First Report of Leaf Spot Caused by Cylindrocladium pauciramosum on Dwarf Willow Myrtle in Italy.

Dwarf willow myrtle (Agonis flexuosa (Willd.) Sweet) cv. Nana, an evergreen ornamental shrub belonging to the Myrtaceae, is grown in Italy as an ornamental potted plant. In November 2008, a widespread new leaf spot disease was noticed on ~80% of 5,000 6-month-old potted plants. Plants were obtained from cuttings and produced by a commercial nursery in Catania Province. Symptomatic leaves showed minute, reddish brown spots that enlarged (3 to 5 mm in diameter) and then darkened, presenting a necrotic center defined by a dark purple halo. Leaf spots were surface disinfested with 0.8% NaOCl and plated on potato dextrose agar. Twenty isolates of the fungus that was consistently isolated from the spots were selected and cultured for 8 days at 25°C 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). Cylindrical conidia were rounded at both ends, straight, one-septate, and ranged from 44 to 60 × 4 to 5 µm. The fungus was tentatively identified as Cylindrocladium pauciramosum based on these morphological characteristics (2). All single-conidium isolates were mated with tester strains of Calonectria pauciramosa C. L. Schoch & Crous, telomorph of C. pauciramosum, on CLA and produced fertile perithecia (4). Perithecia were solitary or in groups, orange to red-brown, subglobose to ovoid, and ranged from 280 to 400 µm long × 180 to 290 µm in diameter. Further confirmation of species was obtained by amplification and sequencing of the intergenic spacer (IGS) region of rDNA, using M13 Forward (-20) and M13 Reverse primers. On the basis of the complete IGS sequence, two primer sets (218F/218R and 106F/106R) were designed and successfully used in a nested-PCR protocol for the detection of C. pauciramosum from tissues of infected plants (3). On the basis of the combination of morphological characters, mating type, and molecular data, the isolates were identified as C. pauciramosum C.L. Schoch & Crous. One representative isolate (DISTEF-Af1) was deposited at Centraalbureau voor Schimmelcultures open fungi collection (Fungal Biodiversity Centre, Utrecht, the Netherlands; CBS 124659). Pathogenicity tests were performed by adding sterile water to CLA cultures of C. pauciramosum from a single-conidium isolate (DISTEF-Af1) and spraying the resulting spore suspension (105 conidia per ml) on the leaf surface of 20 6-month-old A. flexuosa cv. Nana potted plants. The same number of plants served as noninoculated controls. Following inoculation, plants were kept in plastic bags in a growth chamber at 25 ± 1°C. All inoculated plants developed circular, brown leaf spots identical to those observed in the nursery 5 to 7 days after inoculation. Control plants remained symptomless. C. pauciramosum was always reisolated from the infected plants and identified as previously described. Leaf spotting in seedlings of A. flexuosa was previously associated with infections by C. scoparium in Australia (1). To our knowledge, this is the first record in the world of leaf spots caused by C. pauciramosum on A. flexuosa. References: (1) A. L. Bertus. Agric. Gaz. N. S. W. 87:22, 1976. (2) P. W. Crous. Taxonomy and Pathology of Cylindrocladium (Calonectria) and Allied Genera. The American Phytopathological Society, St. Paul MN, 2002. (3) F. Nigro et al. J. Plant Pathol. 88:S22, 2006. (4) G. Polizzi and P. W. Crous. Eur. J. Plant Pathol. 105:407, 1999.

First Report of Southern Blight on Silverbush (Convolvulus cneorum) Caused by Sclerotium rolfsii in Italy.

