Response of black rot in cabbage to spatial distribution of inoculum

Disease progress of black rot in cabbage crop was studied over three years in field plots to compare the effects of uni-focal and multi-focal inoculum applied in equal amounts per plot. Disease progress (plant incidence and leaf incidence) was plotted over time, three dimensional maps were made, and disease aggregation was studied by means of geostatistics, black-black counts and Moran's I statistic. Black rot progress was primarily due to focus expansion. Secondary foci may appear at short distances from the initial focus but they usually merge with the expanding initial focus. Anisotropy occurred occasionally but was of minor importance. Disease proceeds faster in plots with multi-focal inoculation than in those with uni-focal inoculation. Probably, serious epidemics in Dutch cabbage fields originate from large numbers of foci.     

Survival and Extinction of Xanthomonas campestris pv. campestris in Soil

Carry-over of inoculum of X.c. pv. campestris in the soil from one cropping season to the next was studied in field experiments over three years. These studies were supported by laboratory and greenhouse experiments on quantitative assessment of bacteria by bioassay using the Most Probable Number technique, and on recovery rates of bacteria from the soil. The mean recovery rate from artificially infested soil was 58%. Extinction of X.c. pv. campestris in soil infested with infected plant debris proceeded exponentially and extinction rates depended on temperature, as did the decomposition of plant debris. In replicated field plots, over three years, infection foci of black rot disease were established. At harvest time, all plants were chopped and resulting plant debris was rotovated into the soil. The resulting soil infestation was sampled and showed clear infestation foci reflecting the original infection foci of the crop. These infestation foci decreased with time and disappeared after the winter. Follow-up crops remained virtually uninfected. The results show that in The Netherlands good crop and soil management impedes survival of inoculum from one year to the next, so that cabbage can be grown continuously. Polyetic carry-over of inoculum by debris in the soil can be avoided in The Netherlands.     

我国部分省域甘蓝主要产区黑腐病菌鉴定及致病力分析

 为了明确我国甘蓝主栽区黑腐病菌株种类与致病力水平,为优化抗病性鉴定方法、筛选抗病品种及抗病品种的合理布局提供依据,分别采集了10个省域的黑腐病病叶,经过分离纯化与分子鉴定,获得了26个典型甘蓝黑腐病菌株;选用5个甘蓝抗、感品种作为鉴别寄主,以甘蓝黑腐病菌株的悬浮液为接种体,采用喷雾接种法测定致病力。结果表明:来源于不同省域的黑腐病菌株致病力存在显著差异,致病力最强的是YU,病情指数平均值为38.31;致病力最弱的是SH,病情指数平均值为17.03。来源于不同海拔高度的甘蓝黑腐病菌株致病力最强的是G1500,病情指数平均值为37.93;最弱的是G800,病情指数平均值为19.80。根据鉴别寄主抗感反应将26个黑腐病菌株划分为12类致病型,发现来自不同地域的甘蓝黑腐病菌株间存在着显著的致病性分化现象,对甘蓝品种的致病力存在着显著性差异,其中菌株YU致病力最强,菌株YU可作为甘蓝抗病性鉴定的主要致病菌株,指导抗病品种的选育。     

我国部分省域甘蓝主要产区黑腐病菌鉴定及致病力分析

 为了明确我国甘蓝主栽区黑腐病菌株种类与致病力水平,为优化抗病性鉴定方法、筛选抗病品种及抗病品种的合理布局提供依据,分别采集了10个省域的黑腐病病叶,经过分离纯化与分子鉴定,获得了26个典型甘蓝黑腐病菌株;选用5个甘蓝抗、感品种作为鉴别寄主,以甘蓝黑腐病菌株的悬浮液为接种体,采用喷雾接种法测定致病力。结果表明:来源于不同省域的黑腐病菌株致病力存在显著差异,致病力最强的是YU,病情指数平均值为38.31;致病力最弱的是SH,病情指数平均值为17.03。来源于不同海拔高度的甘蓝黑腐病菌株致病力最强的是G1500,病情指数平均值为37.93;最弱的是G800,病情指数平均值为19.80。根据鉴别寄主抗感反应将26个黑腐病菌株划分为12类致病型,发现来自不同地域的甘蓝黑腐病菌株间存在着显著的致病性分化现象,对甘蓝品种的致病力存在着显著性差异,其中菌株YU致病力最强,菌株YU可作为甘蓝抗病性鉴定的主要致病菌株,指导抗病品种的选育。     

