First Report of a Decline and Wilt of Young Olive Trees Caused by Simultaneous Infections of Verticillium dahliae and Phytophthora palmivora in Sicily.

In summer 2008, leaf chlorosis, defoliation, exceptional fruit set, twig dieback, and wilt were observed on 4-year-old olive (Olea europea L.) trees cv. Tonda Iblea in a drip-irrigated orchard in eastern Sicily. Rot of fine roots was associated with these symptoms and on ~15% of symptomatic trees rot extended to the crown and basal stem. Trees declined slowly or collapsed suddenly with withered leaves still attached. Incidence of affected trees was ~10%. A fungus identified as Verticillium dahliae Kleb. was isolated from the xylem of main roots and basal stem. An oomycete identified as Phytophthora palmivora (Butler) Butler was isolated from roots and basal trunk bark. Both pathogens were recovered from symptomatic trees with mean frequency of positive isolations per tree of 80 and 30% for V. dahliae and P. palmivora, respectively. To isolate V. dahliae, wood chips were surface disinfested in 0.5% NaOCl for 1 min and plated onto potato dextrose agar (PDA). The fungus was identified on the basis of microsclerotia, verticillate arrangement of phialides on conidiophores, and hyaline single-celled conidia. Ten monoconidial isolates were characterized by PCR using primer pairs INTND2f/INTND2r and DB19/espdef01 (3). Only 824-bp amplicons, diagnostic of the virulent, nondefoliating V. dahliae pathotype, were obtained. P. palmivora was isolated on selective medium (2) and pure cultures were obtained by single-hypha transfers. Colonies grew on PDA between 10 and 35°C (optimum at 27°C). Chlamydospores and elliptical to ovoid, papillate, caducous (mean pedicel length = 5 µm) sporangia (length/breadth ratio of 1.8) were produced on V8 juice agar. All isolates were paired with reference isolates of P. nicotianae and produced gametangia only with isolates of the A2 mating type. PCR amplicons of a representative isolate generated using primers ITS 6 and ITS 4 (1) were sequenced and found to be identical to those of a reference isolate of P. palmivora (GenBank No. AY208126). Pathogenicity of V. dahliae (IMI 397476) and P. palmivora (IMI 397475) was tested on 6-month-old rooted cuttings of olive cv. Tonda Iblea. Ten cuttings were transplanted into pots with steam-sterilized soil and inoculum of P. palmivora (4% vol/vol) produced on wheat kernels. Ten olive cuttings were inoculated with V. dahliae by injecting the stem with 150 µl of a conidial suspension (107 conidia ml-1) and 10 cuttings were stem inoculated with V. dahliae and transplanted into soil infested with P. palmivora. Controls were 10 noninoculated cuttings transplanted into steam-sterilized soil. Pots were kept in a greenhouse (25 ± 3°C) for 4 months. No aerial symptoms were observed on cuttings transplanted into soil infested with P. palmivora. However, root dry weight was reduced by 40% in comparison with the controls. Cuttings inoculated solely with V. dahliae had a 15% reduction in height compared with the controls but only four cuttings wilted. All cuttings inoculated with P. palmivora and V. dahliae wilted, indicating a synergism between the two pathogens. Controls remained healthy. Each pathogen was reisolated solely from inoculated cuttings and both pathogens were reisolated from cuttings with double inoculations. A similar syndrome 'seca' (drying) was reported in Spain (4). References: (1) D. E. L. Cooke et al. Fungal Genet. Biol. 30:17, 2000. (2) H. Masago et al. Phytopathology 67:425, 1977. (3) J. Mercado-Blanco et al. Plant Dis. 87:1487, 2003. (4) M. E. Sánchez-Hernández et al. Eur. J. Plant Pathol. 104:34, 1998.

First Report of Phytophthora tentaculata Causing Root and Stem Rot of Oregano in Italy.

