First Report of Fusarium oxysporum Causing Wilt on Iceland Poppy (Papaver nudicaule) in Italy

During fall 2011, symptoms of a wilt disease were observed in a commercial nursery near Ventimiglia as well as in the Research Center of Floriculture of Sanremo (northern Italy) on plants of Papaver nudicaule (Iceland poppy) of a local unnamed cultivar. In the commercial nursery, 15 to 20% of plants were affected, while about 3% of plants were affected at the Research Center. Symptoms consisted of chlorosis, premature leaf drop, and foliar wilting, followed by the stem wilting, bending, and eventually rotting from the base. Brown discoloration was observed in the stem vascular tissue. Using Komada's Fusarium-selective agar medium (2), a fungus was consistently and readily isolated from symptomatic vascular tissue of plants collected from both sites. The isolates were purified and subcultured on potato dextrose agar (PDA), on which medium both isolates produced pale violet, abundant, aerial mycelium, felted in old cultures, with pale purple pigments in the agar medium. The isolate generated short monophialides with unicellular, ovoid-elliptical microconidia of 3.9 to 6.7 × 1.4 to 3.0 (average 5.4 × 2.3) μm. On carnation leaf agar (CLA) (1), isolates produced pale orange sporodochia with macroconidia that were 3-septate, slightly falcate with a foot-shaped basal cell and a short apical cell, and 26.0 to 43.5 × 3.1 to 4.4 (average 35.3 × 3.7) μm. Chlamydospores were abundant, terminal, and intercalary, rough walled, mostly singles but sometime in short chains or clusters, and 5.2 to 10.1 μm in diameter. Such characteristics are typical of Fusarium oxysporum (3). The internal transcribed spacer (ITS) region of rDNA was amplified from the isolates using the primers ITS1/ITS4 (4), and sequenced. BLASTn analysis of the 507-bp ITS sequence of one isolate from P. nudicaule collected from the commercial nursery (GenBank Accession No. JX103564) showed an E-value of 0.0 and 100% identity with the ITS sequence of F. oxysporum (HQ649820). To confirm pathogenicity of one of the Iceland poppy isolates, tests were conducted on 2-month-old plants of the same cultivar on which symptoms were first observed. Plants (n = 14) were inoculated by dipping roots in a 1 × 107 CFU/ml conidial suspension of the isolate of F. oxysporum prepared from 10-day-old cultures grown in potato dextrose broth (PDB) on a shaker (90 rpm) for 10 days at 22 ± 1°C (12-h fluorescent light, 12-h dark). Non-inoculated control plants (n = 14) were dipped in sterilized water. All the plants were transplanted into pots filled with steamed potting mix (sphagnum peat/perlite/pine bark/clay at 50:20:20:10), and maintained in a glasshouse at 24 to 28°C. Inoculated plants showed typical symptoms of Fusarium wilt after 10 days. The stems then wilted and plants died. Non-inoculated plants remained healthy. F. oxysporum was reisolated from inoculated plants but not from control plants. The pathogenicity test was conducted twice with the same results. Since Fusarium wilt has not previously been described on Iceland poppy at any location, this is first report of F. oxysporum on P. nudicaule in Italy and anywhere in the world. References: (1) N. L. Fisher et al. Phytopathology 72:151, 1982. (2) H. Komada. Rev. Plant Prot. Res. 8:114, 1975. (3) J. F. Leslie and B.A. Summerell. The Fusarium Laboratory Manual, Blackwell Professional, IA, 2006. (4) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, CA, 1990.

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First Report of Fusarium Wilt of Lavandula pubescens Caused by Fusarium oxysporum in Saudi Arabia

Plant Disease ◽

10.1094/pdis-94-9-1163b ◽

2010 ◽

Vol 94 (9) ◽

pp. 1163-1163 ◽

Cited By ~ 2

Author(s):

K. Perveen ◽

N. Bokhari

Keyword(s):