Symptomless Spread of Blight-inducing Strains of Xanthomonas campestris pv. campestris on Cabbage Seedlings in Misted Seedbeds

Xanthomonas campestris pv. campestris induces two types of symptoms, namely, black rot and blight. Black rot symptoms are V-shaped lesions and black veins on the leaf, and blight symptoms are sudden collapse of interveinal tissues following the lack of veinal necrosis at early stages of infection. These two symptoms can occur simultaneously. However, the tendency to induce either symptom type is strain-dependent. Six strains were evaluated for their rate and pattern of spread in misted seedbeds by using strain-specific monoclonal antibodies and miniplate enrichment/ELISA. Data on pathogen incidence was defined as the presence of the pathogen in or on plants rather than visual symptoms. The results indicated that blight-inducing strains spread to more seedlings than black rot-inducing strains. The high incidences of blight-inducing strains in experimental plots were associated with non-randomness of spatial pattern of pathogen spread, indicating that high incidence is primarily due to the spread from adjacent plants by leaf contact and water splash. Most ELISA-positive seedlings were symptomless, indicating that the sensitivity of the system used in this study was adequate for detection of latent or epiphytic spread.     

Biochemical and molecular characterization of Bacillus amyloliquefaciens, B. subtilis and B. pumilus isolates with distinct antagonistic potential against Xanthomonas campestris pv. campestris

Fifty-one Bacillus isolates were characterized by fatty acid methyl ester (FAME) analysis; universal primer polymerase chain reaction (UP-PCR) fingerprinting; production of secondary metabolites and antagonistic activity against Xanthomonas campestris pv. campestris (causal agent of black rot in cabbage) in vitro and in vivo . Based on FAME analysis and/or PCR fingerprinting, the isolates were clustered into three different groups, named as Bacillus amyloliquefaciens , B . subtilis and B . pumilus . Seed treatment with Bacillus spp. generally reduced germination of seeds and incidence of black rot, but no relationship was found between the results of in vitro and in vivo experiments. The B .  amyloliquefaciens group contained isolates that were generally the most effective at reducing attack of black rot in vivo . The metabolic profiles of these isolates suggested that they produced surfactin, iturin, bacillomycine and/or azalomycin F. Isolates belonging to the B . subtilis group were mostly able to synthesize surfactin and arthrobactin. Surfactin, amphomycin, arthrobactin and valinomycin were generally found in culture extracts of isolates belonging to the B . pumilus group. No effect on growth of the pathogen was detected when the activity of filtered culture extracts and selected metabolites produced by the three different Bacillus species was tested in vitro against X . c . pv. campestris . However, inhibition was seen when bacterial liquid cultures were used. When the ability to colonize cabbage endophytically was examined for seven selected isolates with different antagonistic potential against black rot, it was found that the ability was related to the species and not to the antagonistic activity of the isolates.     

Splash dispersal of conidia of Mycocentrospora acerina in the field

The dispersal of conidia of Mycocentrospora acerina was studied in caraway field trials. A Burkard spore trap, rotorods, inverted Petri dishes containing sucrose agar and rain gauges were used to trap conidia of M. acerina . Sporulation was stimulated by rainfall (2 mm) and moderate temperatures (around 15°C). Solar radiation had a negative effect on sporulation. Hardly any conidia were found in the spore traps on rainless days. Short distance (9 m) spread of M. acerina is mainly caused by splash dispersal of its conidia. Trap plants at 0, 0.1, 1 and 4 m from the inoculum source were readily infected under moist conditions. Beyond 9 m from an inoculum source no infection of caraway trap plants was found. Trap plants at 9 m from an inoculum source were infected in one out of three seasons only. Long distance (>9 m) spread could not be demonstrated by the techniques used in this study. The results suggest that, usually, a caraway field is infected by inoculum sources within that field.     

