scholarly journals First Report of Verticillium Wilt Caused by Verticillium dahliae on Fava Bean in the United States

Plant Disease ◽  
2017 ◽  
Vol 101 (2) ◽  
pp. 384-384 ◽  
Author(s):  
C. L. Blomquist ◽  
S. Rooney-Latham ◽  
N. Barrera
Plant Disease ◽  
2011 ◽  
Vol 95 (7) ◽  
pp. 878-878 ◽  
Author(s):  
A. Garibaldi ◽  
D. Bertetti ◽  
A. Poli ◽  
M. L. Gullino

Coleus verschaffeltii Lem. (synonym C. blumei Benth., Plectranthus scutellaroides (L.) R. Br., and Solenostemon scutellarioides (L.) Codd), a perennial plant belonging to the Lamiaceae family, is used as a bedding plant for public gardens. The most popular cultivars produce speckled leaves of various colors. In October 2010, severe outbreaks of a previously unknown wilt were observed in a public garden at Torino (northern Italy) on 50 8-month-old plants. Plants were sprinkle irrigated. Initial symptoms were withering of leaves starting from the collar and brown streaks in the vascular tissue of roots, crown, and stem. Subsequently, infected tissues wilted and plants became stunted. Early leaf drop was observed and plants appeared bare, keeping few leaves only at the end of stems. Infected plants did not die but they lost the original ornamental aspect. Seventy percent of the plants were affected. Stems of 10 plants were disinfected with 1% sodium hypochlorite. Cross-sections through symptomatic vascular tissues were plated on potato dextrose agar amended with 25 ppm of streptomycin sulfate. After 10 days at 20 to 23°C, a fungus was consistently recovered from 90% of stems. Irregular, black microsclerotia, 29 to 76 × 14 to 52 (average 49 × 28) μm, developed in hyaline hyphae after 15 days of growth. Hyaline, elliptical, single-celled conidia, 3.9 to 7.2 × 1.7 to 2.8 (average 5.1 × 2.2) μm, developed on verticillate conidiophores with three phialides at each node. On the basis of these morphological characteristics, the fungus was identified as Verticillium dahliae (3). The internal transcribed spacer (ITS) region of rDNA was amplified using the primers ITS1/ITS4 (4) and sequenced. BLASTn analysis (1) of the 491-bp segment showed a 99% homology with the sequence of V. dahliae (Accession No. GU461634). The ITS nucleotide sequence of our isolate has been assigned the GenBank Accession No. JF704205. Pathogenicity tests were performed twice using 45-day-old plants obtained from seeds of C. verschaffeltii grown in 1-liter pots containing a 50:20:20:10 steamed mix of peat moss/pumice/pine bark/clay. Roots of 10 healthy plants were immersed in a conidial suspension (1.7 × 107 ml–1) of one culture of V. dahliae isolated from infected plants. Ten plants immersed in sterile water served as controls. Plants were maintained in a glasshouse at daily average temperatures between 20 and 28°C and relative humidity between 50 and 80%. First wilt symptoms and vascular discoloration in the roots, crown, and stems developed 20 days after inoculation. V. dahliae was consistently reisolated from infected vascular tissues of crown and stems of symptomatic plants. Noninoculated plants remained healthy. To our knowledge, this is the first report of Verticillium wilt on C. verschaffeltii in Italy. Verticillium wilt had been previously reported on S. scutellaroides in the United States (2). At this time, the economic importance of Verticillium wilt on C. verschaffeltii in Italy is limited. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) D. Farr et al. Fungi on Plants and Plant Products in the United States. The American Phytopathological Society. St Paul, MN, 1989. (3) G. F. Pegg and B. L. Brady. Verticillium Wilts. CABI Publishing, Wallingford, UK, 2002. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, 1990.


