Reaction of biofortified lettuce genotypes to different strains of Xanthomonas campestris pv. vitians

Lettuce bacterial leaf spot caused by Xanthomonas campestris pv. vitians is an aggressive disease that is difficult to control. So far there are no reports of the reaction of biofortified lettuce genotypes to different isolates of the bacteria. Thus, the objective was to evaluate the aggressiveness of X. campestris pv. vitians, as well as the reaction of biofortified lettuce genotypes to bacterial spot. Two experiments were performed in two distinct seasons (winter and summer), in greenhouse at the Vegetable Experimental Station of the Federal University of Uberlândia (UFU). The experimental design in both experiments was a randomized block design, in a factor scheme of 5 × 4 (five genotypes and four strains), with four repetitions. Were evaluated the severity and the area under the disease progress curve. In general, the biofortified lettuce 'Uberlândia 10000' was more resistant to most bacterial strains in the summer cultivation, and in the winter period UFU 'Crespa 206'. The commercial cultivar Robusta was the most susceptible to the strains during both seasons. The UFU E125 strain was the most aggressive for most genotypes in both seasons.

Biopolymer Xanthan: A New Adjuvant for DNA Vaccines

Abstract DNA vaccines have been evaluated as an option to prevent several diseases. In this study, the capacity of the xanthan biopolymer to improve the DNA vaccines immune response, administered intramuscularly, was evaluated. The experimental vaccines consisted of genes encoding fragments of the proteins LigA and LigB of Leptospira interrogans serogroup Icterohaemorrhagiae serovar Copenhageni strain Fiocruz L1-130. The humoral immune response was evaluated by indirect ELISA. Cytokine expression levels were determined by RT-qPCR. Compared to the control group, the IgG antibody levels of animals immunized with pTARGET/ligAni and pTARGET/ligBrep plasmids associated with xanthan biopolymer were significantly higher than the control group. Additionally, there was a significant increase in IL-17 expression in animals vaccinated with pTARGET/ligBrep and xanthan.

MarR family transcription regulator HpaR and XC0449 coordinately regulate the virulence of Xanthomonas campestris pv. campestris / 生物工程学报

MarR family transcription regulators are ubiquitous among bacteria and archaea. They extensively control multiple cellular processes and elaborately regulate the expression of genes involved in virulence, stress response and antibiotics at translational level. In Xanthomonas campestris pv. campestris, insertional inactivation of MarR family transcription regulator HpaR (XC2827) resulted in significantly decrease in virulence and increase in the production of the extracellular proteases. Here, we reported that the genome of Xcc 8004 encodes nine MarR family transcription regulators. The MarR family transcription regulators, HpaR (XC2827) and XC0449, were heterologous expressed and purified. In vitro MST and Pull-down assay confirmed the physical interaction between HpaR and XC0449. Phenotypical assay determined that deletion of XC0449 resulted in substantial virulence attenuation. In vitro EMSA, in vivo qRT-PCR and GUS activity assay identified that HpaR and XC0449 coordinately act as the transcriptional activator to regulate the expression of the virulence-associated gene XC0705, and eventually control the bacterial virulence and the production of extracellular proteases.

Yield and rheological properties of exopolysaccharide from a local isolate: Xanthomonas axonopodis pv. vesicatoria

