scholarly journals First Report of Cytospora punicae Causing Wood Canker and Branch Dieback of Pomegranate (Punica granatum) in the United States

Plant Disease ◽  
2014 ◽  
Vol 98 (6) ◽  
pp. 853-853 ◽  
Author(s):  
F. Peduto Hand ◽  
R. A. Choudhury ◽  
W. D. Gubler
Keyword(s):  
United States ◽  
Elongation Factor ◽  
Punica Granatum ◽  
The United States ◽  
Internal Transcribed Spacer Region ◽  
Intermittent Light ◽  
First Report ◽  
Consensus Sequences ◽  
Vascular Discoloration ◽  
Branch Dieback

Pomegranates (Punica granatum L.) are an expanding industry in the United States with California growing approximately 32,000 acres with a crop value of over $155 million (1). During June and July of 2012, we observed severe limb and branch dieback in pomegranate orchards cv. Wonderful located in Contra Costa, Kings, and Kern counties of California. Disease symptoms included yellowing of leaves, branch and limb dieback, wood lesions, and canker formation. Dark brown Cytospora-like cultures were consistently isolated from active cankers on potato dextrose agar (PDA) amended with 100 mg l−1 tetracycline hydrochloride. Three isolates (UCCE1223, UCCE1233, and UCCE1234) representative of each orchard were sub-cultured onto PDA and incubated at 22°C under fluorescent intermittent light (12 h light, 12 h dark). Fungal colonies had whitish mycelia that turned olive green to dark brown with maturity and formed globose and dark brown pycnidia after 12 days. Conidia were hyaline, aseptate, allantoid, and (4) 4.5 to 5 (6) × (1) 1.5 (2) μm (n = 180). Pycnidia formed in culture measured (250) 350 to 475 (650) μm in diameter (n = 40). Identification of the isolates was confirmed by sequence comparison of the internal transcribed spacer region (ITS1-5.8S-ITS2) of the rDNA and part of the translation elongation factor 1-α gene (EF1-α) with sequences available in GenBank. Consensus sequences of both genes of all isolates showed 99% homology to the species Cytospora punicae Sacc. (2). All sequences were deposited in GenBank (Accession Nos. KJ621684 to 89). Pathogenicity of the isolates was determined by branch inoculation. In December 2012, 3-year-old branches of P. granatum cv. Wonderful were inoculated by placing 5-mm-diameter mycelium plugs from the growing margin of 14-day-old PDA cultures in fresh wounds made with a 5-mm-diameter cork-borer. Eight branches per isolate were inoculated on eight different trees. Eight control branches were inoculated with non-colonized PDA agar plugs. Inoculations were covered with Vaseline and wrapped with Parafilm to retain moisture. Branches were harvested in August 2013 and examined for canker development and the extent of vascular discoloration spreading downward and upward from the inoculation point. Isolations from the edge of discolored tissue were conducted to fulfill Koch's postulates. C. punicae was re-isolated from 100% of the inoculated branches. Total length of vascular discoloration averaged 30.2 mm in branches inoculated with the three C. punicae isolates and 9 mm in the control branches. No fungi were isolated from the slightly discolored tissue of the controls. To our knowledge, this is the first report of C. punicae as a fungal trunk pathogen of pomegranate trees in the United States. References: (1) California County Agricultural Commissioners' Data, 2010 Crop Year. USDA NASS California field office, retrieved from http://www.nass.usda.gov/Statistics_by_State/California/ Publications/AgComm/201010cactb00.pdf , 2011. (2) P. A. Saccardo. Sylloge Fungorum 3:256, 1884.

scholarly journals First Report of Grapevine Cankers Caused by Lasiodiplodia crassispora and Neofusicoccum mediterraneum in California

Plant Disease ◽  
2010 ◽  
Vol 94 (6) ◽  
pp. 785-785 ◽  
Author(s):  
J. R. Úrbez-Torres ◽  
F. Peduto ◽  
W. D. Gubler
Keyword(s):  
Dna Sequences ◽  
Vascular Tissue ◽  
Elongation Factor ◽  
Fungal Culture ◽  
Internal Transcribed Spacer Region ◽  
First Report ◽  
Consensus Sequences ◽  
Fast Growing ◽  
Agar Plug ◽  
Vascular Discoloration

