First Report of Root Rot and Wilt Caused by Pythium myriotylum on Hemp (Cannabis sativa) in the United States

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 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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First Report of Fusarium brasiliense Causing Root Rot of Dry Bean in the United States

Plant Disease ◽

10.1094/pdis-02-18-0332-pdn ◽

2018 ◽

Vol 102 (10) ◽

pp. 2035 ◽

Cited By ~ 4

Author(s):

J. L. Jacobs ◽

K. Oudman ◽

H. Sang ◽

M. I. Chilvers

Keyword(s):

United States ◽

Root Rot ◽

The United States ◽

Dry Bean ◽

First Report

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

Plant Disease ◽

10.1094/pdis-12-17-1999-pdn ◽

2018 ◽

Vol 102 (10) ◽

pp. 2045-2045 ◽

Cited By ~ 3

Author(s):

J. Beckerman ◽

J. Stone ◽

G. Ruhl ◽

T. Creswell

Keyword(s):

United States ◽

Root Rot ◽

Pythium Ultimum ◽

The United States ◽

First Report ◽

Crown And Root Rot ◽

Industrial Hemp

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First Report of Pythium myriotylum Causing Basal Rot on Echeveria in the United States

Plant Disease ◽

10.1094/pdis-12-15-1412-pdn ◽

2016 ◽

Vol 100 (8) ◽

pp. 1795-1795 ◽

Cited By ~ 1

Author(s):

S. N. Suarez ◽

G. Sanahuja ◽

P. Lopez ◽

A. J. Palmateer

Keyword(s):

United States ◽

The United States ◽

Pythium Myriotylum ◽

First Report ◽

Basal Rot

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First Report of Brown Marmorated Stink Bug (Hemiptera: Pentatomidae) Associated With Cannabis sativa (Rosales: Cannabaceae) in the United States

Journal of Integrated Pest Management ◽

10.1093/jipm/pmz014 ◽

2019 ◽

Vol 10 (1) ◽

Cited By ~ 2

Author(s):

Kadie Elizabeth Britt ◽

Mika K Pagani ◽

Thomas P Kuhar

Keyword(s):

United States ◽

Cannabis Sativa ◽

The United States ◽

Brown Marmorated Stink Bug ◽

Stink Bug ◽

First Report

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First Report of Crown and Root Rot Caused by Pythium myriotylum on Hemp (Cannabis sativa) in Arizona

Plant Disease ◽

10.1094/pdis-12-20-2712-pdn ◽

2021 ◽

Author(s):

Jiahuai Hu

Keyword(s):

