scholarly journals A New Bacterial Leaf Spot Disease of Broccolini, Caused by Pseudomonas syringae pathovar maculicola, in California

Plant Disease ◽  
2001 ◽  
Vol 85 (11) ◽  
pp. 1207-1207 ◽  
Author(s):  
N. A. Cintas ◽  
C. T. Bull ◽  
S. T. Koike ◽  
H. Bouzar
Keyword(s):  
Pseudomonas Syringae ◽  
Leaf Spot ◽  
Similarity Index ◽  
Acid Methyl Ester ◽  
Hypersensitive Reaction ◽  
Nutrient Broth ◽  
Leaf Spot Disease ◽  
Fluorescent Pseudomonad ◽  
Bacterial Leaf Spot ◽  
Initial Symptoms

In 1998, a new disease was detected on 3-week-old commercial broccolini (Brassica oleracea L. var. botrytis × B. alboglabra) transplants in a Salinas Valley, Monterey County, CA greenhouse. Initial symptoms were small (2 to 4 mm diameter) circular to angular, water-soaked spots. As the disease progressed, spots remained relatively small, but turned tan to brown. When diseased tissues were macerated and streaked on King's medium B, a blue-green fluorescent pseudomonad was consistently isolated. Strains were levan positive, oxidase negative, and arginine dihydrolase negative. Strains did not rot potato slices, but induced a hypersensitive reaction on tobacco (Nicotiana tabacum L. ‘Turk’). Fatty acid methyl ester analysis (MIS-TSBA, version 4.10, MIDI Inc., Newark, DE) indicated that strains had a high similarity index (0.82 or higher) to Pseudomonas syringae, and GN (version 3.50, Biolog, Inc., Hayward, CA) profiles also identified strains as P. syringae. The bacterium associated with the disease, therefore, was identified as P. syringae van Hall. Pathogenicity was demonstrated by growing inoculum in nutrient broth shake cultures for 48 h, misting the broth cultures (1×106 CFU/ml) onto broccolini (cv. Aspabrock), and subjecting the plants to 48 h of high humidity. Control plants were misted with sterile nutrient broth. After 4 to 5 days in a greenhouse, leaf spot symptoms developed on all inoculated broccolini plants, and reisolated strains were characterized and found to be P. syringae. Control plants remained symptomless. The results of two sets of pathogenicity tests were the same. Repetitive sequence-based polymerase chain reaction using the BOXA1R primer resulted in identical banding patterns for the broccolini pathogen and for known isolates of P. syringae pv. maculicola from crucifers. In host range testing, P. syringae pv. maculicolawas pathogenic to broccolini plants. The broccolini isolates and P. syringae pv. maculicola isolates had the same pathogenicity results when crucifers and tomatoes were tested as hosts; broccoli and cauliflower (B. oleracea var. botrytis) were infected, and tomato results were variable. These tests suggest that the broccolini pathogen is the bacterial leaf spot pathogen, Pseudomonas syringae pv. maculicola, that occurs on broccoli and cauliflower transplants (1). To our knowledge, this is the first report of this pathogen causing a disease on commercially grown broccolini. Reference: (1) S. T. Koike et al. Plant Dis. 82:727, 1998.

scholarly journals Bacterial Blight on Arugula, a New Disease Caused by Pseudomonas syringae pv. Alisalensis in California

Plant Disease ◽  
2004 ◽  
Vol 88 (12) ◽  
pp. 1384-1384 ◽  
Author(s):  
C. T. Bull ◽  
P. Goldman ◽  
S. T. Koike
Keyword(s):  
Pseudomonas Syringae ◽  
Bacterial Blight ◽  
Acid Methyl Ester ◽  
Hypersensitive Reaction ◽  
Nutrient Broth ◽  
Leaf Spot Disease ◽  
Diagnostic Feature ◽  
Fluorescent Pseudomonad ◽  
Fresh Market ◽  
Banding Patterns

