Characterization of Isolates of Bacterial Blight of Cotton (Xanthomonas campestris pv. malvacearum) from Nicaragua

Seeds from cotton plants infected with Xanthomonas campestris pv. malvacearum were collected in different parts of Nicaragua in 1986. When the seeds were homogenized the pathogen could not always be identified by dilution plating. Therefore, an enrichment of bacteria on the natural host was induced before isolation. The cotton seeds were shaken with water and sand for 2 days and then sown in sand for germination. Rather often the developing cotyledons showed typical water-soaked spots, from which the pathogen could be isolated easily. This new method needed more time but made it possible to detect a low level of bacterial infestation. Altogether, 42 bacterial isolates were obtained. For inoculation experiments suspensions with 5x10⁵ CFU - ml^?1 were infiltrated into cotton leaves. Incubations of inoculated plants in growth chambers, but not in greenhouses, resulted in typical and uniform disease symptoms (water-soaked leaf spots). Nine of the ten cotton differentials tested were highly susceptible to all the 42 bacterial isolates. Since only line 101-102B proved to be resistant, the Nicaraguan isolates of bacterial blight of cotton were characterized as race 18, of the pathogen. The main cotton cultivars grown in Nicaragua (H-373 and G-286) were strongly affected by the isolated bacterial strains. In order to reduce the disease incidence in Nicaragua, the cultivation of resistant cotton varieties is suggested.     

Races of Xanthomonas campestris pv. malvacearum (Smith) Dye, the Causal Organism of Bacterial Blight of Cotton, in Nigeria

Races 6, 7 and 10 of Xanthomonas campestris pv. malvacearum (Smith) Dye, the causal organism of bacterial blight of cotton, were identified among twelve isolates of the pathogen from the three cotton growing zones of Nigeria. Races 7 and 10 were, however, predominant. The races were distinguished by inoculation of eight cotton lines known to differentiate through possession of different resistance genes. Further work is necessary, on a much greater number of isolates, to determine therange and distribution of different virulence types in Nigeria.     

Survival ofXanthomonas campestris pv.vesictoria in pepper seeds and roots in symptomless and dry leaves in non-host plants and in the soil

Summary A population ofXanthomonas campestris pv.vesicatoria developed as endophytes in the leaves and rhizosphere of apparently symptomless plants grown under mist but not under dry conditions. The pathogen survired for long periods on, and could be isolated from, the surface of infested dried seeds, inoculated sandy loam soil, dried leaves, and the rhizosphere of pepper and of other non-host plants. In addition, small numbers of the pathogen survived for 18 months in a field previously cropped with pepper diseased with bacterial scab. Healthy nursery or mature plants developed symptoms while growing in soil containing infested leaves, either buried or placed on the soil surface.     

Gene expression in Brassica campestris showing a hypersensitive response to the incompatible pathogen Xanthomonas campestris pv. vitians

Xanthomonas campestris pv. vitians, a pathogen of lettuce, elicits a hypersensitive response within 12 hours of inoculation into Brassica leaves, characterized by tissue collapse, loss of membrane integrity, vein blockage and melanin production. In contrast, the compatible pathogen, X. c. pv. campestris, has no visible effects on leaves for 48 hours, after which inoculated areas show chlorosis which eventually spreads, followed by rotting.mRNA was prepared from leaves inoculated with suspensions of both pathovars or with sterile medium up to 24 hours following inoculation. In vitro translation of total and poly A⁺ RNA in rabbit reticulocyte lysate in the presence of ³⁵S methionine followed by separation of the polypeptide products by 2D-PAGE, allowed comparison of the effects of these treatments on plant gene expression. Major changes in gene expression were observed as a consequence of the inoculation technique. In addition, after inoculation with X. c. vitians, up to fifteen additional major polypeptides appeared or greatly increased by four hours. Some of these had disappeared by nine hours and several more had appeared. No major polypeptides disappeared or decreased greatly in intensity following inoculation with X. c. vitians.     

In vitro evaluation of sugarcane resistance to gumming disease and of Xanthomonas campestris pv. vasculorum aggressiveness

Sugarcane plantlets were sectioned halfway between the base and the youngest ligule and then inoculated by soaking the wound in a suspension of Xanthomonas campestris pv. vasculorum. The infection caused rapid necrosis of the inoculated leaves, chlorosis of uninoculated leaves, or death of the inoculated plantlet. New tillers sometimes showed chlorosis or white streaks. The effects of the inoculum concentration, the cultivar, and the bacterial strain on symptom severity were determined. The ranking of cultivars depended on the inoculum concentration, and strains were found to differ with regard to aggressiveness. However, cultivars and strains were more effectively classified in greenhouse trials. The poor expression of leaf resistance appeared to limit the use of the in vitro test.     

