scholarly journals Bacterial Leaf Spot Diseases of Leafy Crucifers in Oklahoma Caused by Pathovars of Xanthomonas campestris

Plant Disease ◽  
2000 ◽  
Vol 84 (9) ◽  
pp. 1008-1014 ◽  
Author(s):  
Youfu Zhao ◽  
John P. Damicone ◽  
David H. Demezas ◽  
Carol L. Bender
Keyword(s):  
Pseudomonas Syringae ◽  
Leaf Spot ◽  
Xanthomonas Campestris ◽  
Leaf Surface ◽  
Black Rot ◽  
Local Strains ◽  
Leaf Spots ◽  
Abaxial Leaf Surface ◽  
Polymerase Chain ◽  
Reference Strains

Fields of kale, spinach mustard, and turnip were severely damaged by bacterial leaf spots during 1994 to 1996. Symptoms included circular to angular necrotic lesions with yellow halos and water-soaking on the abaxial leaf surface. Yellow, mucoid strains isolated from leaf spots were identified as Xanthomonas campestris using Biolog. Four strains caused black lesions on stems of cabbage seedlings in an excised cotyledon assay, leaf spots and sunken dark lesions on petioles of spray-inoculated crucifers, and leaf spots on spray-inoculated tomato. These strains were classified as X. campestris pv. armoraciae. Most other strains from leafy crucifers and all strains from a cabbage field caused black rot in the cotyledon assay and in spray-inoculations. Many of these strains also caused leaf spots on collard and kale but not stem and petiole lesions. The strains causing black rot were classified as X. campestris pv. campestris. Cluster analysis of Biolog profiles yielded a small group that contained local strains of both pathovars, and a large group comprised of reference and local strains of each pathovar, and some local, nonpathogenic strains. Five fingerprint groups were identified by rep-polymerase chain reaction using the BOXA1R primer. Local and reference strains of each pathovar occurred in two of the groups. Two pathovars of X. campestris are involved in the leaf spot diseases. Both pathovars were recovered within several fields, and also were recovered along with Pseudomonas syringae pv. maculicola. This is the first report of Xanthomonas leaf spot caused by X. campestris pv. armoraciae in Oklahoma.

scholarly journals Bacterial Leaf Spot of Leafy Crucifers in Oklahoma Caused by Pseudomonas syringae pv. maculicola

Plant Disease ◽  
2000 ◽  
Vol 84 (9) ◽  
pp. 1015-1020 ◽  
Author(s):  
Youfu Zhao ◽  
John P. Damicone ◽  
David H. Demezas ◽  
Vidhya Rangaswamy ◽  
Carol L. Bender
Keyword(s):  
Carbon Source ◽  
Pseudomonas Syringae ◽  
Leaf Spot ◽  
Xanthomonas Campestris ◽  
Bacterial Disease ◽  
Bacterial Leaf Spot ◽  
Local Strains ◽  
Fluorescent Pseudomonas ◽  
Reference Strains

During 1995 and 1996, bacterial leaf spots severely damaged fields of kale, spinach mustard, and turnip in Oklahoma. Symptoms were small, brown, necrotic spots with irregular edges surrounded by chlorotic halos. Lesion margins were often water-soaked on the abaxial surface. The spots enlarged and coalesced, causing extensive leaf yellowing and necrosis. Nineteen strains of a fluorescent Pseudomonas spp. were isolated from symptomatic plants. LOPAT tests and carbon source oxidation using Biolog GN MicroPlates were used to classify the strains as P. syringae. Cluster analysis of carbon source oxidation profiles for the local strains and selected reference strains of P. syringae pv. maculicola and pv. tomato produced one group with 79.5% similarity. In spray inoculations, all local strains caused chlorotic or water-soaked lesions on collards, kale, cauliflower, and tomato. A few local strains caused necrotic lesions on mustard. Most local strains caused one of the three lesion types on turnip and spinach mustard. Reference strains of P. syringae pv. maculicola caused similar symptoms. All but three of the local strains produced coronatine in vitro. The local strains were thus classified as P. syringae pv. maculicola, the cause of bacterial leaf spot of crucifers. Two distinct groups of P. syringaepv. maculicola were identified by repetitive sequence-based polymerase chain reaction (rep-PCR) with both REP and BOXA1R primers. Three subgroups within each group were further identified using the BOXA1R primer. Except for two strains of P. syringae pv. tomato which were pathogenic on crucifers, the pathovars maculicola and tomato had different genetic fingerprints. The pathogen was recovered from seven of ten fields sampled during 1994 to 1996. In five of the fields with P. syringae pv. maculicola, pathovars of Xanthomonas campestris were also isolated from lesions forming a bacterial disease complex. This is the first report of bacterial leaf spot caused by P. syringaepv. maculicola on leafy crucifers in Oklahoma.

