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The genus Grevillea (family Proteaceae) comprises over 300 species and is a popular and widely cultivated group of Australian plants. In the last 3 years, numerous potted grevilleas with symptoms of decline associated with a rot of feeder roots were found in ornamental nurseries in Sicily. Aboveground symptoms were reduced growth, yellowing of foliage, wilt, dieback, and death of the entire plant. The disease was observed on many commercial cultivars and was especially severe on G. alpina (mountain grevillea), G. juniperina (juniper-leaf grevillea), G. lavandulacea (lavender grevillea), and G. rosmarinifolia (rosemary grevillea) as well as the hybrid cultivars Clearview David (G. lavandulacea × rosmarinifolia) and Poorinda Rondeau (G. baueri × lavandulacea), while G. lanigera (woolly grevillea) cv. Mount Tamboritha and G. thelemanniana subsp. obtusifolia appeared resistant. A species of Phytophthora was consistently isolated from rotted roots of symptomatic plants using a selective medium (4), and pure cultures were obtained by single-hypha transfers. The species was identified as P. palmivora (E.I. Butler) E.I. Butler on the basis of morphological and cultural characters. On solid media, all isolates produced elliptical to ovoid, papillate sporangia with a mean length/width ratio of 1.8. Sporangia were caducous with a short pedicel (5 μm) and a conspicuous basal plug. All isolates were heterothallic (mating type A1) and produced oogonia and oospores only when paired with A2 mating type reference isolates of P. nicotianae and P. palmivora. Antheridia were amphyginous. Identification was confirmed by electrophoresis of mycelial proteins in polyacrylamide slab gels (1). The electrophoretic patterns of total soluble proteins and six isozymes (alkaline phosphatase, esterase, fumarase, NAD-glucose dehydrogenase, malate dehydrogenase, and superoxide dismutase) of isolates from grevillea were identical to those of a reference isolate of P. palmivora from Coronilla valentina subsp. glauca (2) but distinct from those of reference strains of eight other papillate species of Phytophthora included in Waterhouse's taxonomic group VI. Koch's postulates were fulfilled using 6-month-old rosemary grevillea plants that were transplanted into pots filled with soil that was artificially infested with chlamydospores (50 per gram of soil) produced in submerged cultures (3) by grevillea isolate IMI 390579. Plants were maintained in a glasshouse at 20 to 28°C and watered to field capacity once a week. One month after transplanting, infected plants showed decline symptoms similar to those of naturally infected plants. Control plants grown in pots containing noninfested soil remained healthy. P. palmivora was reisolated from roots of symptomatic plants. It appears that P. palmivora has become a widespread root pathogen in commercial ornamental nurseries in Italy (2). References: (1) S. O. Cacciola et al. EPPO Bull. 20:47, 1990.D. (2) S. O. Cacciola et al. Plant Dis. 86:327, 2002. (3) J. Y. Kadooka and W. H. Ko. Phytopathology 63:559, 1973. (4) H. Masago et al. Phytopathology 67:425, 1977.

The genus Cuphea (Lythraceae) includes approximately 250 species of annual, evergreen perennials and short shrubs native to Central and South America. During the springs of 2003 and 2004, 10% of the nursery stock of approximately 12,000 potted cigar-flowers (C. ignea A. DC) grown in a screenhouse at a commercial ornamental nursery near Piedimonte Etneo, Sicily, had symptoms of wilt, defoliation, and rapid collapse of the entire plant. These foliar symptoms were associated with a reduced root system, browning of the collar, and dark brown discolored roots. A Phytophthora species was consistently recovered by plating small pieces of rotted roots of symptomatic plants onto selective medium (3); pure cultures were obtained by single-hypha transfers. On potato dextrose-agar (PDA), cardinal temperatures for growth were 10 to 35°C and the optimum was 28 to 30°C. Sporangiophores were umbellate or in a close monoclasial sympodium and mean dimensions of sporangia were 52 × 26 mm, with a mean length/width ratio of 2:1. Sporangia produced on V8 juice agar (VJA) were ellipsoid, fusiform, or limoniform with a tapered base. They were papillate, occasionally bipapillate, caducous, with a long pedicel (as much as 150 μm). All isolates were mating type A1 determined by pairing with A2 reference isolates of P. palmivora (Butl.) Butl. and P. nicotianae Breda de Haan. Oogonia with amphigynous antheridia were formed on VJA after 10 to 15 days at 24°C in the dark. Occasionally, 10 of 15 isolates formed small chlamydospores on VJA. Electrophoretic patterns of total mycelial proteins and four isozymes (acid and alkaline phosphatase, esterase, and malate dehydrogenase) on polyacrylamide slab gels (3) of all Cuphea isolates were very similar to those of reference isolates of P. tropicalis M. Aragaki & J. Y. Uchida from Convolvulus cneorum L. (IMI 391714) and Rhamnus alaternus L., respectively. In addition, the Cuphea isolates were clearly distinct from reference isolates of other species including P. capsici Leon., P. citricola Sawada, P. citrophthora (R. E. Smith & E. H. Smith) Leon., P. nicotianae, and P. palmivora. On the basis of morphological cultural characters and the electrophoretic phenotype, the isolates were identified as P. tropicalis. Internal transcribed spacer (ITS) regions of rDNA sequences (2) confirmed the identification. Koch's postulates were fulfilled by testing three cigar-flower isolates, including isolate IMI 391709, on 10 6-month-old potted cuttings of Cuphea inoculated by applying a 10-ml zoospore suspension (2 × 104 zoospores/ml) to the crowns, incubated for 24 h at 100% relative humidity, and maintained in the greenhouse at 20 to 24°C. After 10 days, crowns and stems were brown and all plants wilted within 20 days. Ten control plants treated with water remained healthy. P. tropicalis was reisolated from infected tissues. The test was repeated with similar results. In Europe, P. tropicalis has been reported on Cyclamen persicum Mill. in Germany (4) and C. cneorum and R. alaternus in Italy (1), indicating a broad host range and spreading in ornamental nurseries. References: (1) S. O. Cacciola et al. Boll. Acc. Gioenia Sci. Nat. 31:57, 1999. (2) S. O. Cacciola et al. For. Snow Landsc. Res. 76:387, 2001. (3) D. C. Erwin and O. K. Ribeiro. Pages 39–41, 138–139 in: Phytophthora Diseases Worldwide. The American Phytopathological Society, St. Paul MN. 1996. (4) W. W. P. Gerlach and A. Schubert. Plant Dis. 85:334, 2001.

