Hypoxic stress inhibits aerobic wound-induced resistance and activates hypoxic resistance to bacterial soft rot

Resistance to bacterial soft rot in potato tubers maintained aerobically was induced by wounding tubers 4 to 24 h prior to inoculation withErwinia carotovora subsp.carotovora. This wound-induced resistance phenomenon was blocked by hypoxic stress. An hypoxia-induced resistance mechanism also was detected: tubers acclimated to a hypoxic environment were resistant to rot when inoculated with aerobically grownE. c. subsp.carotovora and incubated in atmospheres made hypoxic by argon or nitrogen. With increased pretreatment, hypoxia-induced resistance approached the levels of resistance observed in tubers inoculated with aerobic-adapted bacteria and incubated aerobically. Hypoxia-adaptedE. c. subsp.carotovora overcame hypoxia-induced resistance.     

Volatile monitoring as a technique for differentiating betweenE. carotovora andC. sepedonicum infections in stored potatoes

The volatile production characteristics of ring rot (Corynebacterium sepedonicum) and soft rot (Erwinia carotovora) infected potatoes were investigated under controlled conditions in order to assess the monitoring of volatiles as a technique for differentiating between the infections in stored potatoes. Total volatile production/unit time was much greater from soft rot due to more rapid development after infection. Total volatile production during the period required for the development of the soft rot infection was similar to the output detected during the longer period required for the development of the ring rot infection. The volatile profiles of the two pathogens featured a number of commonly shared metabolites, but one compound was unique to the ring rot infections and two compounds were unique to the soft rot infections. There were also consistent disease-specific changes in the relative concentration of many of the metabolites detected in the volatile profiles.     

Interaction ofErwinia spp. andFusarium roseum ‘Sambucinum’ on the Russet Burbank potato

Laboratory and field studies with the Russet Burbank (RB) potato provide evidence for synergism betweenErwinia carotovora var. atroseptica (Ea) andFusarium roseunt ‘Sambucinum’ (Fs). When these pathogens were inoculated together, the severity of tuber rot was significantly greater than when either pathogen was inoculated separately. Similarly, these pathogens interacted to reduce yield. When both organisms were uniformly applied to puncture wounds on potato seed (inoculum suspension consisted of 10⁸ cells/ml Ea and 10⁵ cells/ml Fs), the total yield was reduced by 46% and U.S. #1 yield by 53%. These reductions occurred even though blackleg symptoms (caused by Ea) were negligible (< 1%). In contrast, inoculations withErwinia carotovora var.carotovora (Ec) and Fs did not interact to reduce potato yield. Potato yields were also not influenced when these pathogens (Ea, Ec, Fs) were separately inoculated.Verticillium wilt (Verticillium dahliae) was significantly less when tubers were inoculated with either Ea, Ec, Fs, Ea + Fs, or Ec + Fs than with uninoculated tubers.     

Serogroups ofErwinia carotovora involved in systemic infection of potato plants and infestation of progeny tubers

Enrichment techniques and serological strain typing methods were used to determine the source of stem and progeny tuber populations ofErwinia carotovora (Ec) in 1980 and 1981. Plots were established in a field that had been planted with potatoes since 1972. Seed tubers of cultivars Russet Burbank and Norgold Russet were assayed forErwinia carotovora pv.carotovora (Ecc) andErwinia carotovora pv.atroseptica (Eca) by tuber peeling and direct plating and plating following enrichment in a pectate-based broth. Levels of seed tuber infestation ranged from undetectable to 2.4 × 10⁴ colony-forming units per g peel tissue. Prior to planting Ecc was detected in soil samples from field plots by enrichment in a pectate-based broth. Ec was isolated from symptomless plants with increasing frequencies from 8 weeks following planting regardless of the level of Ec populations in seed tubers. At harvest progeny tuber recontamination was low and not related to levels of seed tuber Ec populations. In 1980, serological typing of representative Ecc isolates indicated serogroups III, XXIX, and unknown strains isolated from the soil but not the seed tubers prior to planting were isolated from stems and progeny tubers. Serogroup XXIX was isolated from soil, seed, stems and progeny tubers in 1981. Throughout both years, Ecc was isolated more frequently than Eca from seed, soil, stems, and progeny tubers. These data emphasize the potential for reinfestation of seed stocks and infection of plants by populations of Ecc that overwinter in soil under Wisconsin conditions.     