Xanthomonas campestris pv.parthenii pathovar nov. incitant of leaf blight of parthenium

A new bacterial leaf blight disease of parthenium (Parthenium hysterophorus L.) is described for the first time. The disease-causing bacterium was isolated and its morphological, physiological and biochemical characters were determined. The pathogenicity of bacterium is apparently limited only to parthenium. The pathogen was identified asXanthomonas campestris pv.parthenii pathovar nov. on the basis of morphological, physiological, biochemical and pathogenic characteristics.     

A new,pathogen-inducible gene of Arabidopsis is expressed in an ecotype-specific manner

In this study we describe a novel gene, which was isolated in an attempt to search for specific plant resistance genes of Arabidopsis against isolates of the phytopathogenic bacterium Xanthomonas campestris pv. campestris. The gene was cloned by differential screening of a genomic library of the Xcc 750-resistant ecotype Col-0, using cDNA populations derived from ecotype Col-0 and the Xcc 750-susceptible ecotype Oy-0. The isolated gene, CXc750, is differentially expressed in ecotypes of Arabidopsis thaliana. In addition, although highly expressed in uninfected plants, gene expression increases in response to pathogen attack. CXc750 potentially codes for a small, basic protein of about 10 kDa. The predicted protein product contains a potential signal leader peptide at the amino-terminal end but no ER retention sequence and no further transmembrane domain. This indicates that the gene product is transported to other compartments or out of the cell.The possible function of CXc750 as a member of the plant defense response system is discussed.     

Effect of culture filtrates ofXanthomonas campestris pv.pelargonii and extracts of geranium stems inoculated withX. c. pv.pelargonii on geranium callus and seedlings

Experiments were designed to determine whetherXanthomonas campestris pv.pelargonii produces a toxin which induces symptoms of bacterial blight in geranium, and is active at the cellular level. Culture filtrates ofX. c. pv.pelargonii were prepared by ethyl acetate extraction and ultrafiltration of the aqueous fraction. Culture filtrates adjusted to several pH values induced maximum disease ratings on geranium seedlings in the pH range 7–10. Geranium callus growth was significantly reduced by the filtrate in the same pH range. An active fraction could also be isolated from diseased tissue. A thin-layer chromatography-callus bioassay system detected toxin activity in the culture filtrate and in extracts of geranium stems inoculated withX. c. pv.pelargonii. Callus growth inhibition was located at R_f = 0.2–0.3 for both sources of toxin. These results suggest thatX. c. pv.pelargonii produces a toxin which causes disease symptoms, is present in diseased tissues, and inhibits callus growth. This opens the possibility of developing resistance to this pathogen by selecting cells insensitive to the toxin and regenerating plants from these cells.     

Transformation of virulence factor among bacterial blight ofXanthomonas campestris pv.malvacearum in plants

Transformation of an avirulent strain of bacterial blight of the cotton pathogenXanthomonas campestris pv.malvacearum (Xcm) to a virulent strain was achieved in plants by using total (i. e. chromosomal and plasmid DNA) as well as chromosomal DNA, but not the plasmid DNA, of a virulent Xcm strain, thereby indicating that virulence was located in the chromosomal replica. The process of transformation began 1 d after inoculation of the DNA preparation with the avirulent strain in plant tissues and was completed within 2 d with a recombination frequency of 10⁻²−10⁻⁴.     

Impact of Host Resistance on the Variability in Virulence of Xanthomonas campestris pv. oryzae

Twenty isolates of Xanthomonas campestris pv. oryzae (Ishiyama) Dye each from susceptible and resistant rice cultivars, were inoculated on the susceptible genotype IR-8 and resistant M Sungsong to measure the impact of host selection pressure on virulence change. Isolates virulent on resistant genotype tended to be less adaptive on susceptible cultivars. While isolates from susceptible genotype showed 2.2 times more general virulence (GV) than specific virulence (SV), isolates of resistant host origin had only 1.3 times GV, indicating that the resistant host plant displays considerable increase in virulence. The SV value increased 1.64 times after one cropping with resistants indicated the potential of the pathogen to change to be slow.     

