scholarly journals First Report of Fusarium oxysporum Causing Wilt on Jade Plant (Crassula ovata) in Italy

Plant Disease ◽  
2011 ◽  
Vol 95 (9) ◽  
pp. 1191-1191 ◽  
Author(s):  
A. Garibaldi ◽  
D. Bertetti ◽  
P. Pensa ◽  
A. Poli ◽  
M. L. Gullino

During summer 2010, symptoms of a wilt disease were observed in a commercial farm in northern Italy on Crassula ovata (jade plant). First symptoms consisted of chlorosis and premature drop of still turgid leaves. As the disease progressed, leaves turned yellow and wilted before dropping off and the stem wilted, bent, and eventually rotted starting from the base. In some cases, the stem broke or the basal portion of the leaf rotted. Brown discolorations were observed in the vascular system. Of 10,000 plants, 65% (cv. Mini) and 5% of 600 plants (cv. Magical Tree) were affected. Premature dropping of leaves was more frequent on cv. Magical Tree. Using the Komada's Fusarium-selective medium, a fungus was consistently and readily isolated from symptomatic vascular tissues of plants belonging to both cultivars. Isolates obtained from both cultivars were purified, subcultured on potato dextrose agar (PDA), and single-spore cultures were obtained. On PDA, both isolates produced pale violet, abundant, aerial mycelium, felted in old cultures, with purple pigments in the agar. The isolates were grown on Spezieller Nährstoffarmer agar for characterization of macroconidia and microconidia (1). Both isolates produced sparse, 3 to 5 septate, nearly straight macroconidia measuring 30 to 56 × 3 to 5 (average 40 × 4) μm with a short apical cell and a foot-shaped basal cell. Sporodochia were not observed. Unicellular, oval-elliptical microconidia measuring 5 to 13 × 3 to 4 (average 8 × 3) μm were produced on short monophialides. Chlamydospores were abundant, single and sometime in pairs, terminal and intercalary, rough walled, and measured 6 to 9 μm. Such characteristics are typical of Fusarium oxysporum (3). The ITS region (internal transcribed spacer) of rDNA was amplified with primers ITS1/ITS4 (4) and sequenced. BLASTn analysis of an isolate from C. ovata cv. Mini (515 bp, Accession No. HQ682196) and C. ovata cv. Magical Tree (509 bp, Accession No. HQ682197) showed an E-value of 0.0 with F. oxysporum. For these sequences, pairwise alignment of EMBOSS (E.B.I. - The European Bioinformatics Institute) revealed identity and similarity of 99.0%. To confirm pathogenicity, tests were conducted on 5-month-old plants of cvs. Mini and Magical Tree. Plants (three per treatment) were inoculated by dipping roots in a 1 × 106 CFU/ml conidial suspension of the two isolates of F. oxysporum prepared from 10-day-old cultures grown on casein liquid medium (2), shaken (90 rpm) for 10 days at 24°C ± 1 (12-h fluorescent light, 12-h dark). Inoculated plants were transplanted into pots filled with steamed mix (sphagnum peat/perlite/pine bark/clay; 50:20:20:10) and maintained in a plant growth chamber at 25 ± 1°C under a regimen of 12 h per day of fluorescent light. Inoculated plants belonging to both cultivars showed typical first symptoms of Fusarium wilt after 13 days. In the following days, leaves dropped, stems wilted, and plants died. Noninoculated plants remained healthy. F. oxysporum was reisolated from inoculated plants. The pathogenicity test was conducted twice. This is, to our knowledge, the first report of F. oxysporum on C. ovata in Italy or worldwide. References: (1) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Professional, Ames, IA, 2006. (2) A. Minuto et al. Phytoparasitica 36:294, 2008. (3) B. A. Summerell et al. Plant Dis. 87:117, 2003. (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.

Plant Disease ◽  
2011 ◽  
Vol 95 (7) ◽  
pp. 877-877
Author(s):  
A. Garibaldi ◽  
P. Pensa ◽  
D. Bertetti ◽  
A. Poli ◽  
M. L. Gullino

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.


