First Report of Fusarium Rot of Garlic Bulbs Caused by Fusarium proliferatum in North Carolina

In August of 2013, garlic bulbs (Allium sativum) of the variety Chesnok Red grown and stored under dry conditions by a commercial producer in Buncombe County showed water-soaked, tan to salmon-pink lesions. Lesions on cloves became soft over time, slightly sunken, and had mycelium near the center of the bulb, which is characteristic of Fusarium rots on garlic (1,2). Approximately 10 to 20% of the bulbs inspected in the drying storage room were affected. Surface-sterilized tissue was excised from the margin of lesions on eight bulbs, plated onto acid potato dextrose agar (APDA), and incubated in the dark at room temperature (21°C). White to light pink colonies with abundant aerial mycelium and a purple pigment were obtained from all samples after 2 to 3 days of incubation. Inspection of colony morphology and reproductive structures under a microscope revealed that isolate characteristics were consistent with Fusarium proliferatum (Matsushima) Nirenberg. Microscopic morphological characteristics of the isolate included hyaline, septate hyphae; slender, slightly curved macroconidia with three to five septae produced in sporodochia; curved apical cell; and club-shaped, aseptate microconidia (measuring 3.3 to 8.3 × 1.1 to 1.3 μm) produced in chains by mono and polyphyalides. To further define the identity of the isolate, the beta-tubulin (Btub), elongation factor 1a (EF1a), and internal transcribed spacer (ITS) regions were amplified and sequenced (3). The resulting sequences were compared against the GenBank nucleotide database by using a BLAST alignment, which revealed that the isolate had 100% identity with F. proliferatum for the Btub, EF1a, and ITS regions (GenBank Accession Nos. AF291055.1, JX118976.1, and HF930594.1, respectively). Sequences for the isolate were deposited in GenBank under accessions KJ128963, KJ128964, and KJ128965. While there have been other reports of F. proliferatum causing bulb rot of garlic in the United States (1), to our knowledge, this is the first report in North Carolina. The finding is significant since F. proliferatum can produce a broad range of mycotoxins, including fumonisins, when infecting its host, which is a concern for food safety in Allium crops. References: (1) F. M. Dugan et al. Plant Pathol. 52:426, 2003. (2) L. J. du Toit and F. M. Dugan. Page 15 in: Compendium of Onion and Garlic Diseases and Pests. H. F. Schwartz and S. K. Mohan, eds. The American Phytopathological Society, St. Paul, MN, 2008. (3) T. J. White et al. Page 315 in: 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 redolens Causing Root Rot of Soybean in Minnesota

Plant Disease ◽

10.1094/pdis-94-8-1069b ◽

2010 ◽

Vol 94 (8) ◽

pp. 1069-1069 ◽

Cited By ~ 10

Author(s):

J. C. Bienapfl ◽

D. K. Malvick ◽

J. A. Percich

Keyword(s):