The genus Convolvulus has more than 200 species that are encountered in temperate to tropical climates all around the world. Convolvulus cneorum L., also known as silverbush, is a perennial shrub native to southern Europe (Sicily and Croatia) with dense, silver foliage and masses of large, circular, white flowers. During July of 2009, a widespread blight was observed on approximately 10% of 12,000 4-month-old potted silverbush plants. The plants were obtained from cuttings and produced by a commercial nursery in eastern Sicily, Italy. Symptomatic plants initially had sunken, tan lesions at the ground level that developed into typical southern blight. Circular and crescent-shaped patches were observed on the masses of weeds on the surface of the containers where silverbush were grown. At the soil line, white mycelia and small (1 to 2 mm in diameter), brown, spherical sclerotia with internally differentiated rind, cortex, and medulla characteristic of Sclerotium rolfsii Sacc. were observed. Crown and stem lesions were surface disinfested (1% NaOCl) for 1 min, rinsed in sterile water, and placed on potato dextrose agar. Isolation consistently yielded colonies of Sclerotium rolfsii (teleomorph Athelia rolfsii (Curzi) Tu & Kimbrough) with typical sclerotia produced within 6 to 7 days (2). Pathogenicity tests were performed on 20 plants by placing 10 sclerotia obtained from 10-days-old cultures in the soil below the crown portion on each of 5-month-old healthy cuttings of silverbush. The same number of plants served as noninoculated controls. All plants were maintained in a growth chamber at 25 ± 1°C and enclosed for 7 days in polyethylene bags. Plants were then moved to a greenhouse where temperatures ranged from 24 to 28°C. Symptoms of southern blight developed after 7 to 20 days on all inoculated plants. Control plants remained symptomless. S. rolfsii was reisolated from symptomatic plants. S. rolfsii was reported for the first time in Sicily in 2004 in an ornamental nursery (1). To our knowledge, this is the first report in the world of S. rolfsii on silverbush and it is the first outbreak of southern blight on Convolvulus species. The high susceptibility of silverbush to the pathogen could be a limiting factor for the cultivation in nursery of this indigenous wildflower plant. References: (1) G. Polizzi et al. Plant Dis. 88:310, 2004. (2) Z. K. Punja and A. Damiani. Mycologia 88:694, 1996.

First Report of Leaf Spot and Shoot Blight Caused by Cylindrocladium scoparium on Mallee Honeymyrtle in Italy.

In December of 2008, a widespread disease was observed on several blocks of approximately 15,000 plants (6-month to 2-year-old) of mallee honeymyrtle (Melaleuca acuminata F. Muell.). The plants were grown in two nurseries in eastern Sicily where high diffusion of diseases caused by Cylindrocladium pauciramosum and C. scoparium was previously detected. The plants exhibited leaf spots, defoliation, and apical blight of shoots. Crown rot and root rot were not present. Leaf spots were detected on all plants, whereas shoot blight was observed on approximately 3% of the plants. A Cylindrocladium sp. was consistently isolated from the diseased portions of plants onto potato dextrose agar. To determine the species, 20 single-conidia isolates of the fungus obtained from symptomatic tissues from different blocks and nurseries were cultured on carnation leaf agar (CLA) for 7 days at 25°C under 12-h light/dark conditions. Mycelia and spores growing on the carnation leaves were examined with a light microscope and the isolates were identified as C 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 isolates to mate with South African and Italian opposite tester strains of C. scoparium (2,4) confirmed the identification of all the isolates. Koch's postulates were fulfilled by inoculating 30 6-month-old plants of mallee honeymyrtle with a spore suspension (105 conidia per ml) of one isolate of the pathogen (DISTEF-MA1) obtained from 14-day-old single-spore colonies grown on CLA at 24°C under fluorescent cool white lights on a 12-h light/dark regimen. 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 mallee honeymyrtle plants was used as uninoculated controls. Leaf spots, defoliation, and apical shoot blight identical to those observed in the nurseries appeared within 4 to 25 days. No symptoms were detected on the control plants. C. scoparium was reisolated from the artificially infected tissues and was identified as previously described. The isolate used in the pathogenicity proof was deposited at the Fungal Biodiversity Centre, Centraalbureau voor Schimmelcultures (Accession No. CBS 124658). In Italy, C. scoparium was detected for the first time on Pistacia lentiscus in 2005 (3). Another report confirmed the spread of the pathogen in Sicilian ornamental nurseries (4). To our knowledge, this is the first record in the world of C. scoparium causing disease on mallee honeymyrtle. The data demonstrate the high susceptibility of this species to the C. scoparium leaf spot and twig blight especially when environmental conditions (heavy rains and mild temperatures) are conductive to the infections. 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. (4) G. Polizzi et al. Plant Dis. 91:769, 2007.

