First Report of Fusarium commune Causing Damping-off, Seed Rot, and Seedling Root Rot on Soybean (Glycine max) in the United States

During 2007 to 2009, symptomatic and asymptomatic soybean plants were collected from fields in 18 Iowa counties. Fusarium isolates were recovered from surface-sterilized root tissue on peptone PCNB agar (2). Single-spore isolates were transferred to synthetic low nutrient agar (SNA) overlain with pieces (1 × 2 cm) of sterile filter paper, and to potato dextrose agar (PDA), and placed in the dark for 10 to 14 days for morphological identification (4). Twenty-three isolates were identified as Fusarium commune K. Skovg., O'Donnell & Nirenberg, previously in the F. oxysporum species complex (4). Colonies on PDA had white, fluffy, aerial mycelium with magenta to violet pigmentation in the medium. On SNA, macroconidia, chlamydospores, and microconidia on monophialides and polyphialides were consistent with the species description (4). Identification of all 23 isolates was confirmed by DNA sequencing of the translation elongation factor (EF1-α) gene, using ef1 and ef2 primers, and the mitochondrial small subunit (mtSSU), using primers MS1 and MS2 (4) [GenBank accessions for two representative isolates: EF1-α (JX289892 and JX289893), and mtSSU (JX289894 and, JX289895)]. Pathogenicity of two representative isolates of F. commune was tested on soybean (cv. AG2403) in a greenhouse, in water baths set at 18°C, using autoclaved soil mixed with infested sand-cornmeal inoculum (3). The experiment entailed a completely randomized design (CRD) with five replications (single plant/150 ml cone) per treatment, and was conducted three times. Dry root and shoot weights, and root rot severity (visual estimate of percent root rot on the entire root system) were evaluated after 6 weeks. Mean seedling emergence in soil infested with F. commune was 47 and 40% for the two isolates; in contrast, non-inoculated control plants had 100% emergence. There were significant differences in root (P < 0.0001) and shoot (P < 0.0001) weights, and root rot severity (P < 0.0001), between inoculated and non-inoculated plants. Seedlings that emerged were severely stunted and had dark brown lesions. F. commune was reisolated from infected roots of inoculated plants, but not from non-inoculated plants. Pathogenicity of both isolates to soybean (cv. MN1805) was also tested using a petri dish assay, in which eight seeds were placed on a plate with a 4-day-old culture growing on 2% water agar (1). Plates were rated 7 days later for seed germination, seed rot, and lesion development, using an ordinal scale (1). The experiment entailed a CRD with three replicate plates/treatment, and was conducted three times. Germination of inoculated seeds ranged from 37.5 to 75.0%, and germinated seedlings had dark brown lesions on the taproots. There was a significant difference between isolates in the petri dish assay (P = 0.0030); one isolate was less aggressive, but both isolates resulted in significantly more disease than on the non-inoculated control plants, which had 100% germination and no symptoms (P < 0.0001). F. oxysporum is a known soybean pathogen (1), but isolates of F. commune may have been misidentified as F. oxysporum in previous studies. To our knowledge, this is the first report of F. commune as a pathogen on soybean in the U.S.A. References: (1) K. E. Broders et al. Plant Dis. 91:727, 2007. (2) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Oxford, UK, 2006. (3) G. P. Munkvold and J. K. O'Mara. Plant Dis. 86:143, 2002. (4) K. Skovgaard et al. Mycologia. 94:630, 2003.

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First Report of Fusarium armeniacum Causing Seed Rot and Root Rot on Soybean (Glycine max) in the United States

Plant Disease ◽

10.1094/pdis-05-12-0429-pdn ◽

2012 ◽

Vol 96 (11) ◽

pp. 1693-1693 ◽

Cited By ~ 10

Author(s):

M. L. Ellis ◽

M. M. Díaz Arias ◽

L. F. Leandro ◽

G. P. Munkvold

Keyword(s):