Differentiation between Xanthomonas campestris pv. graminis (ISPP List 1980), pv. phleipratensis (ISPP List 1980) emend., pv. poae Egli and Schmidt 1982 and pv. arrhenatheri Egli and Schmidt 1982, by Numerical Analysis of Phenotypic Features and Protein Gel Electrophoregrams

A numerical analysis of 257 phenotypic features of 45 bacterial isolates from grasses, revealed three phenons corresponding to (i) X. campestris pv. graminis (ISPP List 1980), (ii) X. campestris pv. phleipratensis (ISPP List 1980) and (iii) X. campestris pv. poae Egli and Schmidt 1982 and X. campestris pv. arrhenatheri Egli and Schmidt 1982. In each phenon, the strains clustered together regardless of the geographical origin of the isolates orthe year of isolation. Polyacrylamide gel electrophoresis of soluble proteins and host range studies, revealed four groups corresponding to the pathovars mentioned above. The four pathovars constitute definite biological entities that can be differentiated by phenotypic, gel electrophoretic and host range features.     

Bacterial Streak Disease of Maize (Zea mays L.) in South Africa

Bacterial streak disease of maize is currently causing some concern among breeders in South Africa. The causal organism of this previously undescribed disease was successfully isolated and its pathogenicity established using KoCH's postulates. Standard physiological and biochemical tests used to identify phytopathogenic bacteria indicated that the bacterium is a Xanthomonas campestris pathovar. Comparisons between this organism and other recognized X. campestris pathovars of the Poaceae indicated that apart from some minor differences the maize streak pathogen is physiologically similar to X. campestris pv. holcicola. However, in repeated reciprocal inoculation experiments all attempts to induce disease symptoms in sorghum with the maize streak pathogen were unsuccessful. Conversely, X. campestris pv. holcicola did produce symptoms in maize leaves. In all the maize cultivars tested the symptoms produced by the maize streak pathogen were, however, always considerably more severe than those caused by X. campestris pv. holcicola. Notwithstanding its physiological similarity to X. campestris pv. holicola it would appear that on the grounds of host specificity the maize streak pathogen warrants new pathovar status. The name X. campestris pv. zeae is proposed.     

DNA fingerprinting of Indian isolates of Xanthomonas oryzae pv. oryzae

 A high level of genetic polymorphism was detected among Indian isolates of Xanthomonas oryzae pv. oryzae using hypervariable probes such as a microsatellite oligonucleotide, probe (TG)₁₀, a human minisatellite probe, pV47, an avirulence gene probe, avrXa10 and a repeat clone, pBS101. These DNA probes detected multiple loci in the bacterial genome generating complex DNA fingerprints and differentiated all of the bacterial isolates. Analysis of fingerprints indicated that pV47, (TG)₁₀ and pBS101 have a lower probability of identical match than avrXa10 and therefore are potential probes for DNA fingerprinting and variability analysis of Xanthomonas oryzae pv. oryzae pathogen populations. Cluster analysis based on hybridization patterns using all of the above probes showed five groups at 56% similarity. Studies on the methylation patterns of isolates representing the three important races of X. oryzae pv. oryzae indicated more methylation in the most virulent isolate, suggesting a possible role of methylation in pathogenicity. Received: 8 December 1996 / Accepted: 20 December 1996     

Identification of Xanthomonas campestris pv. Juglandis from (Persian) English Walnut Nursery Trees in South Africa

Bacterial isolates producing yellowish colonies on Nutrient Agar were recovered from symptoms of suspect walnut blight disease on leaves of nursery trees in the southwestern Cape Province of South Africa. The isolates were identified by pathogenicity tests on leaves of walnut and plum trees in the greenhouse. Fifteen isolates from four cultivars at two nurseries produced typical lesions of blight on walnut and one isolate. typical lesions of bacterial spot disease on plum leaves. Cluster analysis was done on 28 characteristics recorded from colony growth. colour. form. and elevation on four different culture media, and starch hydrolysis on a semi-selective medium for the isolation of Xanthomonas campestris pv. juglandis. Total DNA of the isolates was digested with restriction endonuclease Spel and resolved by contour-clamped homogeneous electric field (CHEF) electrophoresis. Two phenotypic clusters were distinguished among the 15 South African and one reference strain of X.c.pv. juglandis at the 54%S_sm level. The isolate which induced disease symptoms on plum grouped with reference strains of Xanthomonas campestris pv. pruni in a third cluster. Two-thirds of the isolates were not characterized on the semi-selective medium for X.c. pv. juglandis. DNA restriction fragment banding patterns were similar for most isolates of X.c.juglandis in the same phenotypic cluster. However, DNA banding patterns were non-distinct for some isolates with similar phenotypic characters. Phenotypic characteristics and DNA restriction fragment banding patterns of the isolates were not correlated with geographical origin or cultivar specificity.     

Effect of phenolic substances on pigmentation and pathogenicity of Xanthomonas campestris pv. malvacearum

Summary Phenols inhibited the growth as well as pigment production by Xanthomonas campestris pv. malvacearum (Xcm). Albino (white) colonies were formed at 0.005M concentration of certain phenois, which retained their white colour on successive transfers on phenol containing media indefinitely. Both, albino and yellow isolates were pathogenic and maintained their race number.     