Oregano (Origanum vulgare L.; Lamiaceae) is cultivated for culinary and medicinal purposes and as an ornamental. In October of 2007, 1- to 2-year-old potted plants of oregano showed symptoms of decline associated with root and basal stem rot in a nursery in Liguria (northern Italy) that produces 1 million to 1.5 million potted aromatic plants per year. Aboveground symptoms included leaf russeting and chlorosis, wilt, defoliation and dieback of twigs, browning of the basal stem, and subsequent collapse of the entire plant. Approximately 80% of the plants died within 30 days after the appearance of the first symptoms on the canopy. Approximately 20% of a stock of 30,000 oregano plants was affected. Stocks of other aromatic species, such as mint, lavender, rosemary, and sage, appeared healthy. A Phytophthora species was consistently isolated from symptomatic stems and roots of oregano plants on BNPRAH selective medium (2). Ten pure cultures were obtained by single-hypha transfers, and the species was identified as Phytophthora tentaculata Kröber & Marwitz by morphological criteria and sequencing of the internal transcribed spacer (ITS) region of rDNA using the ITS 4 and ITS 6 universal primers for DNA amplification. Isolates from oregano formed stoloniferous colonies with arachnoid mycelium on potato dextrose agar and had a growth rate of 2 to 3 mm per day at 24°C with optimum, minimum, and maximum temperatures of 24, 8, and 34°C, respectively. Sporangia formed in soil extract solution and were papillate and spherical or ovoid to obpyriform with a length/breadth ratio of 1.3:1. Few sporangia were caducous and all had a short pedicel (<5 µm). Hyphal swellings and chlamydospores were produced in sterile distilled water and corn meal agar, respectively. All isolates were homothallic and produced globose terminal oogonia (mean diameter of 34 µm) with one or occasionally two paragynous, monoclinous, or diclinous antheridia. Amphigynous antheridia were also observed. The sequence of the ITS region of the rDNA (GenBank No. FJ872545) of an isolate from oregano (IMI 395782) showed 99% similarity with sequences of two reference isolates of P. tentaculata (Accession Nos. AF266775 and AY881001). To test for pathogenicity, the exposed root crowns of 10 6-month-old potted plants of oregano were drench inoculated with 10 ml of a suspension of 2 × 104 zoospores/ml of isolate IMI 395782. Sterile water was pipetted onto the roots of 10 control plants. All plants were maintained in 100% humidity at 22 to 24°C in a greenhouse under natural light and watered once a week. Within 3 weeks after inoculation, all inoculated plants developed symptoms identical to those observed in the nursery and died within 30 to 40 days after the appearance of the first symptoms. Control plants remained healthy. P. tentaculata was reisolated solely from symptomatic plants. P. tentaculata has been reported previously on several herbaceous ornamental plants (1,3). However, to our knowledge, this is the first report of this species on O. vulgare. Root and basal stem rot caused by P. tentaculata is the most serious soilborne disease of oregano reported in Italy so far. References: (1) G. Cristinzio et al. Inf. Fitopatol. 2:28, 2006. (2) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society, St. Paul, MN, 1996. (3) H. Kröber and R. Marwitz. Z. Pflanzenkr. Pflanzenschutz 100:250, 1993.

Blight Caused by Sclerotium rolfsii on Potted Ornamental Citrus in Sicily.

