Soybean Sudden Death Syndrome Causal Agent Fusarium brasiliense Present in Michigan

Sudden death syndrome (SDS), caused by members of Fusarium solani species complex (FSSC) clade 2, is a major and economically important disease in soybean worldwide. The primary causal agent of SDS isolated to date in North America has been F. virguliforme. In 2014 and 2016, SDS symptoms were found in two soybean fields located on the same farm in Michigan. Seventy Fusarium strains were isolated from roots of the SDS-symptomatic soybeans in two fields. Phylogenetic analysis of partial sequences of elongation factor-1α, the nuclear ribosomal DNA intergenic spacer region, and the RNA polymerase II beta subunit revealed that the primary FSSC species isolated was F. brasiliense (58 and 36% in each field) and the remaining Fusarium strains were identified as F. cuneirostrum, F. phaseoli, an undescribed Fusarium sp. from FSSC clade 2, and strains in FSSC clade 5 and FSSC clade 11. Molecular identification was supported with morphological analysis and a pathogenicity assay. The soybean seedling pathogenicity assay indicated that F. brasiliense was capable of causing typical foliar SDS symptoms. Both root rot and foliar disease severity were variable by strain, just as they are in F. virguliforme. Both FSSC 5 and FSSC 11 strains were also capable of causing root rot, but SDS foliar symptoms were not detected. To our knowledge, this is the first report of F. brasiliense causing SDS in soybean in the United States and the first report of F. cuneirostrum, F. phaseoli, an as-yet-unnamed Fusarium sp., and strains in FSSC clade 5 and FSSC clade 11 associated with or causing root rot of soybean in Michigan.

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First Report and Confirmed Distribution of Soybean Sudden Death Syndrome Caused by Fusarium virguliforme in Southern Michigan

Plant Disease ◽

10.1094/pdis-94-9-1164b ◽

2010 ◽

Vol 94 (9) ◽

pp. 1164-1164 ◽

Cited By ~ 9

Author(s):

M. I. Chilvers ◽

D. E. Brown-Rytlewski

Keyword(s):

Sudden Death ◽

Root Rot ◽

Field Trial ◽

The United States ◽

Sudden Death Syndrome ◽

Fusarium Virguliforme ◽

Koch’S Postulates ◽

Interveinal Chlorosis ◽

Koch's Postulates ◽

Β Tubulin

Leaf lesions and root rot symptoms typical of soybean sudden death syndrome (SDS) caused by Fusarium virguliforme O'Donnell & T. Aoki were observed in commercial soybeans (Glycine max (L.) Merr.) in southern Michigan. Leaf symptoms ranged from chlorotic spots to severe interveinal chlorosis and necrosis, no foliar pathogens were noted. In 2008, isolates were collected from Berrien and St. Joseph counties. In 2009, isolates were collected from Cass, St. Joseph, Van Buren, Allegan, and Monroe counties. Pieces of roots with root rot symptoms were washed prior to surface disinfestation with 70% ethanol for 30 s and 0.5% NaOCl for 1 min and incubated on water agar (WA) in petri plates amended with 50 μg/ml of chloramphenicol for the production of sporodochia. Alternatively, spores were collected directly from nondisinfested roots expressing blue sporodochia. Single-spore cultures were derived by streaking macroconidia with a bacterial loop onto 3% WA + chloramphenicol and incubated overnight. With a dissecting microscope, single germinated macroconidia were collected with a sterile 0.2-mm-diameter insect pin and transferred to potato dextrose agar (PDA). Cultures on PDA grew slowly and developed blue-to-purple masses of sporodochia typical of F. virguliforme descriptions and similar to a representative isolate, Mont-1, grown alongside (1,2). Size of macroconidia from the six representative isolates, one from each county (including isolates derived from surface-disinfested and nondisinfested roots), and Mont-1 were determined to be within the range for F. virguliforme (42 to 56 × 5 to 6 μm), with an average of four septa per macroconidia. Identity of the representative isolates was confirmed by partial DNA sequencing of both strands of the internal transcribed spacer (ITS) region of the ribosomal RNA gene, translation elongation factor 1-α, and β-tubulin loci. All six representative isolates were identical in each of the three loci and matched with 100% similarity F. virguliforme accessions in GenBank and Fusarium-ID database searches, except for the β-tubulin locus in which a single nucleotide insertion was noted (Accession Nos. HM453328–HM453330). Sequences were 98 to 99% similar to other SDS Fusarium spp. not yet recorded in the United States. Koch's postulates were performed in the greenhouse according to Malvick and Bussey (3). Infested sorghum seed (~20 g) was placed 2 cm below soybean seed of susceptible cv. Williams 82 in plastic pots. Noninfested sorghum seed was used as a negative control and sorghum infested with Mont-1 as a positive control. Chlorotic spots developed 2 weeks after establishing the trial, and 3 to 4 weeks postinoculation, severe SDS symptoms of foliar interveinal chlorosis and necrosis and severe root rot developed. Koch's postulates were completed by reisolating F. virguliforme from a subset of infected plants. In addition, an isolate of F. virguliforme collected in 2008 was used to inoculate a 2009 field trial in East Lansing, MI with no history of SDS. Typical SDS symptoms developed in the field trial and F. virguliforme was isolated from a symptomatic plant that was identified as described above. Despite being reported across the majority of soybean-producing states, to our knowledge, this is the first confirmation and distribution report for SDS in Michigan. References: (1) T. Aoki et al. Mycoscience 46:162, 2005. (2) G. L. Hartman et al. Plant Dis. 81:515, 1997. (3) D. K. Malvick and K. E. Bussey. Can. J. Plant Pathol. 30:467, 2008.