Beginning in 1995, a leaf spot disease has occasionally developed on the leafy crucifer arugula (Eruca vesicaria subsp. sativa) that is grown in coastal California as a fresh market commodity used mostly in bagged salad mixes. Initially, symptoms consist of small (<2 mm in diameter), angular, water-soaked spots that are visible from both sides of the leaf. The spots later enlarge, remain angular in shape, and turn brown to tan. A purple margin sometimes occurs around the spots. An important diagnostic feature is that this disease closely resembles downy mildew infections that have not produced sporangia (3). A blue-green fluorescent pseudomonad was consistently isolated from both types of lesions on King's medium B. Strains were levan positive, oxidase negative, and arginine dihydrolase negative. Strains did not rot potato slices but induced a hypersensitive reaction on tobacco (Nicotiana tabacum L. cv. Turk). These data indicated that the bacteria belonged to Lelliot's LOPAT group 1 (4). This was confirmed with data from fatty acid methyl ester analysis (MIS-TSBA version 4.10; MIDI, Inc., Newark, DE), which indicated that the strains were highly similar (similarity > = 0.758) to Pseudomonas syringae. Amplification of repetitive bacterial sequence-based polymerase chain reaction (rep-PCR) was used to determine the relationship between the P. syringae strains isolated from arugula and two common crucifer pathogens, P. syringae pv. maculicola and P. syringae pv. alisalensis (1). Using the BOXA1R primer, banding patterns for the arugula strains and the P. syringae pv. alisalensis pathotype were similar, differing by only one band. In contrast, the banding patterns of the arugula strains differed significantly from those of P. syringae pv. maculicola. Additionally, the arugula isolates were sensitive to a bacteriophage originally isolated for its ability to lyse P. syringae pv. alisalensis (1). Previously, the pathogen from arugula was reported to be P. syringae pv. maculicola (2). It is the intent of this disease note to clarify this identification. We completed Koch's postulates by confirming pathogenicity on arugula (cv. Rocket Salad). The strains were grown as nutrient broth shake cultures for 48 h at 24°C, adjusted to 108 CFU/ml, and misted onto 2- to 3-week old plants. Control plants were misted with sterile nutrient broth. After 4 to 5 days in a greenhouse (24 to 26°C), large, angular leaf lesions developed on all inoculated arugula plants. Strains were reisolated from symptomatic tissue and identified as P. syringae pv. alisalensis. Control plants remained symptomless. Similar methods confirmed that the host range of the arugula isolates were identical to that of P. syringae pv. alisalensis. The arugula and P. syringae pv. alisalensis isolates caused disease on broccoli (Brassica oleracea var. botrytis cvs. Patriot and Titleist), broccoli raab (B. rapa subsp. rapa cv. Sorento), and oats (Avena sativa cv. Montezuma), while P. syringae pv. maculicola caused disease on broccoli only. Pathogenicity tests were conducted two times with identical results. This confirms that the bacterial blight that has been occurring on commercial plantings of arugula is caused by P. syringae pv. alisalensis. References: (1) N. A. Cintas et al.Plant Dis. 86:992, 2002. (2) S. T. Koike et al. Plant Dis. 80:464, 1996. (3) S. T. Koike. Plant Dis. 82:1063, 1998. (4) R. A. Lelliott, J. Appl. Bacteriol. 29:470, 1966.

scholarly journals Report of Bacterial Leaf Spot on Collards and Turnip Leaves in Ohio

Plant Disease ◽  
2002 ◽  
Vol 86 (2) ◽  
pp. 186-186 ◽  
Author(s):  
M. L. Lewis Ivey ◽  
S. Wright ◽  
S. A. Miller
Keyword(s):  
Pseudomonas Syringae ◽  
Leaf Spot ◽  
Xanthomonas Campestris ◽  
Similarity Index ◽  
Acid Methyl Ester ◽  
Bacterial Suspension ◽  
Water Control ◽  
Bacterial Leaf Spot ◽  
Turnip Leaves ◽  
Negative Controls

In 2000, circular water-soaked lesions typical of bacterial leaf spot were observed on leaves of collards (Brassica oleracea L. var. viridis) throughout commercial fields in northwest Ohio. Light brown, rectangular, water-soaked lesions were observed on turnip leaves (Brassica rapa L.). Bacterial streaming from lesions on both crops was observed microscopically. Cream colored, fluorescent colonies were isolated from diseased tissues on Pseudomonas F medium, and eight representative colonies (four from collards and four from turnip) were selected and purified. Fatty acid methyl ester analysis was performed on all of the isolates. Two from collards and two from turnip were identified as Pseudomonas syringae pv. maculicola (mean similarity index = 0.82 [MIDI Inc., Newark, DE]). DNA extracts from pure cultures of the P. syringae pv. maculicola strains were used as template in a polymerase chain reaction (PCR) assay with primers derived from the region of the coronatine gene cluster controlling synthesis of the coronafacic acid moiety found in P. syringae pv. tomato and P. syringae pv. maculicola (CorR and CorF2) (D. Cuppels, personal communication). DNA from P. syringae pv. tomato strain DC3000 and P. syringae pv. maculicola strain 88–10 (2) served as positive controls, while water and DNA from Xanthomonas campestris pv. vesicatoria strain Xcv 767 were used as negative controls. The expected 0.65-kb PCR product was amplified from three of four strains (two from turnip and one from collards) and the positive control DNA, but not from the negative controls. Pathogenicity tests were performed twice on 6-week-old turnip (‘Forage Star’, ‘Turnip Topper’, ‘Turnip Alamo’, ‘Turnip 7’), collard (‘Champion’) and mustard (Brassica juncea L. ‘Southern Giant Curl’) seedlings using the three PCR-positive strains. Premisted seedlings were spray-inoculated separately with each of the three strains (2 × 108 CFU/ml, 5 ml per plant) and a water control. Greenhouse temperatures were maintained at 20 ± 1°C. For both tests, all strains caused characteristic lesions on all of the crucifer cultivars within 5 days after inoculation; the control plants did not develop symptoms. To satisfy Koch's postulates, one of the turnip strains was reisolated from ‘Turnip Topper’ plants, and the collard strain was reisolated from ‘Champion’ plants. The three original and two reisolated strains induced a hypersensitive response in Mirabilis jalapa L. and Nicotiana tabacum L. var. xanthia plants 24 h after inoculation with a bacterial suspension (1 × 108 CFU/ml). The original and reisolated strains were compared using rep-PCR with the primer BOXA1R (1). The DNA fingerprints of the reisolated strains were identical to those of the original strains. To our knowledge, this is the first report of bacterial leaf spot on commercially grown collards and turnip greens in Ohio. References: (1) B. Martin et al. Nucleic Acids Res. 20:3479, 1992. (2) R. A. Moore et al. Can. J. Microbiol. 35:910, 1989.