Root Rot ◽

Fusarium Species ◽

Apical Cell ◽

Elongation Factor ◽

Morphological Characteristics ◽

Aerial Mycelium ◽

Pathogenic Fungi ◽

The United States ◽

First Report ◽

Root Necrosis

Multiple Fusarium species have been found in association with soybean (Glycine max) plants exhibiting root rot in the United States (3). Soybean plants that lacked apparent foliar symptoms, but exhibited 2- to 5-mm brown, necrotic taproot lesions and lateral root necrosis were observed in Minnesota in one field each in Marshall and Otter Tail counties in July of 2007, as well as in one field in Marshall County in July of 2008. Sampling was conducted as part of a study investigating root rot in major soybean-production areas of Minnesota. Plants were arbitrarily dug up at the R3 growth stage. Root systems were washed, surface disinfested in 0.5% NaOCl for 3 min, rinsed in deionized water, and dried. Fusarium isolates were recovered from root sections with necrotic lesions embedded in modified Nash-Snyder medium (1). One resulting Fusarium colony from one plant per county was transferred to half-strength acidified potato dextrose agar (PDA) and carnation leaf agar (CLA) to examine morphological characteristics (4). Culture morphology on PDA consisted of flat mycelium with sparse white aerial mycelium. On CLA, thick-walled macroconidia with a hooked apical cell and a foot-shaped basal cell were produced in cream-colored sporodochia. Macroconidia ranged from 32.5 to 45.0 μm long. Microconidia were oval to cylindrical with 0 to 1 septa, ranged from 7.5 to 11.25 μm long, and were produced on monophialides. Chlamydospores were produced abundantly in chains that were terminal and intercalary in the hyphae of 4-week-old cultures. Morphological characteristics of the three isolates were consistent with descriptions of F. redolens (2,4). The identity of each isolate was confirmed by sequencing the translation elongation factor 1-α (TEF) locus (4). BLAST analysis of the TEF sequences from each isolate against the FUSARIUM-ID database resulted in a 100% match for 17 accessions of F. redolens (e.g., FD 01103, FD 01369). Each F. redolens isolate was tested for pathogenicity on soybean. Sterile sorghum grain was infested with each isolate and incubated for 2 weeks. Sterile sorghum was used for control plants. Soybean seeds of cv. AG2107 were planted in 11.4-cm pots ~1 cm above a 25-cm3 layer of infested sorghum or sterile sorghum. Two replicate pots containing four plants each were used per treatment and the experiment was repeated once. Root rot was assessed 28 days after planting. Each F. redolens isolate consistently caused taproot necrosis on inoculated plants, whereas control plants did not exhibit root necrosis. Isolations were made from roots of inoculated and control plants and the isolates recovered from inoculated plants were identified as F. redolens based on morphological characteristics and TEF sequences. Fusarium species were not isolated from control plants. To our knowledge, this is the first report of F. redolens causing root rot of soybean; however, it is possible F. redolens has been found previously and misidentified as F. oxysporum (2,4). Results from inoculations suggest that F. redolens may be an important root rot pathogen in Minnesota soybean fields. References: (1) J. C. Bienapfl et al. Acta Hortic. 668:123, 2004. (2) C. Booth and J. M. Waterston. No. 27 in: CMI Descriptions of Pathogenic Fungi and Bacteria. CMI, Kew, England, 1964. (3) G. L. Hartman et al. Compendium of Soybean Diseases. 4th ed. The American Phytopathological Society, St. Paul, MN, 1999. (4) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Ames, IA, 2006.

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Fusarium Canker of Bitternut Hickory Caused by Fusarium solani in the North-Central and Northeastern United States

Plant Disease ◽

10.1094/pdis-09-11-0766 ◽

2012 ◽

Vol 96 (3) ◽

pp. 455-455 ◽

Cited By ~ 1

Author(s):

J.-H. Park ◽

J. Juzwik

Keyword(s):

United States ◽

Fusarium Solani ◽

Apical Cell ◽

Elongation Factor ◽

Morphological Characteristics ◽

Aerial Mycelium ◽

The United States ◽

Eastern United States ◽

Gene Sequences ◽

Inner Bark

Multiple annual cankers were observed on the upper main stems of bitternut hickory (Carya cordiformis) exhibiting top dieback in Indiana, Iowa, Minnesota, New York, Ohio, and Wisconsin during a 2006 to 2008 survey of declining hickory. The top-killed trees had normal-sized, green leaves below and the cankers were oval, sunken, and bounded by heavy callus that seemed to arrest further canker expansion. Fusarium solani was consistently isolated from the margins of inner bark lesions or discolored sapwood of the cankers. When cultured on potato dextrose agar, the isolates grew rapidly with abundant aerial mycelium. On carnation leaf agar, thick-walled macroconidia with 4 to 5 septa were produced in cream, blue-green, or blue sporodochia. Macroconidia were generally cylindrical with a blunt or rounded apical cell and a rounded or foot-shaped basal cell. Microconidia were oval to kidney shaped with 0 to 1 septa and were produced in false heads on elongate monophialides. Chlamydospores were formed singly or in pairs. These morphological characteristics are consistent with descriptions of F. solani (2). The identities of 42 representative isolates were confirmed by sequencing the translation elongation factor (tef) 1-α gene. BLAST analysis of the sequences from each isolate against the GenBank and FUSARIUM-ID database found 98 to 100% similarities to F. solani isolates (GenBank Accession Nos. DQ246841, DQ247025, DQ247282, and DQ247436 and FUSARIUM-ID isolate FD01041). Two haplotypes (BB and BC) were distinguished based on the tef 1-α gene sequences that differed by 10 bp. Pathogenicity tests were conducted with two isolates of each haplotype on asymptomatic C. cordiformis (12 to 21 cm in diameter) in forest stands. In May 2009 in Wabasha County, MN, 0.1-ml spore suspensions (1 × 104 macroconidia/ml) or sterile water was placed in one of three holes (0.6 cm in diameter) drilled to the cambium of 12 trees. The holes were sealed with moist cotton and moldable putty. A duplicate trial, but with BB and BC isolates from Wisconsin, was initiated in Chippewa County, WI in June 2009. The extent of inner bark necrosis was assessed 13 months after inoculation in both sites. Inoculations with F. solani in Minnesota resulted in inner bark lesions with average lengths of 20 and 30 mm for the BB and BC haplotypes, respectively. In Wisconsin, BB and BC haplotypes caused inner bark lesions with average lengths of 34 and 38 mm, respectively. While sunken or open cankers were found for all the BC isolate inoculations, relatively small and callus-bounded cankers were found for BB isolate inoculations. All control wounds were callus-closed with average wound lengths of 12 and 23 mm in Minnesota and Wisconsin, respectively. The same haplotype of F. solani used for inoculation was recovered from each canker as confirmed by analysis of tef 1-α gene sequences. F. solani was not obtained from control wounds. To our knowledge, this is the first report of a canker caused by F. solani on bitternut hickory (1). The same fungus has been previously reported to cause cankers on stems of other hardwood tree genera in the eastern United States and Canada. We hypothesize that numerous main-stem cankers caused by F. solani lead to top dieback of bitternut hickory. 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) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Ames, IA, 2006.

