D. Aiello

Evaluation of Trichoderma harzianum strain T22 as biological control agent of Calonectria pauciramosa

The objective of this research was to evaluate Trichoderma harzianum strain T22 as a biocontrol agent of collar and root rot caused by different Calonectria pauciramosa isolates. Thus, the microsclerotia-forming ability and virulence of twenty C. pauciramosa isolates were assessed. Microsclerotia production varied partially among the isolates and dual culture with T22 on carnation leaf agar revealed isolates with both high and low microsclerotia-forming ability. Inoculation tests on red clover (Triflolium pratense) demonstrated its susceptibility to the pathogen. On red clover, the degree of virulence and T22 effects in controlling infections were highly variable among the isolates tested. A nursery trial performed on Feijoa sellowiana seedlings confirmed previous results, clearly indicating virulence variability among C. pauciramosa isolates. For three isolates tested in nursery trial, T22 effectiveness in controlling infection was inversely related to their degree of virulence. Overall, T. harzianum strain T22 showed good antagonist activity in reducing microsclerotia production on carnation leaf and the incidence and severity of collar and root rot on both selected hosts. This data could be crucial in developing integrated pest management strategies in ornamental plant nurseries.

First Record of Crown and Root Rot Caused by Cylindrocladium pauciramosum on Brush Cherry in Italy

Brush cherry (Eugenia myrtifolia Sims; synonym Syzygium paniculatum Gaertn.) is a woody evergreen ornamental plant belonging to the Myrtaceae family. This plant is a very common species in Sicilian landscapes. In June of 2008, a new blight disease was detected in a commercial nursery located in Sicily (Italy) in a stock of 10,000 2-year-old E. myrtifolia cv. Newport potted plants obtained from cuttings. The disease was randomly distributed, affecting approximately 2% of the plants. Twig dieback, followed in some cases by plant death, was associated with crown and root rot. Roots were necrotic and crown tissue was brown. A Cylindrocladium sp. was consistently isolated from symptomatic roots, crowns, and lower stems of affected plants on potato dextrose agar petri dishes. Phytophthora isolates were not found associated with symptomatic tissues with BNPRAH (benomyl, nystatin, pentachloronitrobenzene, rifampicin, ampicillin, and hymexazol) selective medium. Five Cylindrocladium isolates were selected and subsequently identified as single-conidial colonies on carnation leaf agar. Isolates were incubated at 25°C under near-UV light with a 12-h_light/dark regimen and examined after 7 days (1). Isolates were characterized by having conidiophores terminating in obpyriform to broadly ellipsoidal vesicles and conidia hyaline, one septate, and straight with rounded ends, (50-) 54 to 55 (-59) × (3.5-) 4 to 6 μm. These characteristics, as well as their ability to produce perithecia when mated with Italian tester strains of Cylindrocladium pauciramosum, allowed their identification as C. pauciramosum C.L. Schoch & Crous (1,4). One mating type (MAT1-1) was found to be present on this host and a representative fungal isolate (DISTEF-Em3) was deposited at the Centraalbureau voor Schimmelcultures (CBS) open fungi collection (Fungal Biodiversity Centre, Utrecht, NL) with Accession No. 123917. Pathogenicity of the isolate DISTEF-Em3 was confirmed by applying 10 ml of a spore suspension (105 conidia per ml) to the crowns of 20 potted 4-month-old E. myrtifolia cv. Newport cuttings that were subsequently maintained in a greenhouse (23 to 25°C and 80 to 90% relative humidity). The same number of untreated plants was used as control. After 4 weeks, only four treated plants developed symptoms identical to those observed in the nursery. Control plants remained symptomless. C. pauciramosum was always reisolated from the infected plants and the identification of the isolate was made as previously described. Pathogenicity tests were repeated twice. In Italy, C. pauciramosum is a widespread pathogen in nurseries that causes extensive losses on young ornamental plants belonging to several genera (2,3). On the basis of the limited spread of the disease in the nursery and the pathogenicity tests, we consider Cylindrocladium crown and root rot of brush berry as a minor disease. However, this work demonstrated the susceptibility of brush cherry to C. pauciramosum that could lead to more extensive losses in association with other biotic or abiotic stresses. To our knowledge, this is the first record of crown and root rot caused by C. pauciramosum on brush cherry. References: (1) P. W. Crous. Taxonomy and Pathology of Cylindrocladium (Calonectria) and Allied Genera. The American Phytopathological Society, St. Paul, MN, 2002. (2) G. Polizzi et al. Plant Dis. 85:803, 2001. (3) G. Polizzi et al. Plant Dis. 90:1459, 2006. (4) C. L. Schoch et al. Plant Dis. 85:941, 2001.