Effect of acetylsalicylic acid on soft rot produced byErwinia carotovora subsp.carotovora in potato tubers under greenhouse conditions

Summary The efficiency of acetylsalicylic acid (ASA) in inducing localised acquired resistance against infection byErwinia carotovora subsp.carotovora was evaluated by treating potato tubers with ASA at three concentrations. Three days after treatments, tubers were inoculated withE. carotovora subsp.carotovora by wound inoculation or by irrigation with a bacterial suspension. Experiments were performed for two consecutive years. Statistical analysis revealed that treatment of tubers by immersion in ASA solutions at low concentrations induced a significant reduction in the soft rot incidence. Wounding of the tubers was the most effective inoculation method and ASA at concentration of 0.0125% (w/v), pH 7 was more efficient than at 0.025 or 0.05%. No phytotoxicity of such treatment was observed.     

Survival ofErwinia carotovora in Wisconsin soils

Erwinia carotovora var.atroseptica (van Hall) Dye andE. carotovora var.carotovora (Jones) Dye were detected in agricultural soils in Wisconsin using baiting and enrichment techniques. These soft rot bacteria could not be detected in soils of potato fields prior to planting of crops in the spring period using standard soil dilution plating techniques on a crystal violet pectate medium, however. A procedure involving incubation of samples in pectate enrichment broth followed by preparation of smears on slides and treatment with a fluorescent antibody stain specific forE. carotovora var.atroseptica was the most sensitive of the methods tested for detecting the blackleg pathogen. Erwinia carotovora was isolated more frequently during the spring from fields in which potatoes had been grown the previous year than from fields in which other crops had been grown. It was also isolated from potato tubers and stems that had overwintered in the field. The presence ofE. carotovora could not be detected in root zone samples of weed plants using the dilution plating method.     

Penetration of potato tuber lenticels by bacteria in relation to biological control of blackleg disease

Summary In a glasshouse experiment three potato cultivars were grown to maturity in wet or dry compost. The number of lenticels per tuber was greater with larger tubers but was unaffected by the cultivar or soil moisture regime. The frequency of lenticel penetration by a water soluble stain, safranin O, remained high as tubers developed to maturity in wet soil. In dry soil penetration declined markedly as the tubers matured. When tubers, harvested just before the onset of haulm senescence, were inoculated withErwinia carotovora ssp.atroseptica, the frequency of lenticel rotting was less than the rate of stain penetration, indicating that additional factors were involved in the bacterial infection process. Inoculating mature tubers with bacteria and incubating them in anaerobic conditions frequently resulted in rotting of the lenticels. Comparable tubers incubated in aerobic conditions showed no rotting or invasion by bacteria. The results are discussed in relation to the biological control of blackleg.     

Suppression of bacterial soft rot in potato tubers by application of kasugamycin

An immersion in aqueous solutions of Kasumin®, containing kasugamycin at 10 to 320 mg/L (ppm) for 1 to 320 sec delayed or prevented the development of soft rot on inoculated (Erwinia carotovora subsp.carotovora) tuber disks incubated over wet filter paper at 24C for 24 to 72 hr. Treatment efficacy decreased as inoculum level and incubation period increased. Short immersion periods were not as effective as longer ones. When stored tubers were inoculated withE. c. carotovora and incubated up to 5 days in fog chamber at 20 C, immersion treatments in 20 to 400 ppm kasugamycin either had no effect or increased soft rot development. By contrast, when the stored tubers were cut into sections before inoculation and treatment, immersion in 300 ppm reduced soft rot development on the cut surfaces from 83% (inoculated controls) to 3.2%. With non-inoculated sections, treatment with 25 ppm reduced the SAD from 13.3 to 0.6%. Wash or rinse treatment of freshly harvested tubers in 40 to 160 ppm produced a significant reduction in the severity of bacterial soft rot among tubers incubated for 72 to 96-hr. The 160-ppm treatment delayed disease onset by at least 24 hr. Pre-washing freshly harvested tubers with clean or chlorinated water prior to antibiotic treatment did not consistently improve the efficacy of the kasugamycin treatment.     

The role of phenols in potato tuber resistance against soft rot byErwinia carotovora ssp.carotovora

Summary Increasing doses of nitrogenous fertilizer increased storage rots in six potato cultivars of which cvs Kufri Sindhuri and Kufri Lalima rotted less and were found to contain high amounts of total phenols compared to the more susceptible cultivars Kufri Jyoti and Kufri Badshah. Nine phenolic acids were identified in the peel and the pulp of tubers and four of these and of other phenolic extracts from tubers suppressed the growth ofErwinia carotovora ssp.carotovora. Research Publication No. 5811, Experiment Station, G.B.P.U.A. & T., Pantnagar 263 145, India.     