Plant Disease ◽  
2008 ◽  
Vol 92 (9) ◽  
pp. 1367-1367 ◽  
Author(s):  
A. Garibaldi ◽  
D. Bertetti ◽  
M. L. Gullino

Rudbeckia fulgida (common name orange coneflower) is an herbaceous perennial (Asteraceae) grown in full sun in perennial borders in gardens. At the end of the summer of 2007, in a public garden located in Turin (northern Italy), symptoms of vascular wilt and stunting were observed on approximately 80% of the plants grown in a mixed border. Initial symptoms were yellowing of external leaves and brown or black streaks in the vascular tissue of roots, crown, and leaves. A fungus was consistently and readily isolated on potato dextrose agar from symptomatic vascular tissue previously disinfested in 1% sodium hypochloride. Ovoid, dark microsclerotia, 41 to 108 μm, developed in hyaline hyphae after 10 days of growth at 23°C (12 h of light and 12 h of dark). Hyaline, elliptical, single-celled conidia, 3.2 to 7.3 × 2.1 to 3.7 (average 4.7 × 2.8) μm, developed on verticillate conidiophores. On the basis of these morphological characteristics, the fungus was identified as Verticillium dahliae (4). The internal transcribed spacer (ITS) region of rDNA was amplified using primers ITS4/ITS6 and sequenced. BLASTn analysis (1) of the 530 bp showed a 100% homology with the sequence of V. dahliae. The nucleotide sequence has been assigned GenBank Accession No. EU 627007. Healthy, 30-day-old R. fulgida plants were grown in a steam-disinfested mix of sphagnum peat:pomix:pine bark:clay (50:20:20:10) infested with a conidial suspension (1.5 × 106/ml) of three isolates of V. dahliae isolated from infected plants. Noninoculated plants served as controls. Plants (16 per treatment) were grown in pots (3 liter vol) and maintained in a glasshouse at temperatures between 22 and 25°C and relative humidity between 50 and 70%. First wilt symptoms and vascular discoloration in the roots, crown, and veins developed 17 days after inoculation. Noninoculated plants remained healthy. The pathogenicity tests were carried out twice. To our knowledge, this is the first report in Italy of Verticillium wilt on R. fulgida. Wilts caused by V. dahliae on R. laciniata in Poland (3) and V. albo-atrum on R. hirta in the United States (2) were previously reported. The importance and economic impact of this disease is currently limited but may increase because of the popularity of Rudbeckia spp. in private and public parks. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997. (2) D. F. Farr et al. Fungi on Plants and Their Products in the United States. The American Phytopathological Society, St Paul, MN, 1989. (3) B. Leski. Rocz. Nauk Roln. 253, 1974. (4) G. F. Pegg and B. L. Brady. Verticillium Wilts. CABI Publishing, Wallingford, UK, 2002.


Plant Disease ◽  
2002 ◽  
Vol 86 (1) ◽  
pp. 74-74
Author(s):  
R. T. McMillan ◽  
W. R. Graves ◽  
T. F. Wood ◽  
R. M. Leahy

Mature akee trees, Blighia sapida K. Koenig, in a local south Florida commercial orchard had wilt and dieback symptoms during spring 1999. A fungus isolated from the gray xylem root tissue on V8 agar was identified as Verticillium dahliae Klebahn at the Division of Plant Industry of the Florida Department of Agriculture and Consumer Services. Twenty akee seedlings were transplanted into 3.85-liter plastic pots and grown in a greenhouse at a daytime temperature of 28°C and nighttime temperature of 23°C. When plants were approximately 25 cm high, a 15-cm knife was used to sever roots in the four quadrants of each pot. Inoculum was made from a 2-week-old culture of V. dahliae on V8 agar and blended with 160 ml of sterile water, and 15 ml of this slurry was poured into the disturbed soil of each of 10 treated plants. A plate of uninoculated V8 agar was applied, as above, to 10 control plants. Plants were kept in the greenhouse. After 6 weeks, inoculated plants showed symptoms of leaf wilt, dieback and plant death. No symptoms were seen on control plants. V. dahliae was isolated directly from the gray vascular tissue of inoculated plants. The inoculation experiment was repeated three times, fulfilling Koch's postulates. To our knowledge, this is the first report of Verticillium dieback on B. sapida in the United States.