Saudi Arabia ◽

Fusarium Oxysporum ◽

Vascular Tissue ◽

Agricultural Research ◽

Indian Agricultural Research Institute ◽

Pathogenicity Test ◽

Conidial Suspension ◽

Potential Threat ◽

King Saud University ◽

First Report

In November 2008, a wilt of lavender (Lavandula pubescens) seedlings was observed in the greenhouse at King Saud University, Riyadh, Saudi Arabia. Affected seedlings were wilted and the root system was poorly developed. Diseased stems developed a dark coloration that extended down to the roots. Vascular tissue of the affected seedlings appeared red or brown. Isolations consistently yielded a fungus growing from the discolored stem tissue when placed on potato dextrose agar. The macroscopic characteristics of the colony, as well as microscopic structures, were used to identify the fungus as Fusarium oxysporum (2). Oval to elliptical microconidia without septa and originating from short phialides were used to distinguish the species from F. solani (1). The fungus was authenticated by the ITCC (Indian Type Collection Centre), Indian Agricultural Research Institute, New Delhi, India, and given I.D. No. 7532.09. For conducting further experiments, healthy seedlings of L. pubescens were obtained from the botanical garden of the King Saud University and grown in steam-sterilized soil. Healthy seedlings of lavender were inoculated using a root-dip method with a conidial suspension (1 × 107 CFU/ml) of one strain of F. oxysporum obtained from infected plants. Inoculated seedlings were then transplanted into steam-sterilized soil. Plants inoculated with sterilized water (1 ml per plant) served as control treatments. Wilt symptoms and vascular discoloration in the roots and crown developed within 20 days on all plants inoculated with the pathogen, while control plants remained asymptomatic. F. oxysporum was consistently reisolated from symptomatic plants. The pathogenicity test was conducted twice. To our knowledge, this is the first report of F. oxysporum on L. pubescens in Saudi Arabia or elsewhere in the world, and this newly identified disease may be a potential threat to commercial production of lavender. References: (1) J. F. Leslie and B. A. Summerell. Page 212 in: The Fusarium Laboratory Manual. Blackwell Publishing Professional, Hoboken, NJ, 2006. (2) P. C. Nelson et al. Clin. Microbiol. Rev. 7:479, 1994.

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First Report of Fusarium Wilt Caused by Fusarium oxysporum on Strawberry in Spain

Plant Disease ◽

10.1094/pdis-93-3-0323b ◽

2009 ◽

Vol 93 (3) ◽

pp. 323-323 ◽

Cited By ~ 22

Author(s):

F. T. Arroyo ◽

Y. Llergo ◽

A. Aguado ◽

F. Romero

Keyword(s):

Fusarium Oxysporum ◽

Fusarium Wilt ◽

Potato Dextrose Agar ◽

Fluorescent Light ◽

Pathogenicity Test ◽

Conidial Suspension ◽

Soilless Culture ◽

Fragaria Ananassa ◽

Pcr Assay ◽

First Report

In the spring of 2007, wilted and dead strawberry plants (Fragaria × ananassa Duch. cvs. Camarosa and Ventana) were observed in a soilless culture system in Huelva, southwestern Spain. Approximately 8% of the plants in the field died. Isolations from necrotic crowns and roots and necrotic flowers were made on potato dextrose agar after disinfestation in 0.6% NaOCl for 30 s. Colonies with light purple mycelia and beige or orange reverse colony colors developed after 9 days of incubation at 25°C. Colonies produced abundant microconidia, macroconidia, and chlamydospores. Microconidia were hyaline and oval-ellipsoid to cylindrical (5.9 to 9.2 × 2.1 to 3.4 μm). Macroconidia were 3 to 5 septate and fusoid-subulate with a pedicellate base (28.8 to 37.3 × 3.2 to 4.3 μm). Morphology and growth matched descriptions of Fusarium oxysporum Schlechtend emend. Snyder & Hansen (2). A PCR assay for amplification of r-DNA using primers PFO2 and PFO3 established the identity of the isolate as F. oxysporum (1). To confirm the pathogenicity of the fungus, roots of 30-day-old strawberry cvs. Camarosa and Ventana (20 plants each) were inoculated by dipping the roots into a conidial suspension (107 conidia per ml) for 15 min. The inoculated plants were transplanted into plastic pots containing sterilized peat and maintained at 25°C and 100% relative humidity in a growth chamber with a daily 12-h photoperiod of fluorescent light. The pathogenicity test was conducted twice. Within 30 days, all inoculated plants developed wilt symptoms similar to that observed in the field and eventually 75% of the plants died. No symptoms were observed on plants dipped in distilled water. The fungus was successfully reisolated from crowns, roots, and necrotic flowers, fulfilling Koch's postulates. To our knowledge, this is the first report of the occurrence of Fusarium wilt caused by F. oxysporum on strawberry plants in Spain. References: (1) V. Edel et al. Mycol. Res. 104:518, 2000. (2) W. C. Snyder and H. N. Hansen. Am. J. Bot. 27:64, 1940.

