First Report of Fusarium redolens Causing Root Rot of Soybean in Minnesota

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

Plant Disease ◽

10.1094/pdis-01-14-0040-pdn ◽

2014 ◽

Vol 98 (7) ◽

pp. 1009-1009 ◽

Cited By ~ 2

Author(s):

L. M. Quesada-Ocampo ◽

S. Butler ◽

S. Withers ◽

K. Ivors

Keyword(s):

North Carolina ◽

Apical Cell ◽

Elongation Factor ◽

Morphological Characteristics ◽

Aerial Mycelium ◽

The United States ◽

Fusarium Proliferatum ◽

First Report ◽

Its Regions ◽

Dry Conditions

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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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 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 proliferatum Causing Root Rot on Soybean (Glycine max) in the United States

Plant Disease ◽

10.1094/pdis-04-11-0346 ◽

2011 ◽

Vol 95 (10) ◽

pp. 1316-1316 ◽

Cited By ~ 18

Author(s):

M. M. Díaz Arias ◽

G. P. Munkvold ◽

L. F. Leandro

Keyword(s):

United States ◽

Root Rot ◽

Morphological Characteristics ◽

Dry Weight ◽

The United States ◽

Growth Stages ◽

Species Identity ◽

First Report ◽

Soybean Roots ◽

Soybean Diseases

Fusarium spp. are widespread soilborne pathogens that cause important soybean diseases such as damping-off, root rot, Fusarium wilt, and sudden death syndrome. At least 12 species of Fusarium, including F. proliferatum, have been associated with soybean roots, but their relative aggressiveness as root rot pathogens is not known and pathogenicity has not been established for all reported species (2). In collaboration with 12 Iowa State University extension specialists, soybean roots were arbitrarily sampled from three fields in each of 98 Iowa counties from 2007 to 2009. Ten plants were collected from each field at V2-V3 and R3-R4 growth stages (2). Typical symptoms of Fusarium root rot (2) were observed. Symptomatic and asymptomatic root pieces were superficially sterilized in 0.5% NaOCl for 2 min, rinsed three times in sterile distilled water, and placed onto a Fusarium selective medium. Fusarium colonies were transferred to carnation leaf agar (CLA) and potato dextrose agar and later identified to species based on cultural and morphological characteristics. Of 1,230 Fusarium isolates identified, 50 were recognized as F. proliferatum based on morphological characteristics (3). F. proliferatum isolates produced abundant, aerial, white mycelium and a violet-to-dark purple pigmentation characteristic of Fusarium section Liseola. On CLA, microconidia were abundant, single celled, oval, and in chains on monophialides and polyphialides (3). Species identity was confirmed for two isolates by sequencing of the elongation factor (EF1-α) gene using the ef1 and ef2 primers (1). Identities of the resulting sequences (~680 bp) were confirmed by BLAST analysis and the FUSARIUM-ID database. Analysis resulted in a 99% match for five accessions of F. proliferatum (e.g., FD01389 and FD01858). To complete Koch's postulates, four F. proliferatum isolates were tested for pathogenicity on soybean in a greenhouse. Soybean seeds of cv. AG2306 were planted in cones (150 ml) in autoclaved soil infested with each isolate; Fusarium inoculum was applied by mixing an infested cornmeal/sand mix with soil prior to planting (4). Noninoculated control plants were grown in autoclaved soil amended with a sterile cornmeal/sand mix. Soil temperature was maintained at 18 ± 1°C by placing cones in water baths. The experiment was a completely randomized design with five replicates (single plant in a cone) per isolate and was repeated three times. Root rot severity (visually scored on a percentage scale), shoot dry weight, and root dry weight were assessed at the V3 soybean growth stage. All F. proliferatum isolates tested were pathogenic. Plants inoculated with these isolates were significantly different from the control plants in root rot severity (P = 0.001) and shoot (P = 0.023) and root (P = 0.013) dry weight. Infected plants showed dark brown lesions in the root system as well as decay of the entire taproot. F. proliferatum was reisolated from symptomatic root tissue of infected plants but not from similar tissues of control plants. To our knowledge, this is the first report of F. proliferatum causing root rot on soybean in the United States. References: (1) D. M. Geiser et al. Eur. J. Plant Pathol. 110:473, 2004. (2) G. L. Hartman et al. Compendium of Soybean Diseases. 4th ed. The American Phytopathologic Society, St. Paul, MN, 1999. (3) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Oxford, UK, 2006. (4) G. P. Munkvold and J. K. O'Mara. Plant Dis. 86:143, 2002.

