations of the disease and sampling of the causal agent started in spring 2010, in the area where this crop is typically cultivated, Bronte and Adrano (Catania province) and later extended to the Agrigento and Caltanissetta provinces. Isolations from the margins of twig, branch and stem cankers of declining plants resulted in fungal colonies with the same morphology. Pathogenicity tests on 5-year-old potted plants of Pistaciavera grafted on P.terebinthus reproduced similar symptoms to those observed in nature and the pathogen was confirmed to be a coloniser of woody plant tissue. Comparison of our isolates with the type of the apparently similar Asteromellapistaciarum showed that our isolates are morphologically and ecologically different from A.pistaciarum, the latter being a typical member of Mycosphaerellaceae. Asteromellapistaciarum is lectotypified, described and illustrated and it is considered to represent a spermatial morph of Septoriapistaciarum. Multi-locus phylogenies based on two (ITS and LSU rDNA) and three (ITS, rpb2 and tub2) genomic loci revealed isolates of the canker pathogen to represent a new species of Liberomyces within the Delonicicolaceae (Xylariales), which is here described as Liberomycespistaciae sp. nov. (Delonicicolaceae, Xylariales). The presence of this fungus in asymptomatic plants with apparently healthy woody tissues indicates that it also has a latent growth phase. This study improves the understanding of pistachio decline, but further studies are needed for planning effective disease management strategies and ensuring that the pathogen is not introduced into new areas with apparently healthy, but infected plants.

Effects of Sublabeled Rates of Dazomet and Metam-Sodium Applied Under Low-Permeability Films on Calonectria Microsclerotia Survival.

Infested soil is the primary inoculum source for Calonectria spp. for initiating disease in ornamental and forestry crops. The effects of dazomet and metam-sodium on survival of microsclerotia of 28 isolates belonging to 19 Calonectria spp. were evaluated in this study under nursery conditions. Two experiments with exotic Calonectria spp. in plastic containers in a greenhouse and three trials with endemic species in field plots were performed during different seasons. The containers and plots were artificially infested with Calonectria microsclerotia differentiated on carnation leaf tissues. Basamid (dazomet) was applied at 100, 160, 200, 400, and 500 kg/ha, while Divapan (metam-sodium) was applied at 250, 350, 400, 700, and 1,000 liters/ha in both the containers and plots. The fumigants were applied under virtually and totally impermeable films. Fungal survival was evaluated after 21 days using leaf tissues collected from treated soil and plated on potato dextrose agar, and the ability of microsclerotia to cause infection was tested on red clover. The survival of Calonectria inocula and microsclerotia decreased with increasing fumigant rates. In the greenhouse trials, where Basamid was applied at 200, 400, and 500 kg/ha and Divapan at 400, 700, and 1,000 liters/ha, no viable microsclerotia were recovered for 14 exotic Calonectria spp., whereas viable inocula of Calonectria hongkongensis, C. naviculata, and C. sulawesiensis were retrieved from the fumigated plots. Low rates of Basamid (100 and 160 kg/ha) and Divapan (250 and 350 liters/ha) were less effective at reducing Calonectria viability and, for these treatments, the rate of microsclerotia survival was highly variable among the different isolates and species. Furthermore, totally impermeable film significantly enhanced fumigant performance. Relative to endemic Calonectria spp., all of the treatments killed microsclerotia of C. polizzii and C. pauciramosa independent from fumigant, rate, and film. This research demonstrated the possibility of reducing the application rates by up to 160 kg/ha for Basamid and 400 liters/ha for Divapan under low-permeability films (virtually impermeable film or totally impermeable film) for eradicating or reducing the primary inoculum of Calonectria spp. in soil.

Pleiocarpon gen. nov. and a new species of Ilyonectria causing basal rot of Strelitzia reginae in Italy.

During 2015, a new basal rot disease was observed on potted plants of Strelitzia reginae in an ornamental nursery located in eastern Sicily. Isolations from symptomatic parts of these diseased plants consistently yielded cylindrocarpon-like isolates. Multigene analyses of the partial gene regions of 28S large subunit nrDNA, ß-tubulin, histone H3, translation elongation factor 1-alpha, internal transcribed spacer region and intervening 5.8S nrRNA gene, and RNA polymerase II second largest subunit genes, supported by morphological characters supported the recognition of a new genus, Pleiocarpon based on P. strelitziae sp. nov., and a new Ilyonectria species, described here as I. strelitziae sp. nov. The pathogenicity of both I. strelitziae and P. strelitziae were confirmed on young plants cultivated under controlled conditions in a growth chamber. Both cylindrocarpon-like fungi were pathogenic to S. reginae and reproduced symptoms similar to those observed in the nursery. Of the two species, P. strelitziae was more aggressive than I. strelitziae, resulting in the death of all inoculated plants.

Integrated Management for the Reduction of Calonectria Infections in Ornamental Nurseries.

Chemical control represents the main effective strategy for managing Calonectria diseases in ornamental nurseries. The occurrence of fungicide-resistant strains and the European Directive on "Sustainable Use of Pesticides" has forced ornamental plant growers to establish effective integrated pest management strategies to control Calonectria infections. Here, three nursery experiments were performed to detect the best combinations of fungicides and biological control agents (BCA) to control both leaf spot, caused by six Calonectria spp. on bottlebrush and metrosideros, and stem rot, caused by Calonectria morganii on Dodonaea plants. Overall, the cyprodinil + fludioxonil mixture alone or combined with bioformulates containing Bacillus, Trichoderma, and Streptomyces spp. provided the best performance in reducing leaf spot and stem rot caused by Calonectria spp., followed by the mixture of boscalid + pyraclostrobin. Although BCA alone provided disease suppression significantly lower than the controls in most cases, these treatments were, on average, the least effective in controlling Calonectria infections. Otherwise, there were no significant increases in efficacy with fungicides plus BCA over fungicides alone. Thus, the application of boscalid + pyraclostrobin and cyprodinil + fludioxonil mixtures may also be used in large-scale applications to reduce Calonectria diseases because they effectively managed leaf and stem infections. Our comprehensive research applied previously acquired information on Calonectria disease management in nurseries, resulting in important data that affects integrated plans to fight these pathogens in accordance with European legislation.