Several species in the Botryosphaeriaceae family cause perennial cankers in the vascular tissue of grapevines and are responsible for the disease known as bot canker in California (3). Tissue from grapevine vascular cankers from samples submitted to our laboratory in the summer of 2009 were plated onto potato dextrose agar (PDA) amended with 0.01% tetracycline hydrochloride. Lasiodiplodia crassispora (Burgess & Barber) and Neofusicoccum mediterraneum (Crous, M.J. Wingf. & A.J.L. Phillips) were identified based on morphological and cultural characters as well as analyses of nucleotide sequences. L. crassispora isolates were characterized by a fast-growing, white mycelium that turned dark olivaceous with age on PDA. Conidia from pycnidia formed in cultures were thick walled and pigmented with one septum and vertical striations when mature. Conidia measured (25.8–) 27.5 to 30.5 (–33.4) × (12.1) 14.3 to 16.8 (–18.2) μm (n = 60). Pycnidia contained septate paraphyses. N. mediterraneum was characterized as having moderately fast-growing, light green mycelia on PDA. Pycnidia formation was induced with pine needles placed on 2% water agar. Conidia from pycnidia were hyaline, ellipsoidal, thin walled, unicellular, and measured (18.2–) 20.5 to 27.8 (–29) × (5.1) 5.9 to 6.5 (–7.2) μm (n = 60). DNA sequences of the internal transcribed spacer region (ITS1-5.8S-ITS2), part of the β-tubulin gene (BT2), and part of the translation elongation factor 1-α gene (EF1-α) from L. crassispora (UCD23Co, UCD24Co, and UCD27Co) and N. mediterraneum (UCD695SJ, UCD719SJ, UCD720SJ, UCD749St, and UCD796St) grapevine isolates from California were amplified and sequenced. Consensus sequences from L. crassispora and N. mediterraneum from California showed 99 to 100% homology with L. crassispora and N. mediterraneum isolates previously identified and deposited in GenBank (1,2). Sequences from the examined DNA regions of all isolates were deposited at GenBank (GU799450 to GU799457 and GU799473 to GU799488). Pathogenicity tests using three isolates per species were conducted on detached dormant canes of cv. Red Globe. Ten canes per isolate were inoculated by placing a 7-day-old 5-mm-diameter agar plug from each fungal culture into a wound made with a drill on the internode (4). Twenty shoots were inoculated with noncolonized PDA plugs for negative controls. Six weeks after inoculations, necrosis was measured from the point of inoculation in both directions. One-way analysis of variance was performed to assess differences in the extent of vascular discoloration and means were compared using Tukey's test. L. crassispora isolates caused an average necrotic length of 21.1 mm, which was significantly lower (P < 0.05) than the average necrotic length of 35.6 mm caused by the N. mediterraneum isolates. Reisolation of L. crassispora and N. mediterraneum from necrotic tissue was 100% for each species. The extent of vascular discoloration in infected canes was significantly greater (P < 0.05) than in control inoculations (8 mm) from which no fungi were reisolated from the slightly discolored tissue. To our knowledge, this is the first report of L. crassispora and N. mediterraneum as pathogens of Vitis vinifera and as a cause of grapevine cankers in California. References: (1) T. I. Burgess et al. Mycologia 98:423, 2006. (2) P. W. Crous et al. Fungal Planet. No. 19, 2007. (3) J. R. Úrbez-Torres and W. D. Gubler. Plant Dis. 93:584, 2009. (4) J. R. Úrbez-Torres et al. Am. J. Enol. Vitic. 60:497, 2009.

scholarly journals First Report of Calosphaeria pulchella Associated with Branch Dieback of Sweet Cherry Trees in California

Plant Disease ◽  
2010 ◽  
Vol 94 (9) ◽  
pp. 1167-1167 ◽  
Author(s):  
F. P. Trouillas ◽  
J. D. Lorber ◽  
F. Peduto ◽  
J. Grant ◽  
W. W. Coates ◽  
...  
Keyword(s):  
Sweet Cherry ◽  
Prunus Avium ◽  
Large Diameter ◽  
The United States ◽  
Internal Transcribed Spacer Region ◽  
First Report ◽  
Agar Plug ◽  
Vascular Discoloration ◽  
Branch Dieback ◽  
Cherry Trees

California is the second largest sweet cherry producer in the United States with approximately 10,800 ha and an average annual crop value of approximately $150 million. Perennial canker diseases constitute major threats to the cherry industry productivity by reducing tree health, longevity, and yields. During the course of summer 2006, we observed severe limb and branch dieback of sweet cherry (Prunus avium L.) in San Joaquin, San Benito, Contra Costa, and Stanislaus counties of California. Isolation from diseased branches repeatedly yielded the fungus Calosphaeria pulchella (Pers.: Fr.) J. Schröt. (1,2). Cankers and vascular necroses had developed in tree limbs and branches, generally initiating from the heart wood and later spreading into the sapwood. External symptoms of disease may be unapparent throughout the early stages of infection, particularly in large diameter shoots. Older infections often appeared as wilted leaves. Branches and trunks affected with cankers from which C. pulchella was isolated also generally bore perithecia of C. pulchella beneath the periderm. Perithecia were nonstromatic and arranged in dense, circinate groups, with elongated necks converging radially and fissuring the periderm. Asci were unitunicate, clavate, and 45 to 55 × 5 to 5.5 μm. Ascospores were allantoid to suballantoid, hyaline, and 5 to 6 × 1 μm. Colonies on potato dextrose agar (PDA) were dark pink to red in their center with a white margin. Conidia were hyaline, allantoid to oblong-ellipsoidal, and (3–) 4 to 6 (–9) × 1.5 to 2 (–2.5) μm. Identification of C. pulchella isolates also was confirmed by sequence comparison in GenBank database using the internal transcribed spacer region (ITS1-5.8S-ITS2) of the rDNA. Sequences of California isolates shared 100% similarity with C. pulchella reference isolate CBS 115999 (EU367451) (2). ITS sequences of the California isolates used in this study were deposited into GenBank (Nos. HM237297 to HM237300). Pathogenicity of four isolates recovered from the margin of active cankers was determined by branch inoculations. In December 2006, 2- to 4-year-old twigs of P. avium cv. Bing were inoculated with a 5-mm cork borer to remove bark and by placing an agar plug from the growing margin of 8-day-old colonies directly into the fresh wound, mycelium side down. Ten branches per isolate were inoculated. Ten control shoots were inoculated with noncolonized agar plugs. Inoculations were covered with vaseline and wrapped with Parafilm to retain moisture. Branches were harvested in July 2007 and taken to the laboratory to be examined for canker development, and the extent of vascular discoloration in each branch was assessed. Isolations from the edge of discolored tissue were conducted to fulfill Koch's postulates. After 8 months, C. pulchella was reisolated from 100% of the inoculated branches. Length of vascular discoloration averaged 62.5 mm in branches inoculated with the four C. pulchella isolates and 16.5 mm in the control twigs. No fungi were reisolated from the slightly discolored tissue of the controls. To our knowledge, this study constitutes the first report of C. pulchella as a pathogen of sweet cherry trees in California. References: (1) M. E. Barr. Mycologia 77:549, 1985. (2) U. Damm et al. Persoonia 20:39, 2008.

