scholarly journals First Report of a Bacterial Disease on Creeping Bentgrass (Agrostis stolonifera) Caused by Acidovorax spp. in the United States

Plant Disease ◽  
2010 ◽  
Vol 94 (7) ◽  
pp. 922-922 ◽  
Author(s):  
P. R. Giordano ◽  
J. M. Vargas ◽  
A. R. Detweiler ◽  
N. M. Dykema ◽  
L. Yan

In June of 2009, a golf course putting green sample of creeping bentgrass (Agrostis stolonifera L.) cv. Penn G-2 from a golf club in North Carolina was submitted to the Michigan State University Turfgrass Disease Diagnostic Laboratory for diagnosis. The sample exhibited symptoms of general wilt, decline, and characteristic necrosis from the leaf tips down. Fungal pathogens were ruled out when no phytopathogenic fungal structures were observed with microscopic examination of infected tissue. Symptoms appeared similar to those of annual bluegrass affected by bacterial wilt caused by Xanthomonas translucens pv. poae. Bacterial streaming was present in all of the cut infected tissue of the Penn G-2 bentgrass sample when observed with a microscope. To isolate the causal agent, cut leaf tissue (1- to 3-mm tips) exhibiting bacterial streaming was surface disinfected for 1 min in 10% sodium hypochlorite solution and rinsed for 1 min with sterile distilled water. Leaf blades were placed into Eppendorf microtubes with 20 μl of sterile phosphate-buffered saline (PBS) solution (pH 7) and macerated with a sterile scalpel. Serial dilutions up to 1 × 10–4 were performed in sterile PBS; 10 μl of each suspension was plated onto nutrient agar (NA) (Becton Dickinson, Sparks, MD) and incubated at room temperature for 5 days. Pure cultures of three commonly observed single bacterial colonies growing on plates from serial dilutions were made on NA medium. These pure cultures were grown for 5 days and used to inoculate three replicates of 5-week-old Penn G-2 plants that had uniformly filled in 8.5-cm-diameter pots grown under greenhouse conditions. Uninfected Penn G-2 creeping bentgrass plants were inoculated with 1 ml of 1.3 × 109 CFU/ml of bacterial suspension by adding drops of the suspension to blades of sterile scissors used to cut the healthy plants. Of the three different bacterial cultures selected to inoculate healthy plants, only one resulted in slight browning of leaf tips just 2 days after inoculation. The symptoms progressed, and by 5 days after inoculation, browning, twisting and leaf dieback to the sheath were observed. When leaf tips of the inoculated plants were cut, bacterial streaming was observed. Isolation of the bacterium from inoculated Penn G-2 plants was performed to fulfill Koch's postulates. Once isolated, a single bacterial colony was identified by 16S rDNA sequencing (Microcheck Inc. Northfield, VT). 16S rDNA sequencing results indicated that the causal agent of bacterial infection was a member of the Acidovorax genus, with a 100% sequence match to Acidovorax avenae subsp. avenae (2). The same nonflorescent, aerobic, gram-negative bacterium has been consistently isolated from inoculated plants exhibiting symptoms thus far. A member of the Acidovorax genus has also been identified as a pathogen of creeping bentgrass in Japan (1). To our knowledge, this is the first report of a bacterial disease affecting creeping bentgrass caused by Acidovorax spp. in the United States. References: (1) N. Furuya et al. J. Fac. Agric. Kyushu Univ. 54:13. 2009. (2) N. Schaad et al. Syst. Appl. Microbiol. 31:434. 2008.