Plant Disease ◽  
2009 ◽  
Vol 93 (8) ◽  
pp. 841-841 ◽  
Author(s):  
L. Zhang ◽  
G. L. Zhang ◽  
X. Qian ◽  
G. Y. Li

Verticillium dahliae Kleb. causes Verticillium wilt in large numbers of crops all over the world. Common symptoms caused by the pathogen include yellowing, wilting of leaves, and discoloration in vascular tissue of the stem. In June 2007, symptoms of Verticillium wilt were observed in a grapevine (Vitis vinifera) field in the Shihezi Region of Xinjiang. To isolate the pathogen, stem segments (0.5 cm long) were surface sterilized with 1% HgCl2 for 1 min and then dipped in 70% ethanol for 10 s. The sterilized tissues were rinsed with sterile water and incubated in the dark for 7 days at 25°C on potato dextrose agar (PDA) medium. The fungus growing from the diseased tissue showed dark colonies that produced verticillate conidiophores with two to three layers with colorless, ovoid, unicellular conidia and small, black microsclerotia, which are characteristics of V. dahliae (3). To confirm its identity, ribosomal DNA fragments (regions ITS1, 5.8S rDNA, and ITS4) were PCR amplified with primer pair ITS1/ITS4 (4) and sequenced (GenBank Accession No. FJ475122). Sequential analysis revealed that the rDNA region of the fungus isolated from grapevine was identical to that of a Greek strain of V. dahliae (GenBank Accession No. AF104926). Furthermore, the specific fragment (1,500 bp) of nondefoliating pathotype of V. dahliae (1) was PCR amplified from 24 grapevine isolates of V. dahliae collected in Xinjiang, indicating that the V. dahliae pathogen from Xinjiang is a nondefoliating pathotype. To verify the causal role of the isolated fungus, pathogenicity assays were conducted on 1-year-old seedlings of the Centennial seedless cultivar. Trimmed roots were submerged in a conidial suspension (1 × 106 conidia/ml) for 30 min and sterile tap water was used as a control. The seedlings were transplanted into a pot containing 2:1 sterile mixture of peat/perlite (vol/vol). Plants were grown in a greenhouse at 25°C. Six Verticillium isolates were found to cause the same symptoms as in fields 50 days after inoculation. V. dahliae was successfully reisolated from the stems of inoculated plants. Control seedlings inoculated in sterile tap water remained healthy. Because grapevine (Vitis vinifera) is an economically important crop for fruit and winemaking material in Xinjiang, Verticillium wilt poses a threat. The disease has been previously reported in the United States (2), but to our knowledge, this is the first report from China. References: (1) E. Pérez Artés et al. Eur. J. Plant Pathol. 106:507, 2000. (2) W. C. Schnathorst and A. C. Goheen. Plant Dis. Rep. 61:909, 1977. (3) H. C. Smith. N. Z. J. Agric. Res. 8:450, 1965. (4) T. J. White et al. PCR Protocols. Academic Press, San Diego, CA, 1990.


2012 ◽  
Vol 102 (11) ◽  
pp. 1071-1078 ◽  
Author(s):  
Zahi K. Atallah ◽  
Karunakaran Maruthachalam ◽  
Krishna V. Subbarao