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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 culmorum Causing Maize Stalk Rot in China

Plant Disease ◽

10.1094/pdis-07-21-1442-pdn ◽

2021 ◽

Author(s):

Laikun Xia ◽

Yanyong Cao ◽

Jie Wang ◽

Jie Zhang ◽

Shengbo Han ◽

...

Keyword(s):

Fusarium Species ◽

Apical Cell ◽

Fusarium Culmorum ◽

Aerial Mycelium ◽

Henan Province ◽

Population Diversity ◽

Pathogenicity Test ◽

Ear Rot ◽

First Report ◽

Stalk Rot

Maize stalk rot has become one of the most important diseases in maize production in China. From 2017 to 2019, a survey was conducted to determine the population diversity of Fusarium species associated with maize diseases in 18 cities across Henan Province. Maize stalk rot with an incidence of more than 20% that caused yield losses up to 30% was observed on maize variety Zhengdan958, which was grown in two continuous maize fields in Zhumadian City, Henan Province. The stem tissues from the boundary between diseased and healthy pith were chopped into small pieces (3 × 8 mm), disinfected (70% ethanol for 1 min) and then placed onto potato dextrose agar (PDA) amended with L-(+)-Lactic-acid (1 g/L) and incubated at 25°C for 4 days. Colonies on PDA produced fluffy, light yellow aerial mycelium and purple to deep brick red pigment at 25°C (Fig 1A, 1B). On carnation leaf agar (CLA), macroconidia in orange sporodochia formed abundantly, but microconidia were absent. Macroconidia were short and thick-walled, had 3 to 5 septa, a poorly developed foot cell and rounded apical cell (Fig 1C). These characteristics matched the description of Fusarium culmorum (Leslie and Summerell 2006) and isolates DMA268-1-2 and HNZMD-12-7 were selected for further identity confirmation. Species identification was confirmed by partial sequences of three phylogenic loci (EF1-α, RPB1, and RPB2) using the primer pairs EF1/EF2, CULR1F/CULR1R, and CULR2F/CULR2R, respectively (O'Donnell et al., 1998). The consensus sequences from the two isolates were deposited in GenBank (MZ265416 and MZ265417 for TEF, respectively; MZ265412 and MZ265414 for RPB1, respectively; MZ265413 and MZ265415 for RPB2). BLASTn searches indicated that the nucleotide sequences of the three loci of the two isolates revealed 99% to 100% similarity to those of F. culmorum strains deposited in the GenBank, Fusarium-ID, and MLST databases (Supplementary Table 1~3). Pathogenicity test was conducted at the flowering-stage using Zhengdan958 and Xundan20 plants according to previously described method (Zhang et al., 2016; Cao et al., 2021; Zhang et al., 2021). The second or third internodes of thirty flowering plants were drilled to make a wound approximately 8 mm in diameter using an electric drill. Approximately 0.5 mL inoculum (125 mL colonized PDA homogenized with 75 mL sterilized distilled water) was injected into the wound and sealed with Vaseline and Parafilm to maintain moisture and avoid contamination. Sterile PDA slurry was used as a control. Thirty days after inoculation, the dark-brown, soft rot of pith tissues above and below the injection sites were observed, and some plants were severely rotten and lodged (Fig 1D, 1E). These symptoms were similar to those observed in the field. No symptoms were observed on control plants. The same pathogen was re-isolated from the inoculated stalk lesions but not from the control, thereby fulfilling Koch's postulates. To our knowledge, this is the first report of F. culmorum as the causal agent of stalk rot on maize plants in China. Also, this fungus has been reported to cause maize ear rot in China (Duan et al. 2016) and produce mycotoxins such as trichothecenes, nivalenol, and zearalenone that cause toxicosis in animals (Leslie and Summerell 2006). The occurrence of maize stalk rot and ear rot caused by F. culmorum should be monitored due to the potential risk for crop loss and mycotoxin contamination.

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First Report of Lasiodiplodia theobromae Associated with Decline of Grapevine Rootstock Mother Plants in Spain

Plant Disease ◽

10.1094/pdis-92-5-0832b ◽

2008 ◽

Vol 92 (5) ◽

pp. 832-832 ◽

Cited By ~ 13

Author(s):

A. Aroca ◽

R. Raposo ◽

D. Gramaje ◽

J. Armengol ◽

S. Martos ◽

...