United States ◽

Root Rot ◽

Aerial Mycelium ◽

The United States ◽

Root Tissue ◽

Colony Morphology ◽

Ordinal Scale ◽

First Report ◽

Seed Rot ◽

Slow Growing

In a survey for Fusarium root rot, soybean plants were sampled from eight counties across Iowa in 2008 to 2009. Fusarium isolates were recovered from surface-sterilized symptomatic and asymptomatic root tissue by culturing on peptone PCNB agar (2). Single-spore isolates were transferred to carnation leaf agar (CLA) and potato dextrose agar (PDA) for morphological identification; 11 isolates were identified as F. armeniacum (Forbes, Windels, and Burgess) Burgess and Summerell (previously F. acuminatum ssp. armeniacum) (2). Colonies on PDA produced white aerial mycelium, red to apricot pigment in agar, and bright orange sporodochia in the center of the culture. Some isolates produced a pionnotal form of slow-growing colonies with little aerial mycelium and abundant orange sporodochia. On CLA, macroconidia in orange sporodochia on carnation leaves and chlamydospores formed abundantly, but microconidia were absent (2). Species identity for the 11 isolates was confirmed by sequencing of the elongation factor gene (EF1-α) using ef1 and ef2 primers (4) (reference sequences deposited in GenBank JX101763 and JX101764). Pathogenicity of seven F. armeniacum isolates was tested using surface-sterilized soybean seed, cv. AG2403, in a petri dish assay with 3-day-old cultures on 2% water agar (1). Germination, seed rot, and lesion development were scored 7 dai using an ordinal scale (1). The experiment was a completely randomized design (CRD), had three replicate plates per isolate, and was conducted twice. All seven isolates were pathogenic on soybean, though variation in aggressiveness was observed among isolates (P < 0.0001) related to colony morphology on PDA. Seed germination was 0 to 40% when inoculated with four isolates showing white fluffy aerial mycelium on PDA. Seedlings were severely stunted with dark brown lesions covering a majority of the root system. When inoculated with three isolates showing the pionnotal form of slow-growing mycelium, germination was 70 to 100%, with few small brown lesions (~5 to 10 mm) on the roots. Noninoculated controls showed 100% germination and no symptoms. Pathogenicity was also tested in a growth chamber assay at 18°C using autoclaved soil mixed with an infested sand-cornmeal inoculum (3). Data for dry root and shoot weights and root rot severity (visually scored on a % scale) were collected at 6 weeks. The CRD experiment had five replications (single plant in a cone containing 150 ml infested soil), and was conducted twice. Root symptoms and similar variation in aggressiveness among isolates (based on colony morphology) was observed in inoculated plants. Isolates differed significantly for effects on root weight (P = 0.0125), shoot weight (P = 0.0035), and root rot severity (P = 0.0158). F. armeniacum was reisolated from infected root tissue, but not from noninoculated controls. Recovered isolates maintained their original colony morphology. F. armeniacum was previously reported in Minnesota on symptomless corn (2), but it has not been reported on soybean and its pathogenicity has not been established on any crop. To our knowledge, this is the first report of F. armeniacum as a pathogen on soybean in the United States. References: (1) K. E. Broders et al. Plant Dis. 91:727, 2007. (2) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Oxford, UK, 2006. (3) G. P. Munkvold and J. K. O'Mara. Plant Dis. 86:143, 2002. (4) K. O'Donnell et al. Proc. Natl. Acad. Sci. 95:2044, 1998.

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First Report of Ilyonectria macrodidyma Causing Root Rot of Olive Trees (Olea europaea) in California

Plant Disease ◽

10.1094/pdis-04-12-0330-pdn ◽

2012 ◽

Vol 96 (9) ◽

pp. 1378-1378 ◽

Cited By ~ 8

Author(s):

J. R. Úrbez-Torres ◽

F. Peduto ◽

W. D. Gubler

Keyword(s):