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First report of Diaporthe ueckerae causing stem canker on soybean (Glycine max L.) in Colombia

Plant Disease ◽

10.1094/pdis-04-21-0718-pdn ◽

2021 ◽

Author(s):

Nathali López-Cardona ◽

YUDY ALEJANDRA GUEVARA ◽

Lederson Gañán-Betancur ◽

Carol Viviana Amaya Gomez

Keyword(s):

Management Strategies ◽

Elongation Factor ◽

Morphological Characteristics ◽

Aerial Mycelium ◽

Stem Canker ◽

Growth Stages ◽

Internal Transcribed Spacer Region ◽

First Report ◽

Glycine Max L ◽

Soybean Plants

In October 2018, soybean plants displaying elongated black to reddish-brown lesions on stems were observed in a field planted to the cv. BRS Serena in the locality of Puerto López (Meta, Colombia), with 20% incidence of diseased plants. Symptomatic stems were collected from five plants, and small pieces (∼5 mm2) were surface sterilized, plated on potato dextrose agar (PDA) and incubated for 2 weeks at 25°C in darkness. Three fungal isolates with similar morphology were obtained, i.e., by subculturing single hyphal tips, and their colonies on PDA were grayish-white, fluffy, with aerial mycelium, dark colored substrate mycelium, and produced circular black stroma. Pycnidia were globose, black, occurred as clusters, embedded in tissue, erumpent at maturity, with an elongated neck, and often had yellowish conidial cirrus extruding from the ostiole. Alpha conidia were observed for all isolates after 30 days growth on sterile soybean stem pieces (5 cm) on water agar, under 25ºC and 12 h light/12h darkness photoperiod. Alpha conidia (n = 50) measured 6.0 – 7.0 µm (6.4 ± 0.4 µm) × 2.0 – 3.0 µm (2.5± 0.4 µm), were aseptate, hyaline, smooth, ellipsoidal, often biguttulate, with subtruncate base. Beta conidia were not observed. Observed morphological characteristics of these isolates were similar to those reported in Diaporthe spp. by Udayanga et al. (2015). DNA from each fungal isolate was used to sequence the internal transcribed spacer region (ITS), and the translation elongation factor 1-α (TEF1) gene, using the primer pairs ITS5/ITS4 (White et al. 1990) and EF1-728F/EF1- 986R (Carbone & Kohn, 1999), respectively. Results from an NCBI-BLASTn, revealed that the ITS sequences of the three isolates (GenBank accessions MW566593 to MW566595) had 98% (581/584 bp) identity with D. miriciae strain BRIP 54736j (NR_147535.1), whereas the TEF1 sequences (GenBank accessions MW597410 to MW597412) had 97 to 100% (330-339/339 bp) identity with D. ueckerae strain FAU656 (KJ590747). The species Diaporthe miriciae R.G. Shivas, S.M. Thomps. & Y.P. Tan, and Diaporthe ueckerae Udayanga & Castl. are synonymous, with the latter taking the nomenclature priority (Gao et al. 2016). According to a multilocus phylogenetic analysis, by maximum likelihood, the three isolates clustered together in a clade with reference type strains of D. ueckerae (Udayanga et al. 2015). Soybean plants cv. BRS Serena (growth stages V3 to V4) were used to verify the pathogenicity of each isolate using a toothpick inoculation method (Mena et al. 2020). A single toothpick colonized by D. ueckerae was inserted directly into the stem of each plant (10 plants per isolate) approximately 1 cm below the first trifoliate node. Noncolonized sterile toothpicks, inserted in 10 soybean plants served as the non-inoculated control. Plants were arbitrarily distributed inside a glasshouse, and incubated at high relative humidity (>90% HR). After 15 days, inoculated plants showed elongated reddish-brown necrosis at the inoculated sites, that were similar to symptoms observed in the field. Non-inoculated control plants were asymptomatic. Fungal cultures recovered from symptomatic stems were morphologically identical to the original isolates. This is the first report of soybean stem canker caused by D. ueckerae in Colombia. Due to the economic importance of this disease elsewhere (Backman et al. 1985; Mena et al. 2020), further research on disease management strategies to mitigate potential crop losses is warranted.