In the summer of 2006, 1-year-old apricot (Prunus armeniaca L.) trees with leaf chlorosis, wilting, and defoliation associated with root and crown rot were observed in a nursery in Sicily (Italy). Of 3,000 plants, ~2% was affected. Four Phytophthora spp. (45, 25, 20, and 10% of the isolations of the first, second, third, and fourth species, respectively) were isolated from decayed roots and trunk bark on BNPRAH (3). Axenic cultures were obtained by single-hypha transfers. Isolates of the first species formed petaloid colonies on potato dextrose agar (PDA) and had an optimum growth temperature of 25°C. On V8 agar (VA), they produced persistent, papillate (often bipapillate), ovoid to limoniform sporangia (length/breadth ratio 1.4:1). They did not produce gametangia when paired with A1 and A2 isolates of Phytophthora nicotianae. The second species formed arachnoid colonies, had an optimum growth of 30°C, and produced uni- and bipapillate, ellipsoid, ovoid or pyriform sporangia (length/breadth ratio 1.3:1). All isolates were A2. The third species formed rosaceous colonies on PDA, had an optimum temperature of 28 to 30°C, and produced papillate (sometime bipapillate), ellipsoid or limoniform (length/breadth ratio 2:1), caducous sporangia with a tapered base and a long pedicel (as much as 150 μm). All isolates were A1 type. The fourth species formed petaloid-like colonies on PDA and had an optimum growth of 26 to 28°C. On VA, it produced papillate (sometimes bipapillate), ovoid (length/breadth ratio 1.3:1), and decidous sporangia with a short pedicel ( References: (1) S. O. Cacciola et al. Plant Dis. 90:680, 2006. (2) D. E. L. Cooke et al. Fungal Genet. Biol. 30:17, 2000. (3) H. Masago et al. Phytopathology 67:425, 1977.

Approximately 150,000 potted mandevillas (Apocynaceae) are produced each year in the Etna District of eastern Sicily. Since 2004, leaf chlorosis, wilt, and sudden collapse of the entire plant associated with root and basal stem rot of 6- to 12-month-old potted mandevillas, including Mandevilla × amabilis ‘Alice du Pont’, M. splendens, and M. sanderi ‘Alba’, ‘My Fair Lady’, and ‘Scarlet Pimpernel’, have been observed in six nurseries. Incidence of affected plants varied from 5 to 40%. Four Phytophthora species were consistently isolated from rotted roots and stems on a selective medium (2). Pure cultures of the first species produced colonies with a camellia pattern on potato dextrose agar and grew between 10 and 37°C with an optimum of 27°C. On V8 juice agar they produced ellipsoid to obpyriform (length/breadth [l/b] 1.45:1), nonpapillate sporangia with internal proliferation, coralloid, spherical hyphal swellings and both terminal and intercalary chlamydospores. In dual cultures with A1 and A2 isolates of P. nicotianae, all isolates produced oogonia with amphyginous antheridia only with A2 isolates. Isolates of the second species formed petaloid colonies, had an optimum growth temperature of 25°C, and produced mono- and bipapillate, ovoid to limoniform sporangia (l/b 1.40:1); they did not produce gametangia. Isolates of the third species formed colonies with a slight petaloid pattern and grew between 2 and 30°C with an optimum of 25°C. Sporangia were obpyriform (l/b 1.48:1), nonpapillate, and proliferous. All isolates were A2 mating type. The isolates of the fourth species formed arachnoid colonies, grew between 8 and 38°C with an optimum of 30°C, and produced mono- and bipapillate, ellipsoid, and obpyriform (l/b 1.3:1) sporangia and apical chlamydospores. All isolates were A2 mating type. DNA was extracted from mycelium and amplified by PCR using the ITS 4/ITS 6 primers (1). Blast search of the rDNA-ITS sequence of isolate IMI 397618 (GenBank Accession No. GQ388261) of the first species showed 100% identity with the ITS sequence of an isolate of P. cinnamomi var. parvispora (EU748548). The sequences (GQ463703 and GQ463704) of isolates IMI 397471 and IMI 397472 of the second species showed 99% similarity with the sequences of a P. citrophthora isolate (EU0000631). The sequence of isolate IMI 397473 (GQ463702) of the third species showed 99% similarity with the sequence of a P. cryptogea isolate (AY659443.1), while the sequence of isolate IMI 397474 (GU723474) of the fourth species showed 99% similarity with the sequence of a P. nicotianae isolate (EU331089). The pathogenicity of individual isolates IMI 397618, IMI 397471, IMI 397472, IMI 397473, and IMI 397474 was tested on 3-month-old potted plants (10 plants per isolate) of mandevilla ‘Alice du Pont’ by applying 10 ml of a suspension (2 × 104 zoospores/ml) to the root crown. Plants were maintained at 25°C and 95 to 100% relative humidity. All inoculated plants wilted after 4 weeks, while noninoculated control plants remained healthy. The four Phytophthora spp. were subsequently reisolated only from symptomatic plants. To our knowledge, this is the first report of P. cinnamomi var. parvispora in Italy and on mandevilla worldwide. In recent years, Phytophthora root and stem rot has become the most serious disease of potted mandevillas in Sicily. References: (1) D. E. L. Cooke et al. Fungal Genet. Biol. 30:17, 2000. (2) H. Masago et al. Phytopathology 67:425, 1977.