2015 ◽  
Vol 25 (6) ◽  
pp. 757-761 ◽  
Author(s):  
Ethan T. Parker ◽  
J. Scott McElroy ◽  
Michael L. Flessner

Smooth crabgrass (Digitaria ischaemum) and goosegrass (Eleusine indica) are problematic weeds in creeping bentgrass (Agrostis stolonifera) because of limited herbicide options for postemergence (POST) control and turfgrass injury potential. Metamifop is a herbicide currently being considered for release to markets in the United States but information is lacking on the most effective rates and application timings for smooth crabgrass and goosegrass control in creeping bentgrass. Field trials were conducted in Auburn, AL in 2009 and 2013 to evaluate metamifop rates (200 to 800 g·ha−1) and single or sequential application timings compared with fenoxaprop (51 to 200 g·ha−1) at two different mowing heights. Metamifop applied twice and three times sequentially at 200 g·ha−1 provided the greatest smooth crabgrass (>97%) and goosegrass (>90%) control at rough (1½ inch) and green (1/8 inch) mowing heights without unacceptable creeping bentgrass injury at 56 days after initial treatment. All treatments caused <20% visible injury on creeping bentgrass at both mowing heights except the highest rate of metamifop. Smooth crabgrass control at the green mowing height was greater than at the rough mowing height, especially at lower metamifop rates with a single application.


Plant Disease ◽  
2007 ◽  
Vol 91 (12) ◽  
pp. 1685-1685 ◽  
Author(s):  
J. Latinović ◽  
J. C. Batzer ◽  
K. B. Duttweiler ◽  
M. L. Gleason ◽  
G. Sun

The sooty blotch and flyspeck (SBFS) complex includes more than 30 fungi that blemish the cuticle of apple fruit, causing economic losses in humid regions worldwide (1). In August 2005, we sampled SBFS-infested wild plum (Prunus americana) fruit growing in hedgerows in Iowa. Colonies were categorized according to mycelial type (1), and isolates were made from representative colonies onto acidified water agar (AWA). Plum skins with SBFS signs were excised, pressed, and photographed. DNA was obtained from purified isolates and also from mycelium and fruiting bodies scraped directly from plum fruit skins. Extracted DNA was amplified using primer pair ITS1-F/Myc1-R (ACTCGTCGAAGGAGCTACG) and PCR products were sequenced using primer pair ITS-1F/ITS4. Six sequences were obtained from pure cultures and seven from colonies on plum fruit skin. BLAST analysis of the 470-bp sequences showed 100% homology to five known species in the SBFS complex: Zygophiala cryptogama, Zygophiala wisconsinensis, Pseudocercosporella sp. RH1, and Stomiopeltis spp. RS1 and RS2 (GenBank Accession Nos. AY598854, AY598853, AY5988645, AY598882, and AY598883, respectively). Observations of colony and fruiting structure morphology from cultures on potato dextrose agar (PDA) and colonies on plums confirmed species identity. A modified version of Koch's postulates was conducted to verify that these fungi caused the signs observed on plum and could also infest apple fruit. In June 2006, 1-month-old cultures on PDA were pulverized in a blender with sterile distilled water, passed through four layers of sterile cheesecloth, and transferred to sterile jars. Each isolate was inoculated onto 20 fruit on plum trees (P. americana) on the Iowa State University (ISU) campus and 20 fruit on cv. Golden Delicious apple trees at the ISU Research Station, Gilbert, IA. Each fruit was disinfested with 70% ethanol, air dried, swabbed with inoculum, and covered with a Fuji bag. At harvest, fungal colonies on fruit were reisolated onto AWA. DNA was extracted from pure cultures; when isolations on agar were unsuccessful, DNA was extracted directly from colonies on fruit. PCR was conducted using ITS1-F/Myc1-R, and PCR products were sequenced using ITS1-F/ITS4. All five species were reisolated and sequenced from apple. Pseudocercosporella sp. RH1 and Stomiopeltis sp. RS1 were sequenced from inoculated plums. Although flyspeck, presumably caused by Schizothyrium pomi, was reported on Japanese plum (P. salicina) in Japan (2) and black cherry (P. serotina) in the United States (3), to our knowledge this is the first report of SBFS fungi on plum in the United States and the first confirmation that fungi from plum can produce SBFS signs on apple fruit. Wild plum may therefore act as a reservoir host, providing inoculum for SBFS infestations on apple. References: (1) J. Batzer et al. Mycologia 97:1268, 2005. (2) H. Nasu and H. Kunoh. Plant Dis. 71:361, 1987. (3) T. B. Sutton. Plant Dis. 72:801, 1988.