Keyword(s):

Elongation Factor ◽

Aerial Mycelium ◽

Its Region ◽

Pathogenic Fungi ◽

Vascular Lesions ◽

Malt Extract ◽

Pathogenicity Test ◽

Lasiodiplodia Theobromae ◽

First Report ◽

Mother Plants

A field of Richter 110 rootstock mother plants in Valencia Province (eastern Spain) was surveyed during November 2006 to study the mycoflora of declining plants. Two canes with stunted leaves were collected from a plant with a reduced number of shoots. No cankers or vascular lesions were observed in the collected canes. Six wood chips (1 to 2 mm thick) were taken from one basal fragment (3 to 4 cm long) of each cane, surface sterilized in 70% ethanol for 1 min, and plated on malt extract agar supplemented with 0.5 g L–1 of streptomycin sulfate. Petri dishes were incubated for 7 days at 25°C. A fungus was consistently isolated from all samples that showed the following characteristics: colonies grown on potato dextrose agar (PDA) at 25°C developed a white, aerial mycelium that turned gray after 4 to 6 days and produced pycnidia after 1 month on sterile grapevine slivers of twigs placed on the PDA surface; conidia from culture were ellipsoidal, thick walled, initially hyaline, nonseptate, and measuring 20 to 25 (22.5) × 12 to 14 (13) μm; aged conidia were brown, 1-septate with longitudinal striations in the wall; and pseudoparaphyses variable in form and length were interspersed within the fertile tissue. The fungus was identified as Lasiodiplodia theobromae (Pat.) Griffon & Maubl. from the above characteristics (2). Identity was confirmed by analysis of the nucleotide sequences of the internal transcribed spacer (ITS) region from the rRNA repeat and part of the translation elongation factor 1-alpha (EF1-α) and the β-tubulin (B-tub) genes, as done elsewhere (1,3). BLAST searches at GenBank showed a high identity with reference sequences (ITS: 100%, EF1-α: 97%; B-tub: 99%). Representative sequences of the studied DNA regions were deposited at GenBank (Accession Nos.: ITS: EU254718; EF1-α: EU254719; and B-tub: EU254720). A pathogenicity test was conducted on 1-year-old grapevine plants cv. Macabeo grafted onto Richter 110 rootstocks maintained in a greenhouse. A superficial wound was made on the bark of 10 plants with a sterilized scalpel, ≈10 cm above the graft union. A mycelial plug obtained from the margin of an actively growing fungal colony (isolate JL664) was placed in the wound and the wound was wrapped with Parafilm. Ten additional control plants were inoculated with sterile PDA plugs. All control plants grew normally, and the inoculation wound healed 3 months after inoculation. Plants inoculated with L. theobromae showed no foliar symptoms in the same period, but developed cankers variable in size surrounding the inoculation sites. Vascular necroses measuring 8.4 ± 1.5 cm (mean ± standard error) developed in the inoculated plants that were significantly longer than the controls (0.3 ± 0.2 cm). The pathogen was reisolated from all inoculated plants and no fungus was reisolated from the controls. These results confirmed the pathogenicity of L. theobromae to grapevine and points to a possible involvement of L. theobromae in the aetiology of grapevine decline as previously reported (3,4). To our knowledge, this is the first report of L. theobromae isolated from grapevine in Spain. References: (1) J. Luque et al. Mycologia 97:1111, 2005. (2) E. Punithalingam. No. 519 in: Descriptions of Pathogenic Fungi and Bacteria. CMI, Kew, Surrey, UK, 1976. (3) J. R. Úrbez-Torres et al. Plant Dis. 90:1490, 2006. (4) J. M. van Niekerk et al. Phytopathol. Mediterr. 45(suppl.):S43, 2006.

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First report of Fusarium cf. longipes associated with maize stalk rot in China

Plant Disease ◽

10.1094/pdis-06-21-1149-pdn ◽

2021 ◽

Author(s):

Shengbo Han ◽

Yanyong Cao ◽

Jie Zhang ◽

Jie Wang ◽

Lili Zhang ◽

...