scholarly journals Identification of Isolates that Cause a Leaf Spot Disease of Brassicas as Xanthomonas campestris pv. raphani and Pathogenic and Genetic Comparison with Related Pathovars

2006 ◽  
Vol 96 (7) ◽  
pp. 735-745 ◽  
Author(s):  
J. G. Vicente ◽  
B. Everett ◽  
S. J. Roberts
Keyword(s):  
Plant Species ◽  
Leaf Spot ◽  
Xanthomonas Campestris ◽  
Leaf Spot Disease ◽  
Avirulence Genes ◽  
Spot Disease ◽  
Leaf Spots ◽  
Brassica Spp ◽  
Polymerase Chain ◽  
Differential Hosts

Twenty-five Xanthomonas isolates, including some isolates received as either X. campestris pv. armoraciae or pv. raphani, caused discrete leaf spot symptoms when spray-inoculated onto at least one Brassica oleracea cultivar. Twelve of these isolates and four other Xanthomonas isolates were spray- and pin-inoculated onto 21 different plant species/cultivars including horseradish (Armoracia rusticana), radish (Raphanus sativus), and tomato (Lycopersicon esculentum). The remaining 13 leaf spot isolates were spray-inoculated onto a subset of 10 plant species/cultivars. The leaf spot isolates were very aggressive on several Brassica spp., radish, and tomato causing leaf spots and dark sunken lesions on the middle vein, petiole, and stem. Based on the differential reactions of several Brassica spp. and radish cultivars, the leaf spot isolates were divided into three races, with races 1 and 3 predominating. A differential series was established to determine the race-type of isolates and a gene-for-gene model based on the interaction of two avirulence genes in the pathogen races and two matching resistance genes in the differential hosts is proposed. Repetitive-DNA polymerase chain reaction-based fingerprinting was used to assess the genetic diversity of the leaf spot isolates and isolates of closely related Xanthomonas pathovars. Although there was variability within each race, the leaf spot isolates were clustered separately from the X. campestris pv. campestris isolates. We propose that X. campestris isolates that cause a nonvascular leaf spot disease on Brassica spp. should be identified as pv. raphani and not pv. armoraciae. Race-type strains and a neopathotype strain for X. campestris pv. raphani are proposed.

scholarly journals An Outbreak of a Leaf Spot Disease of Cabbage in Southern Florida Caused by Xanthomonas campestris pv. armoraciae

Plant Disease ◽  
2003 ◽  
Vol 87 (7) ◽  
pp. 873-873 ◽  
Author(s):  
K. Pernezny ◽  
J. B. Jones ◽  
P. D. Roberts ◽  
E. Dickstein
Keyword(s):  
Leaf Spot ◽  
Xanthomonas Campestris ◽  
Acid Methyl Ester ◽  
Sole Source ◽  
Leaf Spot Disease ◽  
Black Rot ◽  
Red Cabbage ◽  
Spot Disease ◽  
Leaf Spots ◽  
Serological Data

