Effect of temperature on differential persistence and insect transmission ofErwinia carotovora var.atroseptica andErwinia carotovora var.carotovora

Adult fruit flies (Drosophila melanogaster Meig.) transmittedErwinia carotovora var.carotovora (Jones) Dye (ECC) andErwinia carotovora var.atroseptica (van Hall) Dye (Eca) from vegetative material inoculated with equal proportions ofEcc andEca to uninoculated potato plants and from inoculated potato plants to sterile vegetative material at 15° and 27°C. The insects transmittedEcc andEca with approximately equal frequencies over an 18 hr period at 15°C. At 27°C the proportion ofEcc transmitted was significantly greater thanEca. At 27°C moreEcc thanEca was present in the inoculated vegetative material and potato stems, on fruit flies visiting these sites, and in injured potato tissues to which flies transmitted the bacteria. Results indicate that temperature is important in determining success ofEcc andEca transmission by insects and the relative persistence of the two organisms in plant tissues after transmission.     

The association ofErwinia carotovora var.atroseptica andErwinia carotovora var.carotovora with insects in Colorado

Ten genera from 9 families of dipterous insects collected in the field in the San Luis Valley of Colorado were contaminated withErwinia carotovora var.atroseptica (Eca) andErwinia carotovora var.carotovora (Ecc).Erwinia-contaminated insects were collected from settling ponds near potato warehouses, potato cull piles on growers’ farms, municipal dumps, lettuce and potato fields. The percentage of contaminated insects reached 14.5% in early spring but decreased as the season progressed. Insects were more commonly contaminated withEca thanEcc early in the season. Later the proportion of contamination byEcc increased while that ofEca decreased. We suggest that insects may play a potentially important role in the epidemiology of potato blackleg especially in the reintroduction ofErwinia intoErwiniafree potato fields.     

Insect transmission ofErwinia carotovora var.carotovora andErwinia carotovora var.atroseptica to potato plants in the field

Adult fruit flies (Drosophila melanogaster Meig.) artificially contaminated withErwinia carotovora var.carotovora (Jones) Dye (Ecc) and/orE. carotovora var.atroseptica (van Hall) Dye (Eca) readily transmitted the bacteria to plants in the field that had been injured by crushing the stem. Injured, inoculated plants developed disease symptoms when maintained at high relative humidities.Erwinia was transmitted to ten-hour-old injuries and they became infected as frequently as freshly made wounds. Insect transmission ofEca, Ecc, and mixtures was greatest during the afternoon, which was the warmest part of the day.Ecc was transmitted significantly less frequently during the cold morning than during afternoon or evening hours. A potato cull pile placed in a commercial potato field attracted a natural insect population which increased during the season. BothEcc andEca were isolated from uninoculated rotting tubers in the cull pile and from insects associated with the pile from May through September. These naturally infested insects transmittedEcc and/orEca from the cull pile to artificially injured field plants during July and August at distances as great as 183 m from the cull pile. NoErwinia was isolated from injured plants in a neighboring control field, which lacked a cull pile, further than 6 m upwind from the cull pile. We suggest that insects are important agents in the epidemiology of potato blackleg and soft rot even in areas with low relative humidities.     

Seasonal incidence and cause of blackleg and a stem soft rot of potatoes in Oregon

Blackleg of potatoes in a center pivot irrigated circle of Kennebec potatoes in Oregon’s Columbia Basin was caused by eitherErwinia carotovora var.atroseptica (Eca) orE. carotovora var.carotovora (Ecc). Eca was recovered only early in the growing season whereasEcc was associated with blackleg symptoms later in the season. A soft rot, atypical of blackleg, was widespread throughout the circle and was most prevalent during the latter part of the growing season. Initially, the vines near the soil surface appeared translucent and watery and were soft and mushy to the touch. The pathogen associated with this symptom was exclusivelyEcc.     

The influence of inoculum density and environment on the development of potato blackleg

Significant increases in pre-emergence seedpiece decay and preemergence shoot infection were observed asErwinia carotovora var.atroseptica inoculum density increased. As inoculum density increased, a point was reached where virtually 100 per cent of the disease was expressed as preemergence decay and no typical postemergence blackleg occurred. Soil temperature and soil moisture markedly affected blackleg incidence, especially at low inoculum densities. Increasing soil temperature (from O°C to 30°C) and soil moisture (from 65 to 100% of field capacity) usually resulted in an increased total blackleg incidence. The effect of soil temperature and soil moisture appeared to be masked at high inoculum densities. Conditions such as low inoculum density and/or cool soil temperatures and low soil moistures which apparently slowed the rate of seedpiece decay usually increased the amount of postemergence blackleg, but also reduced the total amount of disease which occurred. It is suggested that conditions which are not optimal for the pathogen result in increased amounts of typical easily diagnosed postemergence blackleg expression, whereas conditions which favor the pathogen may reduce this phase of the disease but actually increase the total loss by increasing the amount of seedpiece decay and preemergence death of potato shoots.     