Background: The aim of the present study was to evaluate gum productivity of a local strain, Xanthomonas axonopodis pv. vesicatoria, isolated from pepper plant, and its rheological behavior for the first time compared to the standard strain, Xanthomonas campestris DSM 19000 (NRRL B-1459). The influence of operational conditions (agitation rate and inoculum volume) on gum production and rheological properties of gums from the Xanthomonas strains were investigated. Results: The isolated strain of Xanthomonas showed similar xanthan yield compared to the standard strain. Furthermore, this study clearly confirmed that gum yield depended on bacterial strain, agitation rate, and inoculum size. The most suitable conditions for the gum production in an orbital shaker in terms of agitation rate and inoculum size were 180 rpm and 5%, respectively, resulting in an average production of 10.96 and 11.19 g/L for X. axonopodis pv.vesicatoria and X. campestris DSM 19000, respectively. Regarding the rheological properties, Ostwald-de-Waele and power law models were used to describe flow and oscillatory behavior of the gum solutions, respectively. Consistency of the novel gum solution remarkably was much higher than the commercial xanthan gum solution. Flow and oscillatory behavior and their temperature ramps showed that weak gel-like structure could be obtained with less gum concentrations when the novel gum was used. Conclusion: Therefore, yield and technological properties of the aqueous solutions of the exopolysaccharide synthesized by X. axonopodis pv. vesicatoria were observed to be more suitable for industrial production.

Structure of xanthan gum and cell ultrastructure at different times of alkali stress

Abstract The effect of alkali stress on the yield, viscosity, gum structure, and cell ultrastructure of xanthan gum was evaluated at the end of fermentation process of xanthan production by Xanthomonas campestris pv. manihotis 280-95. Although greater xanthan production was observed after a 24 h-alkali stress process, a lower viscosity was observed when compared to the alkali stress-free gum, regardless of the alkali stress time. However, this outcome is not conclusive as further studies on gum purification are required to remove excess sodium, verify the efficiency loss and the consequent increase in the polymer viscosity. Alkali stress altered the structure of xanthan gum from a polygon-like shape to a star-like form. At the end of the fermentation, early structural changes in the bacterium were observed. After alkali stress, marked structural differences were observed in the cells. A more vacuolated cytoplasm and discontinuities in the membrane cells evidenced the cell lysis. Xanthan was observed in the form of concentric circles instead of agglomerates as observed prior to the alkali stress.

Statistical optimization of xanthan gum production and influence of airflow rates in lab-scale fermentor

The present study was undertaken to investigate and optimize the possibility of xanthan gum production by Xanthomonas campestris PTCC1473 in 500ml shake flasks on the second grade date palm. Using an experimental response surface methodology [RSM] coupled with a central composite design [CCD], three major independent variables [nitrogen source, phosphor source and agitation rate] were evaluated for their individual and interactive effects on biomass and xanthan gum production in submerged fermentation. The optimum conditions selected for gum production were 3.15 g.l[-1] for nitrogen source, 5.03 g.l[-1] for phosphor source, and 394.8 rpm for agitation rate. Reconfirmation test was conducted, and the experimental value obtained for xanthan production under optimum conditions was about 6.7[+2]/-0.26 g.l[-1], which was close to 6.51 g.l[-1] as predicted by the model. A higher yield of biomass production was obtained at 13.74 g.l[-1] for nitrogen source, 4.66 g.l[-1] for phosphor source, and 387.42 rpm for agitation rate. In the next stage, scale-up from the shake flasks to the 1-L batch fermentors was carried. By using the optimum conditions for xanthan gum, the biomass and xanthan gum concentrations after 72h in three levels of air flow rate [0.5, 1 and 1.5 vvm] were obtained as 3.98, 5.31 and 6.04 g.l[-1],and 11.32, 15.16 and 16.84 g.l[-1], respectively. Overall, the second grade date palm seemed to exhibit promising properties that can open new pathways for the production of efficient and cost-effective xanthan gum

Establishment of an inducing medium for type III effector secretion in Xanthomonas campestris pv. campestris

