First Report of a Field Outbreak of a Bacterial Leaf Spot of Cantaloupe and Squash Caused by Pseudomonas syringae pv. syringae in Georgia

In March 2000, a leaf spot was reported affecting yellow summer squash (Cucurbita pepo) and cantaloupe (Cucumis melo) in commercial fields in Colquitt, Echols, and Grady counties in Georgia. All of the crops affected were reported within a 10-day period, and average temperatures during that time were 8 to 22.5°C, which is very close to the 50-year normal temperatures for these areas located in southwest Georgia. Incidence in affected fields was 100%. Lesions on squash leaves appeared irregularly shaped, dark, water soaked, somewhat vein restricted, and were 5 to 10 mm in diameter. Lesions on cantaloupe were angular, light tan, and necrotic with a lesion diameter of 2 to 5 mm. A general chlorosis was observed around lesions of both crops. Leaf distortion was observed on squash. Four isolates collected were used in biochemical, pathogenicity, and physiological tests. Gram-negative, rod-shaped bacteria were isolated from diseased tissue from squash and cantaloupes. Bacteria were aerobic, catalase-positive, fluorescent on King's medium B (1), oxidase-negative, nonpectolytic on potato, arginine dihydrolase-negative, utilized sucrose as a carbon source, produced levan, and gave a hypersensitivity response on tobacco (HR). Analysis of fatty acid methyl ester (FAME) profiles using the Microbial identification system (Sherlock version 3.1, Microbial identification system, Newark, DE) characterized representative strains as Pseudomonas syringae (similarity indices 0.65 to 0.80). Upon further characterization, the strains were negative for l (+)-tartarate utilization but utilized l-lactate and betaine and also exhibited ice nucleation activity. These characteristics are consistent with those of Pseudomonas syringae pv. syringae. Squash and cantaloupes were grown in a greenhouse for 4 weeks. Bacteria were grown in nutrient broth, resuspended in sterile tap water, and standardized using a spectrophotometer. Plants were inoculated by infiltrating leaves with 1 ml of bacterial suspensions (1 × 107 CFU/ml) using sterile syringes. Sterile water was used as a negative control, and 1 ml was infiltrated into leaves of squash and cantaloupes. Water-soaked lesions developed in 4 to 6 days on squash and cantaloupes inoculated with bacterial suspensions, and Pseudomonas syringae pv. syringae was isolated from diseased tissue. No symptoms developed on squash and cantaloupes used as negative controls. This outbreak of Pseudomonas syringae pv. syringae did not cause significant economic damage to either crop as symptoms subsided once daily high temperatures reached 28 to 32°C. This disease has been isolated from several cucurbit transplants reared in greenhouses, but to our knowledge, this is the first report of this disease occurring in the field. Reference: (1) E. O. King et al. J. Lab. Clin. Med. 44:301, 1954.

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First Report of Pear Blossom Blast Caused by Pseudomonas syringae pv. syringae in China

Plant Disease ◽

10.1094/pdis-92-5-0832c ◽

2008 ◽

Vol 92 (5) ◽

pp. 832-832 ◽

Cited By ~ 2

Author(s):

L. H. Xu ◽

G. L. Xie ◽

B. Li ◽

B. Zhu ◽

F. S. Xu ◽

...

Keyword(s):