scholarly journals First Report of Bacterial Leaf Spot of Spinach Caused by a Pseudomonas syringae Pathovar in California

Plant Disease ◽  
2002 ◽  
Vol 86 (8) ◽  
pp. 921-921 ◽  
Author(s):  
S. T. Koike ◽  
H. R. Azad ◽  
D. C. Cooksey
Keyword(s):  
Brassica Oleracea ◽  
Beta Vulgaris ◽  
Pseudomonas Syringae ◽  
Leaf Spot ◽  
The United States ◽  
Nutrient Broth ◽  
Bacterial Leaf Spot ◽  
First Report ◽  
Leaf Spots ◽  
Initial Symptoms

In 2000 and 2001, a new disease was observed on commercial spinach (Spinacia oleracea) in the Salinas Valley, Monterey County, CA. Initial symptoms were water-soaked, irregularly shaped leaf spots (2 to 3 mm diameter). As the disease developed, spots enlarged to as much as 1 to 2 cm, were vein-delimited, and turned dark brown. Faint chlorotic halos sometimes surrounded the spots. Death of large areas of the leaf occurred if spots coalesced. Spots were visible from the adaxial and abaxial sides of leaves, and no fungal structures were observed. The disease occurred on newly expanded and mature foliage. No fungi were isolated from the spots. However, cream-colored bacterial colonies were consistently isolated on sucrose peptone agar, and these strains were nonfluorescent on King's medium B. Strains were positive for levan and negative for oxidase, arginine dihydrolase, and nitrate reductase. Strains did not grow at 36°C, did not rot potato slices, but induced a hypersensitive reaction in tobacco (Nicotiana tabacum cv. Turk). These results suggested the bacterium was similar to Pseudomonas syringae. Fatty acid methyl ester (FAME) analysis (MIS-TSBA 4.10, MIDI Inc., Newark, DE) indicated the strains were highly similar (80.1 to 89.3%) to P. syringae pv. maculicola. However, in contrast to P. syringae pv. maculicola, the spinach strains did not utilize the carbon sources erythritol, L+tartrate, L lactate, and DL-homoserine. Pathogenicity of 10 strains was tested by growing inoculum in nutrient broth shake cultures for 48 h, diluting to 106 CFU/ml, and spraying 4-week-old plants of spinach cv. Bossanova. Control plants were sprayed with sterile nutrient broth. After 5 to 8 days in a greenhouse (24 to 26°C), leaf spots identical to those observed in the field developed on cotyledons and true leaves of inoculated plants. Strains were reisolated from the spots and identified as P. syringae. Control plants remained symptomless. The 10 strains were also inoculated on beet (Beta vulgaris), Swiss chard (Beta vulgaris subsp. cicla), cilantro (Coriandrum sativum), and spinach. Spinach showed leaf spots after 8 days; however, none of the other plants developed symptoms. Two strains were inoculated onto spinach cvs. Califlay, Lion, Nordic IV, Polka, Resistoflay, Rushmore, RZ 11, Spinnaker, Springfield, Viroflay, and Whitney. Leaf spot developed on all cultivars, and the pathogen was reisolated. Because the FAME data indicated a similarity between the spinach pathogen and P. syringae pv. maculicola, we inoculated sets of spinach cv. Bolero, cabbage (Brassica oleracea subsp. capitata cv. Grenedere), and cauliflower (Brassica oleracea subsp. botrytis cv. White Rock) with three P. syringae pv. maculicola and three spinach strains. Cabbage and cauliflower developed leaf spots only when inoculated with P. syringae pv. maculicola; spinach had leaf spots only when inoculated with the spinach strains. All inoculation experiments were done twice, and the results of the two tests were the same. To our knowledge, this is the first report of bacterial leaf spot of spinach in California caused by a nonfluorescent P. syringae, and the first record of this disease in the United States. Biochemical characteristics and limited host range of the pathogen indicate the California strains are likely the same as the P. syringae pv. spinaciae pathogen that was reported in Italy (1) and Japan (2). References: (1) C. Bazzi et al. Phytopathol. Mediterr. 27:103, 1988. (2) K. Ozaki et al. Ann. Phytopathol. Soc. Jpn. 64:264, 1998.