First Occurrence of Calonectria Leaf Spot on Mexican Blue Palm in Italy

In April 2006, a new leaf disease occurred in a private garden in eastern Sicily (Italy) on young, 2-year-old seedlings of Mexican blue palm, Brahea armata S. Watson, in the Arecaceae. Symptoms were detected on 80% of seedlings. The leaves had minute, brown spots that enlarged into dark brown, circular or elliptical lesions, 3 to 6 mm in diameter, and with a necrotic, gray center. The lesions sometimes were surrounded by a chlorotic halo, and older leaves had larger chlorotic areas between spots. Conidia, conidiophores, and terminal vesicles were examined from diseased leaves. A Cylindrocladium sp. was consistently isolated from leaf lesions on Oxoid (Basingstoke, Hampshire, England) potato-dextrose agar after surface disinfestations with 0.8% NaOCl. Cylindrocladium isolates were cultured on carnation leaf agar (CLA) using single hyphal tips. Five isolates were established and identified as Calonectria pauciramosa C.L. Schoch & Crous based on obpyriform to broadly ellipsoidal terminal vesicles, conidiophore branching pattern, conidia size (52 × 4.6 μm), perithecium morphology, and ascopores size (36 × 6.8 μm). Perithecia were obtained with C. pauciramosa tester strains from Italy (G87 and G128) and South Africa (U 971 and U 1670) (2,3) that confirmed both mating types to be present. Further confirmation was obtained by internal transcribed spacer (ITS) analysis. The sequence of rDNA ITS1-5.8 S-ITS2 regions, obtained after amplification with primer ITS1 and ITS4, revealed that the Brahea isolates showed total homology with the sequence of the C. pauciramosa (STE-U 971 from soil) (= Cylindrocladium pauciramosum) available in GenBank. Isolate CBS 120619 from Mexican blue palm was deposited at Centraalbureau voor Schimmelcultures. Spray inoculations of 10 2-year-old Mexican blue palm seedlings were performed with a spore suspension of the fungus adjusted to 105 conidia per ml obtained from 14-day-old single-spore colonies on CLA at 24°C under cool white fluorescent irradiation on a 12-h light/dark regimen. In addition, the following species were similarly inoculated using 10 1-year-old plants: Arecastrum romanzoffianum (Cham.) Becc., B. edulis H. Wendl. ex S. Watson, Chamaerops humilis L., Howea forsteriana Becc., Phoenix canariensis Hort. ex Chabaud., Trachycarpus fortunei (Hook.) H. Wendl., and Ravenea rivularis Jumelle & Perrier. Inoculated, and 10 control plants were placed in separate plastic bags in a growth chamber at 25 ± 1°C. After 7 to 10 days, foliar symptoms including flecks and spots developed on both species of Brahea and on Chamaerops humilis, and on these hosts, pathogenicity tests were repeated. Other palm species and control plants remained healthy. C. pauciramosa was consistently reisolated from inoculated plants on the basis of vesicle shape and conidia sizes of the anamorph. Cylindrocladium candelabrum, Cylindrocladium colhounii, Cylindrocladium floridanum, Cylindrocladium parasiticum, Cylindrocladium pteridis, Cylindrocladium scoparium, and Cylindrocladium theae have been reported as leaf spots pathogens of Arecaceae (1). To our knowledge, this is the first occurrence of C. pauciramosa on Mexican blue palm and the first report of the pathogen on Arecaceae. References: (1) P. W. Crous. Taxonomy and Pathology of Cylindrocladium (Calonectria) and Allied Genera. The American Phytopathological Society, St. Paul MN, 2002. (2) P. W. Crous et al. Stud. Mycol. 50:415, 2004. (3) G. Polizzi and P. W. Crous Eur. J. Plant Pathol. 105:407, 1999.