First Report of Crown Rot Caused by Cylindrocladium pauciramosum on Scarlet Honey Myrtle in Italy.

Scarlet honey myrtle (Melaleuca fulgens R. Br.), native to Australia, is an evergreen colorful shrub (Myrtaceae) and grown in Italy as an ornamental plant. During November 2008, a widespread disease was noticed on ~90% of 3,000 6-month-old M. fulgens cv. Red potted plants. Plants were obtained from cuttings and produced by a commercial nursery in Catania Province. Symptomatic plants showed a crown rot and longitudinal sections of tissues revealed a brown discoloration of the basal stem. As a consequence, leaves gradually became necrotic and abscised, followed by death of the entire plant. Root rots and leaf spots were not observed. M. gibbosa, M. ericifolia, M. thymifolia, and M. elliptica, cultivated in the same nursery, did not show disease symptoms. A Cylindrocladium sp. was consistently isolated from the crown and basal stem of symptomatic plants on potato dextrose agar (1). Ten Cylindrocladium isolates obtained from infected basal stems and crowns were selected and cultured for 8 days at 25°C on carnation leaf agar (CLA). Macroconidiophores consisted of a stipe, a penicillate arrangement, and a stipe extension terminating in an obpyriform to ellipsoidal vesicle (6 to 10 µm in diameter). Cylindrical conidia were rounded at both ends, straight, 1-septate, and 42 to 60 × 4 to 5 µm. All single-conidial isolates were mated with opposite tester strains of C. pauciramosum on CLA and produced fertile perithecia (3). Perithecia were solitary or in groups, orange to red-brown, subglobose to ovoid, and 270 to 400 µm high × 180 to 290 µm in diameter. Further confirmation of species was obtained by amplification and sequencing of the intergenic spacer (IGS) region of rDNA with the M13 forward (-20) and M13 reverse primers. On the basis of the complete IGS sequence, two primer sets (218F/218R and 106F/106R) were designed and successfully used in a nested-PCR protocol for the detection of C. pauciramosum from tissues of infected plants (2). On the basis of morphological characters, mating type, and molecular data, the isolates were identified as C. pauciramosum C.L. Schoch & Crous. One representative isolate (DISTEF-MFR2; CBS 124657) was deposited at Centraalbureau voor Schimmelcultures open fungi collection (Fungal Biodiversity Centre, Utrecht, the Netherlands). 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 M. fulgens cv. Red potted plants. The same number of plants served as uninoculated controls. Following inoculation, plants were well irrigated and maintained in a growth chamber at 25 ± 1°C and 90 to 95% relative humidity. All inoculated plants developed crown rot symptoms identical to those observed in the nursery 2 months after inoculation. Control plants remained symptomless. C. pauciramosum was always reisolated from the infected plants and identified as previously described. C. pauciramosum was previously detected in Italy as being responsible for a leaf spot on M. hypericifolia (3). To our knowledge, this is the first record in the world of crown rot of scarlet honey myrtle caused by C. pauciramosum. References: (1) P. W. Crous. Taxonomy and Pathology of Cylindrocladium (Calonectria) and Allied Genera. The American Phytopathological Society, St. Paul MN, 2002. (2) F. Nigro et al. J. Plant Pathol. 88:S22, 2006. (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 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.

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 Stem Blight Caused by Botrytis cinerea on Jasminum officinalis in Italy.