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First Report of Fusarium proliferatum Causing Rot of Garlic Bulbs (Allium sativum) in India

Plant Disease ◽

10.1094/pdis-08-11-0649 ◽

2012 ◽

Vol 96 (2) ◽

pp. 290-290 ◽

Cited By ~ 9

Author(s):

N. Ravi Sankar ◽

Gundala Prasad Babu

Keyword(s):

Sodium Hypochlorite ◽

Allium Sativum ◽

Morphological Characteristics ◽

Aerial Mycelium ◽

Its Region ◽

Fusarium Proliferatum ◽

Distilled Water ◽

Sequence Comparisons ◽

First Report ◽

Plant Pathol

In September 2009, diseased garlic bulbs (Allium sativum L. cv. Yamuna Safed) were received from producers and exporters in Hyderabad, Andra Pradesh, India. From 2009 to 2010, similar symptoms were observed on stored garlic bulbs (cvs. Yamuna Safed and Agrifound White) in Chittoor, Kadapa, and Hyderabad districts. In some locations, approximately 60% of the garlic bulbs were affected. At first, infected bulbs showed water-soaked, brown spots and then the disease progressed as small, slightly depressed, tan lesions. A total of 120 diseased samples were collected from all localities. Infected tissues were surface sterilized in 1% sodium hypochlorite for 2 min, rinsed three times in sterile distilled water, plated on potato dextrose agar (PDA), and incubated at 25°C for 7 days. Resultant fungal colonies were fast growing with white aerial mycelium and violet to dark pigments. Hyphae were septate and hyaline. Conidiophores were short, simple, or branched. Microconidia were abundant, single celled, oval or club shaped, measuring 4.5 to 10.5 × 1.3 to 2.5 μm, and borne in chains from both mono-and polyphialides. Macroconidia were not produced. On the basis of morphological characteristics, the pathogen was identified as Fusarium proliferatum (Matsushima) Nirenberg (2). Identification was confirmed by amplification of the internal transcribed spacer (ITS) region. Genomic DNA was extracted from pure cultures of an isolate, and the ITS region was amplified using the ITS4/5 primer pair. PCR amplicons of approximately 574 bp were obtained from isolates, and sequence comparisons with GenBank showed 99% similarity with F. proliferatum (Accession No. FN868470.1). Sequence from this study was submitted to GenBank nucleotide database (Accession No. AB646795). Pathogenicity tests were conducted with three isolates of the fungus following the method of Dugan et al. (1). Each assay with an isolate consisted of 10 garlic cloves disinfected in 1% sodium hypochlorite for 45 s, rinsed with sterile distilled water, and injured to a depth of 4 mm with a sterile 1-mm-diameter probe. The wounds were filled with PDA colonized by the appropriate isolate from a 5-day-old culture. Ten cloves for each tested isolate received sterile PDA as a control. The cloves were incubated at 25°C for 5 weeks; tests were repeated once. After 17 days, rot symptoms similar to the original symptoms developed on all inoculated cloves and F. proliferatum was consistently reisolated from symptomatic tissue, fulfilling Koch's postulates. No fungi were recovered from control cloves. F. proliferatum has been reported on garlic in the northwestern United States (1), Serbia (4), and Spain (3). To our knowledge, this is the first report of F. proliferatum causing rot disease on garlic bulbs in India. References: (1) F. M. Dugan et al. Plant Pathol. 52:426, 2003. (2) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Oxford, UK, 2006. (3) D. Palmero et al. Plant Dis. 94:277, 2010. (4) S. Stankovic et al. Eur. J. Plant Pathol. 48:165, 2007.