Plant Disease ◽  
2011 ◽  
Vol 95 (4) ◽  
pp. 496-496 ◽  
Author(s):  
A. Garibaldi ◽  
G. Gilardi ◽  
A. Poli ◽  
M. L. Gullino

In the summer of 2009, a wilt of chicory was observed on 25 to 30% of 30-day-old Cichorium intybus L. cv. Clio plants grown outdoors on a commercial farm in Piedmont (northern Italy). Affected plants were chlorotic and stunted with poorly developed root systems compared with healthy plants. Black streaks were observed in the stem and proximal part of the leaf vascular system in wilted plants. Fusarium oxysporum Schltdl. was isolated from symptomatic vascular tissue on a Fusarium-selective medium (1) from 80% of samples. Grown on potato dextrose agar (PDA) for 4 days at 23°C, the colonies, initially white and later pale pink, produced hyaline microconidia that were oval-elliptical and cylindrical in shape measuring 5.6 to 14.9 (average 10.2) × 2.1 to 4.5 (3.0) μm, borne on short monophialides measuring 8.2 to 16.1 (average 13.2) × 2.1 to 4.2 (3.3) μm. Macroconidia were slightly curved, three-septate, with a slightly hooked apical cell and a foot-shaped basal cell measuring 24.9 to 41.6 (average 32.2) × 3.2 to 5.2 (4.3) μm. Chlamydospores were both terminally and intercalary, solitary but also in short chains (2 to 4 elements) measuring 21.1 to 41.0 (average 27.2) μm (2). The internal transcribed spacer (ITS) rDNA region was amplified using the primers ITS1/ITS4 and sequenced. BLASTn analysis of the 527-bp amplicon (GenBank Accession No. HQ644423) obtained had 98% sequence identity with F. oxysporum (GenBank Accession No. FJ605247). The translation elongation factor-1α (EF-1α) gene was amplified using primers EF-1/EF-2 and sequenced (GenBank Accession No. GU564259). The 663-bp fragment had 99% sequence identity with F. oxysporum (GenBank Accession Nos. EU313540, EU313539, and DQ837696). Pathogenicity tests were conducted on 15-day-old chicory plants from two cultivars (Clio and Katia). Thirty-five plants per cultivar were inoculated by dipping their roots in a 1 × 106 CFU/ml suspension of isolate FusCic45B recovered from wilted chicory. Inoculated and noninoculated plants were transplanted into five pots filled with 10 liters of steamed mix (peat/perlite/sand, 60:20:20 vol/vol) and were maintained in a glasshouse at 25 to 27°C. Wilt symptoms and vascular discoloration of the roots, crown, and veins developed 15 days after inoculation on all inoculated plants. Plants of cv. Clio were more susceptible. F. oxysporum was always reisolated from infected plants using the Fusarium-selective medium. All noninoculated plants remained healthy. The pathogenicity test was conducted twice. To our knowledge, this is the first report of wilt caused by F. oxysporum on chicory, C. intybus, in Italy as well as worldwide. References: (1) H. Komada. Rev. Plant Prot. Res. 8:114, 1975. (2) E. Nelson et al. Fusarium Species: An Illustrated Manual for Identification. The Pennsylvania State University Press, University Park, 1983.


Plant Disease ◽  
2004 ◽  
Vol 88 (3) ◽  
pp. 309-309 ◽  
Author(s):  
A. Garibaldi ◽  
A. Minuto ◽  
M. L. Gullino

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.


Plant Disease ◽  
2014 ◽  
Vol 98 (8) ◽  
pp. 1158-1158 ◽  
Author(s):  
Y. H. Liu ◽  
T. Lin ◽  
C. S. Ye ◽  
C. Q. Zhang

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.


Plant Disease ◽  
2009 ◽  
Vol 93 (3) ◽  
pp. 323-323 ◽  
Author(s):  
F. T. Arroyo ◽  
Y. Llergo ◽  
A. Aguado ◽  
F. Romero

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.


Plant Disease ◽  
2002 ◽  
Vol 86 (9) ◽  
pp. 1052-1052 ◽  
Author(s):  
A. Garibaldi ◽  
G. Gilardi ◽  
M. L. Gullino