First record of crown and root rot caused by Cylindrocladium pauciramosum on California lilac in Italy.

California lilacs, or Ceanothus, are evergreen, drought tolerant, colorful ornamental shrubs belonging to the Rhamnaceae family. Ceanothus thyrsiflorus var. repens is the most common species grown in Italy. In October 2005, a severe wilting was noticed on approximately 3,000 potted, 6-month-old plants obtained from cuttings produced by a commercial nursery in Lecco Province (northern Italy). Wilting was always associated with root rot and brown discoloration at the base of the stem. No leaf spots or stem lesions were detected. A Cylindrocladium sp. was isolated consistently from crown, basal stems, and roots of symptomatic plants with potato dextrose agar. Although many crown rots are caused by Phytophthora spp., this pathogen was not found associated with rotten roots and crown plating small pieces of diseased root tissue and crowns onto selective medium. A random selection of five Cylindrocladium sp. isolates was obtained from the infected crown and basal stem. Subsequently, they were identified on carnation leaf agar (CLA) as Cylindrocladium pauciramosum C.L. Schoch & Crous on the basis of their obpyriform to broadly ellipsoidal terminal vesicles, conidiophore branching pattern, conidium, and perithecial morphology, as well as mating type with tester strains of C. pauciramosum selected isolates (1,4). All single-conidial isolates were mated with tester strains DISTEF-G87 (MAT1-1) and DISTEF-G128 (MAT1-2) of C. pauciramosum on CLA, which confirmed both mating types to be present. Two of the isolates were deposited at Fungal Biodiversity Centre, Centraalbureau voor Schimmelcultures (CBS-120145 and CBS-120146). Pathogenicity tests were performed by macerating CLA cultures of C. pauciramosum, obtained from a single-spore isolate and incorporating them on the soil surface of 20 8-month-old C. thyrsiflorus var. repens potted plants. The same number of plants was used as the control. Following inoculation, plants were well irrigated and kept in a growth chamber at 25 ± 1°C. All inoculated plants developed crown rot, basal stem rot, and root rot 25 days after inoculation. Uninoculated control plants remained healthy. C. pauciramosum was reisolated from the artificially inoculated plants. C. pauciramosum causes considerable damage to the ornamental industry in Italy, where projects have been carried out since the first record of the fungus in Europe (3). C. pauciramosum was previously detected on Ceanothus sp. in the United Kingdom (2), where foliar and stem lesions were described. To our knowledge, this is the first record of the pathogen on C. thyrsiflorus var. repens and the first report of wilting due to crown and basal stem rot. References: (1) P. W. Crous. Taxonomy and Pathology of Cylindrocladium (Calonectria) and Allied Genera. The American Phytopathological Society, St. Paul MN, 2002. (2) C. R. Lane et al. Plant Pathol. 55:582, 2006. (3) G. Polizzi and P. W. Crous. Eur. J. Plant Pathol. 105, 407, 1999. (4) C. L. Schoch et al. Plant Dis. 85:941, 2001.