Mimosa [Acacia dealbata Link, syn. Acacia decurrens (Wendl. F.) Wild. var. dealbata (Link) F. Muell., Fabaceae] is an evergreen shrub native to southeastern Australia that is cultivated as an ornamental plant in warm temperate regions of the world. In spring 2010, in a commercial nursery in Liguria (northern Italy), 6- to 10-month-old potted plants of A. dealbata showed symptoms of sudden collapse, defoliation, and wilt associated with root and basal stem rot. An abundant gum exudate oozed from the basal stem. A Phytophthora species was consistently isolated from roots and stem on BNPRAH selective medium (4). On V8 agar (V8A), axenic cultures obtained by single hyphal transfers formed stellate to radiate colonies with aerial mycelium whereas on potato dextrose agar (PDA) the colonies grew more slowly than on V8A and showed stoloniform mycelium and irregular margins. Minimum and maximum growth temperatures on PDA were 10 and 35°C, with the optimum at 30°C. In water, all isolates produced catenulate or single fusiform hyphal swellings and ellipsoid, nonpapillate, persistent sporangia. Dimensions of sporangia were 46.1 to 65.4 × 23.1 to 30.8 μm (mean l/b ratio 2.1). All isolates were A1 mating type and produced spherical oogonia with amphyginous antheridia when paired with A2 mating type of P. drechsleri Tucker on V8A plus β-sytosterol (4). Internal transcribed spacer (ITS) regions of rDNA of the representative Phytophthora isolate IMI 500394 from A. dealbata were amplified and sequenced in both directions with primers ITS6/ITS4. The consensus sequence (GenBank Accession No. JF900371) was 99% similar to sequences of several isolates identified as Phytophthora taxon niederhauserii Z.G. Abad and J.A. Abad (e.g., GQ848201 and EU244850). Pathogenicity tests were performed on 1-year-old potted plants of A. dealbata with isolate IMI 500394. Twenty plants were transplanted into pots (12-cm-diameter) filled with soil infested (4% v/v) with the inoculum of IMI500394 produced on kernel seeds. Plants were kept in a greenhouse with natural light at 25 ± 2°C and watered to field capacity weekly. All inoculated plants showed symptoms of wilt, leaf chlorosis, and basal stem rot within 3 to 4 weeks. Twenty control plants transplanted in autoclaved soil mix remained healthy. P. taxon niederhauserii was reisolated solely from inoculated plants, thus fulfilling Koch's postulates. Since 2003, this pathogen has been found on bottlebrush and rock rose grown in a nursery in Sicily (southern Italy), as well as on Banksia in a nursery in Liguria (2,3). To our knowledge, this is the first report of P. taxon niederhauserii on A. dealbata. P. taxon niederhauserii, recently described as P. niederhauserii sp. nov. (1), is a polyphagous pathogen that was originally reported on arborvitae and ivy in North Carolina in 2001. References: (1) Z. G. Abad et al. Mycologia (in press), 2013. (2) S. O. Cacciola et al. Plant Dis. 93:1075, 2009. (3) S. O. Cacciola et al. Plant Dis. 93:1216, 2009. (4) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society, St. Paul, MN, 1996.