Evaluation of potato clones for resistance to tuber and stem rot induced byErwinia chrysanthemi

Tubers of 149 clones ofSolarium tuberosum subsp.andigena (andigena) from four self-pollinated (S₁) families (Jak 072, 702440, 700718 and Och 5331) obtained from true seed were rated for resistance toErwinia chrysanthemi. The clones were classified into four arbitrary categories based on disease reaction: resistant, intermediate, susceptible, and very susceptible. In family Jak 072, 23% of the clones were resistant and 61% intermediate. In family Och 5331, 22% of the clones were susceptible and 67% very susceptible; and in families 702440 and 700718, 38% and 29% of the clones were intermediate and 45% and 40% were susceptible, respectively. Tubers of 11 clones chosen to represent each one of the four categories were inoculated withE. chrysanthemi (Ech), E. carotovora subsp.carotovora (Ecc) andE. carotovora subsp.atroseptica (Eca). Clones responded similarly toEch andEca. However,Ecc induced significantly more rot thanEch orEca, presumably favored by the temperature of incubation (25°C). Clones Jak 072-17 and Jak 072-18 were resistant to the three bacterial pathogens. Above ground stems of 6 of the 11 selected clones inoculated withEch were susceptible to stem rot. No correlation was found between the response of tubers and above ground stems to tuber and stem rot induced byEch.     

Survival of two varieties ofErwinia carotovora onDrosophila melanogaster meigen andDrosophila busckii Coquillett, (Diptera:Drosophilidae) vectors of potato blackleg in Colorado

Survival time ofErwinia carotovora var.carotovora (Ecc) andE.c. var.atroseptica (Eca), causal agents of potato blackleg and bacterial soft rot, was determined both internally and externally on two known insect vectors,Drosophila melanogaster Meig. andD. busckii Coq. The results indicate that the two bacterial varieties survived internally on both vectors for 48 hours and in some cases up to 72 hours.Ecc andEca survived externally on both vectors for at least 24 hours and in some cases at low levels for 48 hours. The two varieties of bacteria declined in number with elapsed time both internally and externally. We suggest that the vectors enhance survival of the non-spore formingErwinia cells outside the plant host. The enhanced survival, combined with the dispersal ability of the vectors, increases the destructive potential of these bacteria. There appeared to be no differential survival of the two bacterial varieties on the vectors studied that might account for the differential transmission reported by previous workers.     

Potato early dying disease in the Pacific Northwest caused byErwinia carotovora pv.carotovora andE. carotovora pv.atroseptica

Premature death of potato vines is widespread in irrigated potato fields of the Pacific Northwest.Verticillium dahliae is a major cause of early dying in fields with a history of potato production, but in fields new to potato production, or those that have been fumigated and have a lowV. dahliae population in the soil, premature death of vines can also occur.Erwinia carotovora pv.carotovora (Ecc) andE. carotovora pv.atroseptica (Eca) have been implicated in the early dying disease syndrome in these fields. Both bacterial pathogens are associated with symptoms similar to those caused byV. dahliae; i.e., a progressive chlorosis and necrosis of the foliage and vascular discoloration that may extend a short distance up the stem. Greenhouse pathogenicity studies on cv. Russet Burbank cuttings in which symptom development was identical to that produced byV. dahliae have verified thatEcc andEca can cause potato early dying.     

The role ofErwinia carotovora in the epidemiology of potato blackleg I. Relationship ofE. carotorora var.carotovora andE. carotovora var.atroseptica to potato blackleg in Colorado

Isolations from 315 blackleg infected potato stems collected over a two year period in Colorado, demonstrated that both varieties ofErwinia carotovora, E. carotovora var.carotovora andE. carotovora var.atroseptica, are involved in stem infections under field conditions.E. carotovora var.atroseptica was the predominant organism associated with stem infections in all areas sampled.E. carotovora var.carotovora was isolated, with few exceptions, only from plants collected from the warmer areas of the state. The epidemiological implications of the effect of soil temperature on the distribution of these two organisms are discussed.     

Susceptibility of potatoes to bacterial rot and weight loss as a function of wound-healing interval and temperature

A study was conducted to determine the feasibility and advantages of wound-healing potatoes at lower than recommended temperatures. The progress of wound-healing at 6, 9, and 16°C, of tissue taken from Russet Burbank potato tubers, was evaluated by inducing weight loss with high temperature or decay organisms following various curing intervals. Increasing the wound-healing temperature resulted in a greater resistance to weight loss and a decreased susceptibility to infection byErwinia carotovora subsp.carotovora. Resistance to weight loss increased as wound-healing temperature increased following 6 and 16 day intervals. However, weight loss induced after 21 days of healing appeared to level off at 9°C indicating maximum development of a wound barrier. Loss due to rot, after inoculation of fresh cores withE. carotovora, was directly proportional to incubation time and temperature. When wound-healing progress was measured by loss of fresh weight induced by elevated temperatures or rot, the advantages of woundhealing at 16°C compared with 9°C decreased significantly after 21 days of curing. Wound-healing at approximately 9°C for 25 days was found to be the best compromise temperature for keeping rot progress and weight loss at a minimum while allowing the process of healing and maturation to occur.     