scholarly journals Phomopsis Stem Canker: A Reemerging Threat to Sunflower (Helianthus annuus) in the United States

2015 ◽  
Vol 105 (7) ◽  
pp. 990-997 ◽  
Author(s):  
Febina M. Mathew ◽  
Kholoud M. Alananbeh ◽  
James G. Jordahl ◽  
Scott M. Meyer ◽  
Lisa A. Castlebury ◽  
...  
Keyword(s):  
United States ◽  
Helianthus Annuus ◽  
Great Plains ◽  
Phylogenetic Analyses ◽  
Elongation Factor ◽  
Stem Canker ◽  
The United States ◽  
Northern Great Plains ◽  
Type Specimens ◽  
Internal Transcribed Spacer Region

Phomopsis stem canker causes yield reductions on sunflower (Helianthus annuus L.) on several continents, including Australia, Europe, and North America. In the United States, Phomopsis stem canker incidence has increased 16-fold in the Northern Great Plains between 2001 and 2012. Although Diaporthe helianthi was assumed to be the sole causal agent in the United States, a newly described species, D. gulyae, was found to be the primary cause of Phomopsis stem canker in Australia. To determine the identity of Diaporthe spp. causing Phomopsis stem canker in the Northern Great Plains, 275 infected stems were collected between 2010 and 2012. Phylogenetic analyses of sequences of the ribosomal DNA internal transcribed spacer region, elongation factor subunit 1-α, and actin gene regions of representative isolates, in comparison with those of type specimens, confirmed two species (D. helianthi and D. gulyae) in the United States. Differences in aggressiveness between the two species were determined using the stem-wound method in the greenhouse; overall, D. helianthi and D. gulyae did not vary significantly (P ≤ 0.05) in their aggressiveness at 10 and 14 days after inoculation. These findings indicate that both Diaporthe spp. have emerged as sunflower pathogens in the United States, and have implications on the management of this disease.

scholarly journals First Report of a Root and Crown Disease of the Invasive Weed Lepidium draba Caused by Phoma macrostoma

Plant Disease ◽  
2012 ◽  
Vol 96 (1) ◽  
pp. 145-145 ◽  
Author(s):  
A. J. Caesar ◽  
R. T. Lartey ◽  
T.-C. Caesar-Ton-That
Keyword(s):  
United States ◽  
Biological Control ◽  
Patent Application ◽  
The United States ◽  
Root Tissue ◽  
Conidial Suspension ◽  
Internal Transcribed Spacer Region ◽  
First Report ◽  
South Central ◽  
Lepidium Draba