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First Report of Fusarium oxysporum f. sp. lycopersici Race 2 on Tomato in Italy

Plant Disease ◽

10.1094/pdis.1999.83.10.967b ◽

1999 ◽

Vol 83 (10) ◽

pp. 967-967 ◽

Cited By ~ 1

Author(s):

V. M. Stravato ◽

R. Buonaurio ◽

C. Cappelli

Keyword(s):

Fusarium Oxysporum ◽

Fusarium Wilt ◽

Vascular Tissue ◽

Morphological Features ◽

Pathogenicity Test ◽

First Report ◽

Fungal Isolates ◽

Race 1 ◽

Race 2 ◽

I Gene

During the summer of 1997, symptoms of Fusarium wilt were observed on tomato (Lycopersicon esculentum Mill.) cvs. Monica F1 and PS 110, which bear the I gene for resistance to race 1 of Fusarium oxysporum Schlechtend.:Fr. f. sp. lycopersici (Sacc.) W.C. Snyder & H.N. Hans., in two commercial production greenhouses in Latium (Fondi) and one greenhouse in Sardinia (Oristano). Infected plants showed yellowing, stunting, vascular discoloration, and premature death. A fungus from tomato stems with discolored vascular tissue was consistently isolated on potato dextrose agar (PDA) and, based on morphological features, was identified as F. oxysporum. To verify the pathogenicity of four fungal isolates, cv. Bonny Best tomato plants, which do not carry genes for Fusarium wilt resistance, were inoculated by dipping roots of 2-week-old seedlings in a suspension of 105 microconidia per ml for 30 s. Inocula were obtained from 1-week-old fungal cultures grown on PDA. Roots of control plants were dipped in water. Seedlings were transplanted to pots containing peat and river sand (1:1, vol/vol) and placed in a greenhouse at 20 to 25°C. One month after inoculation, all fungal isolates provoked wilting of inoculated plants. No symptoms were observed on control plants. The morphological features of the fungus reisolated from diseased plants were similar to those of the original isolates. Based on the pathogenicity test, we concluded that the fungal isolates belong to F. oxysporum f. sp. lycopersici. To determine the races of the fungal isolates, differential tomato lines VFN8 (I gene for resistance to race 1), Florida MH-1 (I and I2 genes for resistance to races 1 and 2), and I3R (I, I2, and I3 genes for resistance to races 1, 2, and 3) were inoculated with the four fungal isolates, using the same procedure described for the pathogenicity test. Because disease symptoms were detected on VFN8 but not on Florida MH-1 and I3R, we deduced that the fungal isolates belong to F. oxysporum race 2. This is the first report of F. oxysporum f. sp. lycopersici race 2 in Italy. Previous research indicated that race 1 is present in Italy (1). Currently, many commercially acceptable cultivars resistant to races 1 and 2 are available to Italian greenhouse growers. Reference: (1) M. Cirulli. Phytopathol. Mediterr. 4:63, 1965.

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First Report of Crown and Stem Rot of Crested Molded Wax Agave (Echeveria agavoides) caused by Fusarium oxysporum in Italy

Plant Disease ◽

10.1094/pdis-08-12-0803-pdn ◽

2013 ◽

Vol 97 (2) ◽

pp. 288-288

Author(s):

A. Garibaldi ◽

D. Bertetti ◽

P. Pensa ◽

A. Poli ◽

M. L. Gullino

Keyword(s):

