scholarly journals Testing of different methods for identification of bacterial leaf spot (Siringae pv. maculicola (Mcculloch) Young et al.) plant pathogen in cauliflower leaves

2021 ◽  
Vol 1 (25) ◽  
pp. 174-186
Author(s):  
S.I. Prikhodko ◽  
◽  
A.B. Yaremko ◽  
K.P. Kornev ◽  
◽  
...  
Keyword(s):  
Pseudomonas Syringae ◽  
Leaf Spot ◽  
Biochemical Properties ◽  
Analytical Sensitivity ◽  
Bacterial Strains ◽  
Bacterial Leaf Spot ◽  
Plant Quarantine ◽  
Test Kit ◽  
Russian Plant

Pseudomonas syringae pv. maculicola (McCulloch) Young et al. is a pathogen of cauliflower bacterial spot that affects many plants of the Cruciferae family. The need for quality diagnostics of this species arose due to the mandatory phytosanitary inspection of places of production of plant products intended for export. Our research aims at determining a set of methods for the diagnosis of the pathogen of cauliflower bacterial leaf spot and evaluating these methods’ applicability in laboratory practice. The object of the research is P. syringae pv. maculicola – the causative agent of cauliflower bacterial leaf spot.The article presents the test results of two methods for the identification of Pseudomonas syringae pv. maculicola (McCulloch) Young et al. carried out in 2020 at the All-Russian Plant Quarantine Center (VNIIKR). The first method is based on the determination of biochemical properties using the API 20E test kit produced by bioMérieux’s (France); the second one – is a conventional PCR. The type bacterial strain CFBP 1657 obtained by specialists of the All-Russian Plant Quarantine Center (VNIIKR) from French collection of plant-associated bacteria (Cirm-CFBP) was used in the studies. A comparison of the biochemical properties of 23 bacteria of the genus Pseudomonas showed that there are only two characteristics within this test that distinguish P. s. pv. maculicola from other species pathovars: acetoin products and gelatin hydrolysis. Two pairs of primers with different targets in the P. syringae genome were also tested. PCR with PsyF/PsyR primers demonstrated the highest similarity of the obtained fragments with the NCBI database (97.2 %). The analytical sensitivity of PCR with PsyF/PsyR primers in plant and seed extracts was 105 CFU/ml. Determination of analytical specificity with 33 bacterial strains of the genus Pseudomonas revealed cross-reactions with strains of the following species: P. congelans, P. savastanoi pv. phaseolicola, P. savastanoi pv. glycinea, P. syringae pv. coronafaciens, P. syringae pv. syringae. Thus, to differentiate the species by means of PCR with PsyF/PsyR primers, the nucleotide sequence of the obtained amplification products should be additionally determined by Sanger sequencing.

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.

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 Diagnosis of leaf bacterial diseases of coffee reveals the prevalence of halo blight

2021 ◽  
Vol 45 ◽  
Author(s):  
Melina Korres Raimundi ◽  
Ricardo Magela de Souza ◽  
Antônia dos Reis Figueira ◽  
Gustavo Matheus Silva ◽  
Ana Carolina de Paula Santos ◽  
...  
Keyword(s):  
Pseudomonas Syringae ◽  
Leaf Spot ◽  
Biochemical Properties ◽  
Minas Gerais ◽  
Bacterial Leaf Spot ◽  
Bacterial Diseases ◽  
Halo Blight ◽  
Rpod Gene ◽  
Minas Gerais State ◽  
Species Specific