Since 1995, lettuce in coastal California, where more than half of the crop in North America is grown, has consistently suffered from severe outbreaks of Verticillium wilt. The disease is confined to this region, although the pathogen (Verticillium dahliae) and the host are present in other crop production regions in California. Migration of the pathogen with infested spinach seed was previously documented, but the geographic sources of the pathogen, as well as the impact of lettuce seed sparsely infested with V. dahliae produced outside coastal California on the pathogen population in coastal California remain unclear. Population analyses of V. dahliae were completed using 16 microsatellite markers on isolates from lettuce plants in coastal California, infested lettuce seed produced in the neighboring Santa Clara Valley of California, and spinach seed produced in four major spinach seed production regions: Chile, Denmark, the Netherlands, and the United States (Washington State). California produces 80% of spinach in the United States and all seed planted with the majority infested by V. dahliae comes from the above four sources. Three globally distributed genetic populations were identified, indicating sustained migration among these distinct geographic regions with multiple spinach crops produced each year and repeated every year in coastal California. The population structure of V. dahliae from coastal California lettuce plants was heavily influenced by migration from spinach seed imported from Denmark and Washington. Conversely, the sparsely infested lettuce seed had limited or no contribution to the Verticillium wilt epidemic in coastal California. The global trade in plant and seed material is likely contributing to sustained shifts in the population structure of V. dahliae, affecting the equilibrium of native populations, and likely affecting disease epidemiology.


Plant Disease ◽  
2014 ◽  
Vol 98 (11) ◽  
pp. 1584-1584 ◽  
Author(s):  
E. A. Markakis ◽  
N. Kavroulakis ◽  
G. C. Koubouris

Avocado (Persea americana) is an important crop for Chania, Crete, Greece, and is grown on more than 800 ha. In November 2013, 4-year-old trees in a new avocado grove of cv. Hass grafted onto the rootstock ‘Bacon,’ previously planted in citrus trees, showed symptoms of yellowing, leaf fall, twig and branch dieback and vascular tissue discoloration. Disease incidence was estimated at 2.3% (12 out of 530 trees affected). A fungus was consistently and readily isolated from symptomatic vascular tissue, previously surface-disinfested with 95% ethanol, on acidified potato dextrose agar (APDA). After 7 days, slow-growing colonies were transferred to PDA and the growth rate of the fungus was 2.9 mm/day at 24°C in the dark. Microscopic observations revealed hyaline hyphae with many irregular, dark microsclerotia measuring 40 to 200 × 30 to 75 μm (average 94.5 × 50.3 μm) developing after 21 days of growth. Hyaline, elliptical, single-celled conidia measuring 2.8 to 7.5 × 2.5 to 4.3 μm (average 4.8 × 3.1 μm) developed on verticillate conidiophores. For molecular characterization, Verticillium dahliae specific primer pair ITS1-F/ITS2-R that amplifies the rRNA internal transcribed spacer (ITS) region was used (2). Band of expected size was amplified, sequenced, and deposited in GenBank (Accession No. KJ818294). On the basis of morphological characteristics (3) and a BLAST search with 100% identity to the published ITS sequence of a V. dahliae isolate in GenBank (KC834733.1), the fungus was identified as V. dahliae. Five 1-year-old avocado plants of cv. Hass, grafted onto the rootstock ‘Bacon,’ were used for pathogenicity tests. Artificial inoculation was performed by making a 5.0 × 3.5 mm hole in the rootstock trunk, injecting approximately 40 μl of a 2.8 × 107 conidia/ml suspension into the vessels (spores were introduced passively), sealing with Vaseline, and covering with adhesive paper tape. Five control plants were mock inoculated with sterilized distilled water. Disease symptoms that appeared 18 days post artificial inoculation were similar to those observed under natural infection conditions. Thirty-five days post artificial inoculation, disease incidence was 80%, whereas the percentage of positive V. dahliae re-isolations from infected tissues was 95% (96.7 and 93.3% from rootstock and graft, respectively). The extent of vascular tissue discoloration from the point of inoculation ranged from 11 to 62 cm, whereas V. dahliae was successfully re-isolated even from the end of the graft (approximately 60 cm above the initial inoculation point), thus confirming Koch's postulates. Neither symptoms nor positive isolations were observed in control plants. The pathogenicity test was repeated twice with similar results. Verticillium wilt of avocado has been observed in several countries including Argentina, Chile, Ecuador, Israel, Mexico, Morocco, Spain, and the United States (1). To the best of our knowledge, this is the first report of Verticillium wilt on avocado in Greece. This disease could potentially be an increasing problem in areas where young avocado trees are established on land previously planted in vegetable crops. References: (1) J. C. Goud and J. A. Hiemstra. Chapter 3 in: A Compendium of Verticillium Wilt in Trees Species, 1998. (2) E. A. Markakis et al. Eur. J. Plant Pathol. 124:603, 2009. (3) G. F. Pegg and B. L. Brady. Verticillium Wilts. CABI Publishing, Wallingford, UK, 2002.