First Report of Stem Bleeding and Trunk Rot of Kentia Palm Caused by Thielaviopsis paradoxa in Italy

The Kentia palm or Sentry palm (Howea forsteriana (C. Moore & F.v. Muell.) Becc.) is one of the most popular palms in the world. In Italy, it is suitable for growing indoors or outdoors. In September 2006, bleeding was found on the stem of a 20-year-old Kentia palm growing in a field nursery in eastern Sicily. The trunk began to bleed from cracks or fissures of the bark at 5 ft (1.5 m) from the ground level. Tissues surrounding the stem cracks were black. The plant canopy showed no symptoms. Cross-sections revealed brown rot confined to one side of the trunk where the rot was spreading inward from the surface. Infected tissues did not emanate odor of fermented fruit. Internal tissue adjacent to the rotted areas was placed on carrot agar amended with 500 μl of streptomycin sulphate and acidified (lactic acid; pH = 3.6) potato dextrose agar (PDA). A transverse section of affected palm tissues was maintained in a moist chamber for 6 days. Microscopic examinations of isolates obtained on media and sporulation from affected tissues yielded Thielaviopsis paradoxa (De Seyn.) Höhn (2). Endoconidia, measuring 3.9 (range 3 to 6) × 8.2 μm (range 6 to 14) (n = 50), were cylindrical to somewhat oval when mature, hyaline to brown, and smooth walled. Endoconiodophores were usually straight, colorless to pale brown, as much as 150 μm long, with a terminal spore-bearing cell through which spores are borne. Chlamydospores were smooth, thick walled, brown, in chains, and were 8.8 (range 5.5 to 15.0) × 15.8 μm (range 9.0 to 25.0) (n = 50). Kochs postulates were fulfilled by stem inoculation on a 20-year-old double-stem Kentia palm growing in the same field nursery. Ten mycelial plugs (5-mm diameter) obtained from 14-day-old single-spore colonies growing on PDA at 24°C were applied to 10 5-mm-diameter stem wounds. The same number of inoculations was used as a control in the other stem and treated with sterile agar plugs. Following inoculation, the mycelial plugs and the stems were wrapped with Parafilm. After 20 days, stem rots were detected only on the inoculated stem wounds, and stem bleeding was observed after 3 months. The pathogen was reisolated from symptomatic tissues. A Thielaviopsis sp. was previously reported in Florida on Kentia palm as being responsible for a frond necrosis (1). While stem bleeding seems to be a common symptom on coconut (Cocos nucifera L.), it is not commonly reported on other palm species. Only recently, T. paradoxa was detected for first time in Sicily on date palm imported from Egypt (3). To our knowledge, this is the first record of stem bleeding caused by T. paradoxa on palms in Italy, and the first record of stem bleeding and stem rot on Kentia palm. References: (1) S. A. Alfieri Jr. et al. Fla. Dep. Agric. Consum. Serv. Bull. No. 14. 1994. (2) A. R. Chase and T. K. Broschat, eds. Diseases and Disorders of Ornamental Palms. The American Phytopathological Society, St Paul MN, 1991. (3) G. Polizzi et al. Plant Dis. 90:972, 2006.

First Report of Damping-Off and Leaf Spot Caused by Cylindrocladium scoparium on Different Accessions of Bottlebrush Cuttings in Italy