Approximately 140,000 container-grown ornamental citrus plants are produced each year in the province of Catania (eastern Sicily). In the spring of 2006, a severe blight was observed in a commercial nursery in Catania on 2-month-old rooted cuttings of lemon (Citrus limon (L.) Burm.) and calamondin (× Citrofortunella mitis (Blanco) J. W. Ingram & H. E. Moore). Approximately 80% of the nursery stock of 2,000 cuttings was affected. Cuttings were grown in 7.5-cm2 pots made with compressed peat and wood pulp at 28 to 30°C with 95 to 100% relative humidity on benches in a greenhouse, The pot mix was composed of peat, perlite, and soil (2:1:2). Cuttings showed a dark brown necrotic lesion at the base of the stem that extended upward, resulting in chlorosis and wilting of the leaves. An invasive, white, cottony mycelium with a fan-like pattern and numerous, small, brown spherical sclerotia (0.5 to 4.0 mm in diameter) developed on infected tissues, in the potting mix as well as on the pot wall. Herbaceous cuttings collapsed within 2 weeks while woody cuttings gradually died. Symptomatic basal stem sections were disinfected for 1 min in 1% NaOCl, rinsed in sterile water, and plated on acidified (pH 4.5) potato dextrose agar (PDA). Isolations consistently yielded a fungus whose morphological characters corresponded to Sclerotium rolfsii Sacc. On PDA, it produced a septate mycelium with clamp connections and numerous olive brown-to-clove brown sclerotia (1 to 3 mm in diameter). Pathogenicity of two S. rolfsii isolates (IMI 396204 and IMI 396205) from citrus was confirmed on 3-month-old lemon cuttings grown in 10-cm-diameter plastic pots filled with a sterilized mix of peat moss and vermiculite (3:1) (10 cuttings for each isolate). Each pot was inoculated with 15 sclerotia harvested from 6-week-old cultures on PDA and placed on the soil surface around the base of the cutting. Ten noninoculated cuttings served as the control. Cuttings were kept in a growth chamber at 28°C and relative humidity at >95%. All inoculated cuttings showed wilting, blight, and stem rot within 3 weeks after inoculation. White mycelium and sclerotia were produced on the stem base and soil surface. Noninoculated controls remained symptomless. S. rolfsii was reisolated from infected cuttings. The pathogenicity test was repeated once with calamondin cuttings and the results were similar. Blight caused by S. rolfsii is widespread in nurseries of ornamentals in Italy (1). However, to our knowledge, this is the first report of this disease on potted ornamental citrus. Probably high temperature and moisture during rooting were conducive to the disease. References: (1) A. Garibaldi et al. Malattie Delle Piante Ornamentali. Calderini Edagricole, Bologna, Italy, 2000.

Blight of English Ivy (Hedera helix) Caused by Sclerotium rolfsii in Sicily.

English ivy, Hedera helix L. (Araliaceae), an evergreen climbing vine is widely cultivated as an ornamental and foliage plant. In the summer of 2005, a severe blight of ivy plants trained as topiaries and grown in an open field was observed in a nursery near Giarre (eastern Sicily). Foliage of infected plants appeared lighter green and progressively turned bronze and withered. Eventually, the entire plant collapsed. Foliar symptoms were associated with basal stem and root rot. White, cottony mycelium and numerous sclerotia developed externally on the lower stem and on the soil around the affected plants. The disease was randomly distributed, affecting approximately 5% of plants in a stock of 1,500 English ivy plants. Sclerotium rolfsii Sacc. (teleomorph Athelia rolfsii (Curzi) Tu & Kimbrough) was consistently isolated from symptomatic basal stem tissues by disinfecting in 1% NaOCl and plating on potato dextrose agar (PDA) amended with 100 mg/liter of streptomycin sulfate. The isolated fungus was identified on the basis of morphological and cultural characteristics (2). On PDA, it produced a densely, floccose, white mycelium. Mycelium was septate with clamp connections at hyphal septa. Optimum growth temperature was 30 ± 2°C. Numerous small (0.5 to 1.9 mm in diameter) sclerotia developed on the colony surface; they were spherical, occasionally slightly ellipsoidal, quite uniform in size (modal value of the diameter 1.4 mm), with a smooth surface. The surface color of the sclerotia was initially white, turned to pinkish buff, then to olive-brown, and eventually to clove brown as sclerotia matured. Sclerotia were most numerous in the center as well as close to the edge of petri dishes. Pathogenicity of one isolate obtained from infected plants was confirmed by inoculating 10 1-year-old potted English ivy plants by placing mycelium-infested wheat kernels and sclerotia on the soil surface around the collar of each plant. Ten noninoculated plants served as control. Plants were held in a dew chamber for 48 h at 28°C and subsequently placed in a greenhouse where the temperature ranged between 25 and 31°C. Plants showed wilting within 3 weeks after inoculation. Fans of white mycelium and numerous sclerotia were produced on the basal stem of inoculated test plants. Noninoculated controls remained healthy. S. rolfsii was reisolated from infected plants to fulfill Koch's postulates. English ivy has been already reported as a host of S. rolfsii, the causative agent of southern blight in nurseries of ornamentals (1). However, to our knowledge, this is the first report of southern blight on English ivy in Italy. The disease may have been favored by warm summer temperatures and overwatering with a drip irrigation system. References: (1) A. R. Chase. Compendium of Ornamental Foliage Plant Diseases. The American Phytopathological Society, St. Paul, MN, 1992. (2) J. E. M. Mordue. Corticium rolfsii. No. 410 in: Description of Pathogenic Fungi and Bacteria. CMI. Kew, Surrey, UK, 1974.