Pseudomonas Syringae ◽

Similarity Index ◽

Nutrient Agar ◽

Identification System ◽

Pyrus Pyrifolia ◽

Rrna Gene ◽

Bacterial Disease ◽

First Report ◽

Microbial Identification ◽

Microbial Identification System

In the spring of 2006, a new bacterial disease was noted in pear orchards near Hangzhou, Zhejiang Province, China. The disease caused severe blossom blast on pears (Pyrus pyrifolia; cv. Cuiguan). Early symptoms of the disease included blackening of the calyx end of developing fruit, blackening of blossom clusters while leaves of affected blossom clusters appeared normal, or death of clusters consisting of both blossoms and leaves. Later, tips of twigs turned dark brown and died. No bacterial ooze was observed. Twelve bacterial isolates were recovered from ten samples of buds and blossoms. Six isolates were selected for identification. They were similar to those of the reference strains of Pseudomonas syringae pv. syringae LMG5570 and LMG 2230 from Belgium in phenotypic tests on the basis of the Biolog Microbial Identification System (version 4.2; Biolog Inc., Hayward, CA), pathogenicity tests, gas chromatographic analysis of fatty acid methyl esters (FAMEs) using the Microbial Identification System (MIDI Inc., Newark, DE) with aerobic bacterial library (TABA50), and electron microscopy (TEM, KYKY-1000B, Japan). All isolates tested were gram-negative, aerobic rods measuring 1.5 to 2.4 × 0.5 to 0.6 μm with 2 to 4 polar flagella. Fluorescent green diffusible pigment was produced on King's Medium B. Colonies were gray-white and slightly raised with smooth margins on nutrient agar. They produced levan on sucrose nutrient agar. A hypersensitive reaction was observed on tobacco cv. Benshi 24 h after inoculation. All isolates were identified as P. syringae pv. syringae with Biolog similarity index of 0.57 to 0.86 and FAME similarity index of 0.58 to 0.81. Identification as P. syringae pv. syringae was confirmed using 16S rDNA universal primers (2,3): 5′-AGA GTT TGA TCA TGG CTC AG-3′ forward primer, 5′-ACG GTT ACC TTG TTA CGA CTT-3′ reverse primer. The PCR fragments of the three isolates were sequenced and compared with sequences in GenBank. They had 99% similiarity with P. syringae pv. syringae 16S rRNA gene strain NCPPB 3869. Koch's postulates were conducted on buds of the original pear cultivar growing in pots and detached pear blossoms in flasks by spray inoculation with cell suspensions containing 108 CFU/ml of the six isolates at 18 to 22°C with two replications. The bacteria induced symptoms on buds and blossoms similar to those observed in the field. The bacterium was reisolated from symptomatic pear buds and internal ovary tissues. P. syringae pv. syringae was first reported in England as the cause of pear blossom blast in 1914 (1). After searching all the Chinese agricultural databases and major journals (National Knowledge Infrastructure database, Vip Chinese periodical database, Chinese wanfang database, China InfoBank, Scientia Agricultura Sinica, Acta Phytopathologica Sinica, Acta Phytophylacica Sinica, and Journal of Fruit Science), to our knowledge, this is the first report of pear blossom blast caused by P. syringae pv. syringae in China. The disease cycle on pear trees and the control strategies in the regions are being further studied. References: (1) B. P. Barker et al. Ann. Appl. Biol. 1:85, 1914. (2) U. Edward et al. Nucleic Acids Res. 17:7843,1989. (3) B. Li et al. J. Phytopathol. 34:141, 2006.

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First Report of Pseudomonas cichorii Causing Leaf Spot of Vegetable Sponge Gourd in China

Plant Disease ◽

10.1094/pdis-03-13-0312-pdn ◽

2014 ◽

Vol 98 (1) ◽

pp. 153-153 ◽

Cited By ~ 3

Author(s):

B. J. Li ◽

H. L. Li ◽

Y. X. Shi ◽

X. W. Xie

Keyword(s):