In May of 2006, approximately 10,000 cuttings of bottlebrushes (Callistemon cvs. Laevis, Hannah Ray, Kings Park Special, Masotti Mini Red, and Rose Opal with either C. viminalis (Soland. ex Gaertn.) Cheel. [excluded] or C. citrinus (Curtis) Skeels as one parent) grown in a nursery in eastern Sicily (Italy) exhibited severe disease symptoms including damping-off, leaf spots, and collar and root rot. Initially, the infections were detected on approximately 30% of the cuttings, but by late September 2006, 70% of the plants had symptoms. A Cylindrocladium sp. was consistently isolated from the diseased portions of plants onto potato dextrose agar. To determine the species, single-conidial isolates of the fungus were cultured on carnation leaf agar (CLA) for 7 days at 25°C with 12 h of light/dark conditions. Only the mycelia and spores growing on the carnation leaves were examined with a light microscope, and the isolates were identified as Cylindrocladium scoparium Morgan (teleomorph Calonectria morganii Crous, Alfenas & M.J. Wingf.) on the basis of their pyriform to broadly ellipsoidal terminal vesicles, conidiophore branching pattern, and conidia (1). In addition, the ability of the colonies to mate with South African tester strains of C. scoparium (2,3) confirmed the identification. Kochs postulates were fulfilled by inoculating 10 cuttings for each bottlebrush accession with a spore suspension (105 conidia per ml) of one isolate of the pathogen (DISTEF-GCs7) obtained from 14-day-old single-spore colonies grown on CLA at 24°C under fluorescent cool white lights with 12 h of light/dark. Following inoculation, all plants were maintained in plastic bags in a growth chamber at 25 ± 1°C and 90 to 95% relative humidity. The same number of cuttings was used as a control. Damping-off, crown root rot, and leaf spots symptoms identical to those observed in the nursery appeared within 5 to 20 days. No symptoms were detected on the control plants. C. scoparium was reisolated from the artificially infected tissues. The isolate, used in the pathogenicity proof, was deposited at the Fungal Biodiversity Centre, Centraalbureau voor Schimmelcultures (Accession No. CBS 120930). The presence of C. scoparium was detected for the first time in Italy on mastic tree in 2005 (3). To our knowledge, this is the first report of C. scoparium on bottlebrush in Italy and it represents the first information about the susceptibility of these Callistemon cultivar accessions to the fungus and confirms the spread of the pathogen in Sicilian ornamental nurseries. References: (1) P. W. Crous. Taxonomy and Pathology of Cylindrocladium (Calonectria) and Allied Genera. The American Phytopathological Society, St. Paul MN, 2002. (2) P. W. Crous and M. J. Wingfield. Mycotaxon 51:341, 1994. (3) G. Polizzi et al. Plant Dis. 90:1110, 2006.

Severe outbreak of crown rot and root rot caused by Cylindrocladium pauciramosum on strawberry tree in Italy.

From 2006 to 2009, a new disease was noticed in several commercial nurseries in Sicily (Italy) on ~13,000 potted plants of strawberry tree (Arbutus unedo L.) ranging from 3 months to 4 years old. Symptomatic plants showed a crown and root rot and internal brown discoloration of the basal stem. No leaf spots were detected. The percentage of symptomatic plants was variable in the different stocks (1 to 28%). A Cylindrocladium sp. was isolated consistently from symptomatic tissues on potato dextrose agar (PDA). Although many crown rots are caused by Phytophthora or Rhizoctonia spp., plating small pieces of diseased tissue from rotten roots and crowns onto corn meal agar or PDA yielded these pathogens sporadically. Twenty Cylindrocladium isolates obtained from infected basal stems, crowns, and roots were selected and subsequently cultured on carnation leaf agar (CLA). Macroconidiophores consisted of a stipe, a penicillate arrangement of fertile branches, and stipe extension terminating in an obpyriform-to-ellipsoidal vesicle (6 to 10 μm in diameter). Conidia cylindrical, straight, and 1-septate ranged from 40 to 60 × 4 to 5 μm. All single-conidial isolates were mated with tester strains DISTEF-G87 (MAT1-1) and DISTEF-G128 (MAT1-2) of Cylindrocladium pauciramosum (3) on CLA and produced fertile perithecia after 2 months. Perithecia were solitary or in groups, orange to red-brown, subglobose to ovoid, and ranged from 260 to 400 × 180 to 290 μm in diameter. On the basis of the combination of morphological and perithecial characters and mating type the isolates were identified as C. pauciramosum C.L. Schoch & Crous, teleomorph Calonectria pauciramosa C.L. Schoch & Crous (1,4). One representative isolate was deposited at Centraalbureau voor Schimmelcultures (CBS; No. 123918) open fungi collection. Pathogenicity tests were performed by adding sterile water to CLA cultures of C. pauciramosum from a single-conidial isolate and incorporating the resulting spore suspension (105 conidia per ml) on the soil surface of 20 3-month-old strawberry tree potted plants. The same number of plants served as a control. After inoculation, plants were maintained in a growth chamber at 25 ± 1°C and 90 to 95% relative humidity. All inoculated plants developed crown and root rot identical to one observed in the nursery 2 months after inoculation. Control plants remained symptomless. C. pauciramosum was always reisolated from infected plants. This fungus represents a serious threat for ornamental nurseries in Europe. Diseases related to infection of C. pauciramosum are different in relationship to the host tissues and the affected species. On strawberry tree, C. pauciramosum was previously detected in Italy as causal agent of leaf spots (2). To our knowledge, this is the first record of crown and root rot of strawberry tree caused by C. pauciramosum. In contrast with leaf spots, this report could indicate an important limiting factor for cultivation of strawberry tree potted plants in nurseries. The knowledge of the susceptibility of this host to crown and root rot caused by C. pauciramosum can help growers apply specific control strategies in nurseries. References: (1) P. W. Crous. Taxonomy and Pathology of Cylindrocladium (Calonectria) and Allied Genera. The American Phytopathological Society, St. Paul, MN, 2002. (2) G. Polizzi and V. Catara. Plant Dis. 85:803, 2001. (3) G. Polizzi and P. W. Crous. Eur. J. Plant Pathol. 105:407, 1999. (4) C. L. Schoch et al. Plant Dis. 85:941, 2001.