It is well known that the type III secretion system (T3SS) and type III (T3) effectors are essential for the pathogenicity of most bacterial phytopathogens and that the expression of T3SS and T3 effectors is suppressed in rich media but induced in minimal media and plants. To facilitate in-depth studies on T3SS and T3 effectors, it is crucial to establish a medium for T3 effector expression and secretion. Xanthomonas campestris pv. campestris (Xcc) is a model bacterium for studying plant-pathogen interactions. To date no medium for Xcc T3 effector secretion has been defined. Here, we compared four minimal media (MME, MMX, XVM2, and XOM2) which are reported for T3 expression induction in Xanthomonas spp. and found that MME is most efficient for expression and secretion of Xcc T3 effectors. By optimization of carbon and nitrogen sources and pH value based on MME, we established XCM1 medium, which is about 3 times stronger than MME for Xcc T3 effectors secretion. We further optimized the concentration of phosphate, calcium, and magnesium in XCM1 and found that XCM1 with a lower concentration of magnesium (renamed as XCM2) is about 10 times as efficient as XCM1 (meanwhile, about 30 times stronger than MME). Thus, we established an inducing medium XCM2 which is preferred for T3 effector secretion in Xcc.

Establishment of an inducing medium for type III effector secretion in Xanthomonas campestris pv. campestris

It is well known that the type III secretion system (T3SS) and type III (T3) effectors are essential for the pathogenicity of most bacterial phytopathogens and that the expression of T3SS and T3 effectors is suppressed in rich media but induced in minimal media and plants. To facilitate in-depth studies on T3SS and T3 effectors, it is crucial to establish a medium for T3 effector expression and secretion. Xanthomonas campestris pv. campestris (Xcc) is a model bacterium for studying plant-pathogen interactions. To date no medium for Xcc T3 effector secretion has been defined. Here, we compared four minimal media (MME, MMX, XVM2, and XOM2) which are reported for T3 expression induction in Xanthomonas spp. and found that MME is most efficient for expression and secretion of Xcc T3 effectors. By optimization of carbon and nitrogen sources and pH value based on MME, we established XCM1 medium, which is about 3 times stronger than MME for Xcc T3 effectors secretion. We further optimized the concentration of phosphate, calcium, and magnesium in XCM1 and found that XCM1 with a lower concentration of magnesium (renamed as XCM2) is about 10 times as efficient as XCM1 (meanwhile, about 30 times stronger than MME). Thus, we established an inducing medium XCM2 which is preferred for T3 effector secretion in Xcc.

[Comparison of bench-scale production of xanthan by xanthomonas campestris in solid state and submerged fermentation]

Xanthan gum is an important natural biopolymer with numerous applications in various technologies, specially food industry. In this research, microbial production of xanthan by Xanthomonas campestris PTCC1473 from sugarcane molasses and date sugar in submerged fermentation [SmF] and also dried date waste [cake produced after pressing] in solid state fermentation [SSF] were compared. The Plackett-Burmann design [PBD] was used in this study. Chemical composition and characteristics [dried cell weight, nitrogen, moisture, ash and pH] of the substrates were determined. Yeast malt broth [YMB] and yeast malt agar [YMA] were used as maintenance and inoculum preparation media, and incubation was performed in a shaker incubator [at 28degreeC, 72 h and 200 rpm]. The fermentation medium was centrifuged at 5degreeC and 21055 multiplied by g for 50 minutes and the supernatant separated from the pellet for further xanthan extraction. After precipitation of xanthan by isopropanol, resuspension and further purification by centrifuge [at 2056 x g], the xanthan dry weight was determined. The effects of several variables, including the kind and concentration of carbon [date sugar and sugarcane molasses], nitrogen [ammonium nitrate and diammonium phosphate] and phosphorus [KH2PO4], temperature, shaking, and size and age of inoculum, on the yield were determined. The most effective variables were found to be the type of carbon and nitrogen sources in the medium. It can be concluded that both the yield [% w/w of xanthan/consumed sugar] and productivity [g/g.day xanthan/consumed sugar] are higher in SmF [22.4 and 7.46] than in SSF [13.3 and 4.43]. In addition, date extract results in a higher productivity than date waste and sugarcane molasses. The xanthan yield could be increased by changing the composition and physical conditions of the culture medium

