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.

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Collar and Root Rot of Olive Trees Caused by Phytophthora megasperma in Sicily

Plant Disease ◽

10.1094/pdis.2001.85.1.96a ◽

2001 ◽

Vol 85 (1) ◽

pp. 96-96 ◽

Cited By ~ 5

Author(s):

S. O. Cacciola ◽

G. E. Agosteo ◽

G. Magnano di San Lio

Keyword(s):

Root Rot ◽

Radial Growth ◽

Selective Medium ◽

Phytophthora Megasperma ◽

Electrophoretic Patterns ◽

Pure Cultures ◽

Olive Trees ◽

Maximum Temperatures ◽

Single Hypha ◽

Collar And Root Rot

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.

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First Report of Phytophthora palmivora on Grevillea spp. in Italy

Plant Disease ◽

10.1094/pdis.2003.87.8.1006a ◽

2003 ◽

Vol 87 (8) ◽

pp. 1006-1006

Author(s):

S. O. Cacciola ◽

A. M. Pennisi ◽

G. E. Agosteo ◽

G. Magnano di San Lio

Keyword(s):

Mating Type ◽

Glucose Dehydrogenase ◽

Field Capacity ◽

Selective Medium ◽

Phytophthora Palmivora ◽

Width Ratio ◽

Submerged Cultures ◽

Reference Isolate ◽

Electrophoretic Patterns ◽

Pure Cultures

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.

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Development and quantitative measurement of microsclerotia of Verticillium dahliae

Canadian Journal of Botany ◽

10.1139/b72-272 ◽

1972 ◽

Vol 50 (11) ◽

pp. 2097-2102 ◽

Cited By ~ 7

Author(s):

R. Hall ◽

H. Ly

Keyword(s):

Verticillium Dahliae ◽

Mycelial Growth ◽

Quantitative Measurement ◽

Dry Weight ◽

Pure Cultures ◽

Carbon Nitrogen Ratio ◽

Low Glucose ◽

Nitrogen Ratio ◽

Total Dry Weight ◽

Hyaline Material

The development of microsclerotia of Verticillium dahliae from a few swollen hyaline cells on a hypha to a multicellular, pigmented "mature" structure is described and illustrated. A method for quantitatively estimating the amount of pigmented microsclerotial material in pure cultures was developed to study quantitative relations between mycelial growth and production of microsclerotial material in media containing different concentrations of glucose. At low glucose concentrations (0.6 to 10 mg/ml) microsclerotial material continued to increase after total dry weight of the cultures had reached a maximum, suggesting conversion of hyaline to pigmented material. At high glucose concentrations (20 to 60 mg/ml) the patterns of increase in total dry weight, microsclerotial material, and hyaline material were similar over a 4-week incubation period. Maximum production of both pigmented and hyaline materials occurred at a glucose concentration of 30 mg/ml (carbon/nitrogen ratio of 50/1).

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

Plant Disease ◽

10.1094/pdis.2002.86.3.327b ◽

2002 ◽

Vol 86 (3) ◽

pp. 327-327

Author(s):

S. O. Cacciola ◽

A. Pane ◽

S. Davino ◽

F. Raudino

Keyword(s):

Mating Type ◽

Irrigation System ◽

Field Capacity ◽

Industrial Area ◽

Selective Medium ◽

Phytophthora Palmivora ◽

Dual Culture ◽

Trickle Irrigation ◽

First Report ◽

Pure Cultures

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.

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First Report of Phytophthora palmivora as a Pathogen of Olive in Italy

Plant Disease ◽

10.1094/pdis.2000.84.10.1153a ◽

2000 ◽

Vol 84 (10) ◽

pp. 1153-1153 ◽

Cited By ~ 6

Author(s):

S. O. Cacciola ◽

G. E. Agosteo ◽

A. Pane

Keyword(s):