In spring 2001, plants of the lettuce cv. Salad Bowl showing symptoms of a wilt disease were observed in several commercial plastic greenhouses near Bergamo, in northern Italy. Wilted plants were first observed during the spring and summer of 2001 when temperatures were between 26 and 35°C. Symptoms were observed in the same area and the same farms in March 2002, in concomitance with a period of high temperatures. Although the distribution of the disease was generally uniform, symptoms were more severe in the central part of the greenhouses where temperatures were warmest. Symptoms were first observed at thinning, when seedlings (30 days old) appeared wilted. Vascular tissues of affected seedlings appeared red or brown. Affected plants were stunted and developed yellow leaves and brown or black streaks in the vascular system. The vascular streaks in the yellow leaves extended from the crown and were continuous with a red-brown discoloration in the vascular system of the crown and upper taproot. Symptoms were typically not visible on the outside of the crowns or roots. Fusarium oxysporum was consistently and readily isolated from symptomatic vascular tissues on a Fusarium-selective medium (2). Seeds of cv. Salad Bowl were planted in steam-sterilized soil artificially infested with 1 × 104 CFU/g soil of each of two isolates of F. oxysporum obtained from infected plants. Plants grown in noninfested soil served as noninoculated control treatments. Plants (25 per treatment) were grown at 25 to 28°C in growth chambers. Wilt symptoms and vascular discoloration of the roots, crown, and veins developed 20 days after seeding for all plants grown in the infested soil, and 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 lettuce in Europe. A wilt of lettuce attributed to F. oxysporum f. sp. lactucae was reported in Japan in 1967 (3) and in the United States in 1993 (1), in the latter case the incitant organism was described as F. oxysporum f. sp. lactucum. References: (1) J. C. Hubbard and J. S. Gerik. Plant Dis. 77:750, 1993. (2) H. Komada. Rev. Plant Prot. Res. 8:114, 1975. (3) T. Matuo and S. Motohashi. Trans. Mycol. Soc. Jpn. 8:13, 1967.


Plant Disease ◽  
2021 ◽  
Author(s):  
Sixto Velarde Felix ◽  
Victor Valenzuela ◽  
Pedro Ortega ◽  
Gustavo Fierros ◽  
Pedro Rojas ◽  
...  

Chickpea (Cicer aretinium L.) is a legume crop of great importance worldwide. In January 2019, wilting symptoms on chickpea (stunted grow, withered leaves, root rot and wilted plants) were observed in three fields of Culiacan Sinaloa Mexico, with an incidence of 3 to 5%. To identify the cause, eighty symptomatic chickpea plants were sampled. Tissue from roots was plated on potato dextrose agar (PDA) medium. Typical Fusarium spp. colonies were obtained from all root samples. Ten pure cultures were obtained by single-spore culturing (Ff01 to Ff10). On PDA the colonies were abundant with white aerial mycelium, hyphae were branched and septae and light purple pigmentation was observed in the center of old cultures (Leslie and Summerell 2006). From 10-day-old cultures grown on carnation leaf agar medium, macroconidias were falciform, hyaline, with slightly curved apexes, three to five septate, with well-developed foot cells and blunt apical cells, and measured 26.6 to 45.8 × 2.2 to 7.0 μm (n = 40). The microconidia (n = 40) were hyaline, one to two celled, produced in false heads that measured 7.4 to 20.1 (average 13.7) μm × 2.4 to 8.9 (average 5.3) μm (n = 40) at the tips of long monophialides, and were oval or reniform, with apexes rounded, 8.3 to 12.1 × 1.6 to 4.7 μm; chlamydospores were not evident. These characteristics fit those of the Fusarium solani (Mart.) Sacc. species complex, FSSC (Summerell et al. 2003). The internal transcribed spacer and the translation elongation factor 1 alpha (EF1-α) genes (O’Donnell et al. 1998) were amplified by polymerase chain reaction and sequenced from the isolate Ff02 and Ff08 (GenBank accession nos. KJ501093 and MN082369). Maximum likelihood analysis was carried out using the EF1-α sequences (KJ501093 and MN082369) from the Ff02 and Ff08 isolates and other species from the Fusarium solani species complex (FSSC). Phylogenetic analysis revealed the isolate most closely related with F. falciforme (100% bootstrap). For pathogenicity testing, a conidial suspension (1x106 conidia/ml) was prepared by harvesting spores from 10-days-old cultures on PDA. Twenty 2-week-old chickpea seedlings from two cultivars (P-2245 and WR-315) were inoculated by dipping roots into the conidial suspension for 20 min. The inoculated plants were transplanted into a 50-hole plastic tray containing sterilized soil and maintained in a growth chamber at 25°C, with a relative humidity of >80% and a 12-h/12-h light/dark cycle. After 8 days, the first root rot symptoms were observed on inoculating seedlings and the infected plants eventually died within 3 to 4 weeks after inoculation. No symptoms were observed plants inoculated with sterilized distilled water. The fungus was reisolated from symptomatic tissues of inoculated plants and was identified by sequencing the partial EF1-α gene again and was identified as F. falciforme (FSSC 3 + 4) (O’Donnell et al. 2008) based on its morphological characteristics, genetic analysis, and pathogenicity test, fulfilling Koch’s postulates. The molecular identification was confirmed via BLAST on the FusariumID and Fusarium MLST databases. Although FSSC has been previously reported causing root rot in chickpea in USA, Chile, Spain, Cuba, Iran, Poland, Israel, Pakistan and Brazil, to our knowledge this is the first report of root rot in chickpea caused by F. falciforme in Mexico. This is important for chickpea producers and chickpea breeding programs.