Root Rot ◽

Small Subunit ◽

Internal Transcribed Spacer Region ◽

Olive Cultivar ◽

Field Station ◽

First Report ◽

Light Yellow ◽

Significant Difference ◽

Root Necrosis ◽

Olive Trees

The California olive industry produces 99% of the U.S. olive crop, which represented a value of over $113 million in 2010. During the 2008 and 2009 growing seasons, decline of young super-high-density olive cvs. Arbequina, Arbosana, and Koroneiki trees (<4 years old) was observed in orchards throughout Glenn, Yolo, and San Joaquin Counties. Symptomatic trees showed stunted growth and chlorotic leaves with roots having black, sunken, necrotic lesions, which frequently prolonged into the base and crown of the tree. Twenty-five trees were collected from different orchards and necrotic roots as well as infected trunk tissue were plated onto potato dextrose agar amended with 0.01% tetracycline hydrochloride. Cultures were incubated at room temperature (23 ± 2°C) until fungal colonies were observed. In 17 out of 25 trees collected (68%), light yellow fungal colonies were observed from the symptomatic tissue after 7 to 10 days. Colonies turned dark yellow to orange with age and showed an orange-dark brown reverse. Both microconidia (hyaline, ellipsoidal to ovoidal and aseptate (n = 60) (6.5) 11.5 to 13.5 (17.1) × (3) 3.4 to 4.5 (5.6) μm) and macroconidia (hyaline, cylindrical, straight and/or slightly curved with one, two or three septa (n = 60) (12.5) 26.5 to 38.5 (44.1) × (4) 5.5 to 7.5 (8.5) μm) were observed. Culture and conidial morphology were in concordance with previous published description of Ilyonectria macrodidyma (Halleen, Schroers & Crous) P. Chaverri & C. Salgado (1,3,4). Identification to species level was confirmed by sequence comparison of four Californian isolates (UCCE958, UCCE959, UCCE960, and UCCE961) with sequences available in GenBank using the internal transcribed spacer region (ITS1-5.8S-ITS2) of the rDNA (primers ITS1/ITS4), a portion of the β-tubulin gene (BT1a/BT1b), and a partial sequence of the mitochondrial small subunit rDNA (NMS1/NMS2) (4). Fungal sequences of isolates from olive from California (GenBank JQ868543 to JQ868554) showed 99 to 100% homology with previously identified and deposited I. macrodidyma isolates in Genbank for all three genes. Pathogenicity of I. macrodidyma in olive cvs. Arbequina, Arbosan, and Koroneiki was investigated using two fungal isolates (UCCE958 and UCCE960) as reported by Petit and Gubler (4). The roots of 10 1-year-old trees per fungal isolate for each olive cultivar were individually inoculated with 25 ml of a 106 conidia/ml spore suspension and placed in a lath house at the UC Davis field station. Additionally, 10 trees per cultivar were inoculated with sterile water as controls. Six months after inoculation, most of the inoculated olive plants showed chlorotic leaves similar to those observed in commercial orchards. Root necrosis for each cv. was expressed as the percentage of root length having lesions (2). No significant difference was observed between isolates and average root necrosis was 29.4, 35.6, and 38.3% in Koroniki, Arbosana, and Arbequina, respectiveley. I. macrodidyma was recovered from symptomatic roots in each of the cvs. and identified based on morphology. No root rot symptoms were observed in the controls. To our knowledge, this is the first report of I. macrodidyma causing root rot of olive trees not only in California but anywhere in the world. References: (1) P. Chaverri et al. Stud. Mycol. 68:57, 2011. (2) M. Giovanetti and B. Mosse. New Phytol. 84:489, 1980. (3) F. Halleen et al. Stud. Mycol. 50:421, 2004. (4) E. Petit and W. D. Gubler. Plant Dis. 89:1051, 2005.

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First report of Fusarium fujikuroi causing root rot and seedling elongation of soybean in Indiana

Plant Disease ◽

10.1094/pdis-03-21-0570-pdn ◽

2021 ◽

Author(s):

Christopher Detranaltes ◽

Christopher Robert Jones ◽

Guohong Cai

Keyword(s):

