Erwinia carotovora subsp.atroseptica infection promotes verticillium wilt development in potato in Israel

Summary The interaction betweenErwinia carotovora subsp.atroseptica (Eca) andVerticillium dahliae and its effect on symptom development in potato cultivars showing different degrees of resistance to them was examined over two seasons in irrigated fields in a hot, dry climate. Four cultivars were used: Cara, highly resistant to blackleg and tolerant to Verticillium wilt; Pentland Crown also resistant to blackleg but susceptible to Verticillium wilt; Désirée, moderately susceptible to blackleg but tolerant to Verticillium wilt; and Maris Bard, susceptible to both diseases. Seed tubers were inoculated with Eca immediately after harvest in Scotland, to simulate natural infection, and then planted in Israel in soil naturally infested byV. dahliae. In Verticillium tolerant cultivars, Eca infection increased the rate of colonization byV. dahliae and significantly enhanced symptom development, as assessed by suppression of plant height and wilt symptoms. This report is a portion of the Ph.D. thesis of the senior author in Ben-Gurion University of the Negev.     

Effect of applying nitrogen fertilizer to a potato seed crop on the susceptibility of the daughter plants toErwinia carotovora subsp.atroseptica

Summary Seed potatoes of cvs Record and Bintje were grown with 30, 80 or 130 kg N/ha. In the subsequent growing season the seed tubers, which were inoculated