Wilt and Collapse of Cuphea ignea Caused by Phytophthora tropicalis in Italy.

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.

First Report of Brown Rot and Wilt of Fennel Caused by Phytophthora megasperma in Italy.

Fennel (Foeniculum vulgare Mill. var. azoricum (Mill.) Thell.) in the Apiaceae family is native to southern Europe and southwestern Asia. It is an economically important crop in Italy that produces approximately 85% of all fennel worldwide. The main producing regions are Apulia, Campania, Latium, and Calabria. During the late winter of 2004 in the Crotone Province of the Calabria Region, following heavy rains, patches of fennel plants with symptoms of brown, soft rot of the bulb-like structure formed by the thickened leaf bases, development of yellow leaves, stunting, and wilting of the entire plant were observed in fields. A homothallic Phytophthora sp. was isolated consistently from the brownish tissues of the stout stems and leaf bases of symptomatic plants using a selective medium (3). Pure cultures were obtained by single hyphal tip transfers. On potato dextrose agar (PDA), the diameter of oospores varied from 28 to 42 µm (mean = 36.3 ± 0.4). Antheridia were primarily paragynous. Sporangia were not produced on solid media but were formed in sterile soil extract solution. They were nonpapillate, noncaducous, ovoid and obpyriform (25 to 45 × 35 to 60 µm), and internally proliferating. 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 approximately 6 mm per day. On the basis of morphological and cultural characteristics, the isolates were identified as Phytophthora megasperma Drechsler. Electrophoretic patterns of mycelial proteins and four isozymes (acid and alkaline phosphatase, esterase, and malate dehydrogenase) on polyacrylamide gels of the fennel isolates were identical to those of reference isolates of P. megasperma of the BHR (broad host range) group included in P. gonapodyides-P. megasperma Clade 6 (1,3), but distinct from those of the isolates of other nonpapillate species included in Waterhouse's taxonomic group VI. Internal transcribed spacer (ITS) regions of rDNA sequences (2) confirmed that fennel isolates belonged to P. megasperma BHR group. Pathogenicity of a fennel isolate from Calabria (IMI 391711) was confirmed by pouring a zoospore suspension at 2 × 104 zoospores per ml on the soil of 10 3-month-old potted fennel plants. The soil of the inoculated and 10 control seedlings was flooded for 24 h. After 10 days, stems and leaf bases of the seedlings showed a brown rot. Chlorosis and wilting of all seedlings developed after 20 days. Controls inoculated with water did not develop any symptoms. The pathogen was reisolated from typical brown rot and tests were repeated with similar results. To our knowledge, this is the first report of P. megasperma causing disease on fennel. References: (1) S. O. Cacciola et al. For. Snow. Landsc. Res. 76:387, 2001. (2) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society, St. Paul, MN, 1996. (3) H. Masago et al. Phytopathology, 67:425, 1977.

The value of inching technique in evaluating the peroneal nerve entrapment at the fibular head.

The peroneal nerve palsy at the fibular head is quite common but often difficult to diagnose both clinically and electrophysiologically. The purpose of this study was to evaluate the usefulness of the inching in mononeuropathy of the peroneal nerve at the fibular head. Recording from extensor digitorum brevis muscle the nerve was stimulated supramaximally at 1 cm intervals starting 2 cm distal and ending 8 cm proximal to the fibular head. Forty-six patients were examined: the inching was modified in 32 patients. In five of these the motor conduction using conventional method was normal, but the inching was normal or borderline in fourteen patients with reduced conduction velocity across the fibular head. Despite some limitations, the inching can be useful in evaluating patients with suspected palsy of the peroneal nerve at the fibular head.

Nerve refractory period in early polyneuropathies.

The refractory period of the sural nerve was evaluated in 115 patients with suspected polyneuropathy of various origin. They were asymptomatics or with minor, mainly sensitive, disturbances. In all the conventional neurophysiologic examination (motor and sensory velocities, distal latencies and F-waves of the peroneal, ulnar and sural nerves) was normal. The absolute refractory period (ARP) was defined as the shortest interstimulus interval at which a response to the second stimulus could be recorded. The relative refractory period (RRP) was defined as the interstimulus interval at which the responses to the first and to the second stimulus have equal latencies. The ARP was prolonged in 13 patients and the RRP in 28. In all, the refractory period was prolonged in 38 patients (33%). In any but three patients the ARP and the RRP were prolonged together. Thus the refractory period is more sensitive than the conventional neurophysiological examination in detecting early neuropathy. Moreover the ARP was modified in 2/9 patients with Steinert's myotonic dystrophy without clinical or electrophysiological signs of peripheral neuropathy.