Root Rot ◽

Lateral Roots ◽

Small Subunit ◽

The State ◽

Gibberella Fujikuroi ◽

Fusarium Fujikuroi ◽

Pathogenicity Test ◽

Single Spore ◽

New Host ◽

First Report

In summer 2020, 127 soybean (Glycine max (L.) Merr) seedlings (V1-V3 stage) showing reduced vigor or crown lesions were collected at Purdue’s Agronomy Center for Research and Education in West Lafayette, Indiana. Root tissues from two seedlings with necrotic cotyledons and root rot were surface-sterilized and plated on dichloran-chloramphenicol-peptone agar (Andrews and Pitt 1986). Emerging hyphal tips were transferred to potato dextrose agar (PDA). Single-spore cultures were obtained and grown on PDA. Both isolates developed floccose white aerial mycelia with reddish-pink coloration in the media in 2 weeks on the benchtop. On carnation leaf agar, macroconidia formed on orange sporodochia within 2 weeks in darkness at 25C. Macroconidia were 3-5 septate, measuring 26 – 41 × 2.5 – 3.7 μm (avg. 34.8 × 3.2 μm, n=40). Microconidia were abundant in chains and false heads forming on both mono- and polyphialides, and measured 2.5 – 8.75 x 2.5 μm (avg. 5.9 × 2.5 μm, n=40). These characteristics were consistent with species descriptions of F. fujikuroi [Sawada] Wollenw. (teleomorph Gibberella fujikuroi) (Leslie and Summerell 2006). DNA was extracted from mycelium and the following genes were amplified and sequenced: the internal transcriber spacer (ITS) region using ITS1/ITS4 primers (White et al. 1990) (GenBank accessions MW463362/MW463363), the mitochondrial small subunit (mtSSU) rDNA using MS1/MS2 primers (White et al. 1990) (MW465310/MW465307), and the partial translation elongation factor 1-alpha (TEF1α) gene using 983F/1567R primers (Rehner and Buckley 205) (MW475297/MW475298). In GenBank BLAST searches, these sequences showed 100% identity to both F. proliferatum and F. fujikuroi. Species-specific forward primers Fuji1F and Proli1F were then used in combination with reverse primer TEF1R to amplify another region in the TEF1α gene (Amatulli et al. 2012). Proli1F/TEF1R primers failed under a variety of annealing temperatures while Fuji1F/TEF1R primers succeeded, and the products were sequenced (MW475299/MW475300). GenBank BLAST searches revealed 100% identity of both isolates to F. fujikuroi (MT448248.1). A pathogenicity test was conducted with isolate AC13 in the greenhouse following the protocol of (Ellis et al. 2013). Ten seeds (cv. Williams) each were used for inoculation and control, respectively, with one seed per cup. Root rot symptoms similar to those observed in the field were observed 14 days after planting on all inoculated plants but not on controls (VC stage). Infected plants showed symptoms of pre-emergence damping off, reddish-brown lesions on the tap and lateral roots, and root necrosis. Three plants also exhibited hyper-elongation of the stem (12.5, 11.1, and 18 cm, vs controls: avg. 6.8 cm, max. 8.5 cm, stdev 0.78 cm). F. fujikuroi was successfully reisolated from inoculated plants but not from controls and identified as described above. F. fujikuroi has been reported causing soybean root rot in China (Zhao et al. 2020), Korea (Choi et al. 2019), and the state of Kansas (Pedrozo et al. 2015). To our knowledge this is the first report of F. fujikuroi infecting soybeans in the state of Indiana. F. fujikuroi is known to cause elongated seedlings in rice (Leslie and Summerell 2006). Pedrozo et al. (2015) reported that F. fujikuroi isolated from soybean caused seedling elongation in rice but not in soybean. The increased distribution and new host symptomology observed here warrants heightened attention for the control of this pathogen.

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First Report of Spot Blotch and Common Root Rot Caused by Bipolaris sorokiniana on Switchgrass in Tennessee

Plant Disease ◽

10.1094/pdis-12-10-0880 ◽

2011 ◽

Vol 95 (9) ◽

pp. 1195-1195 ◽

Cited By ~ 7

Author(s):

A. L. Vu ◽

M. M. Dee ◽

K. D. Gwinn ◽

B. H. Ownley

Keyword(s):

Root Rot ◽

Panicum Virgatum ◽

Bipolaris Sorokiniana ◽

The United States ◽

Small Subunit ◽

Spot Blotch ◽

Cochliobolus Sativus ◽

Common Root ◽

First Report ◽

Common Root Rot

Light-to-dark brown, irregular-shaped leaf spots, chlorosis, necrotic roots, and severe stunting were observed on ‘Alamo’ switchgrass (Panicum virgatum L.) grown on the campus of the University of Tennessee in December 2007. Symptomatic leaf and root samples were surface sterilized, air dried on sterile filter paper, and plated on 2% water agar amended with 10 mg/liter of rifampicin (Sigma-Aldrich, St. Louis, MO) and 10 μl/liter of 2,4 EC Danitol miticide (Valent Chemical, Walnut Creek, CA). Plates were incubated at 25°C in darkness for 4 days. A sporulating, dematiaceous mitosporic fungus was noted and transferred to potato dextrose agar (PDA). Conidia were ovate, oblong, mostly straight, and olive to brown with three to nine septa. Conidial dimensions were 12.5 × 27.5 (17.5) to 20 × 77.5 (57) μm. Conidia were produced on single, light brown, multiseptate conidiophores that were polytretic, geniculate, and sympodial. Morphological features were as described for Bipolaris sorokiniana (Sacc.) Shoemaker (teleomorph = Cochliobolus sativus) (2,3). Disease assays were conducted with 5-week-old ‘Alamo’ switchgrass grown from surface-sterilized seed. Ten 9 × 9-cm2 with ~20 switchgrass seedlings were sprayed with 2.4 × 105 spores/ml of sterile water. Plants were subjected to high humidity created by enclosure in a plastic bag for 45 h. The bag was removed and plants were incubated at 25/20°C with 50 to 60% relative humidity. During the incubation, plants were maintained in growth chamber with a 12-h photoperiod of fluorescent and incandescent lighting. Foliar leaf spot symptoms appeared 6 to 10 days postinoculation for plants in all 10 replicates and necrotic lesions were observed on roots. Foliar lesions and diseased roots were surface sterilized, plated on water agar, and resultant fungal colonies were identified as B. sorokiniana. The internal transcribed spacer (ITS) and mitochondrial small subunit (SSU) regions of ribosomal DNA from the original isolate, and the isolate recovered from plants in the pathogenicity assay, were amplified with PCR, with primer pairs ITS4 and ITS5 and NMS1 and NMS2. PCR amplicons of ~551 and 571 bp were obtained with the two primer pairs, respectively. Both amplicons were obtained from both isolates and sequenced. Amplicon sequences from the original isolate and re-isolate were identical and the sequences were submitted to GenBank (Accession Nos. HQ611957 and HQ611958). The ITS sequences had 98% homology to 23 B. sorokiniana isolates, including B. sorokiniana strain DSM 62608 (GenBank Accession No. EF187908); SSU sequences had 99% homology to Cochliobolus sativus isolate AFTOL-ID 271 (GenBank Accession No. FJ190589). Spot blotch caused by B. sorokiniana has been reported on switchgrass in Iowa, Nebraska, Pennsylvania, and Virginia (1). To our knowledge, this is the first report of B. sorokiniana causing spot blotch or common root rot of switchgrass in Tennessee, which extends the current known distribution of these diseases. More recently, we isolated B. sorokiniana from switchgrass seed received from commercial sources in the United States, indicating a seedborne transmission. References: (1) D. F. Farr and A. Y. Rossman. Fungal Databases. Systematic Mycology and Microbiology Laboratory, ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ , 15 November 2010. (2) R. F. Nyvall and J. A. Percich. Plant Dis. 83:936, 1999. (3) A. Sivanesan and P. Holliday. CMI Descr. Pathog. Fungi bact. 71:701, 1981.