The exotic rangeland perennial Lepidium draba occurs as a noxious weed in 22 states, mostly in the western United States. Because chemical control measures against this invasive perennial, a member of the Brassicaceae, have not achieved adequate results, biological control is being pursued. While inventories of arthropods that feed on L. draba have been established, little is known of soilborne pathogens for possible use as biological control agents. To address this deficiency, we have surveyed for diseases of L. draba in the United States and Eurasia to identify and test potential biocontrol agents. In intensive surveys for soilborne diseases in a single infestation that is >20 years old in a cattle pasture in south-central Montana, several chlorotic, stunted plants were noted. Roots of chlorotic plants that exhibited elongated fissures from which other soilborne fungi were isolated also had numerous prominent pycnidia embedded in the crown tissue above the lesions. Examination with a dissecting microscope revealed large ostioles made evident by the wide concave inversions in the short necks of the pycnidia. Culture of root tissue on potato dextrose agar resulted in whitish, becoming pale gray colonies, with a dull peach-to-reddish tinge at the margins, with abundant single pycnidia. Conidia in vitro were mainly unicellular, variable shape, subglobose to ellipsoidal, with several guttules averaging 6 × 2.5 μm. These morphological traits are characteristic of Phoma macrostoma, which is regarded as a weak or wound pathogen. The internal transcribed spacer region of rDNA was amplified using primers ITS1 and ITS4 and sequenced. BLAST analysis of the 575-bp fragment showed a 100% homology with the sequence of an isolate of P. macrostoma that has been investigated extensively for commercialization as a biological control agent of various agricultural weeds (1), including wild mustard (GenBank No. DQ474091). The nucleotide sequence has been assigned GenBank No. HM755951. Pathogenicity tests consisted of making four 1.4-mm-diameter holes in five NaOCl (0.1%)-sterilized root sections of L. draba and pipetting ~50 to 100 μl of a 106 CFU/ml conidial suspension into the incisions, incubating the inoculated roots at 20 to 25°C overnight and planting the root sections, one per pot, in an artificial greenhouse potting mix and placing the pots in the greenhouse at 20 to 25°C. Controls were five root sections that were treated similarly except that sterile water was injected. The experiment was repeated. After 10 days, shoots that grew from inoculated roots were chlorotic and shorter than those produced from control roots. P. macrostoma was isolated from tissue of inoculated roots that became blackened distal to the inoculation points. To examine the host range of P. macrostoma on other brassica species, crowns of 2-week-old seedlings of radish, broccoli, cauliflower, broccoli raab, turnip, kohlrabi, cabbage, Chinese cabbage, mustard greens, and canola were injected with 0.5 ml of a 106 CFU/ml conidial suspension. Plants were grown in the greenhouse at 20 to 25°C for 4 weeks after inoculation and examined for symptoms. The experiment was repeated twice. Blackened root tissue with slight chlorosis occurred only on roots of radish and crowns of broccoli, from which P. macrostoma was reisolated. To our knowledge, this the first report of a disease of L. draba caused by P. macrostoma. Reference: (1) K. L. Bailey et al. U.S. Patent Application Serial No. 60/294,475, Filed May 20, 2001.

scholarly journals First Report of Pilidium concavum Causing Tan-brown Rot on Strawberry Nursery Stock in South Carolina

Plant Disease ◽  
2014 ◽  
Vol 98 (7) ◽  
pp. 1010-1010 ◽  
Author(s):  
D. Fernández-Ortuño ◽  
P. K. Bryson ◽  
G. Schnabel
Keyword(s):  
United States ◽  
South Carolina ◽  
Brown Rot ◽  
The United States ◽  
Conidial Suspension ◽  
Strawberry Fruit ◽  
Internal Transcribed Spacer Region ◽  
First Report ◽  
Nursery Stock ◽  
Wide Range

Pilidium concavum (Desm.) Höhn. [synanamorph: Hainesia lythri (Desm.) Höhn.] is an opportunistic pathogen that causes leaf spots and stem necrosis in a wide range of hosts, including strawberry (Fragaria ananassa) (1,2). In October 2013, 24 strawberry plug plants (cv. Chandler) with brown to dark brown necrotic lesions on stolons were obtained from a nursery in Easley, SC. The lesions were oval shaped and varied in length from 2 to 8 mm. The tips of stolons with larger spots had died. To isolate the causal agent, 3 to 5 cm of necrotic stolon tissue was surface disinfected for 1 min with 10% bleach, rinsed with sterile distilled water, air dried, and placed on potato dextrose agar (PDA). After 7 days of incubation at 22°C, pink-orange masses of spores emerged. Single spore colonies on PDA produced a gray to brown colony with whitish aerial mycelium. Numerous discoid to hemisphaerical conidiomata (0.3 to 2.2 mm in diameter) developed with a dark base and exuded a pink, slimy mass that contained many conidia. Conidiophores (10.2 to 47.8 × 0.8 to 2.0 μm) were hyaline, unicellular, cylindrical, and filiform. Conidia (3.0 to 8.5 × 1.0 to 2.9 μm) were aseptate, fusiform, hyaline, and canoe-shaped to allantoid. On the basis of morphology, the pathogen was identified as P. concavum (3). The internal transcribed spacer region ITS1-5.8S-ITS2 was amplified by PCR and sequenced with primers ITS1 and ITS4 (4). The sequence was submitted to GenBank (Accession No. KF911079) and showed 100% homology with sequences of P. concavum. Pathogenicity was examined on strawberry fruit and leaves. Our previous efforts to achieve infection without wounding failed, which is consistent with experiences of other scientists (not cited). Thus, 24 strawberry fruit were wounded (1 cm deep) with a needle once, and submerged for 3 min in a conidial suspension (2 × 106 conidia ml−1). Controls were wounded and submerged in sterile water. After 4 days of incubation at 22°C, characteristic symptoms were observed at the wound site only on inoculated fruit. Detached leaves (about 6 cm in diameter) from 3- to 4-week-old strawberry plants cv. Chandler were surface sterilized and placed right side up in petri dishes (one leaf per dish) containing water agar. Leaves were inoculated at one site with a 50 μl conidial suspension (2 × 106 conidia ml−1) after inflicting a scraping-type injury with a needle to the surface at the point of inoculation. Control leaves received just water. After 7 days of incubation at 22°C, only the inoculated leaves showed symptoms similar to those observed on strawberry stolons. The fungus was re-isolated from symptomatic fruit and leaf lesions and identity was confirmed based on morphological features. The experiments were repeated. To our knowledge, this is the first report of P. concavum causing tan-brown rot on strawberry tissue in South Carolina. Prior to this study, the pathogen has been described from different hosts and countries including Belgium, Brazil, China, France, Iran, Poland, and the United States. Contamination of strawberry nursery stock by P. concavum could become a plant health management issue in the United States, especially if the pathogen is transferred to strawberry production areas. Further information on in-field occurrence of P. concacum is needed. References: (1) J. Debode et al. Plant Dis. 95:1029, 2011. (2) W. L. Gen et al. Plant Dis. 96:1377, 2012. (3) A. Y. Rossman et al. Mycol. Prog. 3:275, 2004. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.