Fusarium Oxysporum ◽

Apical Cell ◽

The United States ◽

Stem Rot ◽

Economic Losses ◽

Pathogenicity Test ◽

Conidial Suspension ◽

Potato Dextrose Broth ◽

First Report ◽

Leaf Tissues

During March 2012, 95% of 24-month-old plants of crested molded wax agave (Echeveria agavoides), a succulent plant belonging to the Crassulaceae family, showed symptoms of a basal stem and leaf rot in a commercial farm near Ventimiglia (northern Italy). Affected plants showed extensive chlorosis from the crown level to the stem apex, followed by yellowing and by the appearance of a water-soaked aspect of stem and leaf tissues. As disease progressed, leaves became brown, wilted, and rotted. Wilting was at first unilateral and later affected the entire plant. Brown discoloration was observed in the vascular system of cut stems and leaves. In some cases, leaves were covered by a whitish-orange mycelium. This produced 3-septate, slightly curved macroconidia with a foot-shaped basal cell and a short apical cell, measuring 27.4 to 39.6 × 3.0 to 4.1 (average 34.2 × 3.7) μm and unicellular, ovoid to elliptical microconidia measuring 4.8 to 11.6 × 1.5 to 3.7 (avg. 7.2 × 2.7) μm. A fungus was consistently isolated from discolored vascular leaf tissues on Komada selective medium. Cultures on potato dextrose agar (PDA) and carnation leaf-piece agar (CLA) were incubated at 24 to 29°C. On PDA, a thin growth of whitish mycelium without pigments in the agar was observed. On CLA, sparse macroconidia, 18.9 to 30.7 × 3.0 to 4.2 (avg. 23.9 × 3.6) μm, microconidia, 4.7 to 7.7 × 1.7 to 3.1 (avg. 6.0 × 2.4) μm, and abundant chlamydospores that were single or paired, terminal and intercalary, rough walled, and 6.8 to 9.5 (avg. 7.7) in diameter were produced. Such characteristics are typical of Fusarium oxysporum (2). Amplification of the internal transcribed spacer (ITS) of the rDNA using primers ITS1/ITS4 (3) yielded a 486-bp band (GenBank Accession No. JX441893). Sequencing and BLASTn analysis of this band showed 100% identity and an E-value of 0.0 with the ITS sequence of F. oxysporum (JN232163). To confirm pathogenicity, five 3-month-old healthy plants of E. agavoides were inoculated by dipping unwounded roots in a conidial suspension (1.0 × 107 CFU/ml) of one isolate of F. oxysporum obtained from affected plants, grown for 10 days in potato dextrose broth. Plants were transplanted into pots filled with steam-sterilized substrate (sphagnum peat-perlite-pine bark-clay 50:20:20:10) and maintained in a glasshouse at 28 to 33°C. Five non-inoculated plants served as a control. Chlorosis and yellowing developed on the inoculated plants 15 days after the inoculation. Basal stem rot and vascular discoloration in the crown and stem developed within 30 days on inoculated plants. A whitish-orange mycelium producing macroconidia covered the affected leaves. Non-inoculated plants remained healthy. F. oxysporum was consistently reisolated from symptomatic plants. The pathogenicity test was conducted twice. A Fusarium sp. has been reported as the causal agent of a stem rot on Echeveria sp. in the U.S. (1). To our knowledge, this is the first report of F. oxysporum on E. agavoides in Italy. The disease is currently present in few nurseries, although it could spread, causing significant economic losses due to the increasing cultivation of E. agavoides in Italy. References: (1) D. F. Farr et al. Fungi on Plants and Plant Products in the United States. APS Press, St. Paul, MN, 1989. (2) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell, Ames, IA, 2006. (3) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, 1990.

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First Report of Fusarium Wilt of Cilantro Caused by Fusarium oxysporum in California

Plant Disease ◽

10.1094/pd-89-1130a ◽

2005 ◽

Vol 89 (10) ◽

pp. 1130-1130 ◽

Cited By ~ 3

Author(s):

S. T. Koike ◽

T. R. Gordon

Keyword(s):

Fusarium Oxysporum ◽

Fusarium Wilt ◽

Vascular Tissue ◽

Fusarium Species ◽

Wilt Disease ◽

Apium Graveolens ◽

Peat Moss ◽

State University ◽

First Report ◽

Vascular Discoloration

Cilantro, or coriander (Coriandrum sativum), is a leafy vegetable in the Apiaceae and is grown commercially in California primarily for use as a fresh herb. During 2002 and 2003 in coastal California (Santa Barbara County), commercial cilantro fields showed symptoms of a wilt disease. Affected plants grew poorly and were stunted. Lower foliage turned yellow with reddish tinges, and plants wilted during warmer times of the day. The main stem, crown, and taproot exhibited vascular discoloration that was reddish to light brown. As disease progressed, plants eventually died. For both years, the disease distribution was limited to isolated small patches (each patch measuring less than 1 m2 in area). A fungus was consistently isolated from symptomatic vascular tissue in crowns and taproots. On the basis of colony and conidial morphology, the isolates were identified as Fusarium oxysporum (2). No other fungi or bacteria were recovered from these plants. To test pathogenicity, suspensions containing 1 × 106 conidia/ml were prepared for five isolates. The roots of 30-day-old cilantro plants of four cultivars (30 plants each of Festival, Leisure, Santo, and LSO 14) were clipped and then soaked in the suspensions for 20 min. The roots of 30 plants of each cultivar were soaked in water as a control. Plants were repotted into new redwood bark + peat moss rooting medium and maintained in a greenhouse setting at 24 to 26°C. After 1 month, 95% or more of the inoculated plants showed yellowing and vascular discoloration symptoms similar to those seen in the field. F. oxysporum was reisolated from all inoculated plants. The four cilantro cultivars did not show differences in disease severity. Control plants showed no symptoms, and the fungus was not recovered from these plants. The experiment was repeated and the results were the same. Experiments also were conducted to determine if cilantro isolates could cause disease in celery (Apium graveolens var. dulce). Celery transplants and cilantro seedlings were prepared and inoculated as described above. However, after 2 months, celery plants did not show any disease symptoms, while the cilantro developed wilt symptoms and eventually died. A Fusarium wilt disease has been reported on coriander in Argentina and India where the pathogen was named F. oxysporum f. sp. coriandrii (1,3). To our knowledge, this is the first report of Fusarium wilt of cilantro in California. References: (1) M. Madia et al. Fitopatologia 34:155, 1999. (2) P. E. Nelson et al. Fusarium species: An Illustrated Manual for Identification. Pennsylvania State University Press, University Park, 1983. (3) U. S. Srivastava. Indian Phytopathol. 22:406, 1969.