scholarly journals Bacterial Blight, a New Disease of Broccoli Caused by Pseudomonas syringae in California

Plant Disease ◽  
2000 ◽  
Vol 84 (3) ◽  
pp. 370-370 ◽  
Author(s):  
S. T. Koike ◽  
N. A. Cintas ◽  
C. T. Bull
Keyword(s):  
Pseudomonas Syringae ◽  
Bacterial Blight ◽  
Acid Methyl Ester ◽  
Hypersensitive Reaction ◽  
Nutrient Broth ◽  
New Disease ◽  
Initial Symptoms ◽  
Salinas Valley ◽  
Bacterial Blight Pathogen ◽  
Dark Green

In 1998 and 1999, a new disease was detected in commercial broccoli (Brassica oleracea var. botrytis) grown in the Salinas Valley, Monterey County, CA. Initial symptoms consisted of large, water-soaked, dark green, angular leaf sections that were bordered by major leaf veins. Diseased areas were as large as 10 × 3 cm. As the disease developed, affected areas turned tan and papery, and leaf margins sometimes became tattered. The numerous small (<1 cm diameter), round to angular spots that also were present retained their size and did not develop into larger lesions. A blue-green fluorescing pseudomonad was consistently isolated from both types of lesions on King's medium B. Strains were levan positive, oxidase negative, and arginine dihydrolase negative. Strains did not rot potato slices but induced a hypersensitive reaction in tobacco (Nicotiana tabacum L. ‘Turk’). Fatty acid methyl ester analysis (MIS-TSBA version 4.10, MIDI, Inc., Newark, DE) indicated that the strains were highly similar (similarity ≥0.843) to Pseudomonas syringae. Biolog GN (version 3.50, Biolog, Inc., Hayward, CA) profiles also identified the strains as P. syringae. Therefore, the bacterium associated with the disease was identified as P. syringae. Pathogenicity of 13 strains was demonstrated by greenhouse tests. The strains were grown as nutrient broth shake cultures for 48 h at 24°C, diluted to 106 CFU/ml, and misted onto broccoli (cvs. Patriot and Titleist) and broccoli raab (B. rapa subsp. rapa cv. Spring). Control plants were misted with sterile nutrient broth. After 4 to 5 days in a greenhouse (24 to 26°C), large angular leaf lesions developed on all inoculated broccoli and broccoli raab plants. Strains were reisolated from symptomatic tissue and identified as P. syringae. Control plants remained symptomless. The results of two sets of pathogenicity tests were the same. Unlike most P. syringae strains, those isolated from broccoli were sensitive to a bacteriophage recovered from a P. syringae pathovar that infects broccoli raab. These results suggest that the broccoli pathogen may be related to the bacterial blight pathogen of broccoli raab (1). This is the first report of this pathogen causing a disease on commercially grown broccoli. Reference: (1) S. T. Koike et al. Plant Dis. 82:727, 1998.

scholarly journals First Report of Bacterial Leaf Spot of Italian Dandelion (Cichorium intybus) Caused by a Pseudomonas syringae Pathovar in California

Plant Disease ◽  
2006 ◽  
Vol 90 (2) ◽  
pp. 245-245 ◽  
Author(s):  
S. T. Koike ◽  
C. T. Bull
Keyword(s):  
Pseudomonas Syringae ◽  
Leaf Spot ◽  
Late Summer ◽  
Hypersensitive Reaction ◽  
Cichorium Intybus ◽  
Nutrient Broth ◽  
Bacterial Strains ◽  
Bacterial Leaf Spot ◽  
First Report ◽  
Leaf Spots

Italian dandelion (Cichorium intybus) is a leafy, nonhead forming chicory plant that is eaten as a fresh vegetable in salads. During the late summer (August through October) of 2002, in the Salinas Valley (Monterey County) in California, a previously unreported disease was found in commercial Italian dandelion fields. Early symptoms were angular, vein delimited, dark, water-soaked leaf spots that measured 2 to 7 mm in diameter. As disease developed, spots retained angular edges but exhibited various irregular shapes. Spots commonly formed along the edges of the leaves; in some cases these spots developed into lesions that measured between 10 and 30 mm long. Spots were visible from adaxial and abaxial sides and were dull black in color. A cream-colored pseudomonad was consistently isolated from leaf spots that were macerated and streaked onto sucrose peptone agar. Fungi were not recovered from any of the spots. Recovered strains were blue-green fluorescent when streaked onto King's medium B agar. Bacterial strains were levan positive, oxidase negative, and arginine dihydrolase negative. Strains did not rot potato slices but induced a hypersensitive reaction on tobacco (Nicotiana tabacum cv. Turk). These data indicated that the bacteria belonged to LOPAT group 1 of Pseudomonas syringae (1). Pathogenicity of six strains was tested by growing inoculum in nutrient broth shake cultures for 48 h, diluting to 106 CFU/ml, and spraying onto 12 6-week-old plants of Italian dandelion cv. Catalogna Special. Untreated control plants were sprayed with sterile nutrient broth. After 10 to 12 days in a greenhouse (24 to 26°C), leaf spots similar to those observed in the field developed on all inoculated plants. Strains were reisolated from the spots and identified as P. syringae. Control plants remained symptomless. These inoculation experiments were done twice and the results were the same. Amplification of repetitive bacterial sequences (repetitive sequence-based polymerase chain reaction [rep-PCR]) demonstrated that all Italian dandelion strains had the same rep-PCR fingerprint, which differed from fingerprints of P. syringae pv. tagetis and P. syringae pv. tabaci. Additionally, toxin specific primers did not amplify tagetitoxin or tabtoxin biosynthesis genes from Italian dandelion strains. To our knowledge, this is the first report of bacterial leaf spot of commercially grown Italian dandelion in California caused by a P. syringae pathovar. Because fields were irrigated with overhead sprinklers, the disease was severe in several fields and as much as 30% of those plantings were not harvested. Reference: (1) R. A. Lelliott et al. J. Appl. Bacteriol. 29:470, 1966.