First Report of Botrytis Blight on Medinilla magnifica and Various Species of Mandevilla and Allamanda in Italy.

Medinilla magnifica Lindl., Mandevilla splendens (Hook.) Woodson, the hybrid Mandevilla × amoena 'Alice du Pont' (pink allamanda), and various species of Allamanda, such as A. cathartica L. and A. blanchetii A. DC. (purple allamanda), are grown in Sicily as ornamentals. After a frost in early December 2001, a sudden wilt of container-grown cuttings of these tropical species was observed in a plastic-covered production greenhouse, with ≈30% of M. magnifica plants and 70% of Mandevilla and Allamanda plants affected. Medinilla plants (≈35 cm high) had been rooted in trays and transplanted individually in 30-cm-diameter pots. Allamanda (recently rooted cuttings) and Mandevilla (well-established) plants showed symptoms ranging from a dark brown rot at the base of stems to a tan-to-brown rot of stem midsection, respectively. Tissues surrounding lesions were water-soaked and covered by gray mycelium. On Allamanda and Mandevilla plants, lesions extended rapidly to lateral branches, and to the petiole and midrib of leaves that became chlorotic and wilted. As stems were girdled, infected plants collapsed, although the roots appeared healthy. Botrytis cinerea Pers.:Fr. was consistently isolated from infected stem pieces surface-disinfested with 1% NaOCl for 1 min and placed on potato dextrose agar (PDA, Oxoid). Morphology and size (6 to 8 × 8 to 12 µm, modal values 7 × 10 µm) of conidia produced on PDA matched those reported for B. cinerea (1). Dark, spherical, and irregularly shaped sclerotia (1 to 6 mm diameter, mean size 3.3 × 2.3 mm) were produced on PDA. Pathogenicity of a single-conidium isolate isolated from M. magnifica was confirmed using two inoculation methods. Twenty 3-month-old cuttings of M. magnifica and pink and purple allamanda were sprayed with a conidial suspension (106 conidia per ml in a 2% glucose solution). A 2% glucose solution was sprayed on 20 control plants. Plants were sealed in transparent plastic bags for 7 days at 15 to 24°C. Typical symptoms developed only on stems of inoculated pink and purple allamanda cuttings 7 days after inoculation. B. cinerea was reisolated from affected tissues. In a separate test, stems of 10 6-month-old plants of M. magnifica and pink allamanda were inoculated by inserting a 3-mm plug taken from 10-day-old sporulating colonies growing on PDA on a superficial cut made with a sterile scalpel. The inoculated wounds were sealed with Parafilm. Ten wounded but noninoculated plants were used as controls. Plants were kept in high humidity at 10 to 20°C. After 10 days, stem necrosis, leaf chlorosis, and wilt were observed on inoculated pink allamanda plants. On inoculated M. magnifica plants, necrotic lesions were observed on stems (45 to 70 mm long and 10 to 18 mm wide) covered by gray mycelium, but the stem was not girdled. B. cinerea was reisolated from infected tissues of inoculated plants to complete Koch's postulates. No lesions developed on noninoculated control plants. To our knowledge, this is the first report from Italy of Botrytis blight on these species. The occurrence of frost may have predisposed these tropical species to infection by B. cinerea. Reference: (1) M. B. Ellis and J. M. Waller. No 431 in: Descriptions of Pathogenic Fungi and Bacteria, CMI, Kew, Surrey, UK, 1974.

First Report of Root and Crown Rot of Sage Caused by Phytophthora cryptogea in Italy.