scholarly journals First Report of Hiemalis begonias Wilt Disease Caused by Fusarium foetens in Canada

Plant Disease ◽  
2010 ◽  
Vol 94 (10) ◽  
pp. 1261-1261 ◽  
Author(s):  
X. L. Tian ◽  
M. Dixon ◽  
Y. Zheng
Keyword(s):  
United States ◽  
Elongation Factor ◽  
Aerial Mycelium ◽  
The United States ◽  
Wilt Disease ◽  
Flowering Plant ◽  
Economic Losses ◽  
First Report ◽  
Fusarium Foetens ◽  
Light Orange

Hiemalis begonias Fotsch (Begonia × hiemalis), a member of the family Begoniaceae, is a commercially important potted flowering plant in Europe and North America. In the spring of 2010, stunted growth and dull green leaves of H. begonias were observed in a commercial greenhouse in southern Ontario, Canada. Symptoms began with dull green foliage, followed by wilted leaves, then the stem base became water soaked with vascular discoloration, and finally, large macroconidial masses of a fungus developed on the collapsed stems and veins. A fungus was consistently isolated from the leaves, stems, and roots of symptomatic plants. Single conidia were isolated from sporodochia and cultured on potato dextrose agar (PDA) and oatmeal agar (OA) for 7 days. Isolates exhibited strong pungent odors on PDA and OA and a brownish orange colony on OA and a light orange colony on PDA. Masses of light orange and hemispherical-shaped conidia and stromata formed on OA. Conidiophores formed from aerial mycelium producing ellipsoidal microconidia without septation. Sporodochia formed on agar surface producing three-septate, slightly curved macroconidia. The cultural and conidial characteristics of the isolates were similar to those of Fusarium foetens Schroers (4). Partial translation elongation factor 1-α (TEF) gene was amplified and sequenced with primers ef1 and ef2. A comparison of a partial sequence has been deposited in GenBank (Accession No. HM748968) and showed a 100% match with F. foetens (2). Inoculations with F. foetens isolates were performed by injecting a 100-μl suspension of 1 × 106 conidia/ml into stems of five healthy plants near the ground or soaking the soil of five healthy 6-week-old H. begonias cv. Golden Edith with 50 ml of suspension. Control plants were similarly injected with sterile water or sown in sterile soil. After 4 weeks, all inoculated plants developed dark, wilting leaves and collapsed stems and veins similar to those observed in the commercial greenhouse. F. foetens was reisolated from diseased plants, and identification was reconfirmed by conidial characteristics and TEF 1-α sequence. Control plants were healthy and symptom free. F. foetens has recently been described in association with a new disease of H. begonias in Europe (3) and the United States (1). F. foetens can cause major economic losses to farmers and marketers of H. begonias in Europe and the United States. To our knowledge, this is the first report of F. foetens causing wilt disease of H. begonias in Canada. References: (1) W.-H. Elmer et al. Plant Dis. 88:1287, 2004. (2) D.-M. Geiser et al. Eur. J. Plant Pathol. 110:473, 2004. (3) R. Schrage. Phytomed. Ges. 33:68, 2003. (4) H.-J. Schroers et al. Mycologia 96:393, 2004.

scholarly journals First Report of Aloe vera Rust Caused by Uromyces aloes in an Ornamental Nursery in the United States

Plant Disease ◽  
2021 ◽  
Author(s):  
Devin Bily ◽  
Ekaterina V. Nikolaeva ◽  
Tracey Olson ◽  
Scott Rebert ◽  
Seogchan Kang ◽  
...  
Keyword(s):  
United States ◽  
South Africa ◽  
Wall Thickness ◽  
Aloe Vera ◽  
Morphological Characteristics ◽  
The United States ◽  
Internal Transcribed Spacer Region ◽  
Plant Host ◽  
First Report ◽  
Plant Nursery