Leaf Spot ◽

Pathogenic Bacteria ◽

Identification System ◽

Pathogenicity Test ◽

Distilled Water ◽

Pseudomonas Cichorii ◽

First Report ◽

Microbial Identification ◽

Pectolytic Activity ◽

Sponge Gourd

A suspect bacterial leaf spot on vegetable sponge gourd (Luffa cylindrical (L.) Roem.) was found in a commercial greenhouse in Pi County, Chengdu City, Sichuan Province, China, in February 2011. Approximately 20 to 30% of plants were affected, causing serious economic loss. Symptoms occurred only on seedlings and consisted of water-soaked, irregularly shaped, black lesions on the surface and margins of cotyledons. A bacterium was consistently isolated on nutrient agar from diseased leaf tissues that had been surface disinfected in 70% ethyl alcohol for 30 s. The bacterium produced small gray colonies with smooth margins, was gram negative, fluoresced on King's B medium, and showed pectolytic activity when inoculated on potato slices. The partial sequences of 16SrRNA gene (1,377 bp) of the bacterium (GenBank Accession No. KC762217), amplified by using universal PCR primers 16SF (5′-AGAGTTTGATCCTGGCTCAG-3′) and 16SR (5′-GGTTACCTTGTTACGACTT-3′), shared 100% similarity with that of Pseudomonas cichorii (GenBank Accession No. HM190228). The vegetable sponge gourd isolate was also identified by using the Biolog Microbial Identification System (version 4.2, Biolog Inc., Hayward, CA) as P. cichorii with the following characteristics (1): negative for arginine dihydrolase, gelatin liquefaction, and N2 production. Positive reactions were obtained in tests for catalase, oxidase, potato rot, utilization of melibiose, and mannitol. Tests were negative for utilization of sucrose, trehalose, D-arabinose, raffinose, cellobiose, and rhamnose. A pathogenicity test was conducted on 4-week-old vegetable sponge gourd plants by spray-inoculation with 108 CFU/ml sterile distilled water on the leaves of 15 vegetable sponge gourd plants and by needle puncture on the stems of 15 other plants with P. cichorii, respectively. Control plants were misted with sterile distilled water or punctured on the stem with a clean needle. Plants were placed in a greenhouse maintained at 28 ± 2°C with relative humidity of 80 to 85%. Symptoms, the same as seen on the original diseased plants, developed after 7 to 10 days on inoculated plants. Control plants remained healthy. The bacterium was readily re-isolated from inoculated plants and identified as P. cichorii using P. cichorii-specific primer hrpla/hrp2a (1). To our knowledge, this is the first report of P. cichorii causing disease on commercially grown vegetable sponge gourd in China. This new finding will provide the basis for developing resources for diagnostics and management, including screening varieties for resistance. References: (1) S. Mazurier et al. J. FEMS Microbiol. Ecol. 49:455, 2004. (2) N. W. Schaad et al., eds. Laboratory Guide for Identification of Plant Pathogenic Bacteria, 3rd ed. APS Press, St. Paul, MN, 2001.

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First Report of Leaf Blight of Onion Caused by Xanthomonas campestris in the Continental United States

Plant Disease ◽

10.1094/pdis.2000.84.2.201c ◽

2000 ◽

Vol 84 (2) ◽

pp. 201-201 ◽

Cited By ~ 13

Author(s):

T. Isakeit ◽

M. E. Miller ◽

L. W. Barnes ◽

E. R. Dickstein ◽

J. B. Jones

Keyword(s):

United States ◽

Xanthomonas Campestris ◽

Acid Methyl Ester ◽

Leaf Blight ◽

Nutrient Agar ◽

Identification System ◽

Sterile Water ◽

First Report ◽

Microbial Identification ◽

Similarity Indices

In March 1998, a leaf blight of onion (Allium cepa L. ‘1015’) was found on many plants in a plot on the Texas A&M Agricultural Experiment Station in Weslaco. The symptoms were longitudinal chlorotic areas on one side of the leaf, containing sunken, elliptical necrotic lesions. Affected leaves ultimately died. Chlorotic lesions were swabbed with 70% ethanol, and tissue from beneath the epidermis was placed in a drop sterile water for 20 min. Drops were streaked on nutrient agar and incubated at 30°C. Isolations yielded gram-negative, rod-shaped bacteria that formed dark yellow, gummy colonies on yeast dextrose carbonate agar medium, hydrolyzed starch, and had a single, polar flagellum. Analysis of fatty acid methyl ester (FAME) profiles, using the Microbial Identification System (MIS, version 4.15; Microbial Identification, Newark, DE), done at the Texas Plant Disease Diagnostic Laboratory, College Station, identified nine isolates as Xanthomonas campestris (similarity indices of 0.31 to 0.54). Tests at the University of Florida supported this identification: FAME profiles using MIS version 3.9 gave similarity indices of 0.89 to 0.95, and profiles using Biolog GN Microplates, MicroLog database release 3.50 (Biolog, Hayward, CA), gave similarity indices of 0.03 to 0.76. Leaves (15 to 20 cm long) of potted onions (cv. 1015 at the five- to six-leaf stage) were infiltrated with a suspension of bacteria (107 CFU per ml), using a needle and syringe. Plants were maintained in mist chamber in a greenhouse at 24°C. Water-soaking and development of pale green color of the inoculated leaf occurred after 2 days, followed by death after 4 days. There were no symptoms on leaves inoculated with sterile water. Pathogenicity tests on four isolates were repeated once. Bacteria were reisolated on nutrient agar from symptomatic tissue but not from controls. In the field plot, disease severity did not increase as season progressed nor were there any symptoms on bulbs. Symptoms were not observed on onion during the 1999 season. X. campestris was first reported on onion from Hawaii (1). This is the first report of this pathogen on onion in the continental United States. Reference: (1) A. M. Alvarez et al. Phytopathology 68:1132, 1978.