scholarly journals First Report of Bacterial Leaf Spot on Snow Lotus Caused by Pseudomonas syringae in China

Plant Disease ◽  
2009 ◽  
Vol 93 (2) ◽  
pp. 204-204 ◽  
Author(s):  
S. F. Zhao ◽  
Y. N. Luo ◽  
H. Y. Zhao ◽  
J. Du ◽  
X. Y. Fang
Keyword(s):  
Pseudomonas Syringae ◽  
Leaf Spot ◽  
Unknown Etiology ◽  
Leaf Spot Disease ◽  
Sterile Water ◽  
Bacterial Leaf Spot ◽  
Saussurea Involucrata ◽  
First Report ◽  
Initial Symptoms ◽  
Snow Lotus

Snow lotus (Saussurea involucrata (Kar. & Kir.) Sch. Bip.) is an economically important medicinal herb increasingly grown in China in recent years. During the summer and autumn of 2005, 2006, and 2007, a necrosis of unknown etiology was observed on leaves in commercial production areas in Xinjiang Province of China. Disease incidence was approximately 40 to 50% of the plants during the 2005 and 2007 growing seasons. Initial symptoms consisted of pronounced water-soaked, dark brown-to-black spots that were 1 to 2 mm in diameter on young, expanding leaves. Later, some leaf spots on older leaves enlarged and coalesced, causing leaf desiccation. Leaf samples were collected in 2005, 2006, and 2007 from the affected hosts. Bacterial streaming was evident from these samples, and 28 strains were isolated on nutrient agar or King's medium B (KMB). All strains were gram negative and fluoresced bluegreen under UV light after 48 h of growth at 28°C on KMB. On the basis of LOPAT tests, the strains were identified as Pseudomonas syringae (1). The identity of two strains was confirmed by sequencing the 16S rDNA gene, which revealed 98% similarity to P. syringae strains in NCBI (Accession Nos. FJ001817 and FJ001818 for XJSNL 111 and 107, respectively). Infiltration of tobacco leaves with bacterial suspensions resulted in typical hypersensitivity reactions within 24 h. Pathogenicity of the strains was confirmed by spray inoculating five snow lotus leaves of a six-leaf stage plant with 108 CFU ml–1 bacterial suspensions in sterile water and five plants sprayed with sterile distilled water served as controls. Inoculated and sterile water-sprayed controls were maintained in the growth chamber with 90% relative humidity for 15 days at 15 ± 2°C. Symptoms similar to the original symptoms were observed on inoculated plants after 2 weeks. No symptoms developed on controls. Bacteria reisolated from inoculated plants were identified as strains of P. syringae. Isolates were deposited at the Key Laboratory for Oasis Crop Disease Prevention and Cure, Shihezi University. Rust caused by Puccinia carthami and leaf spot disease caused by Alternaria carthami of snow lotus have been reported (2,3). To our knowledge, this is the first report of P. syringae as the cause of bacterial leaf spot on snow lotus in China. References: (1) A. Braun-Kiewnick and D. C. Sands. Pseudomonas. Page 84 in: Laboratory Guide for the Identification of Plant Pathogenic Bacteria. 3rd ed. N. W. Schaad et al., eds. The American Phytopathological Society, St. Paul, MN, 2001. (2) S. Zhao et al. Plant Dis. 91:772, 2007. (3) S. Zhao et al. Plant Dis. 92:318, 2008.

scholarly journals First Report of a Leaf Spot Disease of Tobacco Caused by Pseudomonas cichorii in China