Aloe vera (L.) Burm. f. is a tropical evergreen perennial in the family Liliaceae. Native to the Arabian Peninsula, it is sold in Pennsylvania as an ornamental and for its medical and topical purposes due to its high levels of amino acids, anthraquinones, saponins, and vitamins A, B, C, E (Sahu et al. 2013). In February 2020, at an ornamental plant nursery in Lancaster County, Pennsylvania, 5 out of 15 mature A. vera plants in 15 cm pots showed symptoms and signs of rust on the leaves, exhibiting dark-brown erumpent pycnial spots with a chlorotic band surrounding the infected tissue that turned necrotic after three days of incubation at 20°C. Only the telial stage was present. Sori (n=25) were rounded, concentrically arranged, 0.2-3.7 mm, and covered by a brown epidermis. Teliospores (n=40) were amphigenous, orange-brown, globose to ellipsoidal, measuring (29.2) 30.4-36.1 (39.5) × (27.4) 27.6-30.1 (30.5) µm, with a wall thickness of 4-5 µm, and a persistent hyaline pedicel ranging from 5 to 57.1 µm in length and 5.2 to 9.3 µm in width. These measurements were comparable to the descriptions of Uromyces aloes previously reported from India (teliospore size 25-42.5 x 20-30 µm, wall thickness 3-5 µm, and pedicel size 25-95 x 5-6.25 µm), and South Africa (teliospore size 30-44 x 24-32 µm, wall thickness 4-6 µm, and pedicel size 6-20 µm) (Maier et al. 2007; Soni et al. 2011). Based on these morphological traits and the plant host, the causal agent was identified as Uromyces aloes (Cooke) Magnus (Pucciniaceae, Uredinales). The sample was also independently identified as U. aloes by the USDA APHIS PPQ Beltsville lab (Interception # APEMD200552555001) based on morphological characteristics. Teliospores were harvested with a sterile pin, transferred to a 1.5 ml tube with DNA extraction buffer (100 mM Tris-HCL, 10 mM EDTA, 1 M KCl, pH 8) and macerated using a plastic mini-pestle. The DNA was precipitated using isopropanol, washed with 70% ethanol, and reconstituted in 50 µl of PCR-grade water. The segment of the internal transcribed spacer region (ITS) was amplified using ITS4/ITS5 primers (White et al. 1990). The nuclear ribosomal small subunit (18S) was amplified with rust specific primers Rust18S-R (Aime 2006) and NS1 (White et al. 1990). The nuclear ribosomal large subunit (28S) was amplified with primers LR0R and LR7 (Vilgalys et al. 1990). Amplified PCR products were cleaned using ExoSap (Affymetrix, Santa Clara, CA) or QIAquick PCR Purification Kit (Qiagen, Valencia, CA) and sequenced at Penn State Genomics Core Facility. The nucleotide sequences were trimmed, analyzed, and aligned using Geneious 11.1.5 software (Biomatters, Auckland, NZ). The resulting 692-bp segment of the ITS, 1,633-bp segment of the 18S, and the 1,324-bp segment of the 28S regions were deposited in the GenBank database under accession numbers MT136509, MZ146345, and MZ146342, respectively. Based on GenBank BLAST analysis, a 529-bp fragment of our 28S product was found to share 98.87% (523/529) identity with U. aloes isolate WM3290 (DQ917740) from South Africa, with three nucleotide differences and three gaps between the two strains. Comparisons among ITS and 18S sequences could not be made because no ITS or 18S sequence data from U. aloes has previously been deposited in GenBank. To our knowledge, this is the first report of U. aloes from A. vera in the United States. Infected plants were confined inside a greenhouse and have been destroyed. Since the plants were purchased from either Ontario, Canada or Florida, the extent of infection in the United States is unknown.

scholarly journals First Report of Twig and Branch Dieback of English Walnut (Juglans regia) Caused by Neofusicoccum mediterraneum in California

Plant Disease ◽  
2010 ◽  
Vol 94 (10) ◽  
pp. 1267-1267 ◽  
Author(s):  
F. P. Trouillas ◽  
J. R. Úrbez-Torres ◽  
F. Peduto ◽  
W. D. Gubler
Keyword(s):  
Juglans Regia ◽  
Petroleum Jelly ◽  
Internal Transcribed Spacer Region ◽  
First Report ◽  
Fungal Isolates ◽  
Agar Plug ◽  
Vascular Discoloration ◽  
English Walnut ◽  
Branch Dieback ◽  
Olive Green