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First Report of Bacterial Head Rot of Broccoli Caused by Pseudomonas fluorescens in China

Plant Disease ◽

10.1094/pdis-93-11-1219a ◽

2009 ◽

Vol 93 (11) ◽

pp. 1219-1219 ◽

Cited By ~ 2

Author(s):

B. Li ◽

G. L. Wang ◽

Z. Y. Wu ◽

W. Qiu ◽

Q. M. Tang ◽

...

Keyword(s):

Fatty Acid ◽

Pseudomonas Fluorescens ◽

Pathogenic Bacteria ◽

Disease Incidence ◽

Identification System ◽

Rrna Gene ◽

First Report ◽

Microbial Identification ◽

Microbial Identification System ◽

Head Rot

During warm and humid periods in the winters from 2005 to 2008, head rot symptoms on broccoli (cv. Sijilv) (Brassica oleracea L. var italica Planch) were observed in commercial fields in Ningbo, Zhejiang Province, China. In agreement with the report of Cui and Harling (1), water-soaked lesions developed on the buds and then progressed into a brown-black soft rot. Longitudinal sections of the symptomatic inflorescences showed brown discoloration and rotting of the internal tissues. Broccoli production is hampered by the disease, with disease incidence ranging from 65 to 81%. Bacteria were isolated by streaking on nutrient agar (3) and individual colonies formed after 2 to 3 days of incubation at 28°C. Fifteen of thirty isolates induced hypersensitive reactions (HR) on tobacco leaves (Nicotiana tabacum cv. Samsun) within 48 h. All the HR-positive strains were fluorescent on King's medium B and the colonies were smooth, convex, entire, and round. Classical bacteriological tests indicated that the fluorescent strains were gram negative, obligate aerobes, arginine dihydrolase positive, and oxidase positive. Also, the fluorescent strains were positive for the production of levan from sucrose. Five representative strains were further characterized by the Biolog Microbial Identification System, version 4.2 (Biolog Inc., Hayward, CA) and gas chromatography of fatty acid methyl esters (FAME) using the Microbial Identification System (MIDI Inc., Newark, DE) with the aerobic bacterial library (TSBA50). The five strains were identified as Pseudomonas fluorescens with Biolog and FAME similarity indexes of 0.61 to 0.68 and 0.52 to 0.58, respectively. The 16S rRNA gene sequence of broccoli strain PFB-01 (GenBank Accession No. GQ352649) was determined according to Li et al. (2). A subsequent GenBank search showed that this sequence had 98% nucleotide identity with the type strain of P. fluorescens (ATCC 17386T, GenBank Accession No. AF094726). Koch's postulates were completed by the inoculation of broccoli heads (cv. Sijilv) with cell suspensions (107 CFU/ml) of the above five strains by spraying on the surface of subcorymbs. Each treatment had five replicates. All strains induced head rot symptoms similar to those observed in natural infections. No symptoms were noted on the control plants inoculated with sterile water. Bacteria were successfully reisolated from symptomatic heads and confirmed by the cellular fatty acid composition. To our knowledge, this is the first report in China that P. fluorescens is the causal pathogen of bacterial head rot of broccoli. References: (1) X. Cui and R. Harling. Phytopathology 96:408, 2006. (2) B. Li et al. J. Phytopathol. 154:711, 2006. (3) N. W. Schaad et al. Laboratory Guide for Identification of Plant Pathogenic Bacteria. 3rd ed. The American Phytopathological Society. St. Paul, MN, 2001.