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First Report of Fusarium Wilt on Philotheca myoporoides Caused by Fusarium oxysporum in Italy

Plant Disease ◽

10.1094/pdis-03-11-0243 ◽

2011 ◽

Vol 95 (7) ◽

pp. 877-877 ◽

Cited By ~ 1

Author(s):

G. Polizzi ◽

D. Aiello ◽

V. Guarnaccia ◽

A. Vitale ◽

G. Perrone ◽

...

Keyword(s):

Fusarium Oxysporum ◽

Fusarium Wilt ◽

Morphological Characteristics ◽

Plant Production ◽

Pathogenicity Test ◽

Gene Sequences ◽

Pcr Assay ◽

First Report ◽

Gene Coding ◽

Eastern Australia

Philotheca myoporoides (DC.) M.J. Bayly (previously known as Eriostemon myoporoides), commonly called long-leaf waxflower and native to eastern Australia (Rutaceae family), is a hardy compact shrub or small tree occurring in subtropical to cool temperate regions. P. myoporoides is cultivated in Sicily (Italy) for its ornamental appeal. During April of 2010, a widespread wilting was observed on approximately 80% of 2,000 1-year-old, potted long-leaf waxflower plants grown in a commercial nursery near Catania (eastern Sicily, Italy). Internally, symptomatic plants had conspicuous vascular brown discoloration from the crown to the canopy. Diseased crown and stem tissues of 20 plants were surface disinfested for 30 s in 1% NaOCl, rinsed in sterile water, plated on potato dextrose agar (PDA) amended with 100 mg/liter of streptomycin sulfate, and incubated at 25°C. A Fusarium sp. was consistently isolated from affected plant tissues. Colonies with white or light purple aerial mycelia and violet pigmentation on the underside of the cultures developed after 9 days. On carnation leaf agar, 20 single-spore isolates produced microconidia on short monophialides, macroconidia that were three to five septate with a pedicellate base, and solitary and double-celled or aggregate chlamydospores. A PCR assay was conducted on one representative isolate (ITEM 13490) by analyzing sequences of the benA gene (coding β-tubulin protein) and CaM gene (coding calmodulin protein) using the primers reported by O'Donnell et al. (1). The benA gene sequences of ITEM 13490 (GenBank No. FR828825) exhibited an identity of 100% to Fusarium oxysporum f. sp. radicis-lycopersici strain ATCC 52429 (GenBank No. DQ092480). CaM gene sequences of ITEM 13490 (GenBank No. FR828826) exhibited an identity of 99.6% to F. oxysporum strain ITEM 2367 (GenBank No. AJ560774). Morphological characteristics of the 20 isolates, as well as the PCR assay on a representative strain, identified the isolates associated with disease symptoms as F. oxysporum Schlechtend.:Fr. A pathogenicity test was performed by placing two 1-cm2 plugs of PDA from 9-day-old mycelial cultures near the crown on potted, healthy, 2-month-old cuttings of P. myoporoides. Thirty plants were inoculated with strain ITEM 13490 and the same number of plants served as noninoculated controls. All plants were enclosed for 4 days in plastic bags and placed in a growth chamber at 25 ± 1°C. Plants were then moved to a greenhouse where temperatures ranged from 23 to 27°C. First symptoms, which were identical to those observed in the nursery, developed on one plant 15 days after inoculation. Wilting was detected on all plants after 30 days. Control plants remained symptomless. F. oxysporum was successfully reisolated from symptomatic crown and stem tissues and identified as described above, fulfilling Koch's postulates. To our knowledge, this is the first report of F. oxysporum causing disease of P. myoporoides worldwide. Moreover, this pathogen was recently reported in the same nursery on Eremophila sp. (2), confirming the presence of Fusarium wilt as a potential threat to ornamental plant production in this area, and necessitates the innovation and development of disinfection methods for alveolar trays, greenhouses, and various propagation materials to reduce future disease outbreaks. References: (1) K. O'Donnell et al. Mycoscience 41:61, 2000. (2) G. Polizzi et al. Plant Dis 94:1509, 2010.