Plant Disease ◽  
2021 ◽  
Author(s):  
Dayu Lan ◽  
Fangling Shu ◽  
Yanhui Lu ◽  
Anfa Shou ◽  
Wei Lin ◽  
...  
Keyword(s):  
Pseudomonas Syringae ◽  
Leaf Spot ◽  
Leaf Spot Disease ◽  
Pathogenicity Test ◽  
Pseudomonas Cichorii ◽  
Koch’S Postulates ◽  
First Report ◽  
Initial Symptoms ◽  
Wide Range ◽  
Koch's Postulates

Tobacco (Nicotiana tabacum L.), one of the chief commercial crops, is wildly cultivated worldwide. In June 2020 and 2021, an unknown bacterial leaf spot on tobacco was found in Hezhou and Hechi City, Guangxi, China. 30% of the tobacco were affected and the rate of diseased leaves reached about 10% in the field under high temperature and rainstorm. The disease mainly damaged the middle and top leaves of tobacco plants at vigorous growing stage. The initial symptoms were water-soaked spots on the frontal half of a leaf, and then expanded into circular to irregular spots with a yellow halo at the edge. The spots mostly appeared dark brown at high air humidity, while yellow brown at low humidity and exhibited a concentric pattern. In severe cases, the lesions coalesced and the whole leaf was densely covered with lesions, resulting in the loss of baking value. A bacterium was consistently isolated from diseased leaf tissues on nutrient agar (NA). Growth on NA was predominantly grayish white circular bacterial colonies with smooth margins, and the bacterium is rod-shaped, gram-negative and fluorescent on King’s B medium. Seven isolates (ND04A-ND04C and ZSXF02-ZSXF05) were selected for molecular identification and pathogenicity tests. Genomic DNA of the bacterium was extracted and the housekeeping gene of cts (encoding citrate synthase) was amplified with the primers cts-Fs/cts-Rs (forward primer cts-Fs: 5’-CCCGTCGAGCTGCCAATWCTGA-3’; reverse primer cts-Rs: 5’-ATCTCGCACGGSGTRTTGAACATC-3’) (Berge et al. 2014; Sarkar et al. 2004). 409-bp cts gene sequences were deposited in the GenBank database for seven isolates (accession no. OK105110-OK105116). Sequence of seven isolates shared 100% identity with several Pseudomonas cichorii strains within the GenBank database (accession no. KY940268 and KY940271), and the phylogenetic tree of cts genes of the seven isolates clustered with the phylogroup 11 of Pseudomonas syringae (accession no. KJ877799 and KJ878111), which was classified as P.cichorii. To satisfy Koch’s postulates, a pathogenicity test was tested by using a needle to dip a suspension of the bacterium (108 CFU/ml) and pricking three holes in the tobacco leaf. The control plants leaves were needled with sterile water. Each tobacco plant was inoculated with three leaves, and the test was repeated three times. All plants were placed in transparent plastic boxes and incubated in a greenhouse at 25 ± 3°C. The water-soaked spots appeared 24h after inoculation and quickly expanded through leaf veins. Three days after inoculation, all the inoculated leaves showed symptoms similar to those observed in the field. Control plants remained healthy. Only P. cichorii was successfully re-isolated from the lesions, confirming Koch’s postulates. Pseudomonas cichorii can infect eggplant, lettuce, tomatoand other crops, and has a wide range of hosts (Timilsina et al. 2017; Ullah et al. 2015). To our knowledge, this is the first report of P. cichorii causing leaf spot on tobacco in China.

scholarly journals First Report of Bacterial Leaf Spot of Coriander Caused by Pseudomonas syringae pv. coriandricola in China

Plant Disease ◽  
2021 ◽  
Author(s):  
Lei Li ◽  
Yishuo Huang ◽  
Yanxia Shi ◽  
A LI CHAI ◽  
Xuewen Xie ◽  
...  
Keyword(s):  
Pseudomonas Syringae ◽  
Leaf Spot ◽  
Citrate Synthase ◽  
Morphological Characteristics ◽  
Likelihood Method ◽  
Bacterial Suspension ◽  
Leaf Spot Disease ◽  
Distilled Water ◽  
Bacterial Leaf Spot ◽  
First Report