Root Rot ◽

Cannabis Sativa ◽

Morphological Characteristics ◽

Crown Rot ◽

Pathogenicity Test ◽

Light Cycle ◽

Pythium Myriotylum ◽

First Report ◽

Leaf Chlorosis ◽

Query Coverage

During August and September 2020, symptoms of leaf chlorosis, stunting, and wilting were observed on indoor hemp plants (Cannabis sativa L. cv. ‘Wedding Cake’) in a commercial indoor facility located in Coolidge, Arizona. Plants were grown in soilless coconut coir growing medium (Worm Factory COIR250G10), watered with 1.5 to 2.1 liters every 24 h through drip irrigation, and supplemented with 18 h of lighting. About 35% of plants displayed symptoms as described above and many symptomatic plants collapsed. To identify the causal agent, crown and root tissues from four symptomatic plants were harvested and rinsed with tap water. Tissue fragments (approx. 2 to 4 mm in size) were excised from the margins of the stem and root lesions, surface sterilized in 0.6% sodium hypochlorite for 1 min, rinsed well in sterile distilled water, blotted dry, and plated on potato dextrose agar (PDA) and on oomycete-selective clarified V8 media containing pimaricin, ampicillin, rifampicin, and pentachloronitrobenzene (PARP). Plates were incubated at room temperature (21-24 oC). Five isolates resembling Pythium were transferred after 3 days and maintained on clarified V8 media. Morphological characteristics were observed on grass blade cultures (Waterhouse 1967). Grass blades were placed on CV8 inoculated with the isolate. After a 1-day incubation at 25°C, the colonized blades were transferred to 8 ml of soil water extract in a Petri dish. Ten sporangia and oogonia were selected randomly and their diameters were measured under the microscope. Sporangia were mostly filamentous, undifferentiated or inflated lobulate, ranging from 7 to 17 µm in diameter. Knob-like appressoria were observed on branching clusters. Bulbous-like antheridia were formed on branched stalk with 1-8 antheridia per oogonium. Globose oogonia were terminal or intercalary and ranged from 21 to 33 µm in diameter. Globose oospores were mostly aplerotic and ranged from 15 to 21 μm in diameter. Based on these morphological characteristics, isolates were tentatively identified as Pythium myriotylum (Watanabe, 2002). Genomic DNA was extracted from mycelial mats of two isolates using DNeasy Plant Pro Kit (Qiagen Inc., Valencia, CA) according to the manufacturer’s instructions. The internal transcribed spacer (ITS) region of rDNA was amplified with primers ITS1/ITS4 and two identical nucleotide sequences were obtained and deposited under accession number MW380925. A BLASTn search revealed ≥ 98% query coverage and 100% match with sequences HQ237488.1, KY019264.1, and KM434129, which were isolates of P. myriotylum from palm, tobacco, and ginger, respectively. To fulfill Koch’s postulates, pathogenicity tests were conducted with 2 isolates using plants of ‘Wedding Cake’ grown in 12 1.9-liter pots filled with a steam-disinfested potting mix (Sungro Professional Growing Mix). Pots were placed in a plastic container and watered to flooding three times a week. Plants were maintained in a greenhouse with 18 h/10 h day/night supplemental light cycle (15-28 oC). Plants were fertilized weekly with Peters Professional fertilizer at 1mg/ml. Four plants were inoculated with each isolate at three weeks after seed sowing by placing two 5-mm mycelial plugs from active growing 4 days-old cultures on PDA media adjacent to the main root mass at an approximately 3 cm depth. Four plants were inoculated with blank PDA plugs as controls. Symptoms of leaf chlorosis, crown rot and wilting were observed after four weeks while control plants remained symptomless. P. myriotylum was re-isolated from necrotic roots of inoculated plants after surface-sterilization, but not from control plants. The pathogenicity test was repeated once. While P. myriotylum often occurs in warmer regions and has a wide host range of >100 host plant species including numerous economically important crops (Wang et al., 2003), there are only two reports of this pathogen on indoor hemp plants in a greenhouse in Connecticut (McGehee et al., 2019) and in Canada (Punja et al., 2019). This is the first report of P. myriotylum causing root and crown rot of indoor hemp in Arizona. A more careful water management in soilless growth medium to reduce periods of saturation would minimize the risk of Pythium root rot in indoor hemp production.

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First Report of Root Rot Caused by Fusarium solani on Benincasa hispida in the United States

Plant Disease ◽

10.1094/pdis-06-11-0494 ◽

2012 ◽

Vol 96 (2) ◽

pp. 294-294 ◽

Cited By ~ 2

Author(s):

P. Ji ◽

J. Yin ◽

K. L. Jackson

Keyword(s):

United States ◽

Dna Sequences ◽

Root Rot ◽

Morphological Characteristics ◽

The United States ◽

First Report ◽

Benincasa Hispida ◽

Field Samples ◽

Vegetable Farm ◽

Wax Gourd

Root rot was observed on wax gourd (Benincasa hispida (Thunb.) Cogn.) cv. Black Giant in August 2010 in a commercial vegetable farm in southern Georgia. Approximately 5% of the plants were affected and infected roots turned light to dark brown with partial or entire roots affected and the lower leaves became wilted. Symptomatic roots from six plants were surface sterilized with 0.6% sodium hypochlorite and plated on potato dextrose agar (PDA) medium. Pure cultures had white mycelia and spore masses and were obtained from all six plants by subculturing hyphal tips onto PDA. One- to two-celled, oval- to kidney-shaped microconidia and cylindrical macroconidia with two or three cells plus apical and basal cell were produced, which averaged 12.5 × 4 μm and 28 × 4.5 μm, respectively. Microconidia were abundant and macroconidia were sparse on PDA. Single-spore isolates were obtained and identified as a Fusarium sp. by PCR analysis with primers ITS-Fu-f and ITS-Fu-r (1). Genomic DNA of two isolates obtained from different plants was extracted and a portion of the translation elongation factor 1-α (TEF) gene of the isolates was amplified and sequenced (3). When compared with sequences available in the GenBank database, DNA sequences of the two isolates (GenBank Accession No. JF928376) shared 100% sequence identity with F. solani strain FRC S1734 (GenBank Accession No. DQ247527). The fungus was identified as F. solani (Mart.) Sacc. based on molecular analysis and morphological characteristics (2). Oat grains were separately infected with two isolates, BG2a and BG6, and used to inoculate healthy, 3-week-old wax gourd seedlings (cv. Black Giant) under greenhouse conditions (14-h photoperiod, 24 to 30°C). Each seedling was grown in a 10-cm pot containing a commercial potting mix, and five healthy plants were inoculated with each isolate by placing 15 infected oat grains around each plant at a depth of 5 cm in the soil. Five plants treated with noninfected oat grains served as controls. Symptoms identical to those on field samples developed on all inoculated plants 3 weeks after inoculation but not on the control plants. F. solani was reisolated from inoculated symptomatic plants and the identity was confirmed, which completed Koch's postulates. The experiment was repeated one more time under similar conditions. To our knowledge, this is the first report of root rot caused by F. solani on wax gourd in the United States. Wax gourd is an important specialty crop in the southeastern United States and the occurrence of this disease needs to be taken into account in wax gourd production. References: (1) K. A. Abd-Elsalam et al. Afr. J. Biotechnol. 2:82, 2003. (2) C. Booth. Fusarium Laboratory Guide to the Identification of the Major Species. CMI, Kew, England, 1977. (3) D. M. Geiser et al. Eur. J. Plant Pathol. 110:473, 2004.