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First Report of Curvularia Blight of Zoysiagrass Caused by Curvularia lunata in the United States

Plant Disease ◽

10.1094/pdis-92-1-0173b ◽

2008 ◽

Vol 92 (1) ◽

pp. 173-173 ◽

Cited By ~ 2

Author(s):

J. A. Roberts ◽

L. P. Tredway

Keyword(s):

United States ◽

North Carolina ◽

Morphological Characteristics ◽

Microscopic Analysis ◽

The United States ◽

Curvularia Lunata ◽

One Pot ◽

Plastic Container ◽

First Report ◽

Initial Symptoms

Symptoms of an unknown foliar blight have been observed in zoysiagrass (Zoysia matrella, Z. japonica, and hybrids) landscapes in North Carolina since 2002. Disease activity is most common during spring and summer when temperatures are between 21 and 30°C. Affected leaves initially exhibit small, chocolate brown spots, followed by dieback of leaves from the tips, and eventually blighting of entire tillers. Symptoms appear in small, irregular patches as much as 15 cm in diameter, but numerous patches may coalesce to impact large sections of turf. Infected turf appears tan or brown from a distance, but often turns black during periods of wet or humid weather. Microscopic analysis revealed profuse sporulation of Curvularia spp. on the surface of symptomatic leaves. Leaf sections were surface disinfested in 10% Clorox for 1 to 2 min, blotted dry, then plated on potato dextrose agar (PDA) containing 50 mg/l of tetracycline, streptomycin, and chloramphenicol. Twenty-eight fungal isolates were obtained from six locations. Examination of conidia produced in culture revealed 21 isolates of Curvularia, two isolates of Drechslera, one isolate of Nigrospora, and four unidentified sterile fungi. Curvularia isolates were identified to species on the basis of morphological characteristics (1) and ITS-rDNA sequences. Known isolates of C. eragrostidis, C. geniculata, C. inequalis, C. lunata, C. pallescens, and C. trifolii were obtained from the American Type Culture Collection for comparison. All unknown isolates produced conidia that were characteristic of C. lunata (lacking a protuberant hilum, smooth walled, tri-septate, predominantly curved, and mid- or dark brown, average dimensions 17 to 25 × 8 to 12 μm). Colonies on PDA lacked stroma or the zonate appearance indicative of C. lunata var. aeria. The pathogenicity of C. lunata isolates was tested on zoysiagrass cvs. El Toro (Z. japonica) and Emerald (Z. japonica × matrella). Cores (11.4 cm in diameter) of established zoysiagrass were potted in calcined clay (Turface Allsport; Profile Products LLC, Buffalo Grove, IL), and transferred to a greenhouse where the average temperature was 26°C. Five isolates were selected to represent the geographic range of Curvularia blight in North Carolina, and conidia were produced on PDA under continuous fluorescent illumination. Each isolate was inoculated to one pot of each zoysiagrass variety by spraying with 25 ml of a suspension containing 2 × 105 conidia/ml with an airbrush. Inoculated pots were placed in a sealed, nontransparent plastic container for 48 h at 28°C to encourage infection and then transferred back to the greenhouse bench. Pathogenicity tests were repeated four times over time. Isolates ZFB3 and ZFB28 were most virulent with initial symptoms of foliar dieback appearing within 1 week after inoculation. Continued disease progress resulted in necrosis of the entire plant. Other isolates induced symptoms within 2 to 3 weeks after inoculation; however, disease severity was lower as compared with ZFB3 and ZFB28 throughout each experiment. Cvs. Emerald and El Toro were equally susceptible to infection by C. lunata. To our knowledge, this is the first report of Curvularia blight of zoysiagrass in the United States. This disease was previously described in Japan where it is commonly referred to as ‘dog footprint’ (3) and Brazil (2). References: (1) M. B. Ellis. Dematiaceous Hyphomycetes. CMI, Kew, Surrey, UK, 1971. (2) F. B. Rocha et al. Australas. Plant Pathol. 33:601, 2004. (3) T. Tani and J. B. Beard. Color Atlas of Turfgrass Diseases. Ann Arbor Press, Chelsea, MI, 1997.

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First Report of the Cyst Nematode (Globodera tabacum) Complex on Flue-Cured Tobacco in Spain

Plant Disease ◽

10.1094/pdis.2002.86.12.1402c ◽

2002 ◽

Vol 86 (12) ◽

pp. 1402-1402 ◽

Cited By ~ 1

Author(s):

G. Espárrago ◽

I. Blanco

Keyword(s):