Plant Disease ◽  
2013 ◽  
Vol 97 (6) ◽  
pp. 835-835 ◽  
Author(s):  
K. N. Conner ◽  
J. Olive ◽  
L. Zhang ◽  
J. Jacobi ◽  
M. L. Putnam

Bacterial gall symptoms were observed on Loropetalum chinense (R. Br.) Oliv. in two separate commercial nurseries in South Alabama during the spring of 2012. Limb dieback and plant death was first reported by the growers. Plants with dieback symptoms had galling and irregular dark callus formation on the lower stem and lower branches. Galls were small, 0.2 to 1 cm, inconspicuous, and in some cases girdled the stem causing breakage of the main stem. In both locations, 30 to 40% of the crop was affected. Similar symptoms have been observed on L. chinense in nursery and landscape plantings in central Alabama, North Carolina, and Georgia in previous years. Bacterial colonies were isolated from four plants representing two different locations. Isolates were recovered from surface sterilized symptomatic tissue on nutrient agar and King's medium B (KMB). All isolates were gram-negative and fluoresced blue-green under UV light after 48 h of growth at 28°C on KMB. One representative isolate from each site was identified as Pseudomonas savastanoi based on their fatty acid profiles (similarity index of 0.776; MIS-TSBA, version 4.0, MIDI Inc., Newark, DE) and LOPAT tests (2). The identity was confirmed by sequencing a 900-bp portion of the 16S rDNA gene, which revealed 98% similarity to the P. savastanoi type strain in NCBI (Accession No. AB021402). In greenhouse pathogenicity tests, eight Loropetalum liners were inoculated with a bacterial suspension (107 CFU/ml) of each of the two isolates. Plants were inoculated by injecting the suspension into the lower stem after wounding by puncturing with needles or slicing sections of the bark. Controls were inoculated with water. All plants inoculated with the bacteria developed gall symptoms in 8 weeks under 90% relative humidity at 30°C. The bacteria were reisolated from five inoculated plants. DNA was extracted from each isolate, amplified using primer pair 27F/1492R targeting the 16S rDNA gene (1), and sequenced. Sequences (900 bp) from all isolates shared 98 to 99% similarity to P. savastanoi type strain in GenBank (Accession No. AB021402). Nucleotide sequence data reported are available in GenBank under accessions JX915832 to 37. To our knowledge, this is the first report of bacterial gall of L. chinense caused by P. savastanoi in the United States. Given the increasing prevalence of this disease in South Alabama, its confirmation is a significant step toward management recommendations for growers. References: (1) D. J. Lane. 16S/23S rRNA sequencing. Page 115-175 in: Nucleic Acid Techniques in Bacterial Systematics. E. Stackebrandt and M. Goodfellow, eds. John Wiley and Sons, New York, 1991. (2) N. W. Schaad et al. Laboratory Guide for Identification of Plant Pathogenic Bacteria. 3rd ed. The American Phytopathological Society, St. Paul, MN, 2001.


Plant Disease ◽  
2009 ◽  
Vol 93 (1) ◽  
pp. 108-108 ◽  
Author(s):  
J. A. Abad ◽  
M. Bandla ◽  
R. D. French-Monar ◽  
L. W. Liefting ◽  
G. R. G. Clover