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.     

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.     

Effect ofErwinia carotovora subsp.atroseptica on yield of potatoes from cut and uncut seed tubers in Valencia

Summary The effect on the development of blackleg symptoms and on yield of inoculating seed tubers of 15 cultivars withErwinia carotovora subsp.atroseptica (Eca) was investigated. Tubers were planted either whole or as pieces. Eca inoculation reduced the mean yield per plant by 8% for whole tubers and by 12% for cut tubers. There was no correlation between blackleg symptoms and yield loss.     

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.     

Resistance to blackleg and tuber soft rot in sexual and somatic interspecific hybrids with different genetic background

Diploid interspecific hybrids from crosses betweenSolanum tuberosum haploids andS. canasense, S. multidissectum, andS. tarijense, along with tetraploid and hexaploid somatic hybrids S.tuberosum (+)S. commersonii, were screened for resistance to blackleg and tuber soft rot byErwinia carotovora subsp.atroseptica andE. carotovora subsp.carotovora. Among the diploid hybrids, those involving clone mlt la of S.multidissectum and tar 2b ofS. tarijense yielded the highest number of genotypes resistant to both blackleg and tuber soft rot. By contrast, all the hybrids involving clone tar 1lb ofS. tarijense were susceptible to both bacterial diseases. As far as resistance of somatic hybrids is concerned, the most interesting genotype was the tetraploid one which showed resistance to both blackleg and tuber soft rot Significant correlations were found betweenE. carotovora subsp.atroseptica andE. carotovora subsp.carotovora both in resistance to blackleg and in resistance to tuber soft rot, suggesting that resistance to the two subspecies is controlled by the same gene/s or that the relevant genes are linked. On the other hand, correlations between blackleg and tuber soft rot were never significant, indicating that different mechanisms may control resistance in tuber and stems. Alternatively, the resistance could be controlled by the same gene/s under a different spatial/temporal expression pattern. Twelve diploid sexual hybrids and one tetraploid somatic hybrid were selected for resistance to blackleg and/or tuber soft rot and for tuber characteristics.     

Enhanced resistance to bacterial infection byErwinia carotovora subsp.atroseptica in transgenic potato plants expressing the attacin or the cecropin SB-37 genes

Blackleg and soft rot diseases, caused by the bacteriumErwinia carotovora, are among the diseases that cause important losses in culture and storage of potato. In this paper, we introduced bacterial resistance into potato, via genes encoding for proteins with antibacterial activity. For this purpose, potato clones were transformed either with the gene encoding the acidic attacin protein fromHyalophora cecropia, or with the gene encoding the cecropin analog peptide SB37. These clones were evaluated for soft rot and blackleg resistance, after inoculation with the bacterial strainErwinia carotovora subsp.atroseptica T7. Results reported in this paper indicate that a considerable percentage of the potato clones (15–22%) showed increased resistance to bacterial infection, revealed by reduced severity of blackleg or soft rot symptoms. Expression of the transgenes was demonstrated in some of the clones by Northern blot analysis. This is the first report indicating that expression of the gene encoding for an attacin protein and for the cecropin SB-37 peptide in transgenic potato confers increased resistance to bacterial infection.     

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.     

Transmission ofErwinia carotovora var.atroseptica byDrosophila melanogaster Meig. I. Acquisition and transmission of the bacterium

Drosophila melanogaster acquired the potato blackleg bacterium,Erwinia carotovora var.atroseptica from laboratory cultures and infected potato plants and readily transmitted it to healthy wounded plants under greenhouse conditions.D. melanogaster also carried the bacterium both internally and externally. Transmission to the nearest plants occurred most often. The possibility of at least short term survival of the bacterium inside the body of the insect is postulated.     

Evaluation for chipping and tuber soft rot (Erwinia carotovora) resistance in potato clones from unilateral sexual polyploidization (2x × 4x)

The aim of this work is the evaluation of tetraploid potato clones with germplasm fromS. chacoense andS. phureja for chipping and resistance to tuber soft rot (Erwinia carotovora subsp.carotovora). Tetraploid potato clones were obtained via 2n eggs through 2x–4x crosses. Cytological mechanisms were investigated in the diploid hybrid 2n-egg producer used as parent in the unilateral sexual polyploidization scheme; the omission of the second meiotic division and cytokinesis failure at the end of meiosis were identified with a genetic consequence equivalent to second division restitution (SDR). Tuber marketable yield, specific gravity and chip-processing ability evaluated at harvest and after cold storage with and without reconditioning were higher in some clones as compared to theS. tuberosum tetraploid parent. Tuber soft rot resistance, contributed byS. chacoense, was successfully transferred to the4x level. After inoculation withErwinia carotovora subsp.carotovora and estimation of the rotted area, one tetraploid clone proved to be resistant and three partially resistant. The genetic value of exotic germplasm and sexual polyploidization strategy are discussed in relation to potato improvement.