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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 Pythium torulosum Causing Corn Root Rot in Northeastern China

Plant Disease ◽

10.1094/pdis-08-20-1679-pdn ◽

2020 ◽

Author(s):

Xiujun Tang ◽

Shuning Chen ◽

Xiaojing Yan ◽

Huizhu Yuan ◽

Daibin Yang

Keyword(s):

Root Rot ◽

Petri Dish ◽

Soil Samples ◽

Northeastern China ◽

Growth Chamber ◽

Corn Root ◽

First Report ◽

Corn Root Rot ◽

Soil Mix ◽

Pythium Torulosum

In October 2017, we collected five soil samples from each of several fields with a history of severe corn (Zea mays) seedling disease in Heilongjiang province of China. Affected seedlings were wilted with severe root rot, and a high incidence of seedling death was observed in the fields. Corn seeds were seeded in the collected soil samples and grown in a growth chamber for 21 days set at the following incubation temperatures: 21℃/7℃ for 6 days, 10℃/3℃ for 4 days, 16℃/7℃ for 5 days, 20℃/20℃ for 6 days (16 h/8 h, light/dark) (Tang et al. 2019). The corn seedlings in the growth chamber showed the same symptoms observed in the field as mentioned above. Corn root rot samples were collected from several symptomatic plants in the growth chamber to isolate the possible pathogen. Symptomatic roots were washed in 0.5% NaOCl for 2 min, rinsed in sterile water and cut into 1-2 mm segments and then plated on corn meal agar amended with pimaricin (5 μg/ml), ampicillin (250 μg/ml), rifampicin (10 μg/ml), pentachloronitrobenzene (50 μg/ml), and benomyl (10 μg/ml) (PARP+B), which is selective for oomycetes (Jeffers and Martin 1986). After 3 days of incubation in the dark at 25℃, colonies were transferred to 10% V8 juice agar and incubated at 25℃ for 2 weeks. Six isolates were identified as Pythium torulosum based on the morphology of sexual and asexual structures following van der Plaats-Niterink’s key (van der Plaats-Niterink 1981). On 10% V8 juice agar, the hypha were aseptate and colonies had filamentous sporangia with a dendroid or globose structure. The oogonia were globose or subglobose, laevis, terminal, rarely intercalary, ranging from 12-19 (average 16) μm. Antheridia were mostly sessile or brachypodous, and each oogonium was supplied by 1-2 antheridia cells. Oospores were globose, plerotic, ranging from 9-16 (average 13) μm. For the molecular identification, two molecular targets, the internal transcribed spacer (ITS) region of ribosomal DNA and cytochrome c oxidase subunit II (CoII), were amplified and sequenced using universal primer sets DC6/ITS4 (Cooke et al. 2000) and FM58/FM66 (Villa et al. 2006), respectively for one isolate, “copt”. BLAST analyses of a 971 bp ITS segment amplified from copt (GenBank Accession No. MT830918) showed 99.79% identity with a P. torulosum isolate (GenBank Accession No. AY598624.2). For the COⅡ gene of copt, BLAST analyses of a 553 bp segment (GenBank Accession MT843570) showed 98.37% identity with P. torulosum isolate (GenBank Accession No. AB095065.1). Thus, the isolate, copt, was identified as P. torulosum based on morphological characteristics and molecular analysis. To confirm pathogenicity and Koch’s postulates, a pathogenicity test was conducted as described by Zhang et al. (2000). Briefly, a 5 mm culture plug from the P. torulosum isolate, copt, was transferred to a 9-cm petri dish containing 20mL 10% V8 juice agar and incubated in the dark at 25℃ for 7 days. The culture was cut into small pieces and mixed with a sterilized soil mix (40% organic peat substrate, 40% perlite, and 20% soil) at a ratio of one petri dish per 100 g soil mix. Ten corn seeds were planted at a depth of 2 cm in a 500-mL pot containing the inoculated soil mix. The control pots were mock inoculated with plain 10% V8 juice agar. Pots were incubated in a greenhouse at temperatures ranging from 21 to 23℃. There were four replications. After 14 days, corn roots brown and rotted were observed, which was similar to those observed in the field and growth chamber. Control plants remained symptomless and healthy. P. torulosum copt was consistently re-isolated from the symptomatic roots. To our knowledge, this is the first report of P. torulosum causing root rot of corn in Northeastern China. Corn is an important crop in Heilongjiang and the occurrence of root rot caused by this pathogen may be a new threat to corn plants. There is a need to develop management measures to control the disease.