Sages are cultivated as aromatic and ornamental plants in Italy and represent the common name of certain species of Salvia and Phlomis (family Lamiaceae). In Sicily (southern Italy) during the summer of 2001, ≈40% of 1,400 2-year-old landscape plants of S. leucantha Cav. (Mexican bush sage or velvet sage) showed symptoms of stunting, chlorosis, and gradual dieback or sudden wilt, which are associated with root and crown rot. Plants were supplied by a commercial nursery, transplanted from pots in the spring, and irrigated using a trickle system. Phytophthora was isolated consistently from roots and basal stems of symptomatic plants on a BNPRAH medium (2). The species was identified as P. cryptogea Pethybr. & Laff., primarily on the basis of morphological and cultural characteristics. Five representative single-hypha isolates were characterized. On potato dextrose agar, they formed colonies with a slight petaloid pattern. Cardinal temperatures for mycelium growth were 2°C, minimum; 25°C, optimum; and 30 to 35°C, maximum. Hyphal swellings were abundant in aqueous culture. Sporangia were obpyriform, persistent, nonpapillate, and proliferous (2). All isolates were the A1 mating type and formed oogonia, amphigynous antheridia, and oospores in dual cultures with reference isolates of the A2 mating type of P. cryptogea and P. drechsleri. Identification was confirmed by electrophoresis of mycelium proteins on a polyacrylamide slab gel (1). Electrophoretic patterns of total soluble proteins from the sage isolates were identical or very similar to those from 10 reference isolates of P. cryptogea from various hosts, including isolate IMI 180615 (ex-type isolate). Conversely, the electrophoretic pattern of the isolates of P. cryptogea from sage was clearly distinct from those from reference isolates of other species included in Waterhouse's taxonomic group VI. Esterase (EC 3.1.1.2.) zymograms of the sage isolates corresponded to those of isolates of P. cryptogea included in electrophoretic group 2 (1). The pathogenicity of a representative isolate of P. cryptogea from sage was tested in the greenhouse using 4-month-old plants of Mexican bush sage. Inoculum was produced on a mixture of vermiculite and autoclaved oat seeds (4) and mixed with steam-sterilized sandy loam soil at a concentration of 4% (vol/vol). Plants were transplanted in pots (12 cm diameter) filled with infested soil; control plants were grown in pots containing noninfested soil. After transplanting, all pots were placed in shallow trays filled with water for 24 h to saturate the soil. All plants grown in infested soil showed extensive root necrosis and dieback ≈30 days after transplanting, and P. cryptogea was reisolated from roots of symptomatic plants. Control plants did not develop symptoms. Root and crown rot of sage caused by P. cryptogea has been reported previously in California (3). To our knowledge, this is the first report of P. cryptogea on sage in Italy. Root rot caused by P. cryptogea may be a potential problem for commercial cultivation of sage as no serious disease of this plant has been reported in Italy so far. References: (1) S. O. Cacciola et al. EPPO Bull. 20:47, 1990. (2) D. C Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. American Phytopathological Society, St. Paul MN. 1996. (3) S. T. Koike et al. Plant Dis. 81:959, 1997. (4) E. Sánchez-Hernández et al. Plant Dis. 85:411, 2001.

First Report of Phytophthora palmivora on Coronilla valentina subsp. glauca in Italy.