From October to December 2001, a leaf spot disease was observed in numerous commercial fields of red and green cabbage (Brassica oleracea var. capitata L.) in the Everglades Agricultural Area, south and east of Lake Okeechobee and in the environs of Immokalee in southwestern Florida. Discrete water-soaked to greasy appearing spots were observed in the leaf blades with no evidence of marginal V-shaped lesions characteristic of black rot caused by Xanthomonas campestris pv. campestris. Profuse bacterial streaming was observed when cut leaf sections were examined microscopically. A bacterium that formed yellow colonies on nutrient agar was consistently isolated from these lesions. Ten bacteria were isolated, purified, and characterized. All strains were aerobic, gram-negative rods. Strains were positive for esculin hydrolysis, proteolysis in litmus milk, and gelatin liquefaction. Strains were negative for urease production, nitrate reduction, oxidase, and utilization of asparagine as a sole source of carbon and nitrogen. Fatty acid methyl ester analysis indicated a match with Florida library strains of X. campestris pv. raphani (similarity indices 0.605–0.738). Suspensions (2 × 107 CFU/ml in phosphate-buffered saline) of two Oklahoma strains identified as X. campestris pv. armoraciae provided by J. P. Damicone (3) and four representative Florida strains were applied to plants using a hand-held sprayer. Pathogenicity of the strains was tested on three replicate greenhouse-grown plants of the following: green cabbage cv. Market Early; red cabbage cv. Salad Delight; radish cv. Red Silk; tomato cv. Sunny; sweet bell pepper cv. Jupiter; and fresh horseradish roots purchased from a retail grocery chain. A strain of X. campestris pv. campestris originally isolated from Homestead, FL was also included in pathogenicity tests. All Florida and Oklahoma strains produced leaf spots, but no V-shaped lesions, on leaves of green cabbage, red cabbage, radish, tomato, and horseradish. Typical black rot symptoms were observed only in radish and green and red cabbage inoculated with the X. campestris pv. campestris strain. On the basis of these results, we identify the Florida strains as X. campestris pv. armoraciae (1,2,3), recognizing the precedent of X. campestris pv. armoraciae over X. campestris pv. raphani based on extensive genetic and serological data (1). Our strains appear to be more similar to those causing outbreaks on crucifers in Oklahoma (3) than those in Ohio (2), because Florida strains were pathogenic on tomato. References: (1) A. M. Alvarez et al. Phytopathology 84:1449, 1994. (2) F. Sahin and S. A. Miller. Plant Dis. 81:1334, 1997. (3) Y. Zhao et al. Plant Dis. 84:1008, 2000.

scholarly journals First Report of Pseudomonas syringae pv. aptata Causing Bacterial Leaf Spot on Sugar Beet in Serbia

Plant Disease ◽  
2015 ◽  
Vol 99 (2) ◽  
pp. 281-281 ◽  
Author(s):  
V. Stojšin ◽  
J. Balaž ◽  
D. Budakov ◽  
Slaviša Stanković ◽  
I. Nikolić ◽  
...  
Keyword(s):  
Sugar Beet ◽  
Pseudomonas Syringae ◽  
Leaf Spot ◽  
Leaf Surface ◽  
Negative Control ◽  
Gyrb Gene ◽  
Bacterial Suspension ◽  
Bacterial Strains ◽  
Gene Sequences ◽  
Bacterial Leaf Spot

A severe bacterial leaf spot was observed during June and July 2013 on commercial cultivars of sugar beet (Beta vulgaris var. saccharifera) in the Vojvodina Province of Serbia. Serbia is a major sugar beet production area in southeastern Europe, with 62,895 ha and 3 million tons of sugar beet yield in 2013. A foliar leaf spot observed in 25 commercial sugar beet fields surveyed ranged from 0.1 to 40% severity. Symptoms were characterized as circular or irregular, 5- to 20-mm diameter, white to light brown necrotic spots, each with a dark margin. Diseased leaves were rinsed in sterilized, distilled water (SDW) and dried at room temperature, and leaf sections taken from the margin of necrotic tissue were macerated in SDW. Isolations from 48 symptomatic leaves onto nutrient agar with 5% (w/v) sucrose (NAS) produced bacterial colonies that were whitish, circular, dome-shaped, and Levan-positive. Representative isolates (n = 105) were Gram negative; aerobic; positive for catalase, fluorescence on King's medium B, and tobacco hypersensitivity; and negative for oxidase, potato rot, and arginine dehydrolase. These reactions corresponded to LOPAT group Ia, which includes Pseudomonas syringae pathovars (2). Repetitive extragenic palindromic sequence (rep)-PCR was used for genetic fingerprinting the isolates using the REP, ERIC, and BOX primers. Twenty-five different profiles were obtained among the strains. From each profile group, one representative strain was sequenced for the gyrB gene (1). Four heterogenic groups were observed, and representative gyrB gene sequences of each group were deposited in the NCBI GenBank (Accession Nos. KJ950024 to KJ950027). The sequences were compared with those of pathotype strain P. syringae pv. aptata CFBP 1617 deposited in the PAMDB database; one strain was 100% homologous, and the other three were 99% homologous. To fulfill identification of the Serbian sugar beet isolates, gltA and rpoD partial gene sequences were determined (1), and the sequences were deposited as Accession Nos. KM386838 to KM386841 for gltA and KM386830 to KM38683033 for rpoD. The sequences were 100% homologous with those of pathotype strain CFBP 1617. Pathogenicity of each of four representative bacterial strains was tested on 3-week-old plants of the sugar beet cultivars Marinela, Serenada, and Jasmina (KWS, Belgrade, Serbia) and Lara (NS Seme, Novi Sad, Serbia) by atomizing a bacterial suspension of ~106 CFU/ml of the appropriate isolate onto the abaxial leaf surface of three plants per cultivar until water-soaking of the leaf surface was observed. Three plants of each cultivar atomized similarly with P. syringae pv. aptata CFBP 2473 and SDW served as positive and negative control treatments, respectively. Inoculated plants were kept in a clear plastic box at 80 to 100% RH and 17 ± 1°C and examined for symptom development over 3 weeks. For all test isolates and the control strain, inoculated leaves first developed water-soaked lesions 7 days after inoculation (DAI). By 10 to 14 DAI, lesions were necrotic and infection had spread to the petioles. By 21 DAI, wilting was observed on more than 50% of inoculated plants. Negative control plants were symptomless. Bacteria re-isolated onto NAS from inoculated leaves had the same colony morphology, LOPAT results, and gyrB partial gene sequences as described for the test strains. No bacteria were re-isolated from negative control plants. Based on these tests, the pathogen causing leaf spot on sugar beet in Serbia was identified as P. syringae pv. aptata. References: (1) P. Ferrente and M. Scortichini. Plant Pathol. 59:954, 2010. (2) R. A. Lelliott et al. J. Appl. Bacteriol. 29:470, 1966.