Spatial and temporal spread of powdery mildew (Erysiphe pisi) in peas (Pisum sativum) varying in quantitative resistance

A field experiment was conducted to assess the progress in time and spread in space of powdery mildew (caused by Erysiphe pisi ) in pea ( Pisum sativum ) cultivars differing in resistance to the disease. Disease severity (proportion of leaf area infected) was measured in 19 × 23 m plots of cultivars Pania and Bolero (both susceptible) and Quantum (quantitatively resistant). Inoculum on infected plants was introduced into the centre of each plot. Leaves (nodes) were divided into three groups within the canopy (lower, middle, upper) at each assessment because of the large range in disease severity vertically within the plants. Disease severity on leaves at upper nodes was less than 4% until the final assessment 35 days after inoculation. Exponential disease progress curves were fitted to disease severity data from leaves at middle nodes. The mean disease relative growth rate was greater on Quantum than on Pania or Bolero, but it was delayed, resulting in an overall lower disease severity on Quantum. Gompertz growth curves were fitted to disease progress on leaves at lower nodes. Disease progress on Quantum was delayed compared with Pania and Bolero. The average daily rates of increase in disease severity from Gompertz curves did not differ between the cultivars on these leaves. Disease gradients in the plots from the inoculum focus to 12 m were detected at early stages of the epidemic, but the effects of background inoculum inputs and the rate of disease progress meant that these gradients decreased with time as the disease epidemic intensified. Spread was rapid, and there were no statistically significant differences between cultivar isopathic rates (Pania 2.2, Quantum 2.9 and Bolero 4.0 m d⁻¹).     

Growth of Xanthomonas campestris pv. campestris populations at constant and variable temperatures

Quantitative data were collected to describe the relation between temperature and growth of the cabbage black rot pathogen,Xanthomonas campestris pv.campestris (Xcc). Relative growth rates derived from experiments at constant temperatures were used in dynamic simulation of bacterial population development. The relative growth rates were adequate to simulate growth ofXcc populations at constant temperatures but overestimated growth of populations at variable temperatures. This finding gives rise to the hypothesis, that under field conditions, disease development is slower than is expected on the basis of growth parameters obtained from studies with constant temperatures.     

Modelling the spread of Xanthomonas campestris pv. campestris in module-raised brassica transplants

A series of experiments was performed to quantify the rate of dispersal of Xanthomonas campestris pv. campestris in module-raised brassica transplants, in a simulated commercial plant-raising system. Seeds were sown in '308' module seed trays and set out in blocks in the glasshouse. Primary inoculum was introduced as inoculated seeds sown in one or more cells. Trays were watered via an overhead-gantry irrigation system, hand-watered or capillary-watered. Disease symptoms were monitored visually and the presence of the pathogen on samples of plants was monitored by leaf washing, dilution and plating on selective medium. Spread of symptoms was greatest in the gantry-watered trays, was very limited in hand-watered trays and was almost non-existent in capillary-watered trays. Dispersal of bacteria followed a similar pattern, but the proportion of plants contaminated was much greater than the proportion showing symptoms, and approached 100% after six weeks in the gantry-watered trays within 50 plants distance from a single primary infector. Models relating the proportion of plants with symptoms, or contaminated, to the distance from primary infector and time since sowing were fitted to the data. Predictions of the proportions of plants contaminated in commercial-scale blocks of transplants suggested that high levels of disease in the field could be explained by rapid rates of pathogen spread during plant-raising, and that the widely-used tolerance standard for seed health testing (0·01%) should be revised to 0·004%. In addition to seed health testing, control should focus on raising transplants under conditions that minimise the rates of disease spread and pathogen dispersal.     

Growth of Xanthomonas campestris pv. campestris populations at constant and variable temperatures

Quantitative data were collected to describe the relation between temperature and growth of the cabbage black rot pathogen,Xanthomonas campestris pv.campestris (Xcc). Relative growth rates derived from experiments at constant temperatures were used in dynamic simulation of bacterial population development. The relative growth rates were adequate to simulate growth ofXcc populations at constant temperatures but overestimated growth of populations at variable temperatures. This finding gives rise to the hypothesis, that under field conditions, disease development is slower than is expected on the basis of growth parameters obtained from studies with constant temperatures.     