Association of viral,bacterial and fungal pathogens with vascular discoloration of Russet Burbank potato tubers

Russet Burbank potatoes from the 1979 and 1980 crop years, collected from Chicago, IL repack warehouses and retail markets, were sampled for vascular discoloration. The amount of discoloration varied among sampling months and states of origin and decreased from 7.7% in 1979 to 1.6% in 1980. Highest levels of vascular discoloration were detected in December and January samples. Vascular discolored and non-discolored (control) tubers were assayed for the presence of potato leafroll, potato virus X, and beet western yellows viruses by the enzyme-linked immunosorbent assay (ELISA). Tubers were also assayed for 2 subspecies ofErwinia carotovora and forVerticillium albo-atrum andV. dahliae. Potato leafroll virus was detected in 31 of 831 vascular discolored tubers.V. albo-atrum was detected in 1 of 180 discolored tubers. Beet western yellows virus was not detected in discolored or non-discolored tubers. Two subspecies ofE. carotovora and potato virus X were equally common in discolored and non-discolored tubers.     

Evaluation of procedures for detection of pectolyticErwinia spp. on potato tubers

The effectiveness of a tuber incubation method for detection ofErwinia carotovora var.atroseptica andE. carotovora var.carotovora in potato tubers was compared with a lenticel sampling procedure. In the first method, tubers were injured by puncturing lenticels with sterile toothpicks, then wrapped in moist paper towels and polyvinylidene film, and placed in closed chambers flushed with N₂. In later experiments, wrapping tubers in two layers of polyvinylidene film and incubation in air was found to be as effective as the single layer of polyvinylidene and incubation in chambers flushed with N₂. Isolations were made on a selective crystal violet pectate (CVP) medium from homogenized samples of tissue removed from soft rot lesions developing around injured lenticels. In the second method, 10 lenticels/tuber were aseptically removed with a scalpel and homogenized in distilled water; the suspension was plated on CVP. The first method was less tedious and slightly more effective than the lenticel sampling method. In a preliminary survey, these methods were used to detectErwinia infestations in small samples of certified seed potato tubers from Maine, Minnesota, Montana, New York, North Dakota, and Wisconsin. PectolyticErwinia spp. were detected in at least one sample from each state except Montana. The percentage of tubers withErwinia infestations varied from 0–100% among samples. Characterization ofErwinia isolates showed that bothE. carotovora var.carotovora andE. carotovora var.atroseptica were present. PectolyticErwinia spp. on symptomless potato seed tubers may serve as inoculum sources for blackleg and soft rot diseases.     

Detection of viable and non-viable cells ofErwinia carotovora var.atroseptica in inoculated tubers of var. Bintje with enzyme-linked immunosorbent assay (ELISA)

Summary The ELISA technique was unable to distinguish between viable an non-viable cells (killed with ethanol) ofErwinia carotovora var.atroseptica in inoculated tubers during 8 weeks storage at C. The use of this technique to determine latent infections is questioned.     

The resistance of potato plants toErwinia carotovora subsp.atroseptica in relation to their calcium and magnesium content

Summary The effect of adding gypsum (CaSO₄) to growing media on the resistance toErwinia carotovora subsp.atroseptica (van Hall) Dye of potato (Solanum tuberosum L.) stems and mother and daughter tubers was examined in field and glasshouse experiments. The decayin situ of inoculated mother tubers from field plots treated before planting with gypsum was significantly delayed compared with tubers sampled from non-amended plots. Blackleg development was also significantly delayed in the gypsum-amended plots; however, for both mother tubers and stems the increased resistance associated with the application of gypsum later disappeared. Stem resistance tended to be related to tissue calcium and magnesium concentrations which were higher in plots treated with gypsum than in untreated plots. Daughter tubers produced in plots amended with gypsum generally contained a higher concentration of calcium in their cortex and were more resistant to decay byE. carotovora subsp.atroseptica but the effect was not consistent.     

Quaggy Joe: A high-yielding, round, white potato variety for fresh market

The Quaggy Joe potato variety is high-yielding and medium to medium-late maturing with attractive, round-oval, white-skinned, white-fleshed tubers with medium-shallow pink eyes. Its major use is expected to be as a maincrop tablestock variety. Quaggy Joe is resistant to the potato leafroll virus, and is also moderately resistant to early blight (Alternaria solani), dry rot (Fusarium sambucinum), and soft rot (Erwinia carotovora subsp.carotovora). Quaggy Joe tubers have shown very little shatter or blackspot bruising, but sprout earlier than Atlantic. Young plants have shown sensitivity to the herbicide metribuzin.     