Common jasmine (Jasminum officinalis L.) is an evergreen shrub that is native to the Middle and Far East. It is widely grown in Europe as an ornamental plant and in southeastern France for fragrance for the perfume industry. In March of 2008, a previously undescribed disease was observed on potted (6-month- to 3-year-old) common jasmine plants growing in open fields in a nursery of eastern Sicily, Italy. More than 20% of the plants showed disease symptoms. Diseased plants had small to large, brown or black lesions on stem. The lesions expanded rapidly, girdled the stem and caused blight of entire branches, and occasionally killed the plant. Abundant conidia and mycelia were detected on the surface of dead and dying stems under cool and humid conditions, which resulted in a moldy gray appearance. Botrytis cinerea Pers.:Fr. (1) was consistently isolated from affected tissues disinfected for 1 min in 1% NaOCl, rinsed in sterile water, and plated on potato dextrose agar (PDA). Colonies were at first white then became gray after 6 to 7 days when spores differentiated. White sclerotia developed after 8 to 9 days and turned black with age. Size of the conidia produced on 1-month-old culture ranged from 5.0 to 9.5 × 6.5 to 12.5 µm on the basis of 50 spore measurements. Sclerotia were spherical or irregular and ranged from 1.0 to 2.5 × 0.9 to 2.9 mm (average 1.7 × 1.8 mm). Stems of eight 6-month-old common jasmine plants were lightly wounded with a sterile razor and inoculated with 3-mm-diameter plugs of PDA from 10-day-old mycelial cultures, eight similar plants were inoculated with mycelium without wounding, and an equal number of noninoculated plants inoculated with only PDA plugs served as control. After inoculation, plants were enclosed in transparent plastic bags at 20 ± 2°C for 5 days. Stem lesions identical to the ones observed in the nursery were detected on all wounded and on two nonwounded fungus-inoculated plants within 5 to 7 days. Control plants remained healthy. B. cinerea was reisolated from typical lesions. The unusually cool and humid weather conditions recorded in Sicily are supposed to be highly conducive of disease outbreak. Although B. cinerea does not usually kill the plants, under these environmental conditions this disease can cause significant economic loss to ornamental nurseries. To our knowledge, this is the first report of B. cinerea causing stem blight on J. officinalis. Reference: (1) M. B. Ellis. Dematiaceous Hyphomycetes. CAB, Kew, Surrey, England, 1971.

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). Koch's 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.

First Report of Damping-Off Caused by Rhizoctonia solani AG-4 on Lagunaria patersonii in Italy.

Lagunaria patersonii (Adr.) G. Don (cow itch tree) is native to Australia and tolerates salted winds. During July 2007, damping-off of cow itch tree was observed on 4-month-old seedlings growing in a commercial nursery in eastern Sicily, Italy. More than 20% of the seedlings showed disease symptoms. First symptoms consisting of water-soaked lesions at the seedling base that expand rapidly girdle the stem and collapse the seedling in a few days. Diseased tissues were disinfested for 1 min in 1% NaOCl, rinsed in sterile water, plated on potato dextrose agar (PDA) amended with streptomycin sulphate at 100 mg/l, and then incubated at 25°C. A fungus with mycelial and morphological characteristics of Rhizoctonia solani Kühn was consistently yielded. Fungal colonies were initially white, turned brown with age, and produced irregularly shaped, brown sclerotia. Microscopic examination revealed that hyphae had a right-angle branching pattern, were constricted at the base of the branch near the union with main hyphae, and septate near the constriction. Basidia were not observed in the greenhouses or on the plates. Hyphal cells were determined to be multinucleate when stained with 0.5% aniline blue solution and examined at ×400 magnification with a microscope. Anastomosis groups were determined by pairing isolates on 2% water agar in petri plates (3). Pairings were made with tester strains of AG-1 IA, AG-2-2-1, AG-2-2IIIB, AG-2-2IV, AG-3, AG-4, AG-5, AG-6, AG-11. Anastomosis was observed only with tester isolates of AG-4 producing both C2 and C3 reactions. The hyphal diameter at the point of anastomosis was reduced, the anastomosis point was obvious, and cell death of adjacent cells was observed. These results were consistent with other reports on anastomosis reactions (1). The identification of group AG-4 within R. solani has been confirmed by electrophoretic patterns of pectic enzymes (polygalacturonases) in vertical pectin-acrylamide gel stained with ruthenium red (2). Pathogenicity tests were conducted on potted, healthy, 3-month-old seedlings of cow itch tree. Twenty plants were inoculated by placing plugs of PDA from 5-day-old mycelial cultures near the base of the stem. The same number of plants was treated with 1 cm2 PDA plugs as control. Plants were kept at 25°C and 95% relative humidity on a 12-h fluorescent light/dark regimen. Wilt symptoms due to basal stem rot, identical to ones observed in the nursery, appeared 10 days after inoculation and all inoculated plants showed symptoms within 1 month. Control plants remained healthy. The pathogen was reisolated from symptomatic tissues, completing Koch's postulates. To our knowledge, this is the first report in the world of R. solani causing disease on L. patersonii. References: (1) D. E. Carling. Page 37 in: Grouping in Rhizoctonia solani by Hyphal Anastomosis Reactions. Kluwer Academic Publishers, the Netherlands, 1996. (2) R. H. Cruickshank and G. C. Wade. Anal. Biochem. 107:177, 1980. (3) C. C. Tu and J. W. Kimbrough. Mycologia 65:941, 1973.