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First Report ofFusarium solaniBlue Isolate, a Causal Agent of Sudden Death Syndrome of Soybeans, Recovered from Soybean Cyst Nematode Eggs

Plant Disease ◽

10.1094/pd-77-0647c ◽

1993 ◽

Vol 77 (6) ◽

pp. 647C ◽

Cited By ~ 4

Author(s):

P. A. Donald

Keyword(s):

Sudden Death ◽

Soybean Cyst Nematode ◽

Cyst Nematode ◽

Causal Agent ◽

Sudden Death Syndrome ◽

First Report ◽

Nematode Eggs

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First Report of Pythium aphanidermatum Crown and Root Rot of Industrial Hemp in the United States

Plant Disease ◽

10.1094/pdis-09-16-1249-pdn ◽

2017 ◽

Vol 101 (6) ◽

pp. 1038 ◽

Cited By ~ 7

Author(s):

J. Beckerman ◽

H. Nisonson ◽

N. Albright ◽

T. Creswell

Keyword(s):

United States ◽

Root Rot ◽

The United States ◽

Pythium Aphanidermatum ◽

First Report ◽

Crown And Root Rot ◽

Industrial Hemp

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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 Medical Tree Peony Root Rot Caused by Fusarium solani in Tongling, China

Plant Disease ◽

10.1094/pdis-02-12-0142-pdn ◽

2012 ◽

Vol 96 (6) ◽

pp. 909-909 ◽

Cited By ~ 10

Author(s):

M. Guo ◽

Y. M. Pan ◽

Z. M. Gao

Keyword(s):

Fusarium Solani ◽

Root Rot ◽

Tap Water ◽

Blood Stasis ◽

The United States ◽

Effective Control ◽

Conidial Suspension ◽

Tree Peony ◽

First Report ◽

Peony Root

Tree peony bark, a main component of Chinese traditional medicine used for alleviating fever and dissipating blood stasis, is mainly produced in Tongling, China. Recently, tree peony cultivation in this area was seriously affected by root rot, with approximately 20 to 30% disease incidence each year. The disease severely affects yield and quality of tree peony bark. During the past 2 years, we collected 56 diseased tree peony plants from Mudan and Fenghuang townships in Tongling. We found reddish brown to dark brown root rot in mature roots, especially on those with injuries. Plant samples collected were disinfected with 2% sodium hypochlorite and isolations were conducted on potato sucrose agar (PSA). Eleven isolates were obtained and all had white fluffy aerial hypha on PSA. Two types of conidia were produced; the larger, reaphook-shaped ones had three to five septa and the smaller, ellipse-shaped ones had one or no septum. The reaphook-shaped conidia were 20.15 to 37.21 × 3.98 to 5.27 μm and the ellipse-shaped conidia were 6.02 to 15.52 × 2.21 to 5.33 μm in size. Chlamydospores were produced, with two to five arranged together. Biological characteristics of the fungi indicated that the optimum temperature for the mycelial growth on PSA was 25 to 30°C and the optimum pH range was 5.5 to 7.0. The above morphological characteristics point the fungal isolates to be Fusarium solani. To confirm pathogenicity, 30 healthy 1-year-old tree peony seedling plants were grown in pots (25 cm in diameter) with sterilized soil and a conidial suspension from one isolate (FH-1, 5 × 105 conidia/ml) was used for soil inoculation. Inoculated seedlings were maintained at 28°C in a greenhouse with a 12-h photoperiod of fluorescent light. Seedlings inoculated with distilled water were used as controls. After 3 weeks, the roots were collected and rinsed with tap water. Dark brown lesions were observed in the inoculated mature roots but not in the control roots. To confirm the identity of the pathogen, F. solani strains were reisolated from the lesions and total genomic DNA was extracted with the cetyltriethylammnonium bromide method from the mycelia of the reisolated strains (1). PCR was performed using the fungal universal primers ITS4 (5′-TCCTCCGCTTATTGATATGC-3′) and ITS5 (5′-GGAAGTAAAAGTCGTAACAAGG-3′) to amplify a DNA fragment of approximately 590 bp. The purified PCR products were sequenced (Invitrogen Co., Shanghai, China) and shared 100% sequence identity with each other. A comparison of the sequence (JQ658429.1) by the Clustal_W program (2) with those uploaded in GenBank confirmed with the fungus F. solani (100% sequence similarity to isolate S-0900 from the Great Plains of the United States; EU029589.1). To our knowledge, this is the first report of F. solani causing medical tree peony root rot in China. The existence of this pathogen in China may need to be considered for developing effective control strategies. References: (1). C. N. Stewart et al. Biotechniques 14:748, 1993. (2). J. D. Thompson et al. Nucleic Acids Res. 22:4673, 1994.