Plant Disease ◽  
2008 ◽  
Vol 92 (4) ◽  
pp. 655-655 ◽  
Author(s):  
A. Garibaldi ◽  
A. Minuto ◽  
M. Shiniti Uchimura ◽  
M. L. Gullino

In 2006, gerbera (Gerbera jamesonii) plants, cvs. Basic, Xena, and Olimpia grown for cut flower production in two greenhouse farms in the region of Guarapuava, Paraná, Brazil, exhibited symptoms of a wilt disease. Affected plants (approximately 20, 60, and 50% on cvs. Basic, Xena, and Olimpia, respectively) were stunted and developed yellow leaves unilaterally with initially brown and eventually black streaks in the vascular system. The first symptoms occurred 2 months after transplanting during the hottest period of the summer with an average air temperature of 27°C. Vascular streaks in the yellow leaves were continuous with a brown discoloration in the vascular system of the crown and upper taproot. Occasionally, the leaves of affected plants turned red. A Fusarium sp. was consistently and readily isolated onto a Fusarium-selective medium from symptomatic vascular tissue sampled from the crown of infected plants. Colonies were identified as Fusarium oxysporum on the basis of colony and conidia morphology (1) after subculturing on potato dextrose agar. Since F. oxysporum f. sp. chrysanthemi has been previously reported on Chrysanthemum morifolium, Argyranthemum frutescens, and gerbera (4), pathogenicity tests were carried out by using one monoconidial isolate obtained from wilted plants and one Italian isolate (F. oxysporum f. sp. chrysanthemi MASS 6). The isolates of F. oxysporum were grown in casein hydrolysate in shake culture (90 rpm) for 10 days at 25°C with 12 h of fluorescent light per day. Healthy rooted plants of 30-, 20-, and 45-day-old C. morifolium (cv. Captiva), A. frutescens (cv. Stella 2000), and gerbera (cvs. Jaska, Dalma, and Excellence), respectively, were inoculated by separately dipping roots into a conidial suspension (5 × 107 conidia/ml) of the two isolates of F. oxysporum. Plants were transplanted (one plant per pot) into pots (3.5 liter vol). Noninoculated plants served as control treatments. Plants (15 per treatment) were grown in a glasshouse at an average day temperature of 32°C and night temperature of 23°C (minimum 21°C and maximum 43°C). Wilt symptoms and discoloration of the vascular system in roots, crown, and petioles developed within 29 days on C. morifolium, 26 days on A. frutescens, and 14 days on gerbera. Noninoculated plants remained healthy. F. oxysporum was consistently reisolated from infected plants. The pathogenicity test was carried out twice. Gerbera wilt caused by F. oxysporum f. sp. chrysanthemi was recently reported in Italy (2) and Spain (3). Currently, the wilt of gerbera in the area of Paraná is limited to two farms. To our knowledge, this is the first report of the disease in Brazil as well as in South America. References: (1) C. Booth. Fusarium. CMI, Kew, UK, 1977. (2) A. Garibaldi et al. Plant Dis. 88:311, 2004. (3) A. Garibaldi et al. Plant Dis. 91:638, 2007. (4) A. Minuto et al. J. Phytopathol. 155:373, 2007.


Plant Disease ◽  
2009 ◽  
Vol 93 (10) ◽  
pp. 1078-1078 ◽  
Author(s):  
A. Garibaldi ◽  
G. Gilardi ◽  
M. Troisi ◽  
M. L. Gullino

In the summer of 2007, a wilt of endive (Cichorium endivia) cv. Myrna plants was observed on several commercial farms near Alessandria in northern Italy. Approximately 40% of the plants on each farm were symptomatic. Affected plants were stunted and yellow and their root systems were poorly developed. Basal leaves eventually wilted. Black streaks were observed in the proximal part of the leaf vascular system. Fusarium oxysporum was consistently isolated from symptomatic vascular tissue on a Fusarium-selective medium (1). To confirm identification of the pathogen, the translation elongation factor 1α and the ribosomal DNA intergenic spacer (IGS) region were partially sequenced (2). Genomic DNA was extracted from mycelia growing on potato dextrose agar. Amplification of the EF-1α region generated a sequence of 648 bp; the IGS amplicon was 2,500 bp. The EF-1α sequence (GenBank Accession No GQ398152) was 99% similar to the sequence of a F. oxysporum strain isolated from soil and a strain pathogenic on cotton plants (GenBank Accession No. EU246574). The IGS sequence (GenBank Accession No GQ398153) was 97% similar to the sequence of a F. oxysporum strain (GenBank Accession No. EF661647). Pathogenicity tests were conducted on 15-day-old endive plants (cv. Myrna). Ten plants were inoculated by dipping their roots in a 1 × 106 CFU/ml suspension of one of the isolates recovered from a wilted endive plant. Inoculated and noninoculated plants were transplanted into pots filled with steamed soil and maintained in a glasshouse at 23 to 28°C. Wilt symptoms and vascular discoloration of the roots, crown, and veins developed 60 days after inoculation. F. oxysporum was consistently reisolated from infected plants. Noninoculated plants remained healthy. The pathogenicity test was conducted twice. To our knowledge, this is the first report of wilt caused by F. oxysporum on endive in Italy. References: (1) H. Komada. Rev. Plant Prot. Res. 8:114, 1975. (2) G. Mbofung et al. Phytopathology 97:87, 2007.