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Fusarium Wilt of African Daisy (Osteospermum sp.) Caused by Fusarium oxysporum in Italy

Plant Disease ◽

10.1094/pdis.2004.88.3.309a ◽

2004 ◽

Vol 88 (3) ◽

pp. 309-309 ◽

Cited By ~ 4

Author(s):

A. Garibaldi ◽

A. Minuto ◽

M. L. Gullino

Keyword(s):

Fusarium Oxysporum ◽

Vascular Tissue ◽

Vascular System ◽

Selective Medium ◽

Northern Italy ◽

Wilt Disease ◽

Pathogenicity Test ◽

Conidial Suspension ◽

Average Temperature ◽

Brown Discoloration

During the fall of 2002, African daisy (Osteospermum sp.) plants showing symptoms of a wilt disease were observed in a commercial, nonheated glasshouse in Albenga in northern Italy. Wilted plants were first observed when outside temperatures were between 15 and 28°C. Symptoms were first observed on seedlings 40 days after they had been transplanted into pots. The vascular tissues of affected plants appeared brown. These plants were stunted and developed yellowed leaves with brown or black streaks in the vascular system. The vascular streaks in the yellow leaves extended from the crown and were continuous with a brown discoloration in the vascular system of the crown and upper taproot. Fusarium oxysporum was consistently and readily isolated from symptomatic vascular tissue onto a Fusarium-selective medium (1). Healthy, rooted, 40-day-old plants were inoculated by root-dip with a conidial suspension (1 × 107 CFU/ml) of three isolates of F. oxysporum obtained from infected plants and transplanted into pots filled with steam-sterilized soil. Noninoculated plants served as control treatments. Plants (10 per treatment) were grown in a glasshouse at an average temperature of 25°C (minimum of 12°C and maximum of 39°C). Wilt symptoms and vascular discoloration in the roots, crown, and veins developed within 20 days on each inoculated plant, while noninoculated plants remained healthy. F. oxysporum was consistently reisolated from infected plants. The pathogenicity test was conducted twice. To our knowledge, this is the first report of F. oxysporum on Osteospermum sp. in Italy or elsewhere in the world. Reference: (1) H. Komada. Rev. Plant Prot. Res. 8:114, 1975.

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First Report of Verticillium Wilt caused by Verticillium dahliae Kleb. on New Zealand Spinach (Tetragonia tetragonioides) in Italy

Plant Disease ◽

10.1094/pdis-07-12-0678-pdn ◽

2013 ◽

Vol 97 (1) ◽

pp. 145-145

Author(s):

A. Garibaldi ◽

S. Rapetti ◽

P. Martini ◽

L. Repetto ◽

D. Bertetti ◽

...

Keyword(s):