Pseudocercospora timorensis. [Descriptions of Fungi and Bacteria].

1987 ◽  
Author(s):  
S. Little
Keyword(s):  
Sweet Potato ◽  
Geographical Distribution ◽  
Leaf Spot ◽  
Ipomoea Batatas ◽  
Leaf Surface ◽  
Solomon Islands ◽  
Leaf Spots ◽  
Brown Leaf Spot ◽  
Water Splash ◽  
Brown Leaf

Abstract A description is provided for Pseudocercospora timorensis. Information is included on the disease caused by the organism, its transmission, geographical distribution, and hosts. HOSTS: Ipomoea batatas (sweet potato), I. biloba, I. campanulata, I. cordofana, I. muricata, I. peltata, I. setifera.DISEASE: Leaf spot or brown leaf spot of sweet potato. Small circular lesions first form on the leaf borders and tips before spreading over the leaf surface. These leaf spots enlarge becoming brown to dark brown in colour with a verruculose surface. The larger leaf veins may delimit the spots. GEOGRAPHICAL DISTRIBUTION: Africa: most countries; Asia: Hong-Kong, India, Indonesia, Malaysia, Taiwan; Australasia: Fiji, Papua New Guinea, Solomon Islands; North America: West Indies (St Lucia). TRANSMISSION: Presumably by wind-borne and water-splash dispersed conidia.

scholarly journals Occurrence and Diversity of Xanthomonas campestris pv. campestris in Vegetable Brassica Fields in Nepal

Plant Disease ◽  
2010 ◽  
Vol 94 (3) ◽  
pp. 298-305 ◽  
Author(s):  
Brita Dahl Jensen ◽  
Joana G. Vicente ◽  
Hira K. Manandhar ◽  
Steven J. Roberts
Keyword(s):  
Xanthomonas Campestris ◽  
Mustard Seed ◽  
Black Rot ◽  
Brassica Spp ◽  
Mustard Plant ◽  
Race 1 ◽  
Polymerase Chain ◽  
Race 4 ◽  
Xanthomonas Campestris Pv Campestris ◽  
Vegetable Brassica

Black rot caused by Xanthomonas campestris pv. campestris was found in 28 sampled cabbage fields in five major cabbage-growing districts in Nepal in 2001 and in four cauliflower fields in two districts and a leaf mustard seed bed in 2003. Pathogenic X. campestris pv. campestris strains were obtained from 39 cabbage plants, 4 cauliflower plants, and 1 leaf mustard plant with typical lesions. Repetitive DNA polymerase chain reaction-based fingerprinting (rep-PCR) using repetitive extragenic palindromic, enterobacterial repetitive intergenic consensus, and BOX primers was used to assess the genetic diversity. Strains were also race typed using a differential series of Brassica spp. Cabbage strains belonged to five races (races 1, 4, 5, 6, and 7), with races 4, 1, and 6 the most common. All cauliflower strains were race 4 and the leaf mustard strain was race 6. A dendrogram derived from the combined rep-PCR profiles showed that the Nepalese X. campestris pv. campestris strains clustered separately from other Xanthomonas spp. and pathovars. Race 1 strains clustered together and strains of races 4, 5, and 6 were each split into at least two clusters. The presence of different races and the genetic variability of the pathogen should be considered when resistant cultivars are bred and introduced into regions in Nepal to control black rot of brassicas.