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First Report of Fire Blight Disease Caused by Erwinia amylovora on Rockspray (Cotoneaster horizontalis) in Turkey

Plant Disease ◽

10.1094/pdis-01-12-0002-pdn ◽

2012 ◽

Vol 96 (11) ◽

pp. 1690-1690

Author(s):

K. K. Bastas ◽

F. Sahin

Keyword(s):

Erwinia Amylovora ◽

Disease Incidence ◽

Acid Methyl Ester ◽

Late Summer ◽

Control Measures ◽

Bacterial Suspension ◽

Identification System ◽

Bacterial Strains ◽

First Report ◽

Microbial Identification

In the late summer and early winter of 2008 and 2009, leaf and shoot blight and cankers with reddish and brownish necrotic tissue on mature branches of Cotoneaster horizontalis were investigated in landscape areas of Konya province in Turkey. Disease incidence was estimated at 2%. Bacteria were consistently isolated from the lesions on leaves and shoots on nutrient sucrose agar medium. Twelve representative bacterial strains were isolated and characterized as gram-negative, rod-shaped, mucoid, fermentative, yellow-orange on MS medium, positive for levan formation and acetoin production, no growth at 36°C, positive for gelatin hydrolysis, and negative for indole, urease, oxidase, arginine dehydrolase, reduction of nitrate, and acid production from lactose and inositol (2). Two reference strains of Erwinia amylovora (EaP28 and NCPPB 2791) obtained from the culture collection unit of Selcuk University were used as positive controls. All strains induced a hypersensitive response in tobacco (Nicotiana tobaccum cv. White Burley). All strains were identified as E. amylovora on the basis of amplification of a 1 kb DNA fragment with a species-specific primer set, A/B (1) by PCR, and fatty acid methyl ester profiles determined by Sherlock Microbial Identification System software (TSBA 6 v. 6.00; Microbial ID, Newark, DE) with similarity indices ranging from of 83 to 96%. Pathogenicity tests were performed by injecting 20 μl of a bacterial suspension (108 CFU ml–1) into the shoot tips of 3-year-old C. horizontalis seedlings. Leaf and shoot blighting symptoms were observed within 10 to 15 days, but no symptoms were observed on control plants treated with sterile water. The bacterium was reisolated from the lesions on leaves and shoots and identified as described above. To our knowledge, this is the first report of E. amylovora on cotoneaster in Turkey. Control measures are needed to prevent any further spread of the bacterium to new landscape areas. References: (1) S. Bereswill et al. Appl. Environ. Microbiol. 58:3522, 1992. (2) A. L. Jones and K. Geider. Page 40 in: Laboratory Guide for Identification of Plant Pathological Bacteria, 2001.

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First Report of Fire Blight Disease on Blackberry in Turkey

Plant Disease ◽

10.1094/pdis-12-11-1073-pdn ◽

2012 ◽

Vol 96 (12) ◽

pp. 1818-1818

Author(s):

K. K. Bastas ◽

F. Sahin

Keyword(s):