New Zealand ◽

Verticillium Wilt ◽

Verticillium Dahliae ◽

Vascular Tissue ◽

Spinacia Oleracea ◽

Morphological Characteristics ◽

Conidial Suspension ◽

Potato Dextrose Broth ◽

First Report ◽

Tetragonia Tetragonioides

Tetragonia tetragonioides (New Zealand spinach, Aizoaceae) is an Australasian annual species that occurs naturally in Italy, where it is cultivated for the edible young shoots and succulent leaves. In September 2011, a previously unknown wilt was observed in 10 private gardens, each 0.1 to 0.5 ha, near Castellaro, Northern Italy, on 7-month-old New Zealand spinach plants. Leaves wilted, starting from the collar and moving up the plant, and vascular tissues showed brown streaks in the roots, crowns, and stems. Diseased plants were stunted with small, chlorotic leaves. Infected stems and leaves then wilted, and plants often died. Of about 500 plants, 30% were affected. Stems of 10 diseased plants were disinfected with 1% NaOCl for 1 min. Sections of symptomatic vascular tissue were plated on potato dextrose agar. After 3 days at 23 ± 1°C, colonies developed that were white and turned a grey to dark green color. Irregular, black microsclerotia (32.0) 63.1 ± 16.8 μm (106.1) × (18.7) 39.1 ± 12.3 μm (65.8) developed in hyaline hyphae after 8 days. Hyaline, elliptical, single-celled conidia (2.7) 3.8 ± 0.6 μm (4.8) × (1.9) 2.6 ± 0.5 μm (3.5) developed on verticillate conidiophores with three phialides at each node. Based on these morphological characteristics, the fungus was identified as Verticillium dahliae (1). The internal transcribed spacer (ITS) region of rDNA was amplified for one isolate using the primers ITS1/ITS4 (3) and sequenced (GenBank Accession No. JX308315). BLASTn analysis of the 479-bp segment showed 100% homology with the ITS sequence of a V. dahliae isolate (AB551206). Pathogenicity tests were performed twice using 60-day-old plants of T. tetragonioides. Unwounded roots of eight plants were dipped for 1 min in a conidial suspension (5 × 107 conidia/ml) of one isolate of V. dahliae obtained from the original infected New Zealand spinach plants, and grown in potato dextrose broth. The inoculated plants were transplanted into 2-liter pots (1 plant/pot) containing steamed potting mix (sphagnum peat-perlite-pine bark-clay; 50:20:20:10) and maintained in a growth chamber at 20 to 24°C and 50 to 80% RH. Eight plants immersed in sterile water served as a control treatment. Wilt symptoms were observed 30 days after inoculation, with vascular discoloration in the roots, crowns and stems. V. dahliae was reisolated consistently from infected tissues, but not from the control plants that remained healthy. Pathogenicity was also tested using the same method on plants of four cultivars (five plants/cultivar) of Spinacia oleracea (Matador, Asti, Merlo Nero, and America). Wilt symptoms developed on all cultivars and V. dahliae was reisolated from each inoculated plant. No fungal colonies were reisolated from control plants, which remained healthy. To our knowledge, this is the first report of Verticillium wilt caused by V. dahliae on T. tetragonioides in Italy, as well in Europe. V. dahliae was reported on T. tetragonioides in Canada (2). At this time, the economic impact of Verticillium wilt on New Zealand Spinach in Italy is limited, although the use of this vegetable in Italy is increasing. References: (1) G. F. Pegg and B. L. Brady. Verticillium Wilts. CABI Publishing, Wallingford, UK, 2002. (2) M. J. Richardson. Page 387 in: An Annotated List of Seed-Borne Diseases, Fourth Edition. International Seed Testing Association, Zurich, Switzerland, 1990. (3) T. J. White et al. Page 315 in: PCR Protocols. A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.

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First Report of Fusarium Wilt in Blueberry (Vaccinium corymbosum) Caused by Fusarium oxysporum in China

Plant Disease ◽

10.1094/pdis-02-14-0167-pdn ◽

2014 ◽

Vol 98 (8) ◽

pp. 1158-1158 ◽

Cited By ~ 1

Author(s):

Y. H. Liu ◽

T. Lin ◽

C. S. Ye ◽

C. Q. Zhang

Keyword(s):

Fusarium Oxysporum ◽

Infected Plant ◽

Morphological Characteristics ◽

Vaccinium Corymbosum ◽

Morphological Characterization ◽

Internal Transcribed Spacers ◽

Fluorescent Light ◽

Pathogenicity Test ◽

Conidial Suspension ◽

First Report

Blueberry (Vaccinium corymbosum) production is developing quickly in China with about 20,000 ha presently cultivated. In 2010 in Lin'an, Zhejiang Province, plants developed an apparently new disease of blueberry (cv. Duke) with symptoms consisting of wilting of foliage, stunting of plants, and reduced fruit yields. Internal vascular and cortical tissues of plant crowns showed a brown to orange discoloration. Approximately 3% of the plants in the commercial plantings were affected and eventually died after 50 to 60 days. Infected plant samples (stems and roots) collected from different fields were surface sterilized with 1.5% sodium hypochlorite for 2 min, rinsed in water, plated on 2% potato dextrose agar (PDA), and incubated at 25°C in the dark for 1 week. Single conidium cultures were consistently isolated and cultured on acidified PDA (APDA) for morphological characterization (1,2). Colonies were light with purple mycelia, and beige or orange reverse colony colors developed after 7 days incubation at 25°C. Colonies producing abundant microconidia and macroconidia. Microconidia were hyaline and oval-ellipsoid to cylindrical (3.9 to 9.6 × 1.1 to 3.4 μm). Macroconidia were 3 to 5 septate and fusoid-subulate with a pedicellate base (28.6 to 37.5 × 3.3 to 4.2 μm). Morphology and development of macroconidia and microconida were consistent with a description of Fusarium oxysporum Schltdl (1,2). The ribosomal internal transcribed spacers ITS1 and ITS2 of eight isolates were amplified using primers ITS1/ITS4 on DNA extracted from mycelium and nucleotide sequences showed 100% similarity to that of F. oxysporum. To confirm pathogenicity, 20 blueberry plants (cv. Duke) were inoculated by dipping the roots into a conidial suspension (107 conidia per ml) for 30 min. The inoculated plants were transplanted into pots containing sterilized peat and maintained at 25°C and 100% relative humidity in a growth chamber with a daily 12-h photoperiod of fluorescent light. The pathogenicity test was conducted twice. Within 40 days, all inoculated plants developed wilt symptoms similar to that observed in the field. No symptoms were observed on plants dipped into distilled water. The fungus was successfully re-isolated from crowns and roots cultured on APDA, exhibiting morphological characteristics identical to F. oxysporum (1,2), confirming Koch's postulates. To our knowledge, this is the first report of blueberry wilt caused by Fusarium. References: (1) P. M. Kirk et al. The Dictionary of the Fungi, 10th edition, page 159. CABI Bioscience, Wallingford, UK, 2008. (2) W. C. Snyder and H. N. Hansen. Am. J. Bot. 27:64, 1940.