Southern Italy ◽

Glucose Dehydrogenase ◽

Selective Medium ◽

Phytophthora Palmivora ◽

Infested Soil ◽

Banding Patterns ◽

First Report ◽

Breadth Ratio ◽

Pathogenicity Tests ◽

Olive Trees

Olive (Olea europea L.) is an economically important crop in Italy and is planted on about 1 million ha. The Apulia, Calabria, and Sicily regions of Southern Italy account for about 70% of the production. Many new plantations have been established during the last 10 years. In summer 1999, 1- to 2-year-old olive trees (cv. Carolea) with decline symptoms were observed in new plantations in Catanzaro Province (Calabria). The symptoms associated with the root rot were leaf chlorosis, defoliation, wilting, twig dieback, and eventual plant collapse. In some cases, more than 40% of the trees were affected. A Phytophthora sp. was isolated consistently from rotted rootlets of diseased trees using a selective medium (2). Single-zoospore isolates were obtained from the colonies. The species isolated from olive roots was identified as P. palmivora (E. Butler) E. Butler on the basis of morphological and cultural characters according to Erwin and Ribeiro (1). All isolates produced papillate sporangia, which were elliptical to ovoid, caducous (mean pedicel length = 5 µm), with a length-breadth ratio of 1.8. In addition, some isolates produced subglobose, non-papillate sporangia with a length-breadth ratio ranging from 1.2 to 1.5. Electrophoresis of mycelial proteins on polyacrylamide gels confirmed that all isolates were pure cultures and that they all belonged to the same species. The electrophoretic banding patterns of total soluble mycelial proteins and eight isozymes (alkaline phosphatase, esterase, fumarase, NAD-glucose dehydrogenase, glucose-6-phosphate dehydrogenase, lactate dehydrogenase, malate dehydrogenase, and superoxide dismutase) of all olive isolates were identical to those of two strains of P. palmivora from parlor palm and from pittosporum. Conversely, they were clearly distinct from the electrophoretic banding patterns of reference isolates of P. cactorum (Lebert & Cohn) Schroter, P. capsici Leonian, P. citrophthora (R. E. Sm. & E. H. Sm.) Leonian, and P. nicotianae van Breda de Haan. All isolates of P. palmivora from olive were of the A1 mating type. The pathogenicity of four P. palmivora isolates from olive, two producing only typical and two producing both types of sporangia, was tested in the greenhouse at 18 to 25°C, using 20 1-year-old rooted cuttings of olive cv. Carolea for each isolate. Twenty noninoculated cuttings were used as a control. The inoculum for pathogenicity tests was produced on autoclaved rice grains moistened with V-8 juice. Cuttings were transplanted to pots filled with soil infested with inoculum at 2% (wt/vol). Control plants were grown in pots containing a mixture of soil and 2% autoclaved rice. After transplanting, all pots were flooded once a week for 24 h by plugging the drain hole of the pot. One month after planting, all the plants in infested soil had died and no difference in virulence was observed among the isolates. Control plants remained healthy. P. palmivora was reisolated from the roots of symptomatic plants. Pathogenicity tests were repeated twice with similar results. In a survey of nurseries in Southern Italy, P. palmivora was recorded frequently from roots of young olive trees suggesting that infections originated from nurseries. This is the first report from Italy of P. palmivora on olive. This species has been described recently as a pathogen of olive in Spain (3). References: (1) D. C. Erwin and O. K. Ribeiro. 1996. Phytophthora Diseases Worldwide. American Phytopathological Society. St. Paul, MN. (2) H. Masago et al. Phytopathology 67:25, 1977. (3) M. E. Sanchez Hernandez et al. Eur. J. Plant Pathol. 104:347, 1998.

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

Plant Disease ◽

10.1094/pd-90-0680a ◽

2006 ◽

Vol 90 (5) ◽

pp. 680-680 ◽

Cited By ~ 7

Author(s):

S. O. Cacciola ◽

D. Spica ◽

D. E. L. Cooke ◽

F. Raudino ◽

G. Magnano di San Lio

Keyword(s):

Selective Medium ◽

Phytophthora Species ◽

Width Ratio ◽

Broad Host Range ◽

Rdna Sequences ◽

Electrophoretic Patterns ◽

Pure Cultures ◽

Length Width ◽

Single Hypha ◽

Cardinal Temperatures

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.