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Relationship Between Sudden Death Syndrome caused by Fusarium virguliforme and Soybean Yield: A Meta-Analysis

Plant Disease ◽

10.1094/pdis-11-19-2441-re ◽

2020 ◽

Vol 104 (6) ◽

pp. 1736-1743

Author(s):

Yuba R. Kandel ◽

Carl A. Bradley ◽

Martin I. Chilvers ◽

Febina M. Mathew ◽

Albert U. Tenuta ◽

...

Keyword(s):

Sudden Death ◽

Fixed Effects ◽

Field Experiments ◽

Crop Protection ◽

The United States ◽

Sudden Death Syndrome ◽

Effect Sizes ◽

Uniform Field ◽

Mixed Effect ◽

Moderator Variables

In total, 52 uniform field experiments were conducted in Illinois, Indiana, Iowa, Michigan, South Dakota, and Wisconsin in the United States and Ontario, Canada from 2013 to 2017 comparing crop protection products against sudden death syndrome (SDS) of soybean. Data were analyzed using meta-analytic models to summarize the relationship between foliar disease index (FDX) and yield. For each study, correlation and regression analyses were performed separately to determine three effect sizes: Fisher’s transformation of correlation coefficients (Z r ), intercept (β0), and slope (β1). Random- and mixed-effect meta-analyses were used to summarize the effect sizes. Study- and location-specific moderator variables FDX (low < 10% and high ≥ 10%), date of planting (early = prior to 7 May, conventional = 7 to 21 May, and late = after 21 May) cultivar (susceptible and partially resistant to SDS), study location, and growing season were used as fixed effects. The overall mean effect sizes of transformed correlation coefficient [Formula: see text] r was −0.41 and different from zero (P < 0.001), indicating that yield was negatively correlated with FDX. The [Formula: see text] r was affected by disease level (P < 0.01) and cultivar (P = 0.02), with a greater effect at higher disease levels and with susceptible cultivars. The mean [Formula: see text] 0 was 4,121 kg/ha and mean [Formula: see text] 1 was −21 kg/ha/% FDX and were different from zero (P < 0.01). Results from these data indicate that, for every unit of FDX increase, yield was decreased by 0.5%. Study locations and year affected the [Formula: see text] 0 , whereas none of the moderator variables significantly affected [Formula: see text] 1.

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Field Response of Glyphosate-Tolerant Soybean to Herbicides and Sudden Death Syndrome

Plant Disease ◽

10.1094/pdis.2001.85.7.773 ◽

2001 ◽

Vol 85 (7) ◽

pp. 773-779 ◽

Cited By ~ 45

Author(s):

S. Sanogo ◽

X. B. Yang ◽

P. Lundeen

Keyword(s):

Sudden Death ◽

Field Experiments ◽

Foliar Application ◽

The United States ◽

Sudden Death Syndrome ◽

North Central ◽

The North ◽

Foliar Symptoms ◽

Field Response ◽

Soybean Plants

Three-year field experiments were conducted to assess the development of sudden death syndrome (caused by Fusarium solani f. sp. glycines) in three soybean cultivars, tolerant (P9344 and A3071) and nontolerant (BSR101), to glyphosate following foliar application of four herbicides (acifluorfen, glyphosate, imazethapyr, and lactofen) commonly applied to soybeans in the north-central region of the United States. Cultivar A3071 is resistant to sudden death syndrome, whereas cultivars P9344 and BSR101 are susceptible to this disease. There was no statistically significant cultivar-herbicide interaction with respect to the severity of foliar symptoms of the disease and the frequency of isolation of F. solani f. sp. glycines from roots of soybean plants. Across all herbicide treatments, the level of sudden death syndrome was lower in the disease-resistant cultivar than in the susceptible ones. There was an increase in the disease levels under application of acifluorfen, glyphosate, and imazethapyr compared with nontreated or lactofen-treated plants. The results obtained indicate that the response of glyphosate-tolerant soybeans to sudden death syndrome is not different from the response of conventional soybeans to this disease following application of the selected herbicides, and the resistance of soybean to sudden death syndrome was not changed with application of glyphosate.