Disease Incidence ◽

Acid Methyl Ester ◽

Negative Control ◽

Culture Collection ◽

Bacterial Suspension ◽

Identification System ◽

Distilled Water ◽

Bacterial Strains ◽

First Report ◽

Microbial Identification

During 2008 and 2009, a new disease on blackberry (Rubus fruticosus cv. Chester) causing leaf and shoot blight and cankers with brown discoloration of necrotic tissues on mature branches was observed in Isparta and Konya provinces of Turkey. Disease incidence was estimated to be 4% for the two years. Isolations were made from lesions on leaves and shoots on nutrient sucrose agar (NSA) medium. Bacteria consistently isolated from the diseased tissues were identified on the basis of biochemical, physiological (2), and molecular tests (1). Eleven representative bacterial strains were gram-negative, rod-shaped, mucoid, fermentative, yellow-orange on Miller and Scroth (MS) medium, positive for levan formation and acetoin production, no growth at 36°C, positive for gelatin hydrolysis, and negative for esculin hydrolysis, indole, urease, catalase, oxidase, arginine dehydrolase, reduction of nitrate, acid production from lactose, and inositol. Two reference strains of Erwinia amylovora (EaP28 and NCPPB 2791) obtained from the culture collection unit of Selcuk University were used as positive controls. All strains induced a hypersensitive response in tobacco (Nicotiana tobaccum cv White Burley) 24 h after inoculation with a 108 CFU/ml bacterial suspension in water. All strains were identified as E. amylovora using the species-specific primers set A/B (1), which amplified a 1-kb DNA fragment in PCR, and fatty acid methyl ester (FAME) profiles determined by Sherlock Microbial Identification System software (TSBA 6 v. 6.00; Microbial ID, Newark, DE) with similarity indices ranging from of 79 to 99%. Pathogenicity was confirmed by injecting bacterial suspensions (108 CFU/ml–1) in sterile distilled water into the shoot tips of 2-year-old R. fruticosus cv. Chester and the first blighting symptoms were observed on leaves within 3 days and also 10 days later after inoculation on shoots. Sterile distilled water was used as a negative control. No symptoms were observed on control plants. All tests were repeated three times. The bacterium was reisolated from inoculated plants and identified as. E. amylovora. To our knowledge, this is the first report of E. amylovora on blackberry in Turkey. Phytosanitary measures are needed to prevent any further spread of the bacterium to new blackberry areas. References: (1) S. Bereswill et al. App. Environ. Microbiol. 58:3522, 1992. (2) A. L. Jones and K. Geider. Lab. Guide for Identification of Plant Pathological Bacteria, 40, 2001.

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First Report of Bacterial Leaf Spot Caused by Pseudomonas cichorii on Schefflera arboricola in Turkey

Plant Disease ◽

10.1094/pdis-93-8-0848b ◽

2009 ◽

Vol 93 (8) ◽

pp. 848-848

Author(s):

Y. Aysan ◽

M. Mirik ◽

F. Sahin

Keyword(s):

Leaf Spot ◽

Disease Incidence ◽

Acid Methyl Ester ◽

Identification System ◽

Original Strain ◽

Bacterial Leaf Spot ◽

Pseudomonas Cichorii ◽

First Report ◽

Leaf Spots ◽

Schefflera Arboricola

In late winter and spring of 2006 and 2008, leaf spots with yellow halos were observed on dwarf schefflera (Schefflera arboricola cvs. Gold Capella, Trinette, and Green Gold) that were grown as potted plants in two commercial ornamental greenhouses in Adana and Mersin, Turkey. Average disease incidence was assessed as 10% during the term of the study. Isolations were made from leaf spots symptoms on King's medium B. Bacteria consistently isolated from diseased tissues formed green fluorescent colonies on the medium. Ten representative bacterial strains were examined and found to be gram negative, rod shaped, and aerobic, levan, pectolytic, and arginine dihydrolase negative, and oxidase positive. They all induced a hypersensitive response in tobacco (Nicotiana tabacum cv. Samsun). All strains were identified as Pseudomonas cichorii with similarity indices of 79 to 99% based on fatty acid methyl ester (FAME) profiles determined by Sherlock Microbial Identification System software (TSBA 6 v. 6.00; Microbial ID, Newark, DE). Pathogenicity of the strains was confirmed on five dwarf schefflera plants by leaf tissue infiltration with bacterial suspensions (107 CFU ml–1) in sterile distilled water. P. cichorii NCPPB 3802 and sterile water were used as positive and negative controls, respectively. The same symptoms as those observed in the commercial greenhouses were observed on dwarf schefflera leaves within 12 to 15 days after inoculation. The bacteria were reisolated from the inoculated plants and identified as the same as the original strain by conventional tests and FAME analysis. Negative control plants remained disease free. Occurrence of bacterial leaf spot caused by P. cichorii on vegetable crops in Turkey (1,3) and dwarf schefflera in other countries (2) has been reported previously, but to our knowledge, this is the first report of the observation of P. cichorii on dwarf schefflera in Turkey. References: (1) Y. Aysan et al. Plant Pathol. 52:782. 2003. (2) A. R. Chase and D. D. Brunk. Plant Dis. 68:73, 1984. (3) F. Sahin et al. Acta Hortic. 695:93, 2005.