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Bud and Root Rot of Windmill Palm (Trachycarpus fortunei) Caused by Simultaneous Infections of Phytophthora palmivora and P. nicotianae in Sicily

Plant Disease ◽

10.1094/pdis-11-10-0823 ◽

2011 ◽

Vol 95 (6) ◽

pp. 769-769 ◽

Cited By ~ 5

Author(s):

S. O. Cacciola ◽

A. Pane ◽

R. Faedda ◽

C. Rizza ◽

F. Badalà ◽

...

Keyword(s):

Root Rot ◽

Rhizosphere Soil ◽

Sprinkler Irrigation ◽

Its Region ◽

Selective Medium ◽

Phytophthora Species ◽

Phytophthora Palmivora ◽

Infested Soil ◽

Tissue Samples ◽

Reference Isolate

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.

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Assessment of Maternal Effects and Genetic Variability in Resistance to Verticillium dahliae in Olive Progenies

Plants ◽

10.3390/plants10081534 ◽

2021 ◽

Vol 10 (8) ◽

pp. 1534

Author(s):

Pedro Valverde Caballero ◽

Carlos Trapero Ramírez ◽

Diego Barranco Navero ◽

Francisco J. López-Escudero ◽

Ana Gordon Bermúdez-Coronel ◽

...

Keyword(s):

Verticillium Wilt ◽

Verticillium Dahliae ◽

Control Method ◽

Integrated Management ◽

Genetic Resistance ◽

Resistance Mechanisms ◽

Plant Diseases ◽

Conidial Suspension ◽

Reciprocal Crosses ◽

Olive Trees

The use of genetic resistance is likely the most efficient, economically convenient and environmentally friendly control method for plant diseases, as well as a fundamental piece in an integrated management strategy. This is particularly important for woody crops affected by diseases in which mainly horizontal resistance mechanisms are operative, such as Verticillium wilt, caused by Verticillium dahliae. In this study, we analyzed the variability in resistance to Verticillium wilt of olive trees in progenies from five crosses: ‘Picual’ × ‘Frantoio’, ‘Arbosana’ × ‘Koroneiki’, ‘Sikitita’ × ‘Arbosana’, ‘Arbosana’ × ‘Frantoio’ and ‘Arbosana’ × ‘Arbequina’ and their respective reciprocal crosses. Additionally, seedlings of ‘Picual’ and ‘Frantoio’ in open pollination were used as controls. In October 2016 and 2018, the fruits were harvested, and seeds germinated. Six-week-old seedlings were inoculated by dipping their bare roots in a conidial suspension of V. dahliae, and disease progress in terms of symptom severity and mortality was evaluated weekly. Additionally, seedling growth was evaluated every two weeks. At the end of the experiment, no significant differences were found for any of the assessed parameters when reciprocal crosses were compared. These results suggest that there is no maternal or paternal effect in regard to the heritability of resistance. In addition, this study identifies the best crosses for obtaining the highest number of resistant genotypes, highlighting the importance of the selection of specific cultivars to optimize the breeding process.

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The Influence of Biochar and Solid Digestate on Rose-Scented Geranium (Pelargonium graveolens L’Hér.) Productivity and Essential Oil Quality

Agronomy ◽

10.3390/agronomy9050260 ◽

2019 ◽

Vol 9 (5) ◽

pp. 260 ◽

Cited By ~ 5

Author(s):

Alessandro Calamai ◽

Enrico Palchetti ◽

Alberto Masoni ◽

Lorenzo Marini ◽

David Chiaramonti ◽

...

Keyword(s):