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. Koch's 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.

First Report of Southern Blight on Firewheel Tree, Bay Laurel, Bird of Paradise, Mediterranean Fan Palm, and Liverwort Caused by Sclerotium rolfsii in Italy.

During the summer of 2006, a widespread blight was observed on 6-month-old potted plants of firewheel tree (Stenocarpus sinuatus Endl.) and 3-month-old potted plants of bay laurel (Laurus nobilis L.) growing in a nursery in eastern Sicily, Italy. On both species, symptomatic plants initially had sunken, tan lesions at ground level where white mycelia and small (1 to 2 mm in diameter), brown, spherical sclerotia typical of Sclerotium rolfsii Sacc. were formed. As the disease progressed, the mycelia extended up the stem and entire plants collapsed. A sudden wilting affecting 4-month-old potted seedlings of bird of paradise (Strelitzia reginae Aiton) and 5-month-old potted seedlings of Mediterranean fan palm (Chamaerops humilis L.) was occasionally detected in other greenhouses of the same nursery. Liverwort (Marchantia polymorpha L.) was abundantly present on the surfaces of the containers where these plants were grown. Circular and crescent-shaped patches as much as 100 cm in diameter were observed on the massed liverwort plants. In these patches, the liverwort died and sclerotia typical of S. rolfsii were dispersed on white mycelial strands. Symptomatic tissues of the ornamental plants and liverwort were surface disinfested in 1% NaOCl for 1 min, rinsed in sterile water, and plated on potato dextrose agar. Tissues consistently yielded S. rolfsii (teleomorph Athelia rolfsii (Curzi) Tu & Kimbrough) and typical sclerotia with internally differentiated rind, cortex, and medulla were produced within 6 or 7 days (3). Pathogenicity tests were performed by placing 30 sclerotia obtained from 10-day-old cultures in the soil below the crown portion on each of 2-month-old healthy seedlings of Stenocarpus sinuatus, L. nobilis, Strelitzia reginae, and C. humilis (20 seedlings per host). In addition, liverwort growing in 10 pots (7 cm in diameter) was inoculated with 30 sclerotia per pot. For each species, the same number of plants or pots served as control. All ornamental plants and liverwort were maintained in a growth chamber at 25 ± 1°C and enclosed for 7 days in polyethylene bags and then moved to a greenhouse where temperatures ranged from 24 to 28°C. The inoculation trial was repeated once. Symptoms of southern blight developed after 5 to 20 days on all inoculated plants of Stenocarpus sinuatus and sporadically (two to five plants) after 20 days on L. nobilis, Strelitzia reginae, and C. humilis. After 5 days, liverwort in all inoculated pots was colonized and plants died within 12 days. Control plants of all species remained symptomless. S. rolfsii was reisolated from symptomatic plants. S. rolfsii was reported for the first time in Sicily in 2004 on ornamental plants (2). Strelitzia reginae was previously reported as a host of Corticium rolfsii (synonym S. rolfsii) in Portugal (1). To our knowledge, this is the first report of S. rolfsii on Stenocarpus sinuatus, L. nobilis, and C. humilis. In addition, this is the first report of the susceptibility of M. polimorpha to S. rolfsii. Liverwort could provide a food source for the fungus in container-grown nursery plants. References: (1) M. R. de Sousa Dias and M. T. Lusas. Bol. Soc. Brot. 53:469, 1980. (2) G. Polizzi et al. Plant Dis. 88:310, 2004. (3) Z. K. Punja and A. Damiani. Mycologia 88:694, 1996.