Effects of inoculum potential, shading and soil temperature on root infection of oil palm seedlings by the basal stem rot pathogen Ganoderma boninense

Ganoderma boninense causes severe losses to oil palm in South East Asia. The disease typically manifests itself as basal stem rot, but there remains controversy over the route of infection and source of inoculum. Using isolates differing in aggressiveness, infection via roots was confirmed; it was also shown that large inoculum provided as Ganoderma -infested palm- or rubber-wood blocks (12 × 6 × 6 cm) is necessary for soil infection of seedlings after 6–8 months. Smaller blocks (3 × 3 × 3 cm) produced rapid (≤ 3 months) infection of roots and lower stem when physically attached to roots. Therefore fragmentation of infested palm wood from a felled, mature plantation before subsequent replanting may provide inoculum. Failure of G. boninense to grow through non-sterile soil or organic debris from frond bases, suggests it is a poor competitor and that roots must contact inoculum directly. Severe disease occurred after 8 months on inoculated seedlings under shade, but not on seedlings exposed to sun. Soil temperatures in sunlight frequently rose above 40°C and reached 45°C, whereas in shade they never exceeded 32°C. Ganoderma boninense is probably inhibited in exposed soil since optimal growth in vitro was 25–30°C, and there was no recovery from 45°C. Soil temperature may explain why symptoms often first appear in mature plantations when canopy formation creates shade. Infection is not peculiar to senescing palms but can occur throughout the growth cycle.     

PCR-based detection of Xanthomonas campestris pathovars in Brassica seed

Xanthomonas campestris is a seedborne bacterium that causes black rot of crucifers. Substantial crop losses may result from the rapid spread of the bacteria under favourable conditions, especially those occurring during seedling production. A PCR-based method has been developed for the rapid and sensitive detection of the pathovars of X. campestris that affect crucifers. Primers were designed to specifically amplify a 619 bp fragment of the hrpF gene from X. campestris . Amplification products were not detected from other Xanthomonas species, or from other pathogenic or epiphytic bacteria occurring on these plants. To avoid false-negative results arising from the presence of amplification inhibitors in plant extracts, primers targeting a 360 bp section of the internal transcribed spacer (ITS) region from Brassica spp. were included in a multiplex PCR. The assay readily detected X. campestris pv. campestris infections in diseased plants and from bacterial colonies isolated on growth media, and was more sensitive and specific than traditional plating methods and a commercially available ELISA. A seed-washing protocol was optimized to allow the detection of a single artificially infected seed among 10 000 healthy seeds using the multiplex PCR.     

Population Dynamics of Fusarium Oxysporum f. Sp. Radicis-lycopersici in Relation to the Onset of Fusarium Crown and Root Rot of Tomato

Fusarium oxysporum f. sp. radicis-lycopersici the causal agent of crown and root rot in tomato comprises two overlapping separate phases: monocyclic and polycyclic. Oversummering inoculum is the source of primary infection (the monocyclic phase) and the spread from plant to plant via root-to-root contact is the source of the secondary infection (the polycyclic phase). In the present work, relationships between initial inoculum density, population dynamics of the pathogen in the root zone of diseased plants, and disease onset were studied. For the monocyclic phase, 55.1% of the variance of disease onset was attributed to the rate of pathogen proliferation in the root zone of plants, and only 12.8% of the variance was attributed to the amount of initial inoculum density. For the polycyclic phase, disease onset was not related to either initial inoculum density or the rate of pathogen proliferation in the root zone. At disease onset, the inoculum density of the pathogen in the root zone of plants infected from oversummering inoculum reached an average of 4.08 log cfu g soil^–1. The inoculum density of the pathogen in the root zone of plants infected by their diseased neighbors was 3.23 log cfu g soil^–1. A large variation in pathogen proliferation rate in the root zone was found among individual plants, suggesting that differences in the level of soil suppressiveness may occur not only between fields, but even in the same field over short distances.     