The genus Coronilla L. (family Fabaceae), which includes several species native to central and southern Europe, such as C. varia L. (axseed or crown-vetch), C. emerus (scorpion senna), and C. valentina L., is used in Italy as a landscape shrub or potted ornamental plant. During the summer of 2001, 80% of approximately 10,000 1-year-old plants of C. valentina subsp. glauca (L.) Batt. used to landscape an industrial area in the Caltanissetta Province (Sicily) showed symptoms of dieback associated with basal stem and root rot. Plants had been transplanted from pots in April and watered using a trickle irrigation system. A species of Phytophthora was isolated consistently from rotted roots and basal stems using BNPRAH selective medium (3). Pure cultures of this fungus were obtained by single-hypha transfers. Ten isolates, each originating from a single plant, were identified as P. palmivora (Butler) Butler on the basis of morphological and cultural characters as described by Erwin and Ribeiro (1). On solid media, including potato dextrose agar, cornmeal agar, and V8-juice agar, all the isolates produced elliptical to ovoid, papillate sporangia with a mean length/breadth ratio of 1.8. Sporangia were caducous with a short pedicel (mean pedicel length = 5 µm) and a conspicuous basal plug. Mating type was determined on V8 agar in dual culture with mating type A1 and A2 of reference isolates of P. nicotianae and P. palmivora. All isolates were heterothallic and produced oogonia and oospores only with reference isolates of the A2 mating type. Antheridia were amphigynous. Electrophoresis of mycelial proteins on polyacrylamide slab gel confirmed that all isolates were pure cultures and belonged to the same species. Koch's postulates were fulfilled using 6-month-old C. valentina subsp. glauca plants that were transplanted into pots filled with soil artificially inoculated with chlamydospores (50 chlamydospores per gram of soil) produced in submerged axenic cultures (2). The plants were maintained in a glasshouse at temperatures ranging from 18 to 28°C, and the pots were watered to field capacity once a week. One month after transplanting, 70% of plants showed dieback symptoms, while control plants, which were grown in pots containing noninoculated soil, remained healthy. The pathogen was reisolated from roots and basal stems of symptomatic plants. These results demonstrate that P. palmivora is the causal agent of dieback of C. valentina subsp. glauca plants. High temperatures in summer and waterlogging of soil due to excess irrigation water could have enhanced disease development. To our knowledge, this is the first report of P. palmivora on a species of Coronilla. P. palmivora is an exotic pathogen, but it is becoming widespread in Italy, where it has been reported from various regions on different hosts, including cyclamen, English ivy, palms, Pittosporum, and olive. References: (1) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society. St Paul, MN, 1996. (2) J. Y Kadooka and W. H. Ko. Phytopathology 63:559, 1973. (3) H. Masago et al. Phytopathology 67:425, 1977.

Non motor off in Parkinson's disease.

Besides the classic motor swings, many non motor fluctuations may occur in Parkinson's disease, but the clinical spectrum and the frequency of these symptoms are not well recognized. A total of 47 parkinsonian outpatients were questioned about any symptoms associated with off state. Nine patients had no fluctuations, 16 referred only to motor fluctuations and 22 to motor fluctuations associated with non motor symptoms. Overall, these patients referred to 54 symptoms (average 2.3/patients, range 1-6). These symptoms were classified as: autonomic (3 difficulty in swallowing, 7 hot, 11 sweat, 2 cold, 1 pallor, 1 abdominal bloating, 1 abdominal pain, 1 abdominal and genital pain, 5 bladder dysfunction, 2 feet oedema); sensory (7 sensory dyspnoea, 1 pain in lower limbs, 1 internal tremor); cognitive (3 depression, 4 anxiety, 2 panic, 1 drowsiness, 1 confusion). In patients without off periods, the length, severity and the average dosages of levodopa were fewer than in patients with fluctuations. No significant differences were found between patients with motor off and patients with associated non motor off regarding age (71.2+/-9.6 years vs 71.6+/-10.7 years), length of the disease (83.2+/-38.5 months vs 95.9+/-58.1 months), the Hoehn-Yahr (3.06+/-0.96 vs 3.02+/-0.96) and Webster (15.5+/-6.99 vs 15.1+/-5.9) scale, the dosages of levodopa (680.9+/-238.9 mg/die vs 679.7+/-289.6 mg/die), the number (2.3+/-1.7 vs 2.8+/-1.5) and length (6.8+/-5.2 h vs 7.2+/-7.1 h) of motor off. The non motor fluctuations were recognized in about 60% of patients with motor fluctuations: usually they were mild and less important than motor off, but sometimes these problems were disabling and led to unnecessary tests and therapies.

Tethered median nerve stress test in the diagnosis of carpal tunnel syndrome.

The sensitivity of the Median Nerve Stress Test (Stress Test) described by La Ban et al. and performed hyperextending for one minute the supinated wrist and the distal interphalangeal joint of the index finger and looking for pain in the proximal forearm was evaluated in 140 arms with Carpal Tunnel Syndrome confirmed electrophysiologically. The Stress Test was positive in 60 hands (42.8%), the Phalen's sign in 79 (56.4%) and the Tinel's sign in 59 (42.1%). Hypoaesthesia to pinprick in the distribution of the median nerve was found in 45 hands (32.1%) and weakness or hypotrophia of thenar eminence in 17 (12.1%). In spite of his low sensitivity, in some cases the Stress Test was the only clinical positive sign and, in addition to electrophysiological examination, may be helpful in clinical practice.

Auditory event-related potentials in Parkinson's disease.