Influence of agitation and aeration in xanthan production by Xanthomonas campestris pv pruni strain 101 / Influencia de la agitación y la aireación en la producción de xantano por Xanthomonas campestris pv. pruni cepa 101

Production, viscosity, and chemical composition of xanthan synthesized by bacterium Xanthomonas campestris pv pruni strain 101 were evaluated in bioreactor systems. During the process, the volumetric oxygen mass transfer coefficient (kLa) and the biomass were determined and the pH was monitored. The cultures were grown in a 3 l bioreactor, with aeration and agitation varying as follows: conditions (A) 300 rpm, 3 vvm and (B) 200 rpm, 2 vvm, at 28 °C. Our results showed that gum production was dependent on kLa, with a maximum yield of 8.15 g/l at 300 rpm, 3 vvm, 54 h of fermentation, kLa 21.4/h, while biomass was not affected. All aqueous solutions of 3% (w/v) xanthans synthesized showed a pseudoplastic behavior. The highest viscosity was reached under the strongest aeration/agitation conditions. All xanthan samples contained glucose, mannose, rhamnose, and glucuronic acid as their main components. The highest agitation and aeration rates used under condition A (300 rpm and 3 vvm) favorably influenced the yield and viscosity of the xanthan produced by bacterium X. campestris pv pruni 101 at different fermentation times.

Se evaluó la producción, viscosidad y composición química del xantano sintetizado por la bacteria Xanthomonas campestris pv. pruni cepa 101 en un fermentador. Durante el proceso se controló el pH y se determinaron el coeficiente de transferencia de masa de oxígeno (kLa) y la producción de masa celular seca. Los cultivos se realizaron en un fermentador de 3 l variando la aireación y la agitación, en las siguientes condiciones: (A) 300 rpm, 3 vvm y (B) 200 rpm, 2 vvm; a 28 °C. Nuestros resultados mostraron que la producción de goma fue dependiente del kLa, con un rendimiento máximo de 8,15 g/l a 300 rpm y 3 vvm a las 54 h de fermentación, kLa de 21,4/h, mientras que la producción de biomasa no se afectó. Todas las soluciones acuosas de xantano al 3% (m/v) sintetizadas presentaron comportamiento pseudoplástico. La mayor viscosidad se alcanzó en la condición de aireación/agitación más intensa. Todas las muestras de xantano contenían glucosa, manosa, ramnosa y ácido glucurónico como constituyentes principales. La mayor tasa de agitación y aireación utilizada en la condición A (300 rpm y 3 vvm) influyó favorablemente en el rendimiento y la viscosidad del xantano producido por la bacteria X. campestris pv. pruni 101 a diferentes tiempos de fermentación.

Avaliação da eficácia da tintura etanólica de guaco (Mikania glomerata) no controle da podridão negra (Xanthomonas campestris pv. campestris) em couve-flor / Effect of alcoholic extract of guaco (Mikania glomerata) on the control of dark rot (Xanthomonas campestris pv. campestris) in cauliflower