California produces 99% of the U.S. English walnut crop with more than 30 cultivars on ~89,000 ha. Production for 2008 was ~436,000 tons with a value of $527 million. In early summer of 2009 and 2010, branch and twig dieback of English walnut (Juglans regia L.) was detected in orchards in Yolo County and submitted to our diagnostic laboratory. Disease symptoms included death of twig tips, branch dieback, wood lesions, and canker formation. Pycnidia were embedded within the bark of dead twigs. Conidia from pycnidia were hyaline, fusoid-ellipsoidal, widest usually in the middle, and 21 to 24 (–27) × 5 to 7 μm (n = 30). Isolations from cankers yielded the fungus Neofusicoccum mediterraneum Crous, M.J. Wingf. & A.J.L. Phillips (1). Fungal colonies of N. mediterraneum grew light olive green to gray on potato dextrose agar, becoming dark olive green with age. Identification of fungal isolates was confirmed by sequence comparison of Californian isolates with ex-type (CBS 121558) sequences in GenBank (3) using the internal transcribed spacer region of the rDNA, a portion of the β-tubulin gene, and part of the translation elongation factor. Sequences of Californian isolates (GenBank HM443604–HM443609) were identical to the ex-type sequences for all three genes. Previous studies in California reported the occurrence and pathogenicity of N. mediterraneum into grapevine (Vitis vinifera L.) (3) and almond trees (Prunis dulcis L.) (2). Inderbitzin et al (2) investigated the host range of N. mediterraneum in California and reported the occurrence of pycnidia on English walnut trees. However, this study did not investigate the pathogenicity of N. mediterraneum on this host. In the current study, the pathogenicity of N. mediterraneum in J. regia cvs. Hartley and Chandler was investigated in an orchard at UC Davis using two fungal isolates. Pathogenicity tests were performed by inoculating eight 2- to 4-year-old branches of mature J. regia trees. Inoculations were made in June 2009 with a 5-mm cork borer to remove bark and placing an 8-day-old 5-mm-diameter agar plug bearing fresh mycelium of the fungal isolates directly into the fresh wound, mycelium side down. An additional eight branches of each cultivar were inoculated with noncolonized agar plugs to serve as controls. Inoculated wounds were covered with petroleum jelly and wrapped with Parafilm to retain moisture. Branches were harvested after 10 months of incubation and checked for canker development. The extent of vascular discoloration was measured in each branch and isolations were made from the edge of discolored tissue to confirm Koch's postulates. Statistical analyses were performed with analysis of variance and Dunnett's t-test to assess significant differences in the extent of vascular discoloration between inoculations with N. mediterraneum and the control. Necrosis length for the two isolates averaged 131.5 mm in Hartley branches and 110 mm in the Chandler branches. Average necrosis lengths in the control branches were 18.5 mm and 16.7 mm, respectively, significantly lower (P < 0.05) than the average necrosis length found in branches inoculated with N. mediterraneum. Fungal recovery was 75% in both varieties. To our knowledge, this study is the first report of N. mediterraneum as a pathogen of J. regia trees in California. References: (1) P. W. Crous et al. Fungal Planet 19, 2007. (2) P. Inderbitzin et al. Mycologia. Online publication. doi:10.3852/10-006, 2010. (3) J. R. Úrbez-Torres et al. Plant Dis. 94:785, 2010.

scholarly journals First Report of Flower Anthracnose Caused by Colletotrichum karstii in White Phalaenopsis Orchids in the United States

Plant Disease ◽  
2012 ◽  
Vol 96 (8) ◽  
pp. 1227-1227 ◽  
Author(s):  
I. Jadrane ◽  
M. Kornievsky ◽  
D. E. Desjardin ◽  
Z.-H. He ◽  
L. Cai ◽  
...  
Keyword(s):  
United States ◽  
Species Complex ◽  
The United States ◽  
Likelihood Method ◽  
Conidial Suspension ◽  
Necrotic Tissue ◽  
Internal Transcribed Spacer Region ◽  
First Report ◽  
Whole Plant ◽  
Pure Cultures

In October 2010, a Colletotrichum species was isolated from white Phalaenopsis flowers growing in a greenhouse in San Francisco, CA. This Phalaenopsis is a common commercial orchid hybrid generated mostly likely from Phalaenopsis amabilis and P. aphrodite. The white petals showed anthracnose-like lesions where necrotic tissue is surrounded by a ring of green tissue. The green halo tissues around the necrotic tissue contain functional chloroplasts. One-centimeter disks were cut around the necrotic sites and surface-sterilized with 95% ethanol and 0.6% sodium hypochlorite. The disks were placed on potato dextrose agar (PDA) medium to establish cultures. Pure cultures were obtained by subculturing hyphal tips onto fresh PDA plates. The generated colonies had white aerial mycelia and orange conidial mass. The color of the reverse colony varies between colorless and pale orange. Microscopic observations identified the conidia as cylindrical, straight, and rounded at both ends. In addition, the conidia were approximately 15.0 to 18.0 μm long and 5.0 to 6.5 μm in diameter. These observed morphological features suggested that these isolates possessed the same characteristics as previously described for Colletotrichum karstii, a species considered as part of the C. boninense species complex (1). Four putative independent Colletotrichum isolates were recovered (DED9596, DED9597, DED9598, and DED9599). To confirm the Colletotrichum isolates as the causative pathogen, healthy white Phalaenopsis flowers (five total) in a whole plant were sprayed with a conidial suspension (approximately 1.2 × 106 conidia/ml) of the isolates and incubated at 20°C and 100% relative humidity with cycles of 16 h light and 8 h of darkness. Approximately 1 ml of conidial suspension solution was used for each flower. The plants were watered regularly and flowers were sprayed with sterile double-distilled water daily. As negative controls, five flowers in a whole plant were sprayed with water. Fifteen to twenty days after inoculation, lesions started to form on the petals sprayed with the putative Colletotrichum isolates. All controls remained healthy. The Colletotrichum-inoculated flowers remained alive and did not die as a result of the infection. This same experiment was repeated and the same results were obtained. DNA was extracted from the necrotic regions of the petals infected by the pure cultures of the four isolates and used to sequence the 18S rRNA ITS (internal transcribed spacer) region. All four isolates gave identical ITS sequences. Analysis of the obtained representative sequences (GenBank Accession No. JQ277352) suggested that the isolated pathogen as C. karstii. Using the published ITS data for the C. boninense species complex (1), a phylogenetic tree was generated via the maximum likelihood method. This created tree places the isolates in the same group as C. karstii. This type of C. karstii infection in Phalaenopsis orchid petals was not documented in the U.S. before, although it has been reported in China and Thailand (2). To our knowledge, this is the first report of infection and green island formation caused by C. karstii on orchid flower in the United States. References: (1) Damm et al. Studies in Mycology 73:1, 2012. (2) Yang et al. Cryptogamie Mycologie 32:229, 2011.