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 First Report of Bacterial Leaf Spot Caused by Pseudomonas syringae pv. aptata on Swiss Chard, Beta vulgaris subsp. vulgaris, in Arizona

Plant Disease ◽  
2021 ◽  
Author(s):  
Marilen Nampijja ◽  
Mike Derie ◽  
Lindsey J. du Toit
Keyword(s):  
Beta Vulgaris ◽  
Pseudomonas Syringae ◽  
Leaf Spot ◽  
Phosphate Buffer ◽  
Negative Control ◽  
Bacterial Suspension ◽  
Bacterial Leaf Spot ◽  
First Report ◽  
Leaf Spots ◽  
Swiss Chard

Arizona is an important region of the USA for winter production of baby leaf crops such as spinach (Spinacia oleracea), table beet (Beta vulgaris subsp. vulgaris Condivita Group), and Swiss chard (B. vulgaris subsp. vulgaris Cicla Group). In the winter of 2019, severe leaf spots were observed at 80% incidence and 40% severity per plant in a 1-ha baby leaf Swiss chard crop of an (unknown cultivar) in Arizona. The lesions were circular to irregular, necrotic, water-soaked, and 1 to 5 mm in diameter. Symptomatic leaf sections (1-cm2) were surface-sterilized with 0.6% NaOCl, rinsed, and macerated in sterilized, deionized water. An aliquot of each macerate was streaked onto King’s B (KB) agar medium. Cream-colored, non-fluorescent colonies typical of Pseudomonas were isolated consistently, and all were non-fluorescent. A dozen isolates selected randomly were all negative for potato soft rot, oxidase, and arginine dihydrolase, and positive for levan production and tobacco hypersensitivity, which is typical of fluorescent P. syringae isolates, but can also include non-fluorescent strains (Lelliot et al. 1966). Three isolates were tested for pathogenicity on the table beet cv. Red Ace and Swiss chard cv. Silverado. Strain Pap009 of P. syringae pv. aptata (Psa), demonstrated previously to be pathogenic on Swiss chard and table beet, served as a positive control strain (Derie et al. 2016; Safni et al. 2016). Each isolate was grown inoculated into medium 523 broth and incubated on a shaker at 175 rpm overnight at 25°C. Each bacterial suspension was adjusted to an optical density (OD) of 0.3 at 600 nm (108 CFU/ml), and diluted in 0.0125M phosphate buffer to 107 CFU/ml. Thirty-day-old seedlings grown in Redi-Earth Plug and Seedling Mix in a greenhouse at 22 to 26°C were inoculated by rubbing the abaxial and adaxial leaf surfaces of each plant with a cotton swab dipped in inoculum to which Carborundum had been added (0.06 g/10 ml). The negative control plants were treated similarly with phosphate buffer with Carborundum. The experiment was set up as a randomized complete block design with 4 replications per treatment and 6 seedlings per experimental unit. In both trials, leaf spots resembling those on the original plants developed on all table beet and Swiss chard plants inoculated with the Arizona isolates and Pap009, but not on negative control plants. Disease severity was greater on Swiss chard (average 39% leaf area with spots) than on table beet (14%). Re-isolates obtained from inoculated seedlings using the same method as the original isolations resembled Psa. Multilocus sequence analysis (MLSA) was carried out for the original three Arizona isolates and the re-isolates using DNA amplified from the housekeeping genes gyrB, rpoD, gapA, and gltA (Hwang et al. 2005; Sarkar and Guttman 2004). Sequence identities of these genes of the Arizona isolates (GenBank accession numbers MW291615 to MW291618 for strain Pap089; MW291619 to MW291622 for Pap095; and MW291623 to MW291626 for Pap096 for gltA, gyrB, rpoD, and gapA, respectively) and the re-isolates ranged from 98 to 100% with those of Psa pathotype strain CFBP 1617 in the PAMDB database (Almeida et al. 2010; Altschul et al. 1997). Based on Koch’s postulates, colony characteristics, and MLSA, Psa was the causal agent of leaf spots in the Arizona Swiss chard crop. To our knowledge, this is the first report of bacterial leaf spot on chard in Arizona. The pathogen could have been introduced on infected seed as Psa is readily seedborne and seed transmitted.