Overwintering of Erwinia carotovora subsp. carotovora in Diseased Tissues in Soil and Its Role as Inoculum for Soft Rot of Chinese Cabbage (Brassica campestris, Pekinensis Group)

To examine whether the pathogenic bacterium, Erwinia carotovora subsp. carotovora, causal agent of soft rot of Chinese cabbage (Brassica campestris L., pekinensis group), can overwinter in plant debris and soil and serve as inoculum the following year, we monitored field populations of rifampicin-resistant, phage-sensitive strains of the bacterium. Chinese cabbage (cv. Matsushima Kohai W1116) were planted in field soil in pots that were sunk into the field on Aug. 2, 1996 and eventually reduced to one plant per pot. Outer petioles of the plants were inoculated with mixture of 13 bacterial strains of E. carotovora subsp. carotovora on Sept.5, 1996. After the soft rot spread throughout the plant, the diseased plant was buried in the potted soil. New seeds were sown in the pots on April 30, 1997, and the disease was observed in June and July. The bacterial strains were re-isolated from the potted soil, diseased tissue and rhizosphere soil by the dilution plating method on modified Drigalski's medium containing 100 ppm rifampicin and by the enrichment technique. In addition to rifampicin resistance, phage sensitivities of some of the re-isolated strains were identical to those of the strains buried in the soil with the diseased plant in the previous year. From these results, some of the 13 strains overwintered in the soil and infested plant tissue and acted as primary inoculum the following year. The frequency of re-isolation varied among the strains, perhaps because of competition among the strains, differences in epidemiological behavior and stabilizing selection among the strains, and the presence of different ecotypes of the organism. Received 21 July 2000/ Accepted in revised form 19 September 2000     

Pathogenicity assays restrict the species Xanthomonas campestris into three pathovars and reveal nine races within X. campestris pv. campestris

The species Xanthomonas campestris (Vauterin) groups bacteria associated with cruciferous plants. In order to clarify and refine the pathovar and race structures within X . campestris , 47 representative strains of six pathovars were characterized for their pathogenicity on a large host range of Brassicaceae, including all original hosts. Three diseases were observed on tested plants: (i) black rot disease on cruciferous plants; it was proposed that all strains causing black rot on at least one cruciferous plant be grouped in the single pathovar X . c . pv. campestris ; (ii) leaf spot disease caused by X . c . pv. raphani on hosts belonging to the Brassicaceae and Solanaceae; the sequenced strain 756C identified as X . c . pv. armoraciae was included in this pathovar and the existence of another leaf spot disease caused by X . c . pv. armoraciae was not supported; and (iii) bacterial blight of garden stocks caused by X . c . pv. incanae . No plants susceptible to X . c . pv. barbareae were found. Strains that did not induce any symptom on cruciferous plants tested, including their original hosts, were removed from the pathovar scheme and were named X . campestris only. Three new races were described in addition to the six races previously described within X . c . pv. campestris . The sequenced strains ATCC 33913 (CFBP 5241) and Xcc 8004 (CFBP 6650) belonged to race 3 and to race 9 (one of the new races described), respectively.     

Biological Control of Black Rot (Xanthomonas Campestris Pv. campestris) of Brassicas with an Antagonistic Strain of Bacillus Subtilis in Zimbabwe

Biological control efficiency of an antagonistic, endophytic strain of Bacillus subtilis (strain BB) was evaluated against three strains of the black rot pathogen, Xanthomonas campestris pv. campestris (Xcc), in four Brassica crops (cabbage, cauliflower, rape and broccoli) grown during three consecutive growing seasons and on two soil types, in two different areas in Zimbabwe. Strain BB controlled the disease caused by strain Xcc B-147 in all Brassica crops during the dry and short rainy seasons. A similar effect was observed in cabbage using the strain Xcc 33908. Biological control was effective in broccoli, but not in cabbage and rape during the main rainy season in clay loam soil and limited biological control effect was still observed when these crops were grown in sandy loam soil. The endophytic colonisation of cabbage roots by strain BB was confirmed by immuno-blotting during the whole growing season. Biological control of black rot with strain BB is discussed in relation to its effect on Xcc strains, Brassica crops and to the effect of weather and soil conditions.     