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 Bacterial Blight of Strelitzia augusta Caused by Pseudomonas syringae pv. lachrymans.

White bird of paradise tree (Strelitzia augusta Thunb.), originally from South Africa, is a tender perennial cultivated as an ornamental plant and is used in gardens in Italy. During February of 2004, a new blight disease was noticed on potted S. augusta at different ages (6 months to 4 years) in several commercial nurseries of eastern Sicily. Field inspections revealed disease incidences as high as 40%. Initial symptoms were small, water-soaked leaf spots that expanded throughout the veins in dark brown streaks. Stem cross sections revealed browning of the vascular tissues, which might involve the entire stem. In some cases, the necrosis extended to the apical bud, causing death of the plant. Thirty explants from infected tissues were washed in sterile water and plated on plate count agar (PCA) from which two types of bacterial colonies were consistently isolated. Pathogenicity tests were performed on S. augusta plants. Twenty-four plants were inoculated (12 per bacterial isolate) using two different procedures: spray with a bacterial suspension (106 CFU/ml) and wounding with an infected needle on the midribs. The same number of noninoculated plants was used as controls. All plants were maintained at 24 to 26°C with 95 to 100% relative humidity until symptoms occurred 4 days later. Just one of the two tested bacterial types was pathogenic. The symptoms were similar to those previously observed in the field. No symptoms were observed in the plants spray inoculated with the bacterial suspension, proving that the bacteria were unable to infect in the absence of a wound. The controls showed no symptoms. Koch's postulates were fulfilled by the reisolation of the infective strain which was sent to the CBS (Centraalbu-reau voor Schimmelcultures) and identified as Pseudomonas syringae pv. Lachrymans/pisi using the Biolog MicroLog3 4.01C program (Biolog Inc., Hayward, CA). Further pathogenicity tests have been carried out on zucchini and pea pods to characterize the pathovar using 48SR1 of P. syringae pv. syringae and B4 of P. syringae pv. pisi as reference strains. Necrotic, sunken, water-soaked spots surrounded by a chlorotic halo, reported in the literature as typical symptoms of P. syringae pv. lachry-mans (Smith & Bryan) Young, Dye & Wilkie (1), were observed on zucchini when inoculated with our strain. Our P. syringae strain did not cause the typical symptoms of P. syringae pv. pisi on inoculated pea pods. The results of the pathogenicity tests and the inability of the P. syringae strain isolated from S. augusta to utilize homoserine, used to discriminate pv. pisi from other pathovars of P. syringae, allowed us to identify the strain as P. syringae pv. lachrymans. Low temperature damage and late transplant may have promoted the spread of the disease in the nurseries. Under these conditions, the economic importance of this disease for the crop can be considered high. To our knowledge, this is first report of P. syringae pv. lachrymans on S. augusta. Reference: (1) K. Pohronezny et al. Plant Dis. Rep. 62:306, 1978.

First Report of Southern Blight Caused by Sclerotium rolfsii on Laurustinus.