Zebra chip (ZC), an emerging disease causing economic losses to the potato chip industry, has been reported since the early 1990s in Central America and Mexico and in Texas during 2000 (4). ZC was subsequently found in Nebraska, Colorado, New Mexico, Arizona, Nevada, California, and Kansas (3). Severe losses to potato crops were reported in the last few years in Mexico, Guatemala, and Texas (4). Foliar symptoms include purple top, shortened internodes, small leaves, enlargement of the stems, swollen axillary buds, and aerial tubers. Chips made from infected tubers exhibit dark stripes that become markedly more visible upon frying, and hence, are unacceptable to manufacturers. Infected tubers may or may not produce plants when planted. The causal agent of ZC is not known and has been the subject of increased investigation. The pathogen is believed to be transmitted by the potato psyllid, Bactericera cockerelli, and the association of the vector with the disease is well documented (3). Following the report of a potential new liberibacter species in solanaceous crops in New Zealand, we sought to identify this liberibacter species in plants with symptoms of the ZC disease. Six potato plants (cv. Russet Norkota) exhibiting typical ZC symptoms were collected in Olton, TX in June of 2008. DNA was extracted from roots, stems, midribs, and petioles of the infected plants using a FastDNA Spin Kit and the FastPrep Instrument (Qbiogene, Inc., Carlsbad, CA). Negative controls from known healthy potato plants were included. PCR amplification was carried out with ‘Candidatus L. asiaticus’ omp primers (1), 16S rDNA primers specific for ‘Ca. L. asiaticus’, ‘Ca. L. africanus’, and ‘Ca. L. americanus’ (1), and 16S rDNA primers OA2 (GenBank Accession No. EU834130) and OI2c (2). Amplicons from 12 samples were directly sequenced in both orientations (McLab, San Francisco CA). PCR amplifications using species-specific primers for the citrus huanglongbing liberibacter were negative. However, 1.1- and 1.8-kb amplicons were obtained with the OA2/OI2C and omp primers, respectively. The sequences for the rDNA were submitted to NCBI GenBank (Accession Nos. EU884128 and EU884129). BLASTN alignment of the 16S rDNA sequences obtained with primers OA2 and OI2c revealed 99.7% identity with a new species of ‘Ca. Liberibacter’ identified in New Zealand affecting potato (GenBank Accession No. EU849020) and tomato (GenBank Accession No. EU834130), 97% identity with ‘Ca. L. asiaticus’, and 94% with ‘Ca. L. africanus’ and ‘Ca. L. americanus’. The neighbor-joining phylogenetic tree constructed using the 16S rDNA fragments delineated four clusters corresponding to each of the liberibacter species. These results confirm that ‘Ca. Liberibacter’ spp. DNA sequences were obtained from potatoes showing ZC-like symptoms, suggesting that a new species of this genus may be involved in causing ZC disease. To our knowledge, this is the first report of the detection of ‘Ca. Liberibacter’ spp. in potatoes showing ZC disease in the United States. References: (1) C. Bastianel et al. Appl. Environ. Microbiol. 71:6473, 2005. (2) S. Jagoueix et al. Mol. Cell. Probes 10:43, 1996. (3) J. E. Munyaneza et al. J. Econ. Entomol. 100:656, 2007. (4) G. A. Secor and V. V. Rivera-Varas. Rev. Latinoamericana de la Papa (suppl.)1:1, 2004.


Plant Disease ◽  
2012 ◽  
Vol 96 (12) ◽  
pp. 1736-1742 ◽  
Author(s):  
Paul R. Giordano ◽  
Arielle M. Chaves ◽  
Nathaniel A. Mitkowski ◽  
Joseph M. Vargas

Bacterial etiolation and decline caused by Acidovorax avenae subsp. avenae is an emerging disease of creeping bentgrass (Agrostis stolonifera) in and around the transition zone, a unique area of turfgrass culture between cool and warm regions of the United States. It is suspected that the disease has been present for many years, although diagnosis of the first occurrence was not reported until 2010. Solicitation of samples from golf courses in 2010 and 2011 was undertaken to investigate the prevalence and dissemination of Acidovorax avenae subsp. avenae on creeping bentgrass. At least 21 isolates from 13 states associated with these outbreaks on golf courses were confirmed as A. avenae subsp. avenae by pathogenicity assays and 16S rDNA sequence analysis at two independent locations. Pathogenicity testing of bacterial isolates from creeping bentgrass samples exhibiting heavy bacterial streaming confirmed A. avenae subsp. avenae as the only bacterium to cause significant disease symptoms and turfgrass decline. Host range inoculations revealed isolates of A. avenae subsp. avenae to be pathogenic on all Agrostis stolonifera cultivars tested, with slight but significant differences in disease severity on particular cultivars. Other turfgrass hosts tested were only mildly susceptible to Acidovorax avenae subsp. avenae infection. This study initiated research on A. avenae subsp. avenae pathogenicity causing a previously uncharacterized disease of creeping bentgrass putting greens 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 ◽  
...  