Detection of <Emphasis Type="Italic">Xanthomonas oryzae</Emphasis> pv. <Emphasis Type="Italic">oryzae</Emphasis> in seeds using a specific TaqMan probe

Xanthomonas oryzae pv. oryzae is the pathogen that causes bacterial leaf blight in rice. Bacterial leaf blight is the main cause for severe rice underproduction in many countries. However, with conventional methods it is difficult to quickly and reliably distinguish this pathogen from other closely related pathogenic bacteria, especially X. oryzae pv. oryzicola, the causal organism of bacterial leaf streak in rice. We have developed a novel and highly sensitive real-time method for the identification of this specific bacteria based on a TaqMan probe. This probe is designed to recognize the sequence of a putative siderophore receptor gene cds specific to X. oryzae pv. oryzae, and can be identified from either a bacterial culture or naturally infected rice seeds and leaves in only 2 h. The sensitivity of the method is 100 times higher than that of the current polymerase chain reaction (PCR) gel electrophoresis method for diagnosis.     

Latent infections of in vitro anthurium caused by Xanthomonas campestris pv. dieffenbachiae

Latent infections of tissue-cultured Anthurium andraeanum Lind. caused by the blight pathogen, Xanthomonas campestris pv. dieffenbachiae (McCulloch & Pirone) Dye, were examined. The pathogen survived in or on callus for over 4 months without producing symptoms in callus or turbidity in the medium. The pathogen survived for more than 1 year on or within stage II shoots without producing symptoms and was successively transferred three times as latently infected shoots were multiplied. The pathogen did not grow or survive for more than 2 weeks in Murashige and Skoog medium lacking plant material. The addition of coconut water enhanced bacterial growth and produced turbidity in culture media. Latently infected in vitro anthuriums may be inoculum sources for subsequent outbreaks of the disease.     

Pepper Tree (Schinus terebenthifolius Radii), a New Host Plant for Xanthomonas campestris pv. mangiferaeindicae

Apigmented bacterial colonies were obtained in Reunion Island from angular leaf lesions on Pepper tree (Schinus terebenthifolius Radii), a member of Anacardiaceae. All isolates were identified as Xanthomonas campestris, using physiological and biochemical tests. These strains were reinoculated to Pepper tree leaves, and Koch postulates were verified. Furthermore, they were inoculated to mango leaves and produced lesions identical to those induced by Xanthomonas campestris pv. mangiferaeindicae, the causal agent of bacterial black spot of mangoes. Apigmented and pigmented strains of X. c. pv. mangiferaeindicae from Mango and Ambarella were pathogenic to Pepper tree. Strains isolated from Pepper tree were compared to X. c. pv. mangiferaeindicae, by means of phenotypic features (utilization of 147 carbon sources) and using a serological assay. A high homology among the strains was observed. Thus, It is concluded that strains isolated from Pepper tree belong to pv. mangiferaeindicae, and that Pepper tree is a host species for X. c. pv. mangiferaeindicae.     

Biological control of rice bacterial blight by Lysobacter antibioticus strain 13-1

Bacterial leaf blight (BB) is a worldwide destructive rice disease caused by pathogen Xanthomonas oryzae pv. oryzae (Xoo). A novel strain of Lysobacter antibioticus, which was isolated from the rhizosphere of rice in Yunnan Province of China, can significantly inhibit the growth of various phytopathogenic bacteria and fungi, especially BB pathogen Xoo. In greenhouse experiments, whole bacterial broth culture (WBC) of strain 13-1 was more effective in reducing BB than other components of the culture, with disease suppression efficiency up to 69.7%. However, bacterial cells re-suspended in water, cell-free culture extracts, and heated cultures also significantly reduced BB severity. Suppression efficiencies ranged from 79.0% to 61.8% for undiluted to 100-fold dilution treatments and from 57.6% to 31.7% when the WBC of strain 13-1 (10⁸ CFU/mL) was applied at 3 days and 7 days prior to pathogen inoculation, respectively. In three field trials, strain 13-1 reduced BB incidence by 73.5%, 78.3%, and 59.1%, respectively. Disease suppression by strain 13-1 varied significantly among different rice cultivars, although efficacy was not directly related to the susceptibility level of the cultivars. Efficacy of biocontrol was also affected by different pathogen isolates, with some isolates of Xoo being more sensitive to 13-1 suppression than others. These results suggest that antibiotics and density of colonization on leaves may be involved for biological control of rice BB by strain 13-1. To our knowledge, this is the first report of L. antibioticus being a potential biocontrol agent for rice bacterial blight.