Laurustinus (Viburnum tinus L.), native to the Mediterranean Region, is an evergreen shrub commonly used as a specimen shrub or small tree or used in border plantings. During August 2003, a blight occurred on 2-year-old-plants of laurustinus growing in pots in a nursery in eastern Sicily (Italy). Disease incidence ranged from 2 to 5% across the field. Symptoms included 3 to 4 cm long lesions and the development of white mycelial strands and brown, 1.0 to 1.8 mm, nearly spherical sclerotia on the crown of plants at the soil line that are typical of Sclerotium rolfsii Sacc. The foliage of infected plants wilted, followed by a sudden collapse of the plant. The fungus was consistently isolated on acidified potato dextrose agar (PDA) (pH 4.5) by plating symptomatic tissues that were surface disinfested (1.2% NaOCl) for 1 min. and rinsed in sterile water. Pathogenicity tests were performed by sprinkling 50 sclerotia, obtained from infected oat kernels (2), on the soil surface around the collar of each of 10 healthy, potted 1-year-old plants of laurustinus. Five of the plants were previously wounded on the crown 1.5 cm above or below the soil line with a sterile needle. Five noninoculated plants served as controls. All plants were maintained at 25 ± 2°C and enclosed for 72 hr in polyethylene bags (90 to 95% relative humidity). Blight symptoms similar to those seen in nursery were observed on inoculated plants 20 to 25 days after inoculation, while no symptoms developed on control plants. Koch's postulates were fulfilled by reisolation of the fungus on acidified PDA from all infected laurustinus plants. S. rolfsii was previously recorded on Prague viburnum (Viburnum × pragense L.) as the causal agent of southern blight (1). To our knowledge, this is the first report of southern blight caused by S. rolfsii on laurustinus. References: (1) A. Hagan. Southern blight on flowers, shrubs, and trees. On-line publication ANR-1157. Alabama A & M, and Auburn University ( www.aces.edu/dept/extcomm/publications/html ). (2) R. Rodriguez-Kabana et al. Plant Dis. Rep. 59:5, 1975.

First Occurrence of Downy Mildew on Boxleaf Veronica Caused by Peronospora grisea in Italy.

Boxleaf veronica (Hebe buxifolia (Benth.) Cockayne & Allan), native to New Zealand, is an annual or perennial shrub widely cultivated in Mediterranean zones. During late spring 2003, after seasonably wet and cool weather, a downy mildew epidemic occurred on potted, overhead irrigated, 1-year-old seedlings at a commercial nursery in eastern Sicily (southern Italy). Infections affected boxleaf veronica at first, but spread to variegated boxleaf (H. buxifolia cv. variegata). Initial symptoms on the upper side of leaves were small, gray brown patches that gradually spread, eventually resulting in necrosis in the center of infected areas. At that point, brown patches were evident on both sides of the leaves. Lesions on the lower leaf surface became covered with a fairly dense, pale gray-to-brown layer of conidia and conidiophores. As the disease progressed, these spots coalesced into large and conspicuous brown lesions. The youngest, most succulent shoots withered and died. The large brown lesions on the leaves were disfiguring and affected 95 to 100% of plants in the nursery. All diseased nursery stock had to be discarded. Oospores were not observed in leaf tissues. The fungus recovered from leaves with abundant gray brown sporulation was identified as Peronospora grisea (Unger) Unger. Microscopic observations revealed conidiophores that branched dichotomously five to seven times with branch ends 6 to 10 × 2 to 3 µm, slightly curved, and tapered to a blunt apex. Hyaline conidia were ellipsoid, brownish when mature, and measured 23 to 27 × 15 to 18 µm (mean = 25.2 × 17.1 µm), falling within the range of those reported for P. grisea (1). Pathogenicity was confirmed by inoculating 10 1-yr-old seedlings (10 cm tall) by gently pressing infected leaves with abundant sporulation onto healthy leaves and maintaining inoculated plants in a humid chamber at 21°C. An equal number of noninoculated plants served as controls. After 9 to 11 days, symptoms similar to those originally observed developed onto inoculated plants, and after 12 to 15 days, grayish mildew grew on leaves. Microscopic examination of the developing mycelium confirmed that leaves were infected with P. grisea. Uninoculated control plants did not develop any symptoms. The disease was also confirmed in this way on variegated boxleaf veronica (H. buxifolia cv. variegata). Downy mildew of Hebe spp. has been recorded in New Zealand, Britain, France, Germany, and more recently, West Sussex (2). Heavy rainfall during the spring of 2003 in eastern Sicily could have favored disease development. To our knowledge, this is the first report of downy mildew of boxleaf veronica and variegated boxleaf veronica caused by P. grisea in Italy. References: (1) S. M. Francis and A. M. Berrie. Peronospora grisea. No. 766 in: Descriptions of pathogenic fungi and bacteria. CMI, Kew, Surrey, UK. 1983. (2) J. M. Whipps and C. A. Linfield. Plant Pathol. 36:216, 1987.