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.


Plant Disease ◽  
2011 ◽  
Vol 95 (1) ◽  
pp. 78-78 ◽  
Author(s):  
E. N. Njambere ◽  
B. B. Clarke ◽  
S. A. Bonos ◽  
J. A. Murphy ◽  
R. Buckley ◽  
...  

Waitea circinata var. circinata was first reported as the causal agent of brown ring patch on annual bluegrass (Poa annua L.) in the United States in 2007 (2). In early April to mid-June of 2009, circular to irregularly shaped yellow rings resembling symptoms of this disease were observed on an annual bluegrass putting green at Rutgers University in North Brunswick, NJ. Severely infected foliage eventually turned brown as the disease progressed. During the same time period, similar disease symptoms were observed on creeping bentgrass (Agrostis stolonifera L.) from a golf course in Bedminster Township, NJ. The disease reappeared in both locations in April of 2010. Five additional samples with similar symptoms on creeping bentgrass and annual bluegrass were received at Rutgers Diagnostic Laboratory from Paramus, Madison, Allamuchy, and Farmingdale, NJ between late April and early May of 2010. Portions of diseased leaf and sheath tissue that displayed symptoms of the disease were disinfested for 1 min in 0.5% NaOCl, rinsed with sterile distilled water, and plated on potato dextrose agar (PDA) amended with 50 mg/liter of streptomycin sulfate. At the first sign of fungal growth, single hyphal tips were transferred to PDA. After 1 week at 25°C, white-to-orange mycelial colonies formed in culture and eventually turned brown with age. Minute sclerotia (≤3 mm), which followed the same color development pattern, formed within 10 days. These features are consistent with those described of W. circinata var. circinata (2,3). The internal transcribed spacer (ITS) region of the ribosomal RNA gene was amplified using primer pair ITS1/ITS4 and sequenced with ITS4 (GenBank Accession Nos. HQ166065 to HQ166071). BLASTn analysis of the ITS sequences showed a 99 to 100% similarity to W. circinata var. circinata sequences deposited in GenBank (1,2). Pathogenicity tests were conducted in 2010 using 6-week-old creeping bentgrass seedlings cv. Declaration inoculated with colonized oat grain that had been autoclaved and then infested with the Bedminster Township isolate. Eight colonized oat grains were uniformly spread around the crowns of seedlings grown in 10-cm-diameter pots. Control plants were treated with autoclaved grain. Plants were incubated at 25°C and high humidity maintained by misting the plants three times per day. Within 3 days postinoculation, foliage near infested grain turned chlorotic. All foliage in pots became completely blighted and spherical orange-brown sclerotia were observed on leaf sheaths by the eighth day. W. circinata var. circinata was consistently reisolated from inoculated plants (as confirmed by isolate morphology and ITS sequencing) but not from control plants. The ITS sequence data, morphological characters of the isolates, and pathogenicity tests demonstrate that W. circinata var. circinata is present in New Jersey. To our knowledge, this is the first report of W. circinata var. circinata infecting turfgrass in New Jersey. References: (1) C. M. Chen et al. Plant Dis. 93:906, 2009. (2) K. A. de la Cerda et al. Plant Dis. 91:791, 2007. (3) T. Toda et al. Plant Dis. 89:536, 2005.