scholarly journals First Report of Severe Outbreaks of Bacterial Leaf Spot of Leafy Brassica Greens Caused by Xanthomonas campestris pv. campestris in South Carolina

Plant Disease ◽  
2008 ◽  
Vol 92 (7) ◽  
pp. 1134-1134 ◽  
Author(s):  
W. P. Wechter ◽  
A. P. Keinath ◽  
J. P. Smith ◽  
M. W. Farnham
Keyword(s):  
South Carolina ◽  
Leaf Spot ◽  
Xanthomonas Campestris ◽  
Leaf Spot Disease ◽  
Black Rot ◽  
Spot Disease ◽  
Leafy Greens ◽  
Brassica Crops ◽  
Xanthomonas Campestris Pv Campestris ◽  
Severe Leaf

Severe outbreaks of leaf spot disease of leafy vegetable brassica crops have occurred from early spring to late fall for at least the past 7 years in Lexington County, South Carolina, the major growing region for leafy greens in the state. Significant economic losses to this disease totaling $1.7 million have been incurred by large and small growers. In 2005, Pseudomonas syringae pv. maculicola was reported as one of the causal organisms of leaf spot disease in South Carolina (2). Investigations during 2006 and 2007 have led to the isolation of another bacterium causing leaf spotting of brassica crops. Symptoms in the field were nearly identical to symptoms caused by P syringae pv. maculicola, i.e., small, brown necrotic spots, often with chlorotic halos that expand and coalesce to cover the leaves. Colonies recovered from diseased tissues were xanthomonad like, nonfluorescent on Pseudomonas Agar F, mucoid on yeast extract dextrose chalk medium, grew at 35°C, hydrolyzed starch, positive for protein digestion, alkaline in litmus milk, and produce acid from arabinose. Sequence data from the 16S rDNA and fatty acid methyl ester analysis gave the best homology to Xanthomonas campestris pv. campestris with a similarity score index of >0.98 and >0.70, respectively, confirming genus and species. Excised-cotyledon assays, used to differentiate between pathovars campestris and armoraciae, confirmed the pathovar as campestris (1). Pathogenicity assays with spray inoculations (1 × 107 CFU/ml) (3) on eight plants each of ‘Topper’ and ‘Alamo’ turnip, ‘Early Jersey Wakefield’ cabbage, and ‘Money maker’ tomato produced leaf-spot symptoms within 10 days in the greenhouse and growth chamber on the turnip and cabbage plants, but not the tomato. X. campestris pv. campestris, which is common throughout the world, also is the causal agent of black rot in brassica. Typical black rot symptoms are seen often in Lexington County fields in summer and are quite different from the leaf spot symptoms observed. Leaf-spotting X. campestris pv. campestris (LS) strains and black rot (BR) strains, recovered from black rot-symptomatic plants lacking leaf spots, from the same fields were compared in greenhouse pathogenicity assays on six plants each of ‘Topper’ turnip and ‘Early Jersey Wakefield’ cabbage. Spray inoculations with 20 individual LS strains and 10 individual BR strains, collected from 2005 to 2007, produced symptoms unique to each group. These symptoms included chlorotic ‘V’-shaped lesions initiating from the leaf margins with black veining when plants were inoculated with BR strains, versus rapid and severe leaf spotting followed by chlorotic ‘V’-shaped lesions typically lacking black-veining 10 to 16 days postinoculation associated with LS strains. Additional inoculation tests gave similar results. To our knowledge, this is the first report of a severe leaf spotting disease of field-grown brassica leafy greens caused by X. campestris pv. campestris in South Carolina. These findings may have importance in differentiation of bacterial leaf spot pathogens in brassica crops. References: (1) A. M. Alvarez et al. Phytopathology 84:1449, 1994. (2) A. P. Keinath et al. Plant Dis. 90:683, 2006. (3) W. P. Wechter et al. Hortic Sci. 42:1140, 2007.

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