Essential Oil ◽

Climate Adaptation ◽

Oil Quality ◽

Dry Weight ◽

Chemical Components ◽

Growth Media ◽

Anaerobic Digestate ◽

Pelargonium Graveolens ◽

Positive Effects ◽

Leaf Chlorosis

In recent years, biochar has generated global interest in the areas of sustainable agriculture and climate adaptation. The main positive effects of biochar were observed to be the most remarkable when nutrient-rich feedstock was used as the initial pyrolysis material (i.e., anaerobic digestate). In this study, the influence of solid anaerobic digestate and biochar that was produced by the slow pyrolysis of solid digestate was evaluated by comparing the differences in the crop growth performances of Pelargonium graveolens. The experiment was conducted in a greenhouse while using three different growth media (i.e., solid digestate, biochar, and vermiculite). The results indicated that: (i) the pyrolysis of solid digestate caused a reduction in the bulk density (−52%) and an increase in the pH (+16%) and electrical conductivity (+9.5%) in the derived biochar; (ii) the best crop performances (number of leaves, number of total branches, and plant dry weight) were found using biochar, particularly for plant dry weight (+11.4%) and essential oil content (+9.4%); (iii) the essential oil quality was slightly affected by the growth media; however, the main chemical components were found within the acceptable range that was set by international standard trade; and, iv) biochar induced the presence of leaf chlorosis in Pelargonium graveolens.

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First Report of Armillaria mellea on a Fern from Italy

Plant Disease ◽

10.1094/pdis.2000.84.5.592c ◽

2000 ◽

Vol 84 (5) ◽

pp. 592-592 ◽

Cited By ~ 3

Author(s):

S. Grasso ◽

A. Pane ◽

S. O. Cacciola

Keyword(s):

Pure Culture ◽

Late Summer ◽

The United States ◽

Selective Medium ◽

Mount Etna ◽

Washington Dc ◽

First Report ◽

Reference Isolate ◽

Ornamental Foliage Plants ◽

Electrophoretic Patterns

Several perennial species of rhizomatous herbaceous ferns are cultivated as ornamental foliage plants. During late summer 1999, in a garden at the foot of Mount Etna, eastern Sicily (Italy), we noted a fern hedge showing patches of withered or stunted plants. The fern was identified as Cyrtomium falcatum (L.f.) C. Presl. (=Polystichum falcatum (L.f.) Diels), a house holly fern or Japanese holly fern, which is an ornamental fern native to East and South Asia. Other woody plants in the immediate vicinity had died over the last few years, including apricot and cedar trees whose stumps had not been removed. A close examination of uprooted ferns revealed the presence of creamy white fan-shaped mycelial mats with an odor typical of Armillaria species that were intermixed with the felt-like tangle formed by the rhizomes and roots of the ferns. In autumn, clumps of honey mushrooms with an annulus grew around patches of the withered fern hedge and in other parts of the same garden. The spore print of the basidiocarp was light cream. Basidiospores (8 to 9 × 5 to 6.5 µm) examined under a microscope were hyaline and apiculate. The fungus was isolated in pure culture from infected rhizomes with the selective medium of Kulman and Hendrix (3). In pure culture on 2% malt agar, the fungus formed ribbon-shaped, contorted, fast-growing rhizomorphs that branched profusely. Mycelial proteins of the isolate were analyzed by both polyacrylamide slab gel and starch gel electrophoreses, as described by Bragaloni et al. (1). The electrophoretic patterns of five isozymes (esterase, glutamic oxalacetic transaminase, phospho-glucomutase, alcohol dehydrogenase, and polygalacturonase) of the isolate from fern were identical to those of the reference isolate of A. mellea (Vahl:Fr.) Kumm. from grapevine. Conversely, the patterns were clearly distinct from those of reference isolates from other species, including A. ostoyae (Romagnesi) Herink, A. bulbosa (Barla) Kile et Watling, and A. cepistipes Velenovsky. Thus, on the basis of cultural, morphological, and biochemical characteristics, the species infecting the fern was identified as A. mellea. This pathogen, very common and widespread on wooded or previously wooded sites, has an extremely wide host range, encompassing both woody and herbaceous plants (2,4). However, this is the first report of A. mellea on a fern in Italy. References: (1) M. Bragaloni and N. Anselmi. Eur. J. For. Pathol. 27:147, 1997. (2) D. F. Farr et al. 1989. Fungi on Plants and Plants Products in the United States. The American Phytopathological Society, St. Paul, MN. (3) E. G. Kulman and F. F. Hendrix. Phytopathology 52:1310, 1962. (4) C. G. Shaw and G. A. Kile. 1991 Armillaria root disease. Agric. Handb. No 691. U.S. Department of Agriculture Forest Service, Washington, DC.

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