Keyword(s):

Sequence Data ◽

Fusarium Species ◽

Apical Cell ◽

Morphological Characteristics ◽

Liaoning Province ◽

Bootstrap Support ◽

Flowering Stage ◽

Single Spore ◽

First Report ◽

Stalk Rot

In a field survey from 2017 to 2019, Fusarium stalk rot symptoms including discolored, disintegrated stalk pith tissues and lodged plants were observed in maize hybrid lines Fuyu1611, Jidan66, and Danyu8439 grown in fields in Anshan (40o49′39′′N, 122 o34′6′′E), Liaoning province. Its incidence ranged from 15% to 20% and caused a yield loss of up to 30%. Infected pieces of stem tissues were dissected and then sterilized with 1% NaOCl for 1 min, 70% ethanol for 1 min, rinsed 3 times with sterilized ddH2O, and dried with filter paper in hood. Three pieces were placed onto Potato dextrose agar (PDA) and incubated at 25 °C for 5 days. The colonies were single-spore subcultured on PDA at 25 °C for 2 weeks (Leslie and Summerell 2006). Morphological features were observed on PDA and carnation leaf agar (CLA). The average mycelial growth rate was 4.5 to 10.3 mm/day at 25 °C on PDA. The colonies produced aerial mycelia, varying from dense white to grayish-rose, and secreted red pigments in the agar (Fig. 1A; 1B). Macroconidia produced on CLA were long and relatively slender, commonly 4- to 7-septate, averaging 85.6 × 5.2 μm, with thick walls and pronounced dorsiventral curvature with a distinctly foot-shaped and elongated basal cell and an apical cell that was whip-like (Fig. 1C). Microconidia were rarely observed on PDA or CLA. Morphological characteristics of the isolates were similar to the features of Fusarium longipes as previously described (Leslie and Summerell 2006). The portions of three phylogenic loci (EF1-α, RPB1, RPB2) were PCR amplified using the primer pairs EF1/EF2 (O'Donnell et al, 1998), lonR1F/lonR1R (5-TTTTCCTCACCAAGGAGCAGATCATG-3 and 5-CCAATGGACTGGGCAGCCAAAACGCC-3) and lonR2F/lonR2R (5-TATACATTTGCCTCCACTCTTTCCCAT-3 and 5-CGGAGCTTGCGTCCGGTGTGGCCGTTG-3) and sequenced. The consensus sequences were submitted to GenBank (MT513215 and MT997083 for TEF, MT513213 and MT997088 for RPB1; MT513214 and MW020572 for RPB2). BLASTn searches indicated that the nucleotide sequences of the three loci of the two isolates shared 94.52% to 99.69% identity with sequences of F. longipes strains deposited in the GenBank, Fusarium-ID and Fusarium MLST databases (Supplementary Table 1, 3, 4). A phylogram inferred via maximum likelihood analysis of the combined EF-1α, RPB1, RPB2 partial sequence data of Fusarium species (Supplementary Table 2) was inferred using the CIPRIES website (https://www.phylo.org). Isolates LNAS-05-A and LNAS-09-A clustered with F. longipes, with 98% bootstrap support (Fig. 2). Pathogenicity tests were conducted on three-leaf-stage seedlings and flowering-stage c.v. Zhengdan958 and B104 plants according to previously described methods (Ye et al., 2013; Zhang et al. 2016) with minor modifications. Three days after the roots of the seedlings were inoculated with 1 × 106 macroconidia solution, the leaves and stems exhibited typical wilt symptoms (Fig. 1D). Twenty flowering-stage maize plants were drilled individually at the second or third node above the soil using an electric drill (Bosch TSR1080-2-Li) to create a hole (8 mm in diameter). An approximately 0.5 mL mycelia plug (125 mL homogenized hyphal mats + 75 mL sterilized ddH2O) was injected into the hole and covered with Vaseline. Sterilized PDA plugs were used as a control. The stalk tissue of the split internodes turned dark brown and the brown area expanded above and below the injection site by 14 dpi. All of the inoculated plants developed characteristic stalk rot symptoms, whereas no symptoms were observed in the controls (Fig. 1E). The pathogen was re-isolated, and its identity was confirmed by sequencing the above mentioned loci. F. longipes was generally regarded as a tropical Fusarium species (Leslie and Summerell 2006). This is the first report that F. cf. longipes can cause stalk rot of maize under filed condition in a temperate, typical corn belt region of China.