Com a prática da irrigação e novos híbridos de couve-flor, é possível produzir durante todo o ano e com alta produtividade. Mas, a cultura tem sido afetada por doenças a exemplo da podridão negra causada por Xanthomonas campestris pv. campestris, fomentando novas pesquisas para seu controle. Com o objetivo de verificar o potencial de Mikania glomerata no controle dessa doença, a tintura etanólica 50 ºGL dessa planta medicinal foi avaliada quanto: atividade antimicrobiana in vitro através do crescimento bacteriano em tubos de ensaio contendo 100, 250, 500 e 1000 mg L-1 da tintura; indução de resistência local ou sistêmica em planta de couve-flor aos 25 dias de idade, em casa de vegetação, através da pulverização de tintura concomitantemente e três dias antes da inoculação com o patógeno, sendo água e calda bordaleza tratamentos controle; atividade de peroxidases em folhas tratadas e não tratadas de couve-flor, colhidas concomitantemente e as 24, 48, 72 h da pulverização da tintura e, após pulverização-inoculção. A tintura etanólica, in vitro, promoveu inibição no crescimento bacteriano, a partir da concentração de 250 mg L-1. Nas concentrações de 500 mg L-1 e 1000 mg L-1 foram observadas, respectivamente, 24 e 38 de inibição do crescimento bacteriano. Nas plantas de couve-flor foi observada redução da doença apenas em folhas tratadas com 100 e 500 mg L-1 de tintura, aplicada concomitantemente à inoculação, comportamento este semelhante ao da calda bordaleza, indicando que o controle através da tintura de guaco é através de atividade antimicrobiana direta. Ficou indicado que a indução de peroxidases ocorreu devido ao processo infeccioso e não em função dos tratamentos com tintura etanólica de guaco. Estes resultados indicam o potencial da tintura de guaco para o controle preventivo da podridão negra da couve-flor.

With the use of irrigation and new hybrids of cauliflower, it is possible to get production during all theyear with hight yield. However, the crop has been affected by diseases, as the dark rot caused by X.campestris pv. campestris. The objective of this research work was to study the potential of Mikaniaglomerata for the control of this disease. Alcoholic extract 50 ºGL of M. glomerata was evaluatedregarding to: in vitro antimicrobial activity through bacterial growth in 100, 250, 500 and 1000 mg L-1 ofthe alcoholic extract; induction of local or systemic resistance in 25 days old cauliflower, with spray ofalcoholic extract concomitantly and three days before the inoculation with the pathogen (water andbordeau mixture were used as control); peroxidases activity in leaves of cauliflower treated and nottreated, and harvested concomitantly, 24, 48 and 72 hours after spraying the alcoholic extract and alsoafter inoculation. The alcoholic extract of M. glomerata showed inhibition of the bacterial growth invitro at the concentrations of 250, 500 and 1000 mg L-1. The concentrations of 500 mg L-1 and 1000 mg L-1inhibited 24% and 38% of the bacterial growth. This inhibition could be due to antibacterial compoundsin the alcoholic extract. An inhibition of the disease in vivo occurred only in the leaves treated with 100and 500 mg L-1 of alcoholic extract when applied concomitantly with the bacteria. This result was similarto bordeau mixture, indicating a control by direct antimicrobial activity. There was no systemic resistenceinduction for all treatments. The peroxidases induction was due to infectious pathogen process and notto the treatments with alcoholic extract. The results indicate the potential of M. glomerata alcoholicextract for the preventive control of cauliflower dark rot disease

Meio semi-seletivo para isolamento de Xanthomonas campestris pv. viticola / Semi-selective medium for isolation of Xanthomonas campestris pv. viticola

O cancro bacteriano causado por Xanthomonas campestris pv. viticola é a fitobacteriose mais importante da videira no Submédio São Francisco. O isolamento de X. campestris pv. viticola de tecidos vegetais infectados é dificultado pela presença de contaminantes bacterianos, entre os quais Microbacterium barkeri. Objetivando-se a formulação de meio de cultura semi-seletivo, 22 isolados de X. campestris pv. viticola foram testados com relação a 30 antibióticos. O meio semi-seletivo NYDAM (extrato de carne 3, peptona 5, glicose 10, extrato de levedura 5, ágar 18 e ampicilina 0,1 em g L-1) inibiu M. barkeri e bactérias fitopatogênicas podendo ser utilizado para isolar X. campestris pv. viticola de hospedeiros com infecção natural em campo.

Technological application of an extracellular cell lytic enzyme in xanthan gum clarification

Uma enzima extracelular celulolítica produzida por Pseudomonas sp. foi ativa sobre células de Xanthomonas campestris mortas pelo calor. A atividade lítica causou a digestão enzimática de goma xantana de X. campestris. A digestão foi eficiente tanto para xantana nativa altamante viscosa (2,0 per center w/v) como para xantana comercial Sigma (2,5 per center w/v). Observações por microscopia eletrônica de varredura demonstraram a ação celulolítica sobre células de X. campestris.