North Carolina ◽

Morphological Characteristics ◽

The United States ◽

Cyst Nematode ◽

Nematode Population ◽

State University ◽

First Report ◽

North Carolina State University ◽

Second Stage ◽

Nicotiana Tabacum L

The Globodera tabacum complex infects tobacco (Nicotiana tabacum L.) fields in the United States. In August 2001, plants of flue-cured tobacco cv. K326 from a field of the La Vera Region of Spain displayed a premature wilting and yellowing of foliage, but the roots looked healthy. In the laboratory under the microscope, nematode cysts were observed on the roots. At harvest in September 2001, soil and root samples were collected to identify the nematode and to quantify the population in the soil. Identification of the nematode was based on morphological characteristics of second-stage juveniles collected from cysts and perineal patterns of cysts recovered from the roots (2). Cysts were collected from roots, and second-stage juveniles were extracted from crushed cysts. The nematode population was extracted from the soil and quantified as described by Barker (1). The nematode population was identified as Globodera tabacum. Soil density of the nematode was 5,307 cysts per liter of soil, 64,286 eggs per liter of soil, and 16,071 second-stage juveniles per liter of soil. To our knowledge, this is the first report of G. tabacum complex in Spain. References: (1) K. R. Barker. Nematode extraction and bioassays. Page 19 in: An Advanced Treatise on Meloidogyne. Vol II, Methodology. K. R. Barker, C. C. Carter, and J. N. Sasser, eds. North Carolina State University Graphics, Raleigh, 1985. (2) R. H. Mulvey and A. Morgan Golden, J. Nematol. 15:1, 1983.

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First report of Fusarium falciforme (FSSC 3+4) causing root rot on chickpea in Mexico

Plant Disease ◽

10.1094/pdis-05-21-1123-pdn ◽

2021 ◽

Author(s):

Sixto Velarde Felix ◽

Victor Valenzuela ◽

Pedro Ortega ◽

Gustavo Fierros ◽

Pedro Rojas ◽

...

Keyword(s):

Fusarium Solani ◽

Root Rot ◽

Species Complex ◽

Elongation Factor ◽

Morphological Characteristics ◽

Aerial Mycelium ◽

Pathogenicity Test ◽

Conidial Suspension ◽

Single Spore ◽

First Report

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.

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First Report of Fusarium mangiferae Causing Mango Malformation in China

Plant Disease ◽

10.1094/pdis-12-11-1030 ◽

2012 ◽

Vol 96 (5) ◽

pp. 762-762 ◽

Cited By ~ 8

Author(s):

R.-L. Zhan ◽

S.-J. Yang ◽

F. Liu ◽

Y.-L. Zhao ◽

J.-M. Chang ◽

...

Keyword(s):

Mangifera Indica ◽

Elongation Factor ◽

Morphological Characteristics ◽

Fungal Growth ◽

Aerial Mycelium ◽

Laboratory Manual ◽

First Report ◽

Normal Number ◽

Mango Malformation ◽

Fusarium Mangiferae

Mango (Mangifera indica L.) malformation caused by Fusarium mangiferae has been reported in many mango-growing regions of the world (3). The disease was also observed in Yunnan and Sichuan provinces of China (1). Typical symptoms in seedlings included loss of apical dominance, hyperplasia and hypertrophy of vegetative buds, shortened internodes, and leaves that were more brittle than those of healthy plants. Inflorescences were abnormally branched and thickened, with panicles producing as much as two to five times the normal number of flowers. Flowers in the malformed inflorescence were much more enlarged and crowded than the generally hypertrophied axes of the panicle, thus producing no fruit or aborting early. To identify the pathogen, samples of malformed and healthy mango seedlings were collected from the affected plantings. For isolation, portions of stems were cut into 3- to 4-mm segments, surface disinfested, dried, and then plated on potato dextrose agar and incubated at 25°C. Within 5 days, white, fluffy, aerial mycelium developed. With the aid of an inverted microscope, single conidia were transferred to carnation leaf agar (CLA) medium. After 10 days of incubation, morphological characteristics were found to be identical to those of F. mangiferae (4). Aerial mycelium was white with no pigmentation observed on potato sucrose agar. Pigmentation on rice medium was pink. On CLA medium, conidia grew in branched conidiophores with false heads bearing monophialides or polyphialides. No conidiospores in chains were observed. Microconidia were ovate to long and oval, 0 to 1 septate, and 3.1 to 10.2 × 1.5 to 2.2 μm. Macroconidia are falculate, 3 to 5 septate, and 18 to 38 × 1.8 to 2.4 μm. Chlamydospores were not observed. Pathogenicity studies were conducted with 7-month-old asymptomatic mango seedlings. These seedlings, except for the controls, were inoculated by injection of the isolated fungus in the axillary or apical bud position. A 1-ml spore suspension (1 × 106 spores/ml) was injected slowly into the stems using a microsyringe with three buds per seedling, for a total of 10 seedlings. Typical malformation symptoms developed within 3 to 4 months, and none of the plants inoculated with sterile water resulted in malformation symptoms. Reisolations from the induced malformed shoots yielded the same fungus, and no fungal growth was observed to be growing from the control plants. To confirm identity of the causal fungus, the gene encoding translation elongation factor 1 alpha (EF-1α) was amplified and sequenced (2). The EF-1α sequence was 660 bp long. The sequence (GenBank Accession No. HM068871) was 99.68% similar to sequences of FD_01167 in the Fusarium ID database. On the basis of symptoms, fungal morphology, the EF-1α region sequence, and pathogenicity testing, this fungus was identified as F. mangiferae. To our knowledge, this is the first report of F. mangiferae causing mango malformation in China. This report will establish a foundation for further study of F. mangiferae and effectively addressing the disease. References: (1) X. H. Chen. Pract. Technol. (in Chinese) 6:5, 1992. (2) D. M. Geiser et al. Eur. J. Plant Pathol. 110:473, 2004. (3) J. Kumar et al. Annu. Rev. Phytopathol. 31:217, 1993. (4) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Ames, IA, 2006.