Cactus pear (Opuntia ficus-indica, Cactaceae), native to Mexico, is a multipurpose crop. About 90% of Italian production of cactus pear fruit is from Sicily. In 2013, a disease of cactus pear was noticed in minor islands of Sicily, Lampedusa and Linosa (Pelagie archipelago), Favignana (Aegadian archipelago), and Ustica, where cactus pear is grown as living fences. Symptoms were on flattened stems functioning as leaves (cladodes) and included radially expanding cankers, up to 20 cm in diameter, concentric, crusty, silvery areas, with minute, black dots (pycnidia erumpent from epidermis) and a leathery, brown halo. A milky to buff colored exudate, caking on contact with air, oozed from active cankers. Cankers coalesced and the cladode wrinkled. Infected plants declined and took a gray, ghostly appearance. The disease was found on 100% of plants in Lampedusa and Linosa, whereas it occurred sporadically in Favignana and Ustica but with an incidence of 100% of infected plants in some sites. It was not found in Sicily. Ten plants per island were randomly chosen and pieces (5 mm) of diseased tissue or single pycnidia were plated onto potato dextrose agar supplemented with 1 mg/ml of streptomycin and incubated at 20°C. A fast-growing fungus with a gray, aerial mycelium was consistently recovered. Optimum temperature for growth was around 25°C. Conidia from pycnidia produced on 1.5% water agar and sterilized pine needles were unicellular, hyaline, smooth, fusoid to ovoid, thin-walled,15.0 ± 1.5 × 5.5 ± 0.6 μm (length/width ratio = 2.9). Four representative isolates, one from each island, were identified according to internal transcribed spacer (ITS), EF1-α, and β-tubulin genes (Dissanayake et al. 2016; Lopes et al. 2016; Phillips et al. 2013). All isolates had identical ITS (GenBank accession nos. MF414730, MF414738, MF414747, and MF414748), β-tubulin (MF414749, MF414757, MF414766, and MF414767), and EF1-α (MF414768, MF414776, MF414785, and MF414786) sequences and 100% identity with the corresponding sequences of a reference isolate (CBS124923) of Neofusicoccum batangarum from Indian almond (Terminalia catappa; Lopes et al. 2016). Two polymorphic bases were identified in ITS and TEF sequences as compared with another reference isolate (ex-type isolate CBS124924) of N. batangarum from the same host (Lopes et al. 2016). Isolates were identified as N. batangarum and deposited at CBS-KNAW Biodiversity Centre (CBS143023, CBS143024, CBS143025, and CBS143026). On May 2015, field-grown cactus pear plants were wound-inoculated with these isolates (3 plants/isolate, 6 cladodes/plant). On each cladode, two holes (5-mm diameter) were made 20 cm apart with a cork-borer, and an agar plug from a 5-day-old colony was inserted into the hole. Three plants inoculated with sterile agar served as a control. Wounds were sealed with the excised tissues. Four days after inoculation (a.i.), a brown halo appeared around the wound and a buff-colored exudate oozed from the hole. Cankers were identical to those on naturally infected plants. Pycnidia emerged 20 days a.i. N. batangarum was reisolated from cankers and identified by sequencing the ITS, EF-α, and β-tubulin regions. Controls were asymptomatic. This disease is most likely the same reported as gummy canker caused by Dothiorella ribis (Somma et al. 1973). A similar disease of a distinct cactus species (Nopalea cochenillifera) incited by N. batangarum has been reported in Brazil (Conforto et al. 2016). This is the first report of N. batangarum in Europe and as a pathogen of cactus pear worldwide.

The taxonomic status of Colletotrichum gloeosporioides sensu lato (s.l.) associated with olive anthracnose is still undetermined and the pathogenic ability of this species complex is controversial. In the present study, isolates obtained from olive and provisionally identified as C. gloeosporioides s.l. on the basis of morphological and cultural features were reclassified using ITS and TUB2 as DNA barcode markers and referred to seven distinct species, recently separated within C. gloeosporioides (C. aenigma, C. gloeosporioides sensu stricto (s.s.), C. kahawae, C. queenslandicum, C. siamense and C. theobromicola) and C. boninense (C. karstii) species complexes. Furthermore, isolates of C. kahawae were ascribed to the subspecies ciggaro by analysing the GS gene. A single isolate, not in either of these two species complexes, was not identified at the species level. In pathogenicity tests on detached olive drupes some of these species, including C. aenigma, C. kahawae subsp. ciggaro, C. queenslandicum, C. siamense and C. karstii, were shown to be weakly pathogenic. Moreover, they were found very sporadically on olive. In contrast, some isolates of C. gloeosporioides s.s. and isolates of C. theobromicola proved to be virulent on both green and ripening olives. This study gives a better insight into both the aetiology and the epidemiology of olive anthracnose and might have implications for biosecurity and quarantine because C. theobromicola has never been reported in major European olive-producing countries.