Effect of wet incubation time and temperature on infection, and of storage time and temperature on soft rot lesion expansion in potatoes inoculated withErwinia carotovora ssp.Carotovora

Summary Infection and lesion expansion studies were carrried out on potato tubers. In the infection study, the tubers were inoculated withErwinia carotovora ssp.carotovora (Ecc) and exposed to incubation temperatures of 10–25 °C for 3–48 h of wetness. In the lesion expansion study, the tubers were exposed to 12 h of wetness at 20 °C to establish infection, and then·stored at temperatures of 4–16 °C for 15–90 d, at 95% RH. The volume of diseased tissue was determined and the data were transformed to proportion of maximum volume diseased (PVD). Infection was close to zero for 3 and 6 h wet incubation time, regardless of incubation temperature, and reached the maximum in 12 h of wetness at 20 °C. Significant lesion expansion occurred at a storage temperature of 16 °C after about 60 d of storage time. Cubic models of infection and lesion expansion potentials explained 95 and 96% of the variations in infection and lesion expansion, respectively.     

Bacterial soft rot potential in washed potato tubers in relation to temperatures of tubers and water during simulated commercial handling practices

The bacterial soft rot potential in potato tubers was affected by differences in temperature between tubers and suspensions ofErwinia carotovora at the time of inoculation by immersion. In general, disease severity after incubation was higher when tuber temperatures were higher as opposed to equal to or lower than that of water containing cells of the bacterium. Disease severity was also greater when tubers were stored at 20° to 23°C in contrast to 4° or 26° to 30°C for 16–18 h prior to immersion. Tubers with a pulp temperature of 8°C were more prone to bruise injury as compared with those at 18°C when struck with a pendulum type bruising instrument. When the injured tubers were inoculated and incubated, most bruises became infected with soft rot bacteria.     

The bionomics ofErwinia carotovora in North Dakota

A study into the ecology ofErwinia carotovora in North Dakota was performed to determine possible sources ofErwinia spp. which recontaminate stem cut derived seed stocks (SCDS).Erwinia carotovora pv.carotovora (Ecc) was found to overwinter in naturally infested agricultural soils that had been planted to potato the previous cropping season. However, this bacterium was only present in the 30–71 cm sampling depth.E. carotovora pv.atroseptica (Eca) and Ecc were also found in the rhizospheres of a few weed and crop species but the frequency of recovery was low. Ecc and other strains not identifiable to pathovar, but notE. chrysanthemi, were readily recovered from a variety of water sources and from sunflowers with stem and head soft-rot symptoms. AllErwinia strains were characterized, but not all could be identified to pathovar. WateringErwinia-free potato plants with water naturally infested withErwinia spp. resulted in progeny tubers contaminated withErwinia. The importance of this information with regard to the recontamination of SCDS is discussed.     

Effect ofErwinia carotovora subsp.atroseptica alone and withAlternaria solani orVerticillium dahliae on disease development and yield of potatoes in Israel

Summary Tubers of 17 cultivars were inoculated withErwinia carotovora subsp.atroseptica (ECA) and planted in 1986 and 1987 in the Negev in randomised plots in three blocks where eitherVerticillium dahliae orAlternaria solani were present or where both were absent. ECA reduced the yield in the control block and exacerbated the yield losses in the disease blocks, but the intensity of symptom expression did not always accord with the extent of yield loss. ECA had a greater effect on plants infected withA. solani than on those withV. dahliae. It was possible to categorise the cultivars as resistant, intermediate or susceptible with respect to ECA and early blight, but resistance toV. dahliae could not be differentiated from foliage maturity.     

Methods for assessing bacterial density on potato tubers

Summary Formulae which determined tuber surface area from tuber dimensions and weight were tested by using samples from three potato cultivars. Surviving populations of a labelled isolate ofErwinia carotovora ssp.atroseptica following planting, and populations of a fluorescent pseudomonad following dusting with a bacterial powder formulation were used to demonstrate the application of these formulae to the calculation of bacterial density on the tuber surface. Results obtained by washing whole tubers using the formulae to calculate the surface area were shown to be comparable in sensitivity with those obtained by dilution-plating of periderm discs of 1cm².     

Factors affecting the accuracy of the tuber incubation test for the detection of contamination of potato stocks byErwinia carotovora

Summary Erwinia carotovora in potato stocks can be detected more efficiently by the tuber incubation method in which the test tubers are first induced to rot under anaerobic conditions (followed by detection isolation of the bacteria on a selective growth medium or by serological techniques) than by direct lenticel sampling. However, the efficiency of detection is affected by both the extent of decay at sampling time and the incubation temperature. Detection ofE. carotovora is poor from extensive lesions developed at temperature of 25 C or above. Furthermore, detection ofE. carotovora var.atroseptica is more frequent than that ofE. carotovora var.carotovora at temperatures below 22 C in tubers contaminated by both organisms, whereas at higher temperatures the reverse is true. Only within a narrow temperature range, close to 22 C is there an equal chance of detecting both varieties.