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First Report of Basal Stem Rot of Apple Cactus (Cereus peruvianus monstruosus) Caused by Fusarium oxysporum in Italy

Plant Disease ◽

10.1094/pdis-03-11-0216 ◽

2011 ◽

Vol 95 (7) ◽

pp. 877-877

Author(s):

A. Garibaldi ◽

P. Pensa ◽

D. Bertetti ◽

A. Poli ◽

M. L. Gullino

Keyword(s):

United States ◽

Fusarium Oxysporum ◽

The United States ◽

Stem Rot ◽

Fluorescent Light ◽

Pathogenicity Test ◽

Conidial Suspension ◽

Single Spore ◽

Basal Stem Rot ◽

First Report

During the summer of 2010, 20% of 7,000 4-month-old plants of apple cactus (Cereus peruvianus monstruosus) showed symptoms of a basal stem rot in a commercial nursery located in Liguria (northern Italy). Affected plants showed yellow orange-to-pale brown color from the crown level to the stem apex and a water-soaked rot was observed on the stem starting from the base. Brown discoloration was observed in the vascular system. Eventually stems bent, plants collapsed and died, and affected tissues dried out. A Fusarium sp. was consistently and readily isolated from symptomatic tissue on Komada selective medium. Isolates were purified and subcultured on potato dextrose agar (PDA). Single-spore cultures on PDA, Spezieller Nährstoffarmer agar (SNA) (3), and carnation leaf-piece agar (CLA) (2) were incubated at 26 ± 1°C (12-h fluorescent light, 12-h dark). On PDA, cultures produced a thick growth of white-to-pink mycelium and pale pink pigments in the agar. On SNA, cultures produced short monophialides with unicellular, ovoid-elliptical microconidia measuring 4.3 to 8.2 × 2.3 to 3.8 (average 6.0 × 2.8) μm. Chlamydospores were abundant, single or paired, terminal and intercalary, rough walled, and 6 to 8 μm in diameter. On CLA, cultures produced orange sporodochia with macroconidia that were 3 to 4 septate, nearly straight with a foot-shaped basal cell and a short apical cell, and measured 31.1 to 51.5 × 4.4 to 3.5 (average 43.2 × 3.8) μm. Such characteristics are typical of Fusarium oxysporum (3). Amplification of the ITS (internal transcribed spacer) of the rDNA using primers ITS1/ITS4 (4) yielded a 498-bp band. Sequencing and BLASTn analysis of this band showed an E-value of 0.0 with F. oxysporum. The nucleotide sequence has been assigned GenBank Accession No. JF422071. To confirm pathogenicity, five 6-month-old healthy plants of C. peruvianus monstruosus were inoculated by dipping roots in a conidial suspension (2.4 × 106 CFU/ml) of F. oxysporum isolated from affected plants. Inoculum was obtained from pure cultures of three single-spore isolates grown for 10 days on casein hydrolysate liquid medium. Roots were not wounded before the inoculation. Plants were transplanted into pots filled with steam-sterilized substrate (sphagnum peat/perlite/pine bark/clay 50:20:20:10). Five noninoculated plants served as a control. Plants were placed in a climatic chamber at 25 ± 1°C (12-h fluorescent light, 12 h-dark). Basal stem rot and vascular discoloration in the crown and stem developed within 30 days on each inoculated plant. Noninoculated plants remained healthy. F. oxysporum was consistently isolated from symptomatic plants. The pathogenicity test was conducted twice. F. oxysporum has been reported on Cereus spp. in the United States (1). To our knowledge, this is the first report of F. oxysporum on C. peruvianus monstruosus in Italy as well as in Europe. Currently, this disease is present in a few nurseries in Liguria. References: (1) D. F. Farr et al. Fungi on Plants and Plant Products in the United States. The American Phytopathological Society, St Paul, MN, 1989. (2) N. L. Fisher et al. Phytopathology 72:151, 1982. (3) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell, Ames, IA, 2006. (4) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, 1990.

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