ABSTRACT The diagnosis of foliar bacterial diseases in coffee (Coffea arabica), such as halo blight (Pseudomonas syringae pv. garcae), bacterial leaf spot (P. syringae pv. tabaci), bacterial blight (P. cichorii), and dark leaf spot (Robbsia andropogonis), is considered a challenge for plant pathologists. The misidentification has been occurring when the diagnosis is solely based on symptoms and biochemical properties. Thus, the objective of this study was to identify and differentiate species and pathovars of Pseudomonas pathogenic to coffee plants, enabling a survey of the occurrence of these bacteria in the main producing regions of Minas Gerais state, Brazil. Firstly, the pathogenicity of the isolates was confirmed by inoculation in C. arabica cv. Catuaí Vermelho IAC 99. Then, biochemical analyses, combined with, repetitive element-polymerase chain reaction (rep-PCR) and phylogeny based on rpoD gene sequences were used to characterize 84 Pseudomonas isolates from coffee crops and nurseries. Based on rpoD-phylogeny, 73 isolates were identified as P. syringae pv. garcae, five as P. syringae pv. tabaci and six as P. cichorii. The rep-PCR results suggest a high genetic variability in populations of Pseudomonas syringae pv. garcae and P. cichorii. This is the first report of the occurrence of bacterial leaf spot (P. syringae pv. tabaci) in the coffee-producing filed in Minas Gerais State. The findings confirmed the prevalence of P. syringae pv. garcae in coffee production fields in the State and the generated knowledge will contribute for the development of species-specific primers for the identification and detection of this pathogen.

scholarly journals Phenotypically and Genotypically Heterogeneous Strains of Pseudomonas syringae Associated With Alfalfa Leaf Spot Disease in Iran

Plant Disease ◽  
2019 ◽  
Vol 103 (12) ◽  
pp. 3199-3208 ◽  
Author(s):  
Maryam Ansari ◽  
S. Mohsen Taghavi ◽  
Sadegh Zarei ◽  
Soraya Mehrb-Moghadam ◽  
Hamzeh Mafakheri ◽  
...  
Keyword(s):  
Host Range ◽  
Pseudomonas Syringae ◽  
Leaf Spot ◽  
Phylogenetic Analyses ◽  
Housekeeping Genes ◽  
Pcr Primers ◽  
Phylogenetic Position ◽  
Leaf Spot Disease ◽  
Bacterial Strains ◽  
Alfalfa Leaf

In this study, we provide a polyphasic characterization of 18 Pseudomonas spp. strains associated with alfalfa leaf spot symptoms in Iran. All of the strains were pathogenic on alfalfa, although the aggressiveness and symptomology varied among the strains. All strains but one were pathogenic on broad bean, cucumber, honeydew, and zucchini, whereas only a fraction of the strains were pathogenic on sugar beet, tomato, and wheat. Syringomycin biosynthesis genes (syrB1 and syrP) were detected using the corresponding PCR primers in all of the strains isolated from alfalfa. Phylogenetic analyses using the sequences of four housekeeping genes (gapA, gltA, gyrB, and rpoD) revealed that all of the strains except one (Als34) belong to phylogroup 2b of P. syringae sensu lato, whereas strain Als34 placed within phylogroup 1 close to the type strain of P. syringae pv. apii. Among the phylogroup 2b strains, nine strains were phylogenetically close to the P. syringae pv. aptata clade, whereas the remainder were scattered among P. syringae pv. atrofaciens and P. syringae pv. syringae strains. Pathogenicity and host range assays of the bacterial strains evaluated in this study on a set of taxonomically diverse plant species did not allow us to assign a “pathovar” status to the alfalfa strains. However, these results provide novel insight into the host range and phylogenetic position of the alfalfa-pathogenic members of P. syringae sensu lato, and they reveal that phenotypically and genotypically heterogeneous strains of the pathogen cause bacterial leaf spot of alfalfa.

scholarly journals Epidemiology and management of bacterial leaf spot on watermelon caused byPseudomonas syringae

Plant Disease ◽  
2017 ◽  
Vol 101 (7) ◽  
pp. 1222-1229 ◽  
Author(s):  
E. A. Newberry ◽  
L. Ritchie ◽  
B. Babu ◽  
T. Sanchez ◽  
K. A. Beckham ◽  
...  
Keyword(s):  
Disease Severity ◽  
Pseudomonas Syringae ◽  
Leaf Spot ◽  
Negative Relationship ◽  
Disease Outbreaks ◽  
Field Trials ◽  
Multiple Regression Model ◽  
Bacterial Leaf Spot ◽  
Progress Curve ◽  
Cool Conditions