scholarly journals First Report of Powdery Mildew Caused by Podosphaera xanthii on Sechium edule in the United States

Plant Disease ◽  
2009 ◽  
Vol 93 (12) ◽  
pp. 1348-1348 ◽  
Author(s):  
R. Singh ◽  
D. M. Ferrin ◽  
M. C. Aime
Keyword(s):  
United States ◽  
Powdery Mildew ◽  
New Orleans ◽  
Dna Analysis ◽  
The United States ◽  
Abaxial Surface ◽  
Internal Transcribed Spacer Region ◽  
Voucher Specimen ◽  
First Report ◽  
Sechium Edule

Sechium edule (Jacq.) Sw., also known as mirliton or chayote, is a perennial, monoecious, cucurbitaceous plant native to Mexico and Central America. It is cultivated worldwide for a variety of uses (4). Mirliton fruit is rich in carbohydrates, has 16 amino acids, and is a traditional staple in New Orleans, LA. During the spring of 2009, the LSU AgCenter's Plant Disease Diagnostic Clinic received diseased mirliton plants from a small commercial grower in New Orleans. Symptoms included yellow, irregular spots on both surfaces of the leaves. Microscopic examination revealed the presence of powdery mildew conidia and conidiophores. Initially, white, cottony mycelial colonies were present on the abaxial surface, but as the disease progressed, white, cottony colonies developed on the adaxial surface, the spots coalesced, and the entire leaf turned yellow and necrotic. Conidia were hyaline, ovoid, borne in long chains with crenate edges, and measured 25.6 to 36.6 μm long (mean = 31.2) × 14.6 to 18.3 μm wide (mean = 17.1). Conidia contained fibrosin bodies and produced a lateral germ tube with a simple appressorium. Conidiophores were erect, simple, unbranched, and measured 54.9 to 76.9 μm long (mean = 66.4) × 11.0 to 14.6 μm wide (mean = 12.9). The cylindrical foot cell had a simple base with basal septum adjacent to the mycelium. No teleomorph was observed. Morphologically, this powdery mildew fits either Podosphaera fusca or P. xanthii so DNA analysis was conducted. We designed Podosphaera-specific primers PFITS-F (5′-CCAACTCGTGCTGTGAGTGT-3′) and PF5.8-R (5′-TGTTGGTTTCTTTTCCTCCG-3′) to amplify and sequence the internal transcribed spacer region (ITS) of the nuclear rDNA. The 331-bp sequence (GenBank Accession No. GQ902939) was identical with haplotype 27 of P. fusca (GenBank Accession No. AB040324) (3), which is now called P. xanthii (1). Pathogenicity tests were conducted by pressing infected leaves against healthy leaves of two vines. A noninoculated vine served as a control. Plants were maintained in a greenhouse at 30°C. Five days after inoculation, yellow, irregular spots appeared on the inoculated vines and white, powdery mildew colonies appeared on the abaxial surface. Spots coalesced and the entire leaf turned yellow 8 days after inoculation and necrotic 12 days after inoculation. No symptoms developed on the controls. On the basis of DNA sequence data, this powdery mildew is identified as P. xanthii sensu (1). Erysiphe cichoracearum has been previously reported to cause powdery mildew on mirlitons in Florida and Hawaii (2). To our knowledge, this is the first report of powdery mildew caused by P. xanthii on mirliton in the United States. A voucher specimen has been deposited in the Bernard Lowy Mycological Herbarium (LSUM 185359). References: (1) U. Braun and S. Takamatsu. Schlechtendalia 4:31, 2000. (2) D. F. Farr and A. Y. Rossman. Fungal Databases. Systematic Mycology and Microbiology Laboratory, SMML, Online publication. USDA-ARS, 2009. (3) T. Hirata et al. Can. J. Bot. 78:1521, 2000. (4) M. Janssens et al. Tropical Crops. ARTS; Field and Vegetable Crops, PTS 130. Bonn, Germany, 2002/03.

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