1970 ◽  
Vol 27 ◽  
pp. 21-24 ◽  
Author(s):  
Mosharrof Hossain

Aeromonas spp infections are probably the most common bacterial disease diagnosed in cultured warm water fish. In the present study, six strains of Aeromonas spp bacteria were isolated from the gourami (Colisa lalia) by 16S rDNA sequencing analyses that are pathogenic to freshwater fish. Among them, three were under Aeromonas veronii species, two were Aeromonas sp ATCC and one was Aeromonas hydrophila. Colisa lalia usually imported in Korea from the South and South-east Asian countries for recreational purposes. However, they are playing important role as a disease vector or carriers. The infected fish of this study frequently have hemorrhages at the base of the fins or on the skin, and gross ulcerative lesions. Internal signs include, fluid in the abdomen, swollen liver and spleen, and the intestine was distended and fluid-filled. In this study, the utility of 16S rDNA sequencing was employed to isolate Aeromonas bacteria from freshwater imported fish are important to environment, veterinary, and clinical purposes. Key word: Bacteria, Colisa lalia, 16S rDNA, Aeromonas   doi:10.3329/ujzru.v27i0.1948 Univ. j. zool. Rajshahi Univ. Vol. 27, 2008 pp. 21-24


Plant Disease ◽  
2009 ◽  
Vol 93 (5) ◽  
pp. 552-552 ◽  
Author(s):  
J. E. Munyaneza ◽  
V. G. Sengoda ◽  
J. M. Crosslin ◽  
G. De la Rosa-Lozano ◽  
A. Sanchez

Zebra Chip (ZC), an emerging disease of potato (Solanum tuberosum L.) first documented in potato fields around Saltillo in México in 1994, has been identified in the southwestern United States, México, and Central America and is causing losses of millions of dollars to the potato industry (4). Recently, this damaging potato disease was also documented in New Zealand (3). This disease is characterized by a striped pattern of necrosis in tubers produced on infected plants, and fried chips processed from these infected tubers are commercially unacceptable (4). Recent studies conducted in the United States and New Zealand have associated ZC with a new species of ‘Candidatus Liberibacter’ vectored by the potato psyllid, Bactericera cockerelli Sulc (1,3,4). A bacterium designated ‘Candidatus Liberibacter psyllaurous’ has recently been identified in potato plants with “psyllid yellows” symptoms that resemble those of ZC (2). To investigate whether liberibacter is associated with ZC in México, 11 potato (cv. Atlantic) tuber samples exhibiting strong ZC symptoms and six asymptomatic tubers were collected from a ZC-affected commercial potato field near Saltillo City, Coahuila, México in September 2008 and tested for this bacterium by PCR. Total DNA was extracted from symptomatic and asymptomatic tubers with cetyltrimethylammoniumbromide (CTAB) buffer (4). DNA samples were tested by PCR using primer pair OA2/OI2c (5′-GCGCTTATTTTTAATAGGAGCGGCA-3′ and 5′-GCCTCGCGACTTCGCAACCCAT-3′, respectively) specific for 16S rDNA and primer pair CL514F/R (5′-CTCTAAGATTTCGGTTGGTT-3′ and 5′-TATATCTATCGTTGCACCAG-3′, respectively) designed from ribosomal protein genes (3). Seven of eleven (63.7%) ZC-symptomatic tubers and one of six (16.7%) asymptomatic potatoes yielded the expected 1,168-bp 16S rDNA and 669-bp CL514F/R amplicons, indicating the presence of liberibacter. Amplicons generated from symptomatic tubers were cloned into pCR2.1-Topo plasmid vectors (Invitrogen, Carlsbad, CA) and one clone of each amplicon was sequenced in both directions (ACGT, Inc., Wheeling, IL). BLAST analysis of the ZC OA2/OI2c sequence (GenBank Accession No. FJ498806) showed 100% identity to liberibacter 16S rDNA sequences amplified from potato psyllids from Dalhart, TX and potato tubers from Garden City, KS (GenBank Accession Nos. EU921627 and EU921626, respectively). The ZC CL514F/R sequence (GenBank Accession No. FJ498807) was 98% identical to analogous rplJ and rplL liberibacter ribosomal protein gene sequences amplified from several solanaceous plants in New Zealand (GenBank Accession Nos. EU834131 and EU935005). The OA2/OI2c sequence was also identical to the 16S rDNA sequence (Genbank Accession No. EU812559) of ‘Ca. Liberibacter psyllaurous’ (2). To our knowledge, this is the first report of ‘Ca. Liberibacter psyllaurous’ associated with ZC-affected potatoes in México. References: (1) J. A. Abad et al. Plant Dis. 93:108, 2009. (2) A. K. Hansen et al. Appl. Environ. Microbiol. 74:5862, 2008. (3) L. W. Liefting et al. Plant Dis. 92:1474, 2008. (4) J. E. Munyaneza et al. J. Econ. Entomol. 100:656, 2007.