Plant Disease ◽  
2013 ◽  
Vol 97 (2) ◽  
pp. 287-287 ◽  
Author(s):  
D. J. Vakalounakis ◽  
E. A. Markakis

During the 2011 to 2012 crop season, a severe leaf spot disease of cucumber (Cucumis sativus) cv. Cadiz was noticed on crops in some greenhouses in the Goudouras area, Lasithi, Crete, Greece. Symptoms appeared in late winter, mainly on the leaves of the middle and upper part of the plants. Initially, small necrotic pinpoint lesions with white centers, surrounded by chlorotic halos, 1 to 3 mm in diameter, appeared on the upper leaf surfaces, and these progressively enlarged to spots that could coalesce to form nearly circular lesions up to 2 cm or more in diameter. Stemphylium-like fructifications appeared on necrotic tissue of older lesions. Severely affected leaves became chlorotic and died. No other part of the plant was affected. Small tissue pieces from the edges of lesions were surface disinfected in 0.5% NaClO for 5 min, rinsed in sterile distilled water, plated on acidified potato dextrose agar and incubated at 22 ± 0.5°C with a 12-h photoperiod. Stemphylium sp. was consistently isolated from diseased samples. Colonies showed a typical septate mycelium with the young hyphae subhyaline and gradually became greyish green to dark brown with age. Conidiophores were subhyaline to light brown, 3- to 10-septate, up to 200 μm in length, and 4 to 7 μm in width, with apical cell slightly to distinctly swollen, bearing a single spore at the apex. Conidia were muriform, mostly oblong to ovoid, but occasionally nearly globose, subhyline to variant shades of brown, mostly constricted at the median septum, 22.6 ± 6.22 (11.9 to 36.9) μm in length, and 15.1 ± 2.85 (8.3 to 22.6) μm in width, with 1 to 8 transverse and 0 to 5 longitudinal septa. DNA from a representative single-spore isolate was extracted and the internal transcribed spacer region (ITS) of ribosomal DNA (rDNA) was amplified using the universal primers ITS5 and ITS4. The PCR product was sequenced and deposited in GenBank (Accession No. JX481911). On the basis of morphological characteristics (3) and a BLAST search with 100% identity to the published ITS sequence of a S. solani isolate in GenBank (EF0767501), the fungus was identified as S. solani. Pathogenicity tests were performed by spraying a conidial suspension (105 conidia ml–1) on healthy cucumber (cv. Knossos), melon (C. melo, cv. Galia), watermelon (Citrullus lanatus cv. Crimson sweet), pumpkin (Cucurbita pepo, cv. Rigas), and sponge gourd (Luffa aegyptiaca, local variety) plants, at the 5-true-leaf stage. Disease symptoms appeared on cucumber and melon only, which were similar to those observed under natural infection conditions on cucumber. S. solani was consistently reisolated from artificially infected cucumber and melon tissues, thus confirming Koch's postulates. The pathogenicity test was repeated with similar results. In 1918, a report of a Stemphylium leaf spot of cucumber in Indiana and Ohio was attributed to Stemphylium cucurbitacearum Osner (4), but that pathogen has since been reclassified as Leandria momordicae Rangel (2). That disease was later reported from Florida (1) and net spot was suggested as a common name for that disease. For the disease reported here, we suggest the name Stemphylium leaf spot. This is the first report of a disease of cucumber caused by a species of Stemphylium. References: (1) C. H. Blazquez. Plant Dis. 67:534, 1983. (2) P. Holliday. Page 243 in: A Dictionary of Plant Pathology. Cambridge University Press, Cambridge, UK, 1998. (3) B. S. Kim et al. Plant Pathol. J. 15:348, 1999. (4) G. A. Osner. J. Agric. Res. 13:295, 1918.


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