ABSTRACT
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.
ABSTRACT
Seashore paspalum (Paspalum vaginatum Sw.) is a newly cultivated C4 turfgrass that has exceptional salinity tolerance and is highly suited for use on golf courses in coastal areas. In October 2008 and June 2009, circular patches of blighted seashore paspalum ranging from 30 cm to >3 m in diameter were observed in fairways, tees, and roughs established with 'Supreme' seashore paspalum at Roco Ki Golf Club in Macao, Dominican Republic. Affected patches were initially chlorotic followed by reddish brown necrosis of leaves and leaf sheaths. Reddish brown-to-gray lesions were also observed on leaf sheaths during the early stages of necrosis. During periods of wet or humid weather from June through October, basidiocarps were produced on necrotic plant tissue and identified as Marasmiellus mesosporus Singer (2). Three isolates were obtained by plating symptomatic leaf sheaths that were surface sterilized with a 0.5% NaOCl solution on potato dextrose agar amended with 50 ppm each of streptomycin, chloramphenicol, and tetracycline (PDA+++). Sequences of the internal transcribed spacer (ITS) region of rDNA, obtained from these three isolates and three stipes of basidiocarps, were identical to each other and 99% similar to a M. mesosporus sequence deposited in the NCBI database (Accession No. AB517375). To confirm pathogenicity, a M. mesosporus isolate obtained from symptomatic plant tissue was inoculated onto 6-week-old P. vaginatum ('Seaspray') planted (0.5 mg seed/cm2) in 10-cm-diameter pots containing a mixture of 80% sand and 20% reed sedge peat. Two weeks prior to inoculation, the isolate was grown on a sterilized mixture of 100 cm3 of rye grain, 4.9 ml of CaCO3, and 100 ml of water. Infested grains were placed 0.5 cm below the soil surface for inoculation. Pots were inoculated with five infested grains or five sterilized, uninfested grains with three replications of each treatment. After inoculation, pots were placed in a growth chamber with a 12-h photoperiod set to 30°C during the day and 26°C at night. Approximately 20% of plants in inoculated pots were necrotic 7 days postinoculation and this increased to 75% by 21 days postinoculation. Diseased plants in inoculated pots exhibited symptoms similar to those observed in the field. Leaves were initially chlorotic with brown lesions on lower leaf sheaths and eventually turned necrotic, reddish brown, and collapsed. Pots receiving uninfested grains were healthy and showed no symptoms on all rating dates. At 21 days postinoculation, basidiocarps were observed emerging from three colonized plants at the base of the oldest leaf sheath near the crown. Three reisolations were made on PDA+++ from stem lesions surface sterilized with a 0.5% NaOCl solution. All reisolations were confirmed as M. mesosporus by culture morphology and ITS sequence data. M. mesosporus was previously reported causing disease on American beachgrass (Ammophila breviligulata Fernald) in North Carolina (1) and recently in Japan (3). The pathogen was initially placed in the genus Marasmius and reported as the cause of the disease Marasmius blight (1). Subsequent morphological observation found that the pathogen belonged in the genus Marasmiellus (2). To our knowledge, this is the first report of M. mesosporus causing Marasmiellus blight on seashore paspalum, a high-amenity turfgrass. References: (1) L. Lucas et al. Plant Dis. Rep. 55:582, 1971. (2) R. Singer et al. Mycologia 65:468, 1973. (3) S. Takehashi et al. Mycoscience 48:407, 2007.
ABSTRACT
Recent collections of bird's nest fungi (i.e. Crucibulum, Cyathus, Mycocalia, Nidula, and Nidularia species) in northern Thailand resulted in the discovery of a new species of Cyathus, herein described as C. subglobisporus. This species is distinct by a combination of ivory-coloured fruiting bodies covered with shaggy hairs, plications on the inner surface of the peridium and subglobose basidiospores. Phylogenetic analyses based on ITS and LSU ribosomal DNA sequences using neighbour-joining, maximum likelihood and weighted maximum parsimony support Cyathus subglobisporus as a distinct species and sister to a clade containing C. annulatus, C. renweii and C. stercoreus in the Striatum group.
ABSTRACT
Chronic lung infection with mucoid Pseudomonas aeruginosa is the major pathologic feature of cystic fibrosis. Previous studies suggested that a failure to produce opsonic antibody to the mucoid exopolysaccharide (MEP; also called alginate) capsule is associated with the maintenance of chronic bacterial infection. Provision of MEP-specific opsonic antibodies has therapeutic potential. To evaluate the ability of MEP to elicit opsonic antibodies, humans were immunized with two lots of MEP vaccine that differed principally in molecular size. Lot 2 had a larger average MEP polymer size. Both vaccines were well tolerated, but lot 1 was poorly immunogenic, inducing long-lived opsonic antibodies in only 2 of 28 vaccinates given doses of 10 to 150 micrograms. In contrast, at the optimal dose of 100 micrograms, lot 2 elicited long-lived opsonic antibodies in 80 to 90% of the vaccinates. The antibodies elicited by both lots enhanced deposition of C3 onto mucoid P. aeruginosa cells and mediated opsonic killing of heterologous mucoid strains expressing distinct MEP antigens. These results indicate that the polymers of MEP with the largest molecular sizes safely elicit opsonic antibodies in a sufficiently large proportion of vaccinates to permit studies of active and passive immunization of cystic fibrosis patients against infection with mucoid P. aeruginosa.