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First Report of Bacterial Leaf Spot on Leafy Brassica Greens Caused by Pseudomonas syringae pv. maculicola in South Carolina

Plant Disease ◽

10.1094/pd-90-0683c ◽

2006 ◽

Vol 90 (5) ◽

pp. 683-683 ◽

Cited By ~ 8

Author(s):

A. P. Keinath ◽

W. P. Wechter ◽

J. P. Smith

Keyword(s):

South Carolina ◽

Pseudomonas Syringae ◽

Leaf Spot ◽

Acid Methyl Ester ◽

The United States ◽

Dna Fingerprint ◽

Bacterial Leaf Spot ◽

First Report ◽

Turnip Greens ◽

Salad Greens

As of 2001, South Carolina ranked second in the United States in acreage of turnip greens (Brassica rapa) and collard (B. oleracea) and third in acreage of mustard (B. juncea). In June 2001, a leaf disease was found on turnip greens (cv. Alamo), mustard (cvs. Southern Giant Curled and Florida Broadleaf), and rape salad greens (B. napus var. napus cv. Essex) on a commercial farm in Lexington County, South Carolina. Symptoms appeared after a heavy rainstorm that included blowing sand. The disease was found in May and June 2002 on three additional farms in the same county on turnip greens cv. Topper and Royal Crown and collard cv. Top Bunch. Symptoms included small tan spots, water soaking, yellowing, and brown necrosis of leaves after spots coalesced on the lower halves of plants. Yellowing was more prevalent on older than on younger leaves. Leaf samples were collected in 2001 and 2002 from the affected hosts on the four farms. Bacterial streaming was evident from these samples and 27 strains were isolated on nutrient agar or King's medium B (KMB). All strains were gram negative and fluoresced bluegreen or yellow under UV light after 48-h growth at 28°C on Pseudomonas agar F (PAF). On the basis of LOPAT tests, the strains were identified as P. syringae (2). All 27 strains were tested for pathogenicity to rape salad greens cv. Essex and then to turnip greens cv. Topper. Plants were grown in peat-vermiculite potting mix in 10-cm-diameter pots in a greenhouse. P. syringae pv. maculicola F41, isolated from turnip in Oklahoma, and P. syringae pv. tomato F33, isolated from tomato in Oklahoma, were included as positive and negative controls along with a noninoculated control. Bacteria were grown on KMB for 48 h at 24°C, and bacterial suspensions were prepared and adjusted to 0.1 optical density at 600 nm. Three-week-old plants were held at 95 to 100% relative humidity (RH) for 48 h before they were sprayed just to runoff with inoculum and then held at 95 to 100% RH for 48 h after inoculation (4). After an additional 5 to 8 days in a greenhouse, nine strains and F41 caused symptoms on both Topper and Essex similar to symptoms observed in the field. No symptoms were observed on noninoculated plants or plants inoculated with F33. On the basis of repetitive sequence-based polymerase chain reactions with the BOXA1R primer, the DNA fingerprint of each of the nine pathogenic strains from South Carolina was nearly identical to that of F41. Bacteria isolated from inoculated, symptomatic turnip leaves had identical LOPAT and BOXA1R profiles to the corresponding original strains. Pathogenic strains had bluegreen fluorescence on PAF, whereas nonpathogenic strains fluoresced yellow. Five pathogenic strains, as well as F41, were further identified to species and pathovar with fatty acid methyl ester profiles as P. syringae pv. maculicola. To our knowledge, this is the first report of P. syringae pv. maculicola from South Carolina. Over the past 10 years, P. syringae pv. maculicola has been found in Oklahoma (4), California (1), and Ohio (3). Bacterial leaf spot has occurred yearly in South Carolina since the initial outbreaks. Currently, it is the disease that causes the greatest yield losses of leafy brassica greens in the state. References: (1) N. A. Cintas et al. Plant Dis. 85:1207, 2001. (2) R. A. Lelliott et al. J. Appl. Bacteriol. 29:470, 1966. (3) M. L. Lewis Ivey et al. Plant Dis. 86:186, 2002. (4) Y. F. Zhao et al. Plant Dis. 84:1015, 2000.

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