Genetic fingerprinting of Iranian Xanthomonas campestris pv. campestris strains inducing black rot disease of crucifers

Northern Iran has one of the largest and most diverse populations of cultivated crucifers in Iran. Symptoms of black rot disease were observed in 40 % of fields. To assess the genetic diversity of Xanthomonas campestris pv. campestris (Xcc) strains, associated with black rot disease, 40 strains were isolated from infected samples of crucifers such as cabbage, radish, cauliflower, turnip and kohlrabi, and were collected from different geographic regions of northern Iran including West and East Azarbayjan and Ardabil provinces. Bacterial strains were characterized by their morphological, biochemical and physiological features and pathogenicity tests. Four races were found in northern Iran (1, 4, 5 and 6) and the majority of the tested strains belonged to either race 4 (45 %) or race 6 (20 %). To examine the distribution of dispersed repetitive DNA, Enterobacterial Repetitive Intergenic Consensus (ERIC), BOX, Repetitive Extragenic Palindromic (REP) and random amplified polymorphic DNA (RAPD) sequences in the genome of Xcc using conserved primers. The different markers produced characteristic banding patterns and the similarity matrices from binary banding data was derived with the similarity for qualitative data program (SIMQUAL). On the basis of the fingerprint patterns generated by the combination data set of both rep-PCR and RAPD, the Xcc strains were differentiated into seven clusters (A–G) at 76 % similarity level. The geographical origin of the Iranian strains does not seem to be correlated with the RAPD and rep-PCR clusters. The clusters seem to be more related to the race of the strains. This is the first study on genetic diversity of Xcc strains inducing black rot disease of crucifers in Iran.     

Clove-transmissibility of <Emphasis Type="Italic">Pseudomonas salomonii</Emphasis>, the causal agent of ‘Café au lait’ disease of garlic

Bacterial blight of garlic, caused by Pseudomonas salomonii, results in leaf and sheath necrosis and sometimes leads to soft rot and plant death. The epidemiology of this bacterial disease, known as ‘Café au lait’ disease, is poorly understood and no resistant cultivars are currently available. To develop control strategies for this disease, we investigated principal sources of inoculum. The pathogen was isolated from bulbs from plants with typical vegetative symptoms of bacterial blight. Subsequent development of typical foliar symptoms on plantlets originating from symptomatic bulbs demonstrated transmission of the pathogen in the planting material. In one of three field experiments the contamination rate of planting stock influenced the disease incidence in field-grown garlic. The importance of planting stock as a source of inoculum was demonstrated here and should be evaluated relative to other potential sources such as crop debris, soil or alternate hosts in order to develop successful control strategies.     

The Distribution and Spread of Sorghum Downy Mildew in Sorghum and Maize Fields in Nigeria and Zimbabwe

Sorghum downy mildew (Peronosclerospora sorghi, SDM) is a damaging disease of sorghum and maize crops in Africa. Runs analysis was used to study the distribution of systemically infected sorghum and maize plants in Nigeria and Zimbabwe. The temporal and spatial development of local lesions of SDM on sorghum in Zimbabwe was investigated by assessing the local lesion symptoms caused by conidia in plots with a single point source of inoculum. With ordinary runs analysis, there was evidence of clustering of disease in some fields in the humid areas of Nigeria and the semi-arid areas of Nigeria and Zimbabwe. Clustering was found in two of the eight runs analyses performed on maize in the humid south of Nigeria, and in only one of the eight runs in Zimbabwe, which was interpreted as a predominance of random infection at the time of assessment and at the spatial scales assessed. Symptoms of local lesions of SDM developed rapidly across plots from an introduced point source of infection. After 9 days-exposure to the source of inoculum, the incidence of diseased leaves was 1.2%, and after 50 days it was 74.5%. A disease gradient which initially developed flattened as the plot became uniformly diseased. The predominant wind direction was NNE, and most rapid spread of disease was towards the SSW and WSW. In conclusion, local lesions can spread rapidly in sorghum crops, suggesting that they may be an important source of conidial inoculum for further local and systemic infections during the growing season.