First Report of Crown Rot and Root Rot Caused by Cylindrocladium pauciramosum on Feijoa (Feijoa sellowiana) in Italy

Feijoa sellowiana (O. Berg) O. Berg is native to South America and grown in Italy as an ornamental plant and for its fruit. During 2006, 2007, and 2008, a severe wilting was noticed on ∼ 10,000 potted 6-month to 3-year-old plants obtained from seeds and produced by three commercial nurseries in Catania Province (southern Italy). Wilting was always associated with root rot and brown discoloration at the base of the stem. Leaf spots or stem lesions were not observed. A Cylindrocladium sp. was isolated consistently from the crown, basal stem, and roots of symptomatic plants on potato dextrose agar (PDA). Ten isolates of a Cylindrocladium sp. were obtained from single-spore colonies made from a 14-day-old culture grown on PDA and subsequently subcultured on carnation leaf agar (CLA). Cultures were incubated at 25°C under near-UV light with a 12-h light/dark regimen and examined after 7 days (1). Only fungal structures occurring on the carnation leaf tissue were examined. Isolates were identified based on morphological traits and mating type responses (2,4). Isolates were mated with opposite mating tester strains of C. pauciramosum (DISTEF-G128 [MAT1-1] and DISTEF-G87 [MAT1-2]) and C. scoparium (DISTEF-GCs7 = CBS 120892 [MAT1-1] and DISTEF-GP0 = CBS 119669 [MAT1-2]) selected from an Italian collection. Plates were parafilmed, sealed in plastic bags, incubated in a chamber at 25 ± 1°C with a photoperiod of 16 h of light and 8 h of darkness, and examined weekly until perithecia developed. Successful crosses were determined after 2 months of incubation and were regarded as those isolate combinations that produced perithecia with viable ascospores. Conidia were observed to be 30 to 60 × 3.5 to 5.0 μm while conidiophores mostly had two to three series of branches and stipes terminated in obpyriform to broadly ellipsoidal vesicles, which is characteristic of C. pauciramosum C.L. Schoch and Crous (teleomorph Calonectria pauciramosa C.L. Schoch and Crous). Fertile perithecia containing viable ascospores were similar to Calonectria pauciramosa. They were obtained only from crosses of the examined isolates with mating type testers of C. pauciramosum. Six isolates (when paired with DISTEF-G128) and four isolates (when mated with DISTEF-G87) provided fertile progeny. No perithecia were obtained in pairings of the 10 isolates with tester strains of C. scoparium. One representative isolate was deposited at the Fungal Biodiversity Centre, Centraalbureau voor Schimmelcultures (DISTEF-AS4 = CBS-120618). Kochs postulates were fulfilled by inoculating 30 6-month-old seedlings of feijoa with a spore suspension (105 conidia per ml) of one isolate of the pathogen (DISTEF-AS4) grown on CLA for 14 days. The same number of seedlings was used as a control. Following inoculation, plants were maintained in a growth chamber at 25 ± 1°C and 90 to 95% relative humidity. Wilting, crown rot, and root rot symptoms identical to those observed in the nurseries appeared within 15 to 50 days. Symptoms were not observed on the control plants. C. pauciramosum was previously reported to cause leaf spots on feijoa (3). To our knowledge, this is the first report of wilting due to crown and root rot caused by C. pauciramosum in this host. The disease is a limiting factor for feijoa cultivation in Sicilian nurseries. References: (1) P. W. Crous et al. Mycologia 84:497, 1992. (2) P. W. Crous and M. J. Wingfield. Mycotaxon 51:341, 1994. (3) G. Polizzi and V. Catara. Plant Dis. 85:803, 2001. (4) C. L. Schoch et al. Mycologia 91:286, 1999.