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First Report of Fusarium proliferatum Causing Fruit Rot of Winter Jujube (Zizyphus jujuba) in Storage in China

Plant Disease ◽

10.1094/pdis-12-11-1035-pdn ◽

2012 ◽

Vol 96 (6) ◽

pp. 913-913 ◽

Cited By ~ 3

Author(s):

M. Zhang ◽

Y. Wang ◽

C. Y. Wen ◽

H. Y. Wu

Keyword(s):

Apical Cell ◽

Morphological Characteristics ◽

Fusarium Proliferatum ◽

Fruit Rot ◽

Distilled Water ◽

First Report ◽

Pale Yellow ◽

Agar Plug ◽

Jujube Fruit ◽

Zizyphus Jujuba

Winter jujube, Zizyphus jujuba Mill., is a Chinese crop with fruit that has an extremely high nutritional value (4). In early November 2010, a severe fruit rot affecting ~20% of 1,000 kg of winter jujube fruit was observed in a storehouse in Zhengzhou, Henan province, China. The same fruit rot symptoms were found in two supermarkets in Zhengzhou in late November 2010 in ~10% of 100 kg of fruit in one supermarket and 25% of 50 kg of fruit in the other. Symptoms first appeared as small, round, pale yellow brown lesions on the fruits, 1 to 3 mm in diameter, then developed into 5- to 10-mm, sunken, brown spots, each with a pale brown margin. Three Fusarium isolates (DZF001 to DZF003) showing similar morphological characteristics were isolated from three specimens (collected from one storehouse and two supermarkets) by surface sterilizing small pieces of necrotic fruit tissue for 1 min in 2% NaOCl, washing the tissue pieces three times with sterile distilled water, and plating the pieces on potato dextrose agar (PDA). Fungal colonies for each isolate were white to light pink, and the adaxial side of each culture was pale yellow. Macroconidia were produced in pale orange sporodochia and were slender, relatively straight, three to five septa, 29.0 to 55.2 × 2.5 to 4.0 μm, with a curved apical cell and a poorly developed basal cell. Microconidia were produced in chains or false heads on synthetic nutrient-poor agar, clavate with a planar base, aseptate, and 4.5 to 8.0 × 2.5 to 3.5 μm. Conidiophores terminated in verticils of two to three phialides or monophialides. Chlamydospores were absent. The cultural and morphological characteristics were similar to those of Fusarium proliferatum (1,2). The identity of the three fungal isolates was confirmed to be F. proliferatum by DNA sequencing of the internal transcribed spacer (ITS) rDNA region (GenBank Accession Nos. JN889713 to JN889715), which were 99 to 100% homologous to those of other F. proliferatum isolates (GU066714, HQ113948, and GU363955); and the elongation factor 1-alpha (EF-1a) gene (JN889713 to JN889715), which was 99% homologous to those of other F. proliferatum isolates (FJ538244, FJ895277, and GQ848536) (3). Pathogenicity tests were conducted on 20 winter jujube fruits using a mycelial plug harvested from the periphery of a 7-day-old colony of strain DZF001, and placed on the surface of the fruit after the inoculated area of the fruit had been surface sterilized with 75% ethanol for 2 min; an equal number of fresh winter jujube fruits treated with non-colonized plugs of PDA served as the control treatment. Each jujube fruit was pricked three times with an insect needle to create three holes close together before inoculation with an agar plug. Each fruit was then enclosed in a clear plastic box with a cup of sterile distilled water to maintain high relative humidity, and held at 25°C. Symptoms similar to those originally observed on the naturally infected fruit were observed 3 days after inoculation, and the same fungus was reisolated from each of the symptomatic fruits; control fruits remained asymptomatic and no fungus was isolated from the control fruit. Koch's postulates were repeated three times with the same results. To our knowledge, this is the first report of F. proliferatum causing rot of winter jujube fruit in China. References: (1) K. Chehri et al. Saudi J. Biol. Sci. 18:341, 2011. (2) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual, Blackwell Publishing, 2006. (3) H. T. Phan. Studies Mycol. 50:261, 2004. (4) J. Sheng et al. Acta Hort. 620:203, 2003.

