Molecular and phenotypic characterization of Xanthomonas campestris pv. campestris causing black rot in Brassica crops in Brazil

Author(s):  
Josefa Neiane Goulart Batista ◽  
Marisa Alvares da Silva Velloso Ferreira ◽  
Alice Maria Quezado-Duval
2021 ◽  
Vol 60 (1) ◽  
pp. 51-62
Author(s):  
Samia LAALA ◽  
Sophie CESBRON ◽  
Mohamed KERKOUD ◽  
Franco VALENTINI ◽  
Zouaoui BOUZNAD ◽  
...  

Xanthomonas campestris pv. campestris (Xcc) causes the black rot of cruciferous plants. This seed-borne bacterium is considered as the most destructive disease to cruciferous crops. Although sources of contamination are various, seeds are the main source of transmission. Typical symptoms of black rot were first observed in 2011 on cabbage and cauliflower fields in the main production areas of Algeria. Leaf samples displaying typical symptoms were collected during 2011 to 2014, and 170 strains were isolated from 45 commercial fields. Xcc isolates were very homogeneous in morphological, physiological and biochemical characteristics similar to reference strains, and gave positive pathogenicity and molecular test results (multiplex PCR with specific primers). This is the first record of Xcc in Algeria. Genetic diversity within the isolates was assessed in comparison with strains isolated elsewhere. A multilocus sequence analysis based on two housekeeping genes (gyrB and rpoD) was carried out on 77 strains representative isolates. The isolates grouped into 20 haplotypes defined with 68 polymorphic sites. The phylogenetic tree obtained showed that Xcc is in two groups, and all Algerian strains clustered in group 1 in three subgroups. No relationships were detected between haplotypes and the origins of the seed lots, the varieties of host cabbage, the years of isolation and agroclimatic regions.


2008 ◽  
Vol 21 (8) ◽  
pp. 1128-1138 ◽  
Author(s):  
Wei Qian ◽  
Zhong-Ji Han ◽  
Jun Tao ◽  
Chaozu He

The gram-negative bacterium Xanthomonas campestris pv. campestris is the causal agent of black rot disease of cruciferous plants. Its genome encodes a large repertoire of two-component signal transduction systems (TCSTSs), which consist of histidine kinases and response regulators (RR) to monitor and respond to environmental stimuli. To investigate the biological functions of these TCSTS genes, we aimed to inactivate all 54 RR genes in X. campestris pv. campestris ATCC 33913, and successfully generated 51 viable mutants using the insertion inactivation method. Plant inoculation identified two novel response regulator genes (XCC1958 and XCC3107) that are involved in virulence of this strain. Genetic complementation demonstrated that XCC3107, designated as vgrR (virulence and growth regulator), also affects bacterial growth and activity of extracellular proteases. In addition, we assessed the survival of these mutants under various stresses, including osmotic stress, high sodium concentration, heat shock, and sodium dodecyl sulfate exposure, and identified a number of genes that may be involved in the general stress response of X. campestris pv. campestris. Mutagenesis and phenotypic characterization of RR genes in this study will facilitate future studies on signaling networks in this important phytopathogenic bacterium.


Plant Disease ◽  
2008 ◽  
Vol 92 (7) ◽  
pp. 1134-1134 ◽  
Author(s):  
W. P. Wechter ◽  
A. P. Keinath ◽  
J. P. Smith ◽  
M. W. Farnham

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.


2017 ◽  
Vol 83 (6) ◽  
pp. 373-381 ◽  
Author(s):  
Hirofumi Nagai ◽  
Noriyuki Miyake ◽  
Shinro Kato ◽  
Daisuke Maekawa ◽  
Yasuhiro Inoue ◽  
...  

1999 ◽  
Vol 30 (3) ◽  
pp. 191-195 ◽  
Author(s):  
Sayonara M.P. Assis ◽  
Rosa L.R. Mariano ◽  
Sami J. Michereff ◽  
Gil Silva ◽  
Elizabeth A.A. Maranhão

Twenty yeast isolates, obtained from cabbage phylloplane, were evaluated for antagonistic activity against Xanthomonas campestris pv. campestris, in field. Plants of cabbage cv. Midori were pulverized simultaneously with suspensions of antagonists and pathogen. After 10 days, plants were evaluated through percentage of foliar area with lesions. Percentage of disease severity reduction (DSR%) was also calculated. Yeast isolates LR32, LR42 and LR19 showed, respectively, 72, 75 and 79% of DSR. These antagonists were tested in seven different application periods in relation to pathogen inoculation (T1=4 d before; T2=simultaneously; T3=4 d after; T4=4 d before + simultaneously; T5=4 d after + simultaneously; T6=4 d before + 4 d after; T7=4 d before + simultaneously + 4 d after). The highest DSRs were showed by LR42 (71%), LR42 (67%), LR35 (69%) and LR19 (68%) in the treatments T7, T4, T5 and T6, which significantly differed from the others. The same yeast antagonists were also tested for black rot control using different cabbage cultivars (Fuyutoyo, Master-325, Matsukaze, Midori, Sekai I and Red Winner). The DSRs varied from 58 to 61%, and there was no significant difference among cultivars.


2000 ◽  
Vol 78 (9) ◽  
pp. 1144-1149 ◽  
Author(s):  
P A Gay ◽  
S Tuzun

The physiological mechanisms associated with resistance of cabbage to black rot disease seem to be associated with the hydathodes. To investigate the role of hydathodes in disease resistance, total peroxidase activities, anionic peroxidase isozyme expression, and lignin deposition were determined in hydathodes of resistant (Hancock and Green Cup), partially resistant (Cheers), and susceptible (Strukton and Perfect Ball) cabbage varieties (Brassica oleracea L.) during pathogenesis with Xanthomonas campestris pv. campestris. Four-week-old plants were petiole-inoculated with a highly virulent strain of X. campestris pv. campestris (FD91L). Hydathodal fluids were collected daily over a 14-day period from infected plants as well as noninfected, mock-inoculated control plants. Hydathodal fluids of resistant varieties had greater peroxidase activity when compared to susceptible ones, with infected plants having higher peroxidase levels than noninfected plants. Isoelectric focusing revealed the presence of four anionic peroxidase isozymes in hydathodal fluids, with the most anionic one (pI of 3.6) accumulating only upon infection. Lignin deposition in and around the hydathodes was associated with the accumulation of this particular isozyme in hydathodal fluids. The evidence suggests that a rapid, systemic response is associated with resistance to the bacterial pathogen.Key words: peroxidases, hydathodes, isozymes, black rot disease, cabbage, Xanthomonas campestris pv. campestris.


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