Coriander (Coriandrum sativum L.) or Chinese parsley is a culinary herb with multiple medicinal effects that are widely used in cooking and traditional medicine. From September to November 2019, symptoms were observed in 2-month-old coriander plants from coriander fields in Lanzhou and Wenzhou, China. The disease developed rapidly under cold and wet climatic conditions, and the infection rate was almost 80% in open coriander fields. Typical symptoms on leaves included small, water-soaked blotches and irregular brown spots surrounding haloes; as the disease progressed, the spots coalesced into necrotic areas. Symptomatic leaf tissue was surface sterilized, macerated in sterile distilled water, and cultured on nutrient agar plates at 28 °C for 48 h (Koike and Bull, 2006). After incubation, six bacterial colonies, which were individually isolated from collected samples from two different areas, were selected for further study. Colonies on NA plate were small, round, raised, white to cream-colored, and had smooth margins. All bacterial isolates were gram-negative, rod-shaped and nonfluorescent on King's B medium. The bacteria were positive for levan production, Tween 80 hydrolysis, and tobacco hypersensitivity but negative for oxidase, potato slice rot test, arginine dihydrolase, ice nucleation activity, indole production and H2S production. The suspension of representative isolate for inoculating of plants was obtained from single colony on King's B medium for 2-3 days at 28 °C. DNA was extracted from bacterial suspensions of YS2003200102 cultured in 20 ml of King’s B medium broth at 28 °C for 1 day. Extraction was performed with a TIANamp Bacterial DNA Kit (TIANGEN, China) according to the manufacturer’s recommendations. The pathogen was confirmed by amplification and sequencing of the glyceraldehyde-3-phosphate dehydrogenase A (gapA) gene, the citrate synthase (gltA) gene, the DNA gyrase B (gyrB) gene and the RNA polymerase sigma factor 70 (rpoD) gene using gapA-For/gapA-Rev, gltA-For/gltA-Rev, gyrB-For/gryB-Rev, rpoD-For/rpoD-Rev primers, respectively (Popović et al., 2019). The sequences of the PCR products were deposited in GenBank with accession numbers MZ681931 (gapA), MZ681932 (gltA), MZ681933 (gyrB), and MZ681934 (rpoD). Phylogenetic analysis of multiple genes (Xu and Miller, 2013) was conducted with the maximum likelihood method using MEGA7. The sequences of our isolates and ten published sequences of P. syringae pv. coriandricola were clustered into one clade with a 100% confidence level. To confirm the pathogenicity of isolate YS2003200102, 2-month-old healthy coriander plants were inoculated by spraying the leaves with a bacterial suspension (108 CFU ml−1) at 28 °C incubation temperature and 70% relative humidity condition, and sterile distilled water was applied as a negative control treatment (Cazorla et al. 2005). Three replicates were conducted for every isolate, and each replicate included 6 coriander plants. After twelve days, only the inoculated leaves with bacterial suspension showed bacterial leaf spot resembling those observed on naturally infected coriander leaves. Cultures re-isolated from symptomatic leaves showed the same morphological characteristics and molecular traits as those initially isolated from infected leaves in the field. This bacterium was previously reported causing leaf spot of coriander in India and Spain (Gupta et al. 2013; Cazorla et al. 2005). To our knowledge, this is the first report of P. syringae pv. coriandricola causing leaf spot disease on coriander in China. Studies are needed on strategies to manage P. syringae pv. coriandricola in crops, because its prevalence may cause yield loss on coriander in China.

scholarly journals First Report of Bacterial Leaf Spot of Rieger Begonia Caused by Xanthomonas campestris pv. begoniae in China

Plant Disease ◽  
2013 ◽  
Vol 97 (2) ◽  
pp. 282-282 ◽  
Author(s):  
L. H. Zhou ◽  
G. H. Ji
Keyword(s):  
Leaf Spot ◽  
Xanthomonas Campestris ◽  
Infection Process ◽  
Leaf Spot Disease ◽  
Leaf Margin ◽  
Bacterial Strains ◽  
Bacterial Leaf Spot ◽  
First Report ◽  
Initial Symptoms ◽  
Phosphate Buffered Saline

Rieger begonia are collectively referred to as a begonia hybrid group. Its global annual sales is 90,000,000 cutting seedlings. It is one of the top ten potted plants. In the summer of 2011, serious outbreaks of a suspected bacterial leaf spot disease were observed on five Rieger begonia cultivars (Dark Britt, Rebecca, Blitz, Barkos, and Borias). These plants were grown for potted cutting seedling production in commercial nurseries located in Shilin county of Yunnan Province, China. The initial symptoms of the disease were small circular or polygonal water-soaked needle spots on leaf margin that later these spots expanded and joined together, forming bigger inverted V-shaped necrotic specks (4). Yellow-pigmented bacterial colonies were consistently isolated from diseased leaves and stems on NA agar medium and incubated at 28°C. Twelve bacterial strains were isolated and used for further studies. All the isolates were Gram-negative, rod-shaped, motile, aerobic, and non-sporing. All of the bacterial strains isolated in the present study were identified as Xanthomonas campestris pv. begoniae (Xcb) based on biochemical and physiological identification (Biolog carbon source utilization analysis) and 16S rDNA sequences analysis and further pathogenicity determination (1). The results show that the sequence homology rate of HT1-1 (GenBank Accession No. JN648097) and X. euvesicatoria (syn. X. campestris pv vesicatoria) (GeneBank Accession No. AM039952) is 99%. This strongly suggests that the Rieger begonia isolates belong to X. campestris pv. begoniae (2). For Koch's postulates, 10 surface-disinfected young leaves from five susceptible Rieger begonia plants (cv. Dark Britt) were inoculated by spraying a phosphate-buffered saline suspension of each bacterial isolate (3.0 × 108 CFU/ml) onto the leaves (3). Controls were inoculated similarly with phosphate-buffered saline solution. All inoculated plants were covered with polyethylene bags for 24 h at 25°C and then put in the greenhouse. After inoculation, water-soaked and necrotic symptoms were observed on inoculated Rieger begonia leaves within 7 to 9 days. No symptoms were observed on controls. Bacteria were reisolated and confirmed to be identical to the original isolates by the methods described above. To our knowledge, this is the first report of Xcb causing leaf spot on Rieger begonia plants in China. The infection process of Xcb on Rieger begonia plants and rapid detection of this pathogen are underway. References: (1) M. R. Gillings et al. PNAS 12:102, 2005. (2) C. L. Oliver et al. Plant Dis. 4:96, 2012. (3) H. Ornek et al. New Dis. Rep. 13:40, 2006. (4) O. Pruvost et al. Plant Dis. 4:96, 2012.