Bacterial leaf spot of watermelon caused by Pseudomonas syringae has been an emerging disease in the southeastern United States in recent years. Disease outbreaks in Florida were widespread from 2013 to 2014 and resulted in foliar blighting at the early stages of the crop and transplant losses. We conducted a series of field trials at two locations over the course of two years to examine the chemical control options that may be effective in management of this disease, and to investigate the environmental conditions conducive for bacterial leaf spot development. Weekly applications of acibenzolar-S-methyl (ASM) foliar, ASM drip, or copper hydroxide mixed with ethylene bis-dithiocarbamate were effective in reducing the standardized area under the disease progress curve (P < 0.05). Pearson’s correlation test demonstrated a negative relationship between the average weekly temperature and disease severity (–0.77, P = 0.0002). When incorporated into a multiple regression model with the square root transformed average weekly rainfall, these two variables accounted for 71% of the variability observed in the weekly disease severity (P < 0.0001). This information should be considered when choosing the planting date for watermelon seedlings as the cool conditions often encountered early in the spring season are conducive for bacterial leaf spot development.

scholarly journals Bacterial Leaf Spot of Celery in California: Etiology, Epidemiology, and Role of Contaminated Seed

Plant Disease ◽  
1997 ◽  
Vol 81 (8) ◽  
pp. 892-896 ◽  
Author(s):  
E. L. Little ◽  
S. T. Koike ◽  
R. L. Gilbertson
Keyword(s):  
Pseudomonas Syringae ◽  
Leaf Spot ◽  
Disease Incidence ◽  
Field Tests ◽  
Hot Water ◽  
Disease Development ◽  
Bacterial Leaf Spot ◽  
Overhead Irrigation ◽  
Celery Seed ◽  
Salinas Valley

Pseudomonas syringae pv. apii, causal agent of bacterial leaf spot (BLS) of celery, was first identified in California in 1989. By 1991, BLS was apparent in all celery-growing areas of the state. Greenhouse-produced transplants were affected most severely, and disease incidence approached 100% in some greenhouses. In this study, sources of inoculum and factors contributing to disease development were investigated in three Salinas Valley greenhouse operations during the 1991, 1992, and 1993 celery transplant seasons (January to August). Epiphytic P. syringae pv. apii was not detected on celery transplants until April or May of each year. Increased epiphytic populations preceded BLS outbreaks, and high-pressure, overhead irrigation favored bacterial infiltration and disease development. In seed-wash assays, P. syringae pv. apii was recovered from 5 of 24 commercial celery seed lots. In field tests, epiphytic P. syringae pv. apii was found on umbels of inoculated celery plants, and seeds from these plants were heavily contaminated with P. syringae pv. apii. Contaminated seed produced seedlings with large epiphytic P. syringae pv. apii populations. Hot-water treatment (50°C for 25 min) eliminated >99.9% of seed contamination. Based on these results, disease management techniques are proposed.

scholarly journals Complete and Draft Genome Sequences of the Cruciferous Pathogens Pseudomonas cannabina pv. alisalensis and Pseudomonas syringae pv. maculicola

2021 ◽  
Vol 10 (17) ◽  
Author(s):  
Takashi Fujikawa ◽  
Yuichi Takikawa ◽  
Yasuhiro Inoue
Keyword(s):  
Pseudomonas Syringae ◽  
Leaf Spot ◽  
Draft Genome ◽  
Leaf Blight ◽  
Bacterial Leaf Blight ◽  
Genome Sequences ◽  
Bacterial Leaf Spot ◽  
Content Type

ABSTRACT Pseudomonas cannabina pv. alisalensis and Pseudomonas syringae pv. maculicola cause bacterial leaf blight and bacterial leaf spot of crucifers (Brassicaceae). Both pathogens are threats to the cultivation of cruciferous crops. Here, we sequenced two strains of each pathogen, which will contribute to the development of countermeasures for the above diseases.

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