[EN] An analysis of incidence of Phytophthora spp. in 732 European nurseries producing forest transplants, larger specimen trees, landscape plants and ornamentals, plus 2525 areas in which trees and shrubs were planted, is presented based on work conducted by 38 research groups in 23 European countries between 1972 and 2013. Forty-nine Phytophthora taxa were recorded in 670 nurseries (91.5%); within these nurseries, 1614 of 1992 nursery stands (81.0%) were infested, although most affected plants appeared healthy. In forest and landscape plantings, 56 Phytophthora taxa were recovered from 1667 of 2525 tested sites (66.0%). Affected plants frequently showed symptoms such as crown thinning, chlorosis and dieback caused by extensive fine root losses and/or collar rot. Many well-known highly damaging host¿Phytophthora combinations were frequently detected but 297 and 407 new Phytophthora¿host associations were also observed in nurseries and plantings, respectively. On average, 1.3 Phytophthora species/taxa per infested nursery stand and planting site were isolated. At least 47 of the 68 Phytophthora species/taxa detected in nurseries and plantings were exotic species several of which are considered well established in both nurseries and plantings in Europe. Seven known Phytophthora species/taxa were found for the first time in Europe, while 10 taxa had not been previously recorded from nurseries or plantings; in addition, 5 taxa were first detections on woody plant species. Seven Phytophthora taxa were previously unknown to science. The reasons for these failures of plant biosecurity in Europe, implications for forest and semi-natural ecosystems and possible ways to improve biosecurity are discused., The authors are grateful to all public and private owners of nurseries and plantings who contributed to this extensive study. T. Jung acknowledges the support by the Regione Autonoma della Sardegna, Visiting Professor Program at the University of Sassari, Italy. The authors also thank the COST Office and the European Council for providing the European COST Actions FP0801 (http://www.cost.eu/domains_actions/fps/Actions/FP0801) and FP 1002 (http://www.cost.eu/domains_actions/fps/Actions/FP1002), the EU projects FOR-THREATS and ISEFOR, and BiodivERsA project RESIPATH as platforms for stimulating discussions on the nursery pathway and possible management solutions.

Goji ( Lycium barbarum ) is a newly introduced crop in Italy. In June 2016, one- to two-year-old goji plants with symptoms of wilt were observed in commercial fields in Calabria, southern Italy. Approximately 10% of plants were affected. …

In June 2009 in a commercial nursery in eastern Sicily (Italy), 3-year-old potted windmill palms (Trachycarpus fortunei (Hooker) H. Wendl.) showed a decline in growth, wilt, droop, and basal rot of the youngest leaves. The rot progressed inward and killed the bud. Initially, older leaves remained green but eventually the entire plant collapsed. Root rot was consistently associated with aboveground symptoms. Two Phytophthora species were consistently isolated from the petiole base, heart, roots, and rhizosphere soil of symptomatic plants on a selective medium (2) and occasionally recovered from roots and rhizosphere soil of asymptomatic plants. Pure cultures were obtained by single-hypha transfers and the two species were identified on the basis of morphological and molecular characters as Phytophthora palmivora and P. nicotianae. Both species were recovered from all symptomatic plants. From multiple tissue samples per plant, we recovered either or both species. On potato dextrose agar (PDA), P. palmivora isolates grew between 10 and 35°C, with the optimum at 27°C. On V8 juice agar, they produced elliptical to ovoid, papillate, caducous sporangia (32 to 78 × 23 to 39 μm) with a mean length/breadth (l/b) ratio of 1.8:1 and a short pedicel (mean pedicel length = 5 μm). Isolates of P. nicotianae produced arachnoid colonies on PDA, grew at 37°C but did not grow at 40°C. Sporangia (29 to 55 × 23 to 45 μm) were spherical to ovoid (l/b ratio 1.3:1), papillate and often bipapillate, and noncaducous. Isolates of both species produced amphigynous antheridia and oogonia only when paired with reference isolates of P. nicotianae of the A2 mating type. The internal transcribed spacer (ITS) region of rDNA of two isolates of P. palmivora (IMI 398987 and IMI 398988) and an isolate of P. nicotianae (IMI 398989) from T. fortunei was amplified with primers ITS6/ITS4 and sequenced (1). Blast analysis of the sequences of isolates IMI 398987 and IMI 398988 (GenBank Accession Nos. HQ596556 and HQ596558) showed 99% homology with the sequence of two reference isolates of P. palmivora (GQ398157.1 and GU258862), while the sequence of isolate IMI 398989 (HQ596557) showed 99% homology with a reference isolate of P. nicotianae (EU331089.1). Pathogenicity of isolates IMI 398987 and IMI 398989 was proved by inoculating separately each isolate on 1-year-old potted plants of T. fortunei (10 plants per isolate). A zoospores suspension (2 × 104 zoospores/ml) was pipetted onto the petiole base of the three central leaves (200 μl per leaf) of each plant. Sterile water was used for control plants. All plants were incubated at 25 ± 2°C with 100% humidity for 48 h and then maintained in a greenhouse at 24 to 28°C. Within 3 weeks, all inoculated plants showed symptoms of bud rot. Control plants remained healthy. P. palmivora and P. nicotianae were reisolated only from inoculated plants. Bud rot of palms caused by P. palmivora was reported previously in Italy (3). However, to our knowledge, this is the first report of simultaneous infections of P. palmivora and P. nicotianae as causal agents of this disease. Outbreak of bud rot may have been favored by overhead sprinkler irrigation. The recovery of P. palmivora and P. nicotianae from rhizosphere soil and roots of asymptomatic plants suggests infested soil was the primary inoculum source. References: (1) D. E. L. Cooke et al. Fungal Genet. Biol. 30:17, 2000. (2) H. Masago et al. Phytopathology 67:425, 1977. (3) A. Pane et al. Plant Dis. 91:1059, 2007.