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First Report of Stem Blight Caused by Calonectria colhounii (Anamorph Cylindrocladium colhounii) on Greenhouse-Grown Blueberries in the United States

Plant Disease ◽

10.1094/pdis-02-11-0117 ◽

2011 ◽

Vol 95 (9) ◽

pp. 1187-1187

Author(s):

J. J. Sadowsky ◽

T. D. Miles ◽

A. M. C. Schilder

Keyword(s):

United States ◽

North Carolina ◽

Root Rot ◽

The United States ◽

Vaccinium Corymbosum ◽

Conidial Suspension ◽

Centraalbureau Voor ◽

First Report ◽

Stem Lesions ◽

Β Tubulin

Necrotic stems and leaves were observed on 2- to 4-month-old, rooted microshoot plants (Vaccinium corymbosum L. ‘Liberty’ and ‘Bluecrop’, V. angustifolium Aiton ‘Putte’, and V. corymbosum × V. angustifolium ‘Polaris’) in a Michigan greenhouse in 2008 and 2009. As the disease progressed, leaves fell off and 80 to 100% of the plants died in some cases. Root rot symptoms were also observed. A fungus was isolated from stem lesions. On potato dextrose agar (PDA), cultures first appeared light tan to orange, then rusty brown and zonate with irregular margins. Chains of orange-brown chlamydospores were abundant in the medium. Macroconidiophores were penicillately branched and had a stipe extension of 220 to 275 × 2.5 μm with a narrowly clavate vesicle, 3 to 4 μm wide at the tip. Conidia were hyaline and cylindrical with rounded ends, (1-)3-septate, 48 to 73 × 5 to 7 (average 60 × 5.5) μm and were held together in parallel clusters. Perithecia were globose to subglobose, yellow, 290 to 320 μm high, and 255 to 295 μm in diameter. Ascospores were hyaline, 2- to 3-septate, guttulate, fusoid with rounded ends, slightly curved, and 30 to 88 × 5 to 7.5 (average 57 × 5.3) μm. On the basis of morphology, the fungus was identified as Calonectria colhounii Peerally (anamorph Cylindrocladium colhounii Peerally) (1,2). The internal transcribed spacer region (ITS1 and ITS2) of the ribosomal DNA and the β-tubulin gene were sequenced (GenBank Accession Nos. HQ909028 and JF826867, respectively) and compared with existing sequences using BLASTn. The ITS sequence shared 99% maximum identity with that of Ca. colhounii CBS 293.79 (GQ280565) from Java, Indonesia, and the β-tubulin sequence shared 97% maximum identity with that of Ca. colhounii CBS 114036 (DQ190560) isolated from leaf spots on Rhododendron sp. in North Carolina. The isolate was submitted to the Centraalbureau voor Schimmelcultures in the Netherlands (CBS 129628). To confirm pathogenicity, 5 ml of a conidial suspension (1 × 105/ml) were applied as a foliar spray or soil drench to four healthy ‘Bluecrop’ plants each in 10-cm plastic pots. Two water-sprayed and two water-drenched plants served as controls. Plants were misted intermittently for 2 days after inoculation. After 7 days at 25 ± 3°C, drench-inoculated plants developed necrotic, sporulating stem lesions at the soil line, while spray-inoculated plants showed reddish brown leaf and stem lesions. At 28 days, three drench-inoculated and one spray-inoculated plant had died, while others showed stem necrosis and wilting. No symptoms were observed on control plants. Fungal colonies reisolated from surface-disinfested symptomatic stem, leaf, and root segments appeared identical to the original isolate. Cy. colhounii was reported to cause a leaf spot on blueberry plants in nurseries in China (3), while Ca. crotalariae (Loos) D.K. Bell & Sobers (= Ca. ilicicola Boedijn & Reitsma) causes stem and root rot of blueberries in North Carolina (4). To our knowledge, this is the first report of Ca. colhounii causing a disease of blueberry in Michigan or the United States. Because of its destructive potential, this pathogen may pose a significant threat in blueberry nurseries. References: (1) P. W. Crous. Taxonomy and Pathology of Cylindrocladium (Calonectria) and Allied Genera. The American Phytopathological Society, St. Paul, MN, 2002. (2) L. Lombard et al. Stud. Mycol. 66:31, 2010. (3) Y. S. Luan et al. Plant Dis. 90:1553, 2006. (4) R. D. Milholland. Phytopathology 64:831, 1974.

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