The gene wxcA of Xanthomonas campestris pv. campestris 8004 strain involved in EPS yield / 生物工程学报

Xanthomonas campestris pv. campestris (Xcc), the pathogenic agent of black rot disease in cruciferous plants, produces large amount of extracellular polysaccharide (EPS), which has found wide applications in industry. For the great commercial value of the xanthan gum, many of the genes involved in EPS biosynthesis have been cloned and the mechanism of EPS biosynthesis also has been studied. In order to clone genes involved in EPS biosynthesis, Xcc wild-type strain 8004 was mutagenized with transposon Tn5 gusA5, and a number of EPS-defective mutants were isolated in our previous work. The Tn5 gusA5 inserted sites of these mutants were located by using thermal asymmetric interlaced PCR, and results showed that two EPS-defective mutants were insertion mutants of the gene wxcA which involved in lipopolysaccharide (LPS) biosynthesis. The gene wxcA involved in lipopolysaccharide biosynthesis but dose not extracellular polysaccharide in others' report. wxcA::Tn5 gusA5 mutant 021C12, the polar mutant, was complemented with recombinant plasmid pLATC8570 harboring an intact wxcA gene in this work, but the yield of EPS of the wxcA::Tn5 gusA5 mutant was not restored. In order to identify the function of wxcA gene of Xcc 8004 strain, the gene wxcA was deleted by gene replacement strategy, and the no-polar mutant of wxcA was obtained. DeltawxcA mutant strain, named Xcc 8570, was confirmed by using both PCR and southern analysis. Beside the LPS biosynthesis of deltawxcA mutant was affected, The EPS yield of deltawxcA mutant strain reduced by 50% as compared with the wild-type strain 8004. DeltawxcA mutant could be complemented in trans with the intact wxcA gene, and the EPS yield of the mutant was restored. The combined data showed that wxcA gene not only involved in LPS biosynthesis but also EPS yield in Xcc 8004 strain.

Mutageneisis of xanthomonas campestris and selection of strains with enhanced xanthan production

Xanthan gum is a microbial polysaccharide of great commercial importance as it has unusual rheological properties in solution and consequent range of applications. In this study, a series of mutants were isolated from Xanthomonas campestris PTCC 1473 by ethyl methanesulfonate mutagenesis. The polysaccharide yield of one mutant, XC1473E2, was 30% better than that of the parent strain. It also showed higher xanthan formation and glucose consumption rates compared to the parent strain. Xanthan produced by the mutant had enhanced viscosity, higher pseudoplasticity and larger molecular weight. Since mutant XC1473E2 appeared white on agar plates, it underwent pigment extraction with methanol. Contrary to the parent strain, the mutant showed no absorption at 443nm, i.e. the wavelength related to yellow pigment. This finding suggested that yellow pigmentation and normal xanthan biosynthesis are not necessarily concurrent. In general, mutant XC1473E2 seems to be a strain with interesting characteristics for use in commercial production of xanthan

Identification and cloning of a novel gene involved in EPS biosynthesis of Xanthomonas campestris pv. campestris / 生物工程学报