Olive (Olea europea L.) is grown on about 154,000 ha in Sicily (southern Italy). In the summer of 1999, a few 3-year-old olive trees with decline symptoms were observed in a recently planted commercial orchard in the Enna province (Sicily). The trees were propagated on wild olive (O. europea L. var. sylvestris Brot.) rootstock. Aerial symptoms, consisting of leaf chlorosis, wilting, defoliation, and twig dieback followed in most cases by plant death, were associated with root rot and basal stem cankers. A Phytophthora sp. was consistently isolated from rotted rootlets and trunk cankers using the BNPRAH (benomyl, nystatin, pentachloronitrobenzene, rifampicin, ampicillin, and hymexazol) selective medium. Pure cultures were obtained by single-hypha transfers. The species isolated from symptomatic olive trees was identified as P. megasperma Drechsler on the basis of morphological and cultural characteristics. All isolates were homothallic, with paragynous antheridia. The diameter of oospores varied from 28 to 42 μm (mean ± SE = 36.3 ± 0.4) when they were produced on potato-dextrose agar (PDA) and from 30 to 43 μm (mean ± SE = 37.8 ± 0.4) when they were produced in saline solution. Sporangia were non-papillate. Optimum and maximum temperatures for radial growth of the colonies on PDA were 25 and 30°C, respectively. At 25°C, radial growth rate was about 6 mm per day. The identification was confirmed by the electrophoresis of mycelial proteins on a polyacrylamide slab gel. The electrophoretic banding patterns of total soluble proteins and three isozymes (esterase, fumarase, and malate dehydrogenase) of the isolate from olive were identical to those of two isolates of P. megasperma obtained from cherry and from carrot in Italy and characterized previously (1). Conversely, they were clearly distinct from the electrophoretic patterns of four isolates of P. megasperma var. sojae Hildebr. from soybean (= P. sojae Kauf. & Ger.), from those of three isolates from asparagus tentatively identified as P. megasperma sensu lato (1) and from those of reference isolates of various species producing non-papillate sporangia, including P. cambivora (Petri) Buisman, P. cinnamomi Rands, P. cryptogea Pethybr. & Laff., P. drechsleri Tucker, and P. erythroseptica Pethybr. Pathogenicity of the isolate from olive was tested in the greenhouse at 18 to 25°C using 18-month-old rooted cuttings of olive cv. Biancolilla. Cuttings were inoculated on the lower stem by inserting a 3-mm plug taken from actively growing colonies on PDA into an incision made with a sterile scalpel. The wound was sealed with waterproof tape. Agar plugs with no mycelium were placed into the stem of cuttings used as a control. The bark was stripped and lesion areas were traced and measured 60 days after inoculation. The isolate from olive produced a brown necrotic lesion (mean size = 500 mm2) around the inoculation wound and was reisolated from the lesion. Conversely, the wound healed up on control plants. P. megasperma has previously been recognized as a pathogen of olive in Greece and Spain (3). However, this is the first report of P. megasperma causing root and collar rot of olive in Italy. References: (1) S. O. Cacciola et al. Inf. Fitopatol. 46:33, 1996. (2) D. C. Erwin and O. K. Ribeiro, 1996. Phytophthora Diseases Worldwide. The American Phytopathological Society, St. Paul, MN. (3) M. E. Sánchez-Hernádez et al. Plant Dis. 81:1216, 1997.

Blue mediterranean fan palm ( Chamaerops humilis var. argentea ) is a shrub-like palm with clumps that are pale silvery-blue in colour. This species is native to the western Mediterranean basin and appreciated as an ornamental in Europe.…

In the summer of 2005, approximately 5% of a nursery stock of 12-month-old potted plants of bower vine (Pandorea jasminoides (Lindl.) K. Schum.) in Sicily (Italy) showed wilt, leaf chlorosis, defoliation, root rot, and collapse of the entire plant. Three Phytophthora spp. (20, 50, and 30% of the isolations of the first, second, and third species, respectively) were isolated from rotted roots on BNPRAH selective medium (2). Single-hypha isolates of the first species formed petaloid colonies on potato dextrose agar (PDA) and had an optimum growth temperature of 25°C (9.3 mm/day); on V8 juice agar, they produced uni- and bipapillate, ovoid to limoniform sporangia with mean dimensions of 45 × 30 μm and a mean length/width (l/w) ratio of 1.4:1. They did not produce gametangia when paired with A1 and A2 isolates of Phytophthora nicotianae. The second species formed arachnoides colonies on PDA, had an optimum growth temperature of 30°C (6.9 mm/day) and produced sporangia that were uni- and bipapillate, ellipsoid, ovoid, or pyriform to spherical (dimensions 44 × 34 μm; l/w ratio 1.3:1). All isolates were A2 mating type and produced amphyginous antheridia and spherical oogonia with smooth walls. The third species formed rosaceous colonies on PDA, had an optimum growth temperature of 28 to 30°C (11.9 mm/day), and produced uni- and bipapillate, ellipsoid or limoniform, caducous sporangia (dimensions 52 × 26 μm; l/w ratio 2.1:1) with a tapered base and a long pedicel (as much as 150 μm). All isolates were A1 type and produced amphigynous antheridia and spherical oogonia with smooth walls. The three species were identified as P. citrophthora, P. nicotianae, and P. tropicalis, respectively. The electrophoretic analysis of the mycelial proteins and four isozymes (1) confirmed the identification. Blast analysis of the sequence of the internal transcribed spacer region of the rDNA of a P. tropicalis isolate from bower vine (GenBank Accession No. EU076731) showed 99% similarity with the sequence of a P. tropicalis isolate from Cuphea ignea (GenBank Accession No. DQ118649). The pathogenicity of three isolates from bower vine, IMI 395552 (P. citrophthora), IMI 395553 (P. nicotianae), and IMI 395346 (P. tropicalis), was tested on 3-month-old potted bower vine plants (10 plants for each isolate) by applying 10 ml of a suspension (2 × 104 zoospores/ml) to the root crown. The plants were maintained at 24°C and 95 to 100% relative humidity. All inoculated plants wilted after 4 weeks. Noninoculated control plants remained healthy. The three Phytophthora spp. were reisolated from symptomatic plants. To our knowledge, this is the first report of Phytophthora root rot of bower vine in Italy. References: (1) S. O. Cacciola et al. Plant Dis. 90:680, 2006. (2) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society, St. Paul, MN, 1996.