scholarly journals First Report of Bacterial Leaf Spot of Swiss Chard Caused by Pseudomonas syringae pv. aptata in California

Plant Disease ◽  
2003 ◽  
Vol 87 (11) ◽  
pp. 1397-1397 ◽  
Author(s):  
S. T. Koike ◽  
D. M. Henderson ◽  
C. T. Bull ◽  
P. H. Goldman ◽  
R. T. Lewellen
Keyword(s):  
Fatty Acid ◽  
Sugar Beet ◽  
Pseudomonas Syringae ◽  
Leaf Spot ◽  
Nutrient Broth ◽  
Pcr Analysis ◽  
Bacterial Leaf Spot ◽  
First Report ◽  
Leaf Spots ◽  
Swiss Chard

From 1999 through 2003, a previously unreported disease was found on commercial Swiss chard (Beta vulgaris subsp. cicla) in the Salinas Valley, (Monterey County) California. Each year the disease occurred sporadically throughout the long growing season from April through September. Initial symptoms were water-soaked leaf spots that measured 2 to 3 mm in diameter. As disease developed, spots became circular to ellipsoid, 3 to 8 mm in diameter, and tan with distinct brown-to-black borders. Spots were visible from the adaxial and abaxial sides. Cream-colored bacterial colonies were consistently isolated from spots. Strains were fluorescent on King's medium B, levan positive, oxidase negative, and arginine dihydrolase negative. Strains did not rot potato slices but induced a hypersensitive reaction on tobacco (Nicotiana tabacum cv. Turk). The isolates, therefore, belong in LOPAT group 1 (1). Fatty acid methyl esters (FAME) analysis (MIS-TSBA version 4.10, MIDI Inc., Newark, DE) gave variable results that included Pseudomonas syringae, P. cichorii, and P. viridiflava with similarity indices ranging from 0.91 to 0.95. BOX-polymerase chain reaction (PCR) analysis gave identical banding patterns for the chard isolates and for known P. syringae pv. aptata strains, including the pathotype strain CFBP1617 (2). The bacteria were identified as P. syringae. Pathogenicity of 11 strains was tested by growing inoculum in nutrient broth shake cultures for 48 h, diluting to 10 × 6 CFU/ml, and spraying onto 5-week-old plants of Swiss chard cvs. Red, White, Silverado, and CXS2547. Untreated control plants were sprayed with sterile nutrient broth. After 7 to 10 days in a greenhouse (24 to 26°C), leaf spots similar to those observed in the field developed on all inoculated plants. Strains were reisolated from the spots and identified as P. syringae. Control plants remained symptomless. To investigate the host range of this pathogen, the same procedures were used to inoculate three strains onto other Chenopodiaceae plants: five cultivars of sugar beet (B. vulgaris), and one cultivar each of spinach (Spinacia oleracea) and Swiss chard. In addition, five chard strains and strain CFBP1617 were inoculated onto two cultivars of sunflower (Helianthus annuus), and one cultivar each of cantaloupe (Cucumis melo), sugar beet, spinach, and Swiss chard. All Swiss chard, cantaloupe, sunflower, and sugar beet plants developed leaf spots after 7 days. The pathogen was reisolated from spots and confirmed to be the same bacterium using BOX-PCR analysis. Spinach and untreated controls failed to show symptoms. All inoculation experiments were done at least twice and the results were the same. The phenotypic data, fatty acid and genetic analyses, and pathogenicity tests indicated that these strains are P. syringae pv. aptata. To our knowledge this is the first report of bacterial leaf spot of commercially grown Swiss chard in California caused by P. syringae pv. aptata. The disease was particularly damaging when it developed in Swiss chard fields planted for “baby leaf” fresh market products. Such crops are placed on 2-m wide beds, planted with high seed densities, and are sprinkler irrigated. This disease has been reported from Asia, Australia, Europe, and other U.S. states. References: (1) R. A. Lelliott et al. J. Appl. Bacteriol. 29:470, 1966. (2) J. L. W. Rademaker et al. Mol. Microbiol. Ecol. Man. 3.4.3:1–27, 1998.

Sign in / Sign up

Export Citation Format

Share Document