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First Report of Alternaria Leaf Spot of Banana Caused by Alternaria alternata in the United States

Plant Disease ◽

10.1094/pdis-01-13-0007-pdn ◽

2013 ◽

Vol 97 (8) ◽

pp. 1116-1116 ◽

Cited By ~ 3

Author(s):

V. Parkunan ◽

S. Li ◽

E. G. Fonsah ◽

P. Ji

Keyword(s):

United States ◽

Alternaria Alternata ◽

Leaf Spot ◽

Morphological Characteristics ◽

The United States ◽

Its Sequencing ◽

Single Spore ◽

First Report ◽

Alternaria Leaf Spot ◽

Leaf Spots

Research efforts were initiated in 2003 to identify and introduce banana (Musa spp.) cultivars suitable for production in Georgia (1). Selected cultivars have been evaluated since 2009 in Tifton Banana Garden, Tifton, GA, comprising of cold hardy, short cycle, and ornamental types. In spring and summer of 2012, 7 out of 13 cultivars (African Red, Blue Torres Island, Cacambou, Chinese Cavendish, Novaria, Raja Puri, and Veinte Cohol) showed tiny, oval (0.5 to 1.0 mm long and 0.3 to 0.9 mm wide), light to dark brown spots on the adaxial surface of the leaves. Spots were more concentrated along the midrib than the rest of the leaf and occurred on all except the newly emerged leaves. Leaf spots did not expand much in size, but the numbers approximately doubled during the season. Disease incidences on the seven cultivars ranged from 10 to 63% (10% on Blue Torres Island and 63% on Novaria), with an average of 35% when a total of 52 plants were evaluated. Six cultivars including Belle, Ice Cream, Dwarf Namwah, Kandarian, Praying Hands, and Saba did not show any spots. Tissue from infected leaves of the seven cultivars were surface sterilized with 0.5% NaOCl, plated onto potato dextrose agar (PDA) media and incubated at 25°C in the dark for 5 days. The plates were then incubated at room temperature (23 ± 2°C) under a 12-hour photoperiod for 3 days. Grayish black colonies developed from all the samples, which were further identified as Alternaria spp. based on the dark, brown, obclavate to obpyriform catenulate conidia with longitudinal and transverse septa tapering to a prominent beak attached in chains on a simple and short conidiophore (2). Conidia were 23 to 73 μm long and 15 to 35 μm wide, with a beak length of 5 to 10 μm, and had 3 to 6 transverse and 0 to 5 longitudinal septa. Single spore cultures of four isolates from four different cultivars were obtained and genomic DNA was extracted and the internal transcribed spacer (ITS1-5.8S-ITS2) regions of rDNA (562 bp) were amplified and sequenced with primers ITS1 and ITS4. MegaBLAST analysis of the four sequences showed that they were 100% identical to two Alternaria alternata isolates (GQ916545 and GQ169766). ITS sequence of a representative isolate VCT1FT1 from cv. Veinte Cohol was submitted to GenBank (JX985742). Pathogenicity assay was conducted using 1-month-old banana plants (cv. Veinte Cohol) grown in pots under greenhouse conditions (25 to 27°C). Three plants were spray inoculated with the isolate VCT1FT1 (100 ml suspension per plant containing 105 spores per ml) and incubated under 100% humidity for 2 days and then kept in the greenhouse. Three plants sprayed with water were used as a control. Leaf spots identical to those observed in the field were developed in a week on the inoculated plants but not on the non-inoculated control. The fungus was reisolated from the inoculated plants and the identity was confirmed by morphological characteristics and ITS sequencing. To our knowledge, this is the first report of Alternaria leaf spot caused by A. alternata on banana in the United States. Occurrence of the disease on some banana cultivars in Georgia provides useful information to potential producers, and the cultivars that were observed to be resistant to the disease may be more suitable for production. References: (1) E. G. Fonsah et al. J. Food Distrib. Res. 37:2, 2006. (2) E. G. Simmons. Alternaria: An identification manual. CBS Fungal Biodiversity Center, Utrecht, Netherlands, 2007.