Plant Disease ◽  
2006 ◽  
Vol 90 (8) ◽  
pp. 1114-1114 ◽  
Author(s):  
J. E. Kaminski ◽  
T. Hsiang

Dead spot, also known as bentgrass dead spot or bermudagrass dead spot, is a relatively new disease of golf course putting greens and is caused by the pathogen Ophiosphaerella agrostis (1). The disease first was reported on a creeping bentgrass (Agrostis stolonifera) putting green in Maryland (2) and since has been identified on putting greens of creeping bentgrass and hybrid bermudagrass (Cynodon dactylon × C. transvaalensis) in the eastern and southern United States (3,4). In June 2004, disease symptoms resembling dead spot were observed on a golf course in southern Ontario. Small (≤3 cm) spots first appeared approximately 14 months after establishment of the sand-based, ‘L-93’ creeping bentgrass putting greens. The disease became more severe during the summer months and patches increased in size to as much as 5 to 8 cm in diameter. Dead spot infection centers remained visible throughout the winter months and the disease again became active during the spring of 2005. Bentgrass tissues growing adjacent to the periphery of active infection centers were orange-red to reddish-brown. Although dark brown ectotrophic hyphae were observed on bentgrass stolons, none were found on the roots. Few new infection centers occurred in 2005 and pseudothecia embedded within necrotic tissue only were observed in small numbers. No mature ascospores were observed when samples were collected during September 2005. A single fungal morphotype consistently was isolated from leaves and stolons with a rose-quartz color when grown for several days on potato dextrose agar. To demonstrate pathogenicity, ‘L-93’ creeping bentgrass seedlings were grown for 28 days in 10-cm-diameter pots containing an autoclaved greens-mix with a mechanical analysis of 94% sand, 5% silt, and 1% clay. Inoculum was prepared by placing mycelia from a hyphal-tipped isolate on an autoclaved mix of seed of tall fescue (Festuca arundinacea) and wheat (Triticum aestivum) bran (50% [vol/vol]), and grown at 24°C for 14 days. The inoculum (5 g) was embedded a few milliliters into the sand in the center of each pot (n = 5), and uninfested inoculum served as the untreated control. Pots were placed in enclosed plastic containers and incubated at room temperature (13 to 26°C) under natural light (replication 1) or under 14 h of light per day from fluorescent lights (replication 2). After 7 days, tissue along the periphery of each inoculation point became covered in a pink mycelium, and newly infected leaves appeared tan or brownish-red. Most plants were dead after 22 to 28 days of incubation. Reisolation of the pathogen from necrotic leaves produced fungal colonies similar in color, morphology, and growth rate to the original isolates. Few pseudothecia developed on infected tissue but were present in large numbers on infested tall fescue seed. Bitunicate asci containing spirally twisted filiform ascospores were observed. Light brown ascospores (n = 50) were 7 to 15 septate and measured 1.9 to 3.6 μm × 60.7 to 147.9 μm. On the basis of field symptoms, morphological characteristics, and pathogenicity tests, the pathogen was identified as O. agrostis. To our knowledge, this is the first report of dead spot on creeping bentgrass in Canada and of O. agrostis outside the United States. References: (1) M. P. S. Câmara et al. Mycologia 92:317, 2000. (2) P. H. Dernoeden et al. Plant Dis. 83:397, 1999. (3) J. E. Kaminski and P. H. Dernoeden. Plant Dis. 86:1253, 2002. (4) J. P. Krausz et al. Plant Dis. 85:1286, 2001.


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