Characterization of Pseudomonas syringae pv. syringae isolated from mango in Sicily and occurrence of copper-resistant strains.

Mango (Mangifera indica) is grown throughout a wide range of frost-free climates and is one of the world’s most important fruit crops. During 2010-2014, severe symptoms of bacterial apical necrosis (BAN) caused by Pseudomonas syringae pv. syringae (Pss) were observed on mango plants throughout all mango-growing areas in Sicily (southern Italy). The causal agent was identified based on phenotypic, genotypic and pathogenicity tests. The genetic variability of strains obtained from different areas and cultivars were assessed by rep- PCR and allowed clear differentiation of the Pss strains isolated from mango from other representative strains of the pathovar. In addition, the respective production of syringomycin, syringopeptin and mangotoxin was checked by in vitro tests and PCR detection. All tested strains showed the presence of mgoA and mgoB of the mangotoxin operon. Some copper- resistant Pss strains showing cusCBA genes were also found in some orchards. This feature could explain the failure of the disease control using copper compounds. Among 71 strains tested using in vitro assay, no copper-sensitive strains were detected, whereas 44 strains (62%) had high resistance, 16 strains (22.5%) were resistant and 11 strains (15.5%) had low resistance to copper sulfate. To our knowledge, this is the first report of copper resistance among Pss strains causing BAN on mango in Sicily.

Emergence of Prochloraz-Resistant Populations of Calonectria pauciramosa and Calonectria polizzii in Ornamental Nurseries of Southern Italy

Management of Calonectria spp. infections in nurseries requires scheduled fungicide applications, particularly with methyl benzimidazole carbamates (MBCs) and sterol demethylation inhibitors (DMIs). Due to rising concerns about the occurrence of MBC resistance in different Calonectria populations and variability in prochloraz efficacy in controlling these pathogens, a detailed study on prochloraz sensitivity distributions of Calonectria isolates belonging to the Calonectria scoparia complex was carried out. In total, 105 isolates collected in two distinct periods (1993 to 1996 and 2005 to 2009) were analyzed for prochloraz sensitivity. Based on DNA sequencing and phylogenetic analyses of β-tubulin, histone H3, and translation elongation factor-1α gene sequences, 69 and 36 isolates were identified as C. pauciramosa and C. polizzii, respectively. The isolates collected more recently (group B) had a reduced prochloraz sensitivity, as indicated by greater values for the effective dose to reduce growth by 50% than those collected earlier (group A). The reduced sensitivity detected in vitro corresponded to partial loss of fungicide efficacy in controlling infections in red clover and feijoa under controlled and semi-field conditions, respectively. Frequent prochloraz application in nurseries for controlling Calonectria spp. infections is discouraged.