Xanthomonas campestris pv. campestris ( Xcc), causative agent of the black rot disease of cruciferous crops worldwide, produces large amount of extracellular polysaccharide( EPS), which has found wide applications in industry. In order to clone genes involved in EPS biosynthesis, Xcc wild-type strain 8004 was mutagenized with transposon Tn5gus A5, and a number of EPS-defective mutants were isolated. The Tn5gusA5 insertion sites in the mutants were analyzed by using thermal asymmetric interlaced PCR(TAIL-PCR), and the corresponding genes were identified by homology blast to the completely sequenced genome of Xcc 8004 strain. A novel gene, waxE, identified from the EPS-defective mutant 151D09, was found to be disrupted by the insertion of Tn5gusA5 in the open reading frame(ORF) with genome coordinates 4478998bp to 4479819bp.This gene showed 52% similarity to the kdtX gene of Serratia marcescens and 50% to the waaE of Klebsiella pneumoniae at amino acid level, with characteristics of glycostransferase 2 family domain. In order to identify the function of waxE gene, waxE gene deletion mutant of Xcc 8004 was constructed by gene replacement strategy in which waxE gene of genome was replaced by kanamycin resistant gene kan. The waxE gene deletion mutant strain, named Xcc 8570, was confirmed by both PCR and southern analysis. The growth rate of the deletion mutant 8570 in rich medium was not affected, but the EPS yield reduced by 35% as compared with the wildtype strain 8004. The deletion mutant could be completmented in trans with plasmid pLATC8976 harboring an intact waxE gene, and the EPS yield of the mutant was restored. The combined data showed that waxE gene involved in EPS biosynthesis in Xcc.

Cloning of Xanthomonas campestris pv. campestris pathogenicity-related gene sequences by TAIL-PCR / 生物工程学报

Southern blot analysis with probe from mini-Tn5 gfp-km transposon indicated that 5 non-pathogenic mutants which were generated by insertion of mini-Tn5 gfp-km mutagenesis contained a single copy of the transposon. Using genomic DNA of each mutant as a template, TAIL-PCR was performed with seven arbitrary degenerate (AD) primers pairing with 3 nested specific primers designed based on the sequence of GFP toward outside in mini-Tn5 gfp-km. After 3-step PCR reactions, the flanking sequence of each mutant was obtained. The PCR product was ligated with pGEM-T EASY vector and then was transformed into E. coli DH5 alpha by electroporation. Positive clones were selected by white/blue colony and plasmid was isolated, then digested with EcoRI. Plasmid was sequenced if its insert was longer than 300 bp. Our results indicated that TAIL-PCR was proved to be a simple and efficient approach in identification of gene using insertion mutagenesis.

Effect of virulence and serial transfers of Xanthonomas campestris on xanthan gum production

The virulence of six Xanthomonas campestris isolates was evaluated using the percentage of lesion area of leaves in Brassica oleraceae host plant, compared to diameter of colonies, xantham production and gum viscosity. In terms of virulence, the isolates belonged to two statiscally different groups: isolates B, UPF and C(7), showed values between 52 and 69(per cent), while isolates CF, C and strain B-1459 gave 0-30 (per cent) of lesion area. Final xanthan concentration, gum viscosity and colony diameter did not correlate with virulence calculated by percentage of lesion area, showing that this parameter is not a good criterium for selection of potential xantham producer isolates. Serial transfers of X. campestris isolates in host plant did not show a significant effect on in vitro production of xanthan or on viscosity levels, suggesting that the increasing interaction between plant and bacteria did not stimulate the increase in xanthan production and viscosity.

Antagonism of Yeasts to Xanthomonas Campestris PV. Campestris on Cabbage Phylloplane in field

Twenty yeast isolates, obtained from cabbage phylloplane, were evaluated for antagonistic activity against Xanthomonas campestris pv. campestris, in field. Plants of cabbage ev. 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 per cent) was also calculated. Yeast isolates LR32, LR42 and LR19 showed, respectively, 72, 75 and 79 (per cent) 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 per cent), LR42 (67 per cent), LR35 (69 per cent) and LR19 (68 per cent) in the treatments T7, T4, T5, and T6, which significantly differed from the others. The same yeast antagonists were also tested for back rot control using different cabbage cultivars (Fuyutoyo, Master-325, Matsukaze, Midori, Sekai I and Red Winner). The DSRs varied from 58 to 61 (per cent), and there was no significant difference among cultivars