We studied 49 patients with Parkinson's disease (PD) by a neuropsychological battery examining the temporo-spatial orientation, short-term memory, comprehension, non-verbal intelligence, long-term memory and anomia and the Auditory Event-Related Potentials. In the patients the latencies of the N100 and N200 waves were prolonged and the amplitude of the P300 wave was reduced compared with controls. No difference was found in the ERP of patients with and without cognitive deficits. Equally, no correlation was found between the ERP, the cognitive impairment, the length or the severity of the disease evaluated by Hoehn-Yahr's and Webster's scales.

F-wave: sample size and normative values.

F-waves were recorded from the abductor digiti minimi in 60 healthy subjects aged between 22 and 65 years. F-waves following 80 supramaximal stimuli were recorded. The F with minimal latency (Fmin), the mean latency (Fmean) and chronodispersion (CD) were calculated for each set of ten stimuli. In each subject the values after 10, 20, 30 etc. stimuli were compared with the "true" values obtained examining all responses. After twenty stimuli (16-20 F-waves) a Fmean with latency within 95% of the "true" value was recorded in the 95% of subjects. After twenty stimuli a Fmin with latency within 95% of the "true" latency was recorded in the 88% of subjects. Usually the Fmin after twenty stimuli is little different from the "true" Fmin (mean 1.4 ms), but in one subject the difference was 4.1 ms. The CD increases according to the number of responses examined and a careful evaluation requires 64-80 F-waves.

Electrophysiological study of peroneal nerve palsy at the fibular head.

The aim of this paper is to compare the potentials from the extensor digitorum brevis (EDB) with those from the tibialis anterior (TA) stimulating the peroneal nerve proximally and distally the fibular head in patients suffering of peroneal nerve palsy at the fibular head. In 8/30 limbs the electrophysiological studies were normal. According to electrophysiological findings the lesion was of demyelinating type in 6/22 patients, axonal in 8/22 and mixed in 8/22. In 12/22 limbs the abnormalities were found recording both for EDB and TA muscles; in 7/22 recording only from EDB and in 3/22 recording only from TA. Thus it is possible that motor fibers supplying the EDB and TA may be damaged differently and is useful evaluate both muscles.

Radial sensory nerve entrapment in carpal tunnel syndrome.

In 100 subjects with Carpal Tunnel Syndrome (CTS) and any other illness, the conduction velocity of the radial sensory nerve (RSN) was studied on the hypothesis that, because of the relation with the tendon of the brachioradialis and extensor carpi radialis longus muscles, microtrauma can injure the RSN. In 13 patients, for a total of 15 limbs involved, the SCV is reduced less than 2.5 SD, in 5 the distal latency is prolonged and in 1 there is both a slowing of SCV and prolonged distal latency. The involvement of RSN is more frequent in the dominant hand. There is no relation between the severity of CTS and damage of RSN or between clinical and electrophysiological data. It therefore seems that the entrapment of RSN is quite frequent and can explain some symptoms and modify the therapy.

Is temporomandibular dysfunction a cranial dystonia? An electrophysiological study.

In 26 patients suffering from temporomandibular dysfunction (TMD) the silent period (ES), which consists of an early (ES1) and a late (ES2) inhibition which interrupts the voluntary electromyographic activity after an electrical stimulus, was recorded from the masseter muscles. Several different patterns were identified: in 8 patients (group I) the ES was normal; in 1 patient (group II) the ES was entirely absent; in 11 patients (group III) only the ES2 was absent and in 6 patients (group IV) ES1 and ES2 were combined. Such results can be explained by hypothesizing several functional states: normal excitability in group I, absolute (group II) or relative (group III) inexcitability and hyperexcitability in group IV. A central origin of TMD can be related either to a "dysregulation" of circuits located in the brainstem which give rise to the ES or to the centers, probably located in the basal ganglia, which control the circuits of the brainstem.

Effects of positional changes on F wave.

Supine and after standing F wave studies were performed in 22 healthy subjects, in 14 patients with L5 root compression and in 11 patients with low back pain. Unlike previous works, the latency of the F minimal wave, F maximal, F mean and F chronodispersion before and after standing are similar. It is possible that the positional changes of the F wave may be of clinical interest only in sub-groups of patients and the usefulness of further studies is stressed.