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First report of Leaf Wilt caused by Fusarium proliferatum and F. brachygibbosum on Date Palm (Phoenix dactylifera) in Tunisia

Plant Disease ◽

10.1094/pdis-08-20-1791-pdn ◽

2020 ◽

Author(s):

Ahmed Namsi ◽

Amal Rabaoui ◽

Mario Masiello ◽

Antonio Moretti ◽

Ahmed Othmani ◽

...

Keyword(s):

Date Palm ◽

Elongation Factor ◽

Aerial Mycelium ◽

Phoenix Dactylifera ◽

Fusarium Proliferatum ◽

Morphological Identification ◽

First Report ◽

Leaf Yellowing ◽

Colony Color

Since 2017, a new leaf wilt syndrome was observed in plantations of date palm in Tunisia. Its incidence increases sharply from year to year, especially in ‘Deglet Nour’ trees, aged between 5 and 15 years. In severe cases, the large number of dried leaves per tree can lead to complete cessation of date production. Symptoms appear on one or more leaves in the center of the crown. Whitening and drying start at the top of the leaflets and proceed to their base, while the midrib remains green. Then the whole leaf dies. Small white-creamy leaflet fragments and roots were collected from five different regions in the Djerid Oases. They were disinfected with diluted bleach (0,8 % NaOCl) and ethanol (80%) (each 2 min), rinsed with sterile distilled water, dried and finally plated in Petri dishes containing Potato Dextrose Agar (PDA) amended with 50mg/l neomycin. After incubation for 7 days at 25ºC±2, emerging fungal colonies were single-spored by serial dilution. They were transferred to PDA, Carnation Leaf Agar (CLA) and Spezieller Nahrstoffarmer Agar (SNA) for morphological identification. Based on the colony color on PDA, conidial morphology and phialide structures on CLA and/or SNA, of the 85 Fusarium isolates, around 90% were identified as F. proliferatum and around 10% as F. brachygibbosum (Leslie and Summerell, 2006). Fusarium proliferatum colonies rapidly developed white aerial mycelium that became purple in old cultures. Microconidia were abundant in the aerial mycelium and formed chains of variable length, on monophialides and polyphialids, a characteristic that distinguishes F. proliferatum from F. verticilloides. Less often, they were observed in false heads. Chlamydospores were absent. On CLA, microconidia were mostly 2 × 15 µm in size, a large number of sickle shaped macroconidia (2 × 25 µm) had one septum, some were larger (2 × 50 µm) with 3 septa and tips at both ends. Molecular identification was carried out based on elongation factor (EF-1α) gene sequencing. The region between the EF1 and EF2 primers (O’Donnell et al., 1998) was ampliﬁed and the sequences were compared to Fusarium reference sequences (GenBank). The sequences of the isolates Fus 1953 (539 bp), Fus 1962 (618 bp), and Fus 1965 (605 bp) shared respectively 100%, 99.51% and 99.51% homology with that of F. proliferatum JF740713.1 and were deposited in GenBank with the following accession numbers: MT630418, MT630419, and MT630420, respectively. The sequences of isolates 7F, 28F, Fus 1955 and Fus 1956 shared 100 % homology with that of F. brachygibbosum (GQ505418.1) while those of Fus 1955 and Fus 1956 showed 99.02 and 98.91 % identity, respectively, with F. brachygibbosum JX118981.1. The sequences of 7F (535 bp), 28F (535 bp), Fus 1955 (608 bp), and Fus 1956 (647 bp) were deposited in GenBank with the following accession numbers: MT630409, MT630410, MT630411, and MT630412, respectively. Two ml suspension of 106 conidia / ml of each isolate was sprayed separately or in combinations on in vitro cloned ‘Deglet Nour’ plants, placed in a greenhouse at 28°±2 °C and 70% R.H.. Isolates of F. proliferatum led to dryness and wilting leaflets after 3 weeks. Fusarium brachygibbosum only induced mild leaf yellowing, while in combination they were more virulent. Fungal isolates of both species were re-isolated and their identity confirmed to be the same of those isolated from leaflets infected in the open field, confirming Koch’s postulates. Control plants lacked symptoms. Fusarium proliferatum is known as date palm pathogen in many countries (Saleh et al. 2017), however, to our knowledge, this is the first report of F. proliferatum and also F. brachygibbosum causing Leaf Wilt symptoms on P. dactylifera in Tunisia.

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