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First Report of Collar and Root Rot Caused by Phytophthora nicotianae on Daphne odora in Italy

Plant Disease ◽

10.1094/pdis-93-8-0848a ◽

2009 ◽

Vol 93 (8) ◽

pp. 848-848

Author(s):

A. Garibaldi ◽

D. Bertetti ◽

M. L. Gullino

Keyword(s):

United States ◽

Root Rot ◽

The United States ◽

Phytophthora Nicotianae ◽

Control Treatment ◽

Peat Moss ◽

Late Winter ◽

American Phytopathological Society ◽

First Report ◽

Daphne Odora

Daphne odora is becoming popular in gardens because of its variegated foliage and fragrant flowers in late winter and early spring. During October of 2008 in a commercial nursery near Maggiore Lake (Verbano-Cusio-Ossola Province) in northwestern Italy, plants of D. odora showed extensive chlorosis and root rot. Diseased plants eventually wilted and died, dropping leaves in some cases. Most frequently, wilted leaves persisted on stems. At the soil level, dark brown-to-black water-soaked lesions that coalesced often girdled the stem. All of the crown and root system was affected. Disease was widespread and severe with 70% of 2,500 potted plants being affected. A Phytophthora-like organism was isolated consistently on a medium selective for oomycetes (4) after disinfestation of lower stem and root pieces of D. odora for 1 min in a solution containing 1% NaOCl. Tissue fragments of 1 mm2 were excised from the margins of the lesions and plated. The pathogen was identified based on morphological and physiological features as Phytophthora nicotianae (= P. parasitica) (2). Sporangia were produced for identification by growing a pure culture in sterilized soil extract solution at neutral pH (obtained by shaking and then centrifuging 300 g of soil in 1 liter of distilled water). They were spherical to ovoid, papillate, and measured 39.2 to 54.5 × 31.7 to 41.7 μm (average 44.8 × 34.5 μm). Papillae measured 2.4 to 4.9 μm (average 3.7 μm). Chlamydospores were spherical with a diameter ranging from 15.8 to 36.1 μm (average 25.4 μm). The internal transcribed spacer (ITS) region of rDNA of a single isolate was amplified using primers ITS4/ITS6 and sequenced. BLAST analysis (1) of the 804-bp segment showed a 100% homology with the sequence of P. nicotianae EF140988. The nucleotide sequence has been assigned GenBank No. FJ843100. Pathogenicity of two isolates obtained from infected plants was confirmed by inoculating 12-month-old plants of D. odora. Both isolates were grown for 15 days on a mixture of 70:30 wheat/hemp kernels and then 80 g/liter of the inoculum was mixed into a substrate containing sphagnum peat moss/pumice/pine bark/clay (50:20:20:10 vol/vol). One plant per 3-liter pot was transplanted into the substrate and constituted the experimental unit. Three replicates were used for each isolate and noninoculated control treatment; the trial was repeated once. All plants were kept in a greenhouse at temperatures from 20 to 25°C. Plants inoculated with isolate no. 1 developed symptoms of chlorosis and root rot within 14 days and then a wilt rapidly followed. Isolate no. 2 was less aggressive causing the same symptoms within 20 days. Control plants remained symptomless. P. nicotianae consistently was reisolated from inoculated plants. Previously, P. nicotianae (= P. parasitica) has been reported in several states of the United States on D. odora (3). To our knowledge, this is the first report of P. nicotianae on D. odora in Italy. The economic importance of the disease is low because of the limited number of farms that grow this crop in Italy, although spread could increase as the popularity of plantings expand. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997 (2) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society, St Paul, MN, 1996. (3) D. F. Farr et al. Fungi on Plants and Products in the United States. The American Phytopathological Society, St Paul, MN, 1989. (4) H. Masago et al. Phytopathology, 67:425, 1977.

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