Plant Disease ◽  
2007 ◽  
Vol 91 (6) ◽  
pp. 770-770 ◽  
Author(s):  
A. Garibaldi ◽  
G. Gilardi ◽  
M. L. Gullino

Lettuce (Lactuca sativa L.) is an important crop used for fresh and processing markets in Italy and is grown on more than 21,000 ha. During October and November of 2006, wilt symptoms were observed on field-grown lettuce, cv. Estelle, in Forlì, Emila Romagna (northeastern Italy) and on cv. Ballerina grown under plastichouses in Piedmont (northwestern Italy). Both lettuce cultivars were of a butterhead type. Affected plants were stunted and developed yellow leaves with brown or black streaks in the vascular tissue. Yellowing started from the external leaves. Discoloration was observed in the vascular tissue of roots, crown, and leaves. A fungus was consistently and readily isolated from symptomatic vascular tissue, previously disinfested in 1% sodium hypochlorite, when cultured on potato dextrose agar (PDA). Microscopic observations revealed hyaline hyphae with many ovoid, dark microsclerotia measuring 32 to 43 × 16 to 26 μm developing after 15 days of growth at 18°C in the dark. Conidiophores showed two verticils of three elements. Conidia were hyaline, elliptical, single celled, and measured 3.5 to 8.5 × 1.8 to 4.3 μm (average 5.5 × 2.5 μm). According to its morphological characteristics, the fungus was identified as Verticillium dahliae (2). Healthy, 20-day-old lettuce plants, cvs. Principessa and Maxima, both belonging to the butterhead type, were separately inoculated by root dip with a conidial suspension (106/ml) of two isolates of V. dahliae isolated, respectively, at Forlì and Torino. Noninoculated lettuce plants served as control treatments. Plants (10 per treatment) were grown in pots (10-liter vol.) in a steam-disinfested peat/perlite/sand (3:1:1 vol/vol) substrate and were maintained in a glasshouse at temperatures ranging between 17 and 22°C and relative humidity ranging between 60 and 70%. First wilt symptoms and vascular discoloration in the roots, crown, and veins developed 40 days after the artificial inoculation. Forty percent of the plants were affected in the case of cv. Maxima and 30% for cv. Principessa. Noninoculated plants remained healthy. The pathogenicity tests were repeated twice. To our knowledge, this is the first report in Italy of Verticillium wilt on lettuce. The disease has been previously reported in Greece (1) and the United States (3). Currently, Verticillium wilt of lettuce seems restricted in Italy to very few farms in the two locations; moreover, its incidence is very low (0.05%). References: (1) E. K. Ligoxigakis et al. Phytoparasitica 30:141, 2002. (2) G. F. Pegg and B. L. Brady. Verticillium Wilts. CABI Publishing, Wallingford, UK, 2002. (3) G. E. Vallad et al. Plant Dis. 89:317, 2005.