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First Report of Fusarium succisae Causing Flower Rot on Thread-leaf Coreopsis

Plant Disease ◽

10.1094/pdis-12-13-1205-pdn ◽

2014 ◽

Vol 98 (7) ◽

pp. 1002-1002 ◽

Cited By ~ 2

Author(s):

H.-W. Choi ◽

S. K. Hong ◽

Y. K. Lee ◽

W. G. Kim

Keyword(s):

Dna Sequences ◽

Production Control ◽

Fusarium Species ◽

Elongation Factor ◽

Morphological Characteristics ◽

Spore Suspension ◽

Sugar Solution ◽

Laboratory Manual ◽

First Report ◽

Blastn Analysis

In July 2010, flower rot of thread-leaf coreopsis (Coreopsis verticillata) was found in a garden in the Icheon City, Korea. The disease affected about 20 to 50% of a 100 m2 area. The disease was characterized by the appearance of pinkish mycelia on the stigmata and inflorescences of flowers. In some cases, flowers failed to bloom or turned brown before opening fully. Fragments (each 5 × 5 mm) of the symptomatic tissue were surface-sterilized with 1% NaOCl for 1 min, and then rinsed twice in sterilized distilled water. The tissue pieces were placed on water agar (WA) and incubated at 25°C for 4 to 6 days. Twenty-two isolates of Fusarium species were obtained from the diseased flowers. All isolates were identified as Fusarium succisae based on their morphological characteristics on carnation leaf agar (CLA) medium and DNA sequences of the translation elongation factor 1-alpha gene (1). Macroconidia and sporodochia were sparsely produced on CLA medium. Microconidia were abundant, borne in false heads, oval or allantoid and sometimes pyriform, and measured 4.2 to 13 × 2.2 to 5.4 μm. Chlamydospores were absent. The EF-1α gene was amplified from three isolates by PCR assay and the amplification products were sequenced (2). The nucleotide sequences obtained were deposited in GenBank with accession numbers KF514658, KF514659, and KF514660. BLASTn analysis showed 99% homology with the EF-1α sequence of F. succisae NRRL13613 (GenBank Accession No. AF160291). Pathogenicity tests were conducted with inoculation of flowers on Coreopsis verticillata. Spore suspension was prepared by flooding 7-day-old cultures on potato dextrose agar with sterilized 2% (w/v) sugar solution. When the plants started to have buds, the isolates were inoculated by placing one drop (20 μl) of spore suspension (1 × 106 spores ml−1) into the buds. Fifteen buds of the plants were arranged into three replications. The control was treated with sterilized 2% sugar solution. Inoculated plants were kept in a greenhouse at 25/20°C (12 h/12 h). Three weeks after inoculation, the symptoms were observed on buds with mycelial production. Control plants had no mycelia on buds. F. succisae was re-isolated from the inoculated flowers. To our knowledge, this is the first report of flower rot of thread-leaf coreopsis caused by F. succisae. References: (1) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Ames, IA, 2006. (2) K. O'Donnell et al. Proc. Nat. Acad. Sci. 95:2044, 1998.

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Root Rot Caused by Species of Fusarium on Brassica carinata in South Dakota

Plant Health Progress ◽

10.1094/php-04-18-0012-rs ◽

2018 ◽

Vol 19 (3) ◽

pp. 188-192

Author(s):

Paul N. Okello ◽

Kristina Petrović ◽

Brian Kontz ◽

Shaukat Ali ◽

Laura F. Marek ◽

...

Keyword(s):

South Dakota ◽

Root Rot ◽

Rating Scale ◽

Fusarium Species ◽

Phylogenetic Analyses ◽

Elongation Factor ◽

The United States ◽

Brassica Carinata ◽

Oilseed Crop ◽

Root Diseases

Brassica carinata is an emerging oilseed crop in the United States, and root diseases caused by Fusarium have the potential to cause yield losses in production. In this study, B. carinata plants were randomly sampled at vegetative and seed development plant stages from South Dakota State University experimental plots. Reddish-brown lesions were observed on roots of sampled plants from which F. acuminatum, F. oxysporum, F. solani, and F. sporotrichioides were recovered. The Fusarium species were identified based on morphology and phylogenetic analyses of the translation elongation factor 1-α gene region. Pathogenicity of the four Fusarium species was evaluated on five B. carinata accessions using a modified inoculum layer method in the greenhouse. At 21 days after inoculation, root rot severity caused by Fusarium on the B. carinata accessions was assessed on a rating scale of 0 to 4 and evaluated using relative treatment effects (RTEs). The F. oxysporum isolate caused significant differences in RTE (P = 0.01) among the B. carinata accessions. However, there were no significant differences in RTE among the B. carinata accessions in response to F. acuminatum (P = 0.82), F. solani (P = 0.76), and F. sporotrichioides (P = 0.47) isolates.

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