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First Report of Seedborne Fusarium thapsinum and its Pathogenicity on Soybean (Glycine max) in the United States

Plant Disease ◽

10.1094/pdis-08-14-0806-pdn ◽

2014 ◽

Vol 98 (12) ◽

pp. 1745-1745 ◽

Cited By ~ 2

Author(s):

R. Pedrozo ◽

C. R. Little

Keyword(s):

United States ◽

The United States ◽

Soybean Plant ◽

Sodium Hypochlorite Solution ◽

Distilled Water ◽

Seedling Mortality ◽

Beta Tubulin ◽

First Report ◽

Significant Difference ◽

Germinated Seeds

A three-year survey from 2010 to 2012 was conducted in Kansas to investigate the identity and diversity of seedborne Fusarium spp. in soybean. A total of 408 soybean seed samples from 10 counties were tested. One hundred arbitrarily selected seeds from each sample were surface-sterilized for 10 min in a 1% sodium hypochlorite solution to avoid contaminants and promote the isolation of internal fusaria. Seeds were rinsed with sterile distilled water and dried overnight at room temperature (RT). Surface-sterilized seeds were plated on modified Nash-Snyder medium and incubated at 23 ± 2°C for 7 days. Fusarium isolates were single-spored and identified by morphological characteristics on carnation leaf agar (CLA) and potato dextrose agar (PDA) (3). From 276 seedborne Fusarium isolates, six were identified as F. thapsinum (2). On CLA, F. thapsinum isolates produced abundant mycelium and numerous chains of non-septate microconidia produced from monophialides. Microconidia were club-shaped and some were napiform. No chlamysdospores were found. On PDA, three of the isolates presented characteristic dark yellow pigmentation and three were light violet. Confirmation of the isolates to species was based on sequencing of an elongation factor gene (EF1-α) segment using primers EF1 and EF2 and the beta-tubulin gene using primers Beta1 and Beta2 (1). Sequence results (~680 bp, EF primers; ~600 bp, beta-tubulin primers) were confirmed by using the FUSARIUM-ID database (1). All isolates matched F. thapsinum for both genes sequenced (Accession No. FD01177) at 99% identity. Koch's postulates were completed for two isolates of F. thapsinum under greenhouse conditions. Soybean seeds (Asgrow AG3039) were imbibed with 2.5 × 105 conidia ml−1 for 48 h. After inoculation, seeds were dried for 48 h at RT. One isolate each of F. equiseti and F. oxysporum were used as the non-pathogenic and pathogenic inoculation controls, respectively. In addition, non-inoculated seeds and seeds imbibed in sterile distilled water (mock) were also used. Twenty-five seeds from each treatment were planted in pots (500 ml) with autoclaved soil and vermiculite (1:1). The experiment was a completely randomized design with three replicates (pots) per isolate. The entire experiment was repeated three times. After 21 days, aggressiveness of both F. thapsinum isolates was assessed using initial stand (%), final stand (%), and seed mortality (% of non-germinated seeds). Both seedborne F. thapsinum isolates caused reduced emergence and final stand, and increased seedling mortality when compared to the non-inoculated and F. equiseti controls (P< 0.0001). No significant difference was observed between F. thapsinum isolates and F. oxysporum. F. thapsinum isolates were re-isolated from wilted seedlings and non-germinated seeds, but not from the control treatments. Typically, F. thapsinum is considered a pathogen of sorghum, but it has also been recovered from bananas, peanuts, maize, and native grasses (3). However, its presence on soybean plant tissues and its pathogenicity has never been reported. To our knowledge, this is the first report of seedborne F. thapsinum and its pathogenicity on soybean in the United States. References: (1) D. M. Geiser et al. Eur. J. Plant Pathol. 110:473, 2004. (2) C. J. R. Klittich et al. Mycologia 89:644, 1997. (3) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Oxford, UK, 2006.