Plant Disease ◽  
2013 ◽  
Vol 97 (6) ◽  
pp. 840-840
Author(s):  
S. Johnson ◽  
C. Miles ◽  
D. A. Inglis

Solanum aethiopicum L., previously S. integrifolium Poir. (4), has been used as a rootstock for commercial, grafted eggplant production throughout Asia (3). In August 2010 and 2011, symptoms of Verticillium wilt were observed on ‘Epic’ eggplant (S. melongena L.) grafted onto S. aethiopicum at two sites with a history of the disease: one in the irrigated, dryland Columbia Basin of eastern Washington near Eltopia, and the other in maritime western Washington near Mount Vernon. Interveinal chlorosis, V-shaped necrotic lesions, and wilting were evident at both sites in both years. Each year, stems of 20 symptomatic plants from each field site were cut at the soil line to a 20-cm length, surface sterilized for 5 min in a 10% bleach solution, rinsed in tap water for 30 s, cut longitudinally, and incubated in moisture chambers for 4 weeks at room temperature in the dark. Microsclerotia that formed in the stems were typical of those produced by V. dahliae. One isolate, ‘MVEgg301’, from an infected stem at Mount Vernon, developed dark microsclerotia and verticillate conidiophores in a radiating pattern, typical of Verticillium on half-strength potato dextrose agar (1/2 PDA) medium. The internal transcribed spacer (ITS) region of rDNA amplified by PCR assay using primers ITS6 and ITS4 from mycelia sampled directly from 1/2 PDA media revealed a 100% match with ITS rDNA sequences of >50 V. dahliae accessions in GenBank (1). Pathogenicity of ‘MVEgg301’ was assessed in two tests. In both, 12 each of non-grafted and grafted (with cv. Epic) S. aethiopicum plants were inoculated with ‘MVEgg301’ by cutting approximately 5 mm off the root tips and dipping the remaining roots in a suspension of 106 conidia/ml for 5 s. Similarly, 12 each of non-grafted and grafted S. aethiopicum plants were cut and dipped similarly in sterile water as controls. Chlorosis, necrosis, and wilting were observed in 11 of the 12 inoculated, non-grafted plants and 8 of the 12 inoculated, grafted plants in Test I. The same symptoms were observed in 10 of the 12 inoculated, non-grafted plants and 10 of the 12 inoculated, grafted plants in Test II. V. dahliae was reisolated and confirmed from symptomatic, inoculated non-grafted and grafted plants using the stem assay and direct PCR assay described above. Chlorosis, necrosis, and wilting were observed in one non-grafted control plant in Test I, and two non-grafted and four grafted control plants in Test II. The symptoms were mild and likely due to nutritional deficiencies; microsclerotia were not observed in any assayed water-inoculated plant stems. Although there are several reports of V. albo-atrum infecting S. aethiopicum in the United States (2), to our knowledge, this is the first report of V. dahliae causing Verticillium wilt on this eggplant species. This finding is significant because S. aethiopicum is used as a rootstock for control of soilborne diseases like Verticillium wilt in commercial grafted eggplant production (3). References: (1) G. Calmin et al. Biotechnol. Biotechnol. Equ. 21:40, 2007. (2) D. F. Farr et al. Fungi on Plants and Plant Products in the United States. American Phytopathological Society, St. Paul, MN, 1989. (3) M. Oda. Food Fert. Technol. Ctr. Ext. Bul. 480:1, 1999. (4) PBI Solanum Project. 2012. Solanaceae Source. Accessed at http://www.nhm.ac.uk/solanaceaesource/ . Natural History Museum, London, 29 Aug. 2012.


2010 ◽  
Vol 11 (1) ◽  
pp. 42 ◽  
Author(s):  
F. Mathew ◽  
B. Kirkeide ◽  
T. Gulya ◽  
S. Markell

Widespread infection of charcoal rot was observed in a commercial sunflower field in Minnesota in September 2009. Based on morphology, isolates were identified as F. sporotrichioides and F. acuminatum. Koch's postulates demonstrated pathogencity of both species. To our knowledge, this is the first report of F. sporotrichoides and F. acuminatum causing disease on Helianthus annuus L. in the United States. Accepted for publication 23 August 2010. Published 15 September 2010.


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