Lantana (Lantana camara L.) is an evergreen shrub in the Verbenaceae. In some countries, this plant has been declared a noxious weed. However, a number of sterile or near-sterile forms are cultivated as attractive flowered potted and garden plants. In early spring 2004, ≈4,000 potted, small trees of lantana grown in a screenhouse in a commercial nursery of ornamentals near Giarre, Sicily, showed symptoms of chlorosis, defoliation, and sudden collapse of the entire plant. These aboveground symptoms were associated with a reduced root system, rot of feeder roots, and brown discoloration of the base of the stem. A Phytophthora sp. was isolated consistently from roots and basal stems of symptomatic plants using the selective medium of Masago et al. (3). Cardinal temperatures for radial growth of pure cultures obtained by single hypha transfer were 2°C minimum, 25°C optimum, and 30 to 35°C maximum. Sporangia produced in the saline solution of Chen and Zentmyer (3) were obpyriform, persistent, and nonpapillate. All isolates were A1 mating type and differentiated oospores with amphigynous antheridia in dual cultures with A2 reference isolates of P. cryptogea Pethybr. & Laff. and P. drechsleri Tucker (3). Electrophoretic patterns of total mycelial proteins (3) of the isolates from lantana were very similar to those of reference isolates of P. cryptogea from different hosts, but clearly distinct from those of reference isolates of other species included in Waterhouse's taxonomic group VI (3). Indeed, isolates from lantana were identified as P. cryptogea on the basis of morphological and cultural characters as well as the electrophoretic phenotype. Sequences of internal transcribed spacer (ITS) regions of rDNA (1) confirmed the identification as P. cryptogea. Pathogenicity of a representative isolate from lantana (IMI 392045) was tested in a screenhouse by transplanting 20 6-month-old rooted cuttings of lantana in pots (12 cm in diameter) filled with infested soil; the soil was prepared by mixing steam-sterilized sandy loam soil at a concentration of 4% (vol/vol) with inoculum produced on a mixture of vermiculite and autoclaved oat seeds. Twenty control plants were transplanted in pots containing noninfested soil. The soil was saturated with water by plugging the pots' drainage holes for 48 h and watering. After 40 days, all plants except the controls showed symptoms of root and foot rot, and P. cryptogea was reisolated from infected tissues. To our knowledge, this is the first report of P. cryptogea on lantana. On this host and other species in the verbena family, only P. nicotianae van Breda de Haan (= P. parasitica Dastur) has been previously reported (2,3,4). A possible cause of the high incidence of this disease in the nursery was waterlogging due to heavy rain and excessive irrigation. References: (1) S. O. Cacciola et al. For. Snow Landsc. Res. 76:387, 2001. (2) M. L. Daughtrey et al. Compendium of Flowering Potted Plant Diseases. The American Phytopathological Society, St. Paul, MN, 1995. (3) D. C Erwin and O. K. Ribeiro. Pages 39–41, 84–95, 138–139 in: Phytophthora Diseases Worldwide. The American Phytopathological Society, St. Paul, MN, 1996. (4) K. H. Lamour et al. Plant Dis. 87:854, 2003.