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First Report of Leaf Spot of Rudbeckia hirta var. pulcherrima Caused by Septoria rudbeckiae in Korea

Plant Disease ◽

10.1094/pdis-02-12-0172-pdn ◽

2012 ◽

Vol 96 (6) ◽

pp. 911-911 ◽

Cited By ~ 1

Author(s):

J. H. Park ◽

S. E. Cho ◽

K. S. Han ◽

H. D. Shin

Keyword(s):

Leaf Spot ◽

The United States ◽

Flowering Plant ◽

Leaf Spot Disease ◽

Morphological Identification ◽

Its Sequence ◽

Conidial Suspension ◽

First Report ◽

Rudbeckia Hirta ◽

Close Phylogenetic Relationship

Rudbeckia hirta L. var. pulcherrima Farw. (synonym R. bicolor Nutt.), known as the black-eyed Susan, is a flowering plant belonging to the family Asteraceae. The plant is native to North America and was introduced to Korea for ornamental purposes in the 1950s. In July 2011, a previously unknown leaf spot was first observed on the plants in a public garden in Namyangju, Korea. Leaf spot symptoms developed from lower leaves as small, blackish brown lesions, which enlarged to 6 mm in diameter. In the later stages of disease development, each lesion was usually surrounded with a yellow halo, detracting from the beauty of the green leaves of the plant. A number of black pycnidia were present in diseased leaf tissue. Later, the disease was observed in several locations in Korea, including Pyeongchang, Hoengseong, and Yangpyeong. Voucher specimens were deposited at the Korea University Herbarium (KUS-F25894 and KUS-F26180). An isolate was obtained from KUS-F26180 and deposited at the Korean Agricultural Culture Collection (Accession No. KACC46694). Pycnidia were amphigenous, but mostly hypogenous, scattered, dark brown-to-rusty brown, globose, embedded in host tissue or partly erumpent, 50 to 80 μm in diameter, with ostioles 15 to 25 μm in diameter. Conidia were substraight to mildly curved, guttulate, hyaline, 25 to 50 × 1.5 to 2.5 μm, and one- to three-septate. Based on the morphological characteristics, the fungus was consistent with Septoria rudbeckiae Ellis & Halst. (1,3,4). Morphological identification of the fungus was confirmed by molecular data. Genomic DNA was extracted using the DNeasy Plant Mini DNA Extraction Kit (Qiagen Inc., Valencia, CA.). The internal transcribed spacer (ITS) region of rDNA was amplified using the ITS1/ITS4 primers and sequenced. The resulting sequence of 528 bp was deposited in GenBank (Accession No. JQ677043). A BLAST search showed that there was no matching sequence of S. rudbeckiae; therefore, this is the first ITS sequence of the species submitted to GenBank. The ITS sequence showed >99% similarity with those of many Septoria species, indicating their close phylogenetic relationship. Pathogenicity was tested by spraying leaves of three potted young plants with a conidial suspension (2 × 105 conidia/ml), which was harvested from a 4-week-old culture on potato dextrose agar. Control leaves were sprayed with sterile water. The plants were covered with plastic bags to maintain 100% relative humidity (RH) for the first 24 h. Plants were then maintained in a greenhouse (22 to 28°C and 70 to 80% RH). After 5 days, leaf spot symptoms identical to those observed in the field started to develop on the leaves inoculated with the fungus. No symptoms were observed on control plants. S. rudbeckiae was reisolated from the lesions of inoculated plants, confirming Koch's postulates. A leaf spot disease associated with S. rudbeckiae has been reported on several species of Rudbeckia in the United States, Romania, and Bulgaria (1–4). To our knowledge, this is the first report of leaf spot on R. hirta var. pulcherrima caused by S. rudbeckiae in Korea. References: (1) J. B. Ellis and B. D. Halsted. J. Mycol. 6:33, 1890. (2) D. F. Farr and A. Y. Rossman. Fungal Databases. Systematic Mycology and Microbiology Laboratory, ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ February 2, 2012. (3) E. Radulescu et al. Septoriozele din Romania. Ed. Acad. Rep. Soc. Romania, Bucuresti, Romania, 1973. (4) S. G. Vanev et al. Fungi Bulgaricae 3:1, 1997.

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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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