Muskmelon (Cucumis melo L.) is very important economically to agriculture in Italy. The Sicily area accounts for ≈40% of the total muskmelon production. Fusarium wilt caused by Fusarium oxysporum f. sp. melonis (Leach & Currence) W.C. Snyder & H.N. Hans. is the most prevalent and damaging disease of muskmelon in Sicily. Use of cultivars with major resistance genes, Fom 1 and Fom 2, is the most effective control measure for combating the disease. During March 1999, severe infections of Fusarium wilt were noted in a commercial muskmelon crop, cv. Firmo F1, grown in plastic tunnels in Syracuse Province (eastern Sicily). The muskmelon seedlings had been transplanted into the tunnels during January 20 days after soil fumigation with methyl bromide. Firmo F1 possesses both Fom 1 and Fom 2 genes. Of 18,000 Firmo F1 plants, ≈6,500 showed symptoms consisting of stunting, vein clearing; leaf yellowing, wilting, and dying; brown necrotic streak; and gummy exudates on the basal portion of vines. A pinkish white mold developed on dead tissues when infected plants were kept at high relative humidity. The pathogenicity of both a single-conidium isolate of F. oxysporum f. sp. melonis from a symptomatic Firmo F1 plant and two isolates of races 0 and 1, recovered previously from other cultivars in Sicily and used as references, was tested with three differential muskmelon cultivars, Charentais T, Doublon, and CM 17187 (1), as well as three commercial cultivars, Ramon, Cassella, and Geamar (possessing Fom 1, Fom 2, and both Fom 1 and Fom 2 resistance genes, respectively). Muskmelon seedlings were inoculated by the root-dip method (3), using a suspension of 5 × 105 conidia per ml. Inoculated seedlings were transplanted to plastic pots filled with sterilized soil and placed in a greenhouse (25 to 30°C). Symptoms were scored 7 to 10 days after inoculation. The isolate from Firmo F1 was pathogenic to all cultivars tested, the race 0 isolate was pathogenic only to cv. Charentais T, and the race 1 isolate was pathogenic only to cvs. Charentais T, Doublon, and Ramon. F. oxysporum was reisolated from symptomatic plants. Based on its pathogenicity and symptomology, the isolate from Firmo F1 was classified as race 1,2y (yellows), according to the nomenclature proposed by Risser et al. (1). Race 1,2 poses a serious threat to muskmelon production in Sicily, because all currently used cultivars are susceptible to the race, and other control measures, such as preplant soil fumigation with methyl bromide and solarization, are not as effective as use of resistant cultivars. Further study is needed to establish which is the prevalent race of F. oxysporum f. sp. melonis in Sicily. This report confirms that race 1,2 occurs in all major muskmelon-production areas in Italy (2). References: (1) G. Risser et al. Phytopathology 66:1105, 1976. (2) G. Tamietti et al. Petria 4:103, 1994. (3) F. L. Wellman. Phytopathology 29:945, 1939.

Root rot caused by Phytophthora cinnamomi Rands is generally recognized to be the most important disease of avocado (Persea americana Miller) wherever this tropical fruit tree is grown (3). The disease was first found in Italy in the spring of 1998. Eight-year-old trees, with symptoms ranging from initial to severe, were observed in an experimental field near Rocca di Caprileone, in Sicily. Few trees showed symptoms of both root rot and collar rot. Infected trees were of 13 commercial cultivars. Trees were grafted on two different rootstocks: Hass seedlings and G6 seedlings. G6 is a selection reported to have some field resistance to P. cinnamomi infections (2). However, no correlation was observed between symptom severity and rootstock. P. cinnamomi was isolated on BNPRAH selective medium (4) from trunk bark, feeder roots, and rhizosphere soil of diseased trees, and from roots of symptomless trees. The isolates, identified primarily on the basis of morphological and cultural characteristics, formed rosaceous colonies on potato dextrose agar (PDA) and on corn meal agar (CMA) coralloid-type mycelium, with abundant hyphal swellings, which were typically spherical and in clusters. Chlamydospores were either terminal or intercalary, and often occurred in characteristic grapelike clusters. Sporangia, which were produced in saline solution (1), were broadly ellipsoidal or ovoid, persistent, non-papillate and proliferous. The identification was confirmed by the electrophoresis of mycelial proteins on polyacrylamide slab gel. The electrophoretic patterns of total soluble proteins and eight isozymes (AKP [alkaline phosphatase], EST [esterase], FUM [fumarase], GLC [NAD-glucose dehydrogenase], G6PD [glucose-6-phosphate dehydrogenase], LDH [lactate dehydrogenase], MDH [malate dehydrogenase], and SOD [superoxide dismutase]) of the isolates from avocado were identical to those of two strains of P. cinnamomi, used as reference (isolate 70473 from International Mycological Institute, U.K., and an isolate from myrtle from the Institute of Plant Pathology, University of Catania, Italy). Conversely, the electrophoretic phenotype of the P. cinnamomi isolates from avocado was clearly distinct from those of reference strains of eight other species included in Waterhouse's taxonomic group VI. Pairings with isolates of a known mating type of P. cinnamomi, P. cryptogea, and P. drechsleri revealed that all the isolates from avocado were A2 mating type. It is possible that P. cinnamomi had been introduced into the experimental field on infected symptomless nursery trees. In Italy, root rot caused by P. cinnamomi could have a significant impact on commercial avocado plantings extending over about 20 ha. Moreover, this polyphagous pathogen may be a threat to other crops as well as to forest trees. References: (1) D. W. Chen and G. A. Zentmyer. Mycologia 62:397, 1970. (2) M. D. Coffey. Plant Dis. 71:1046, 1987. (3) D. C. Erwin and O. K. Ribeiro. 1996. Phytophthora Diseases Worldwide. American Phytopathological Society, St. Paul, MN. (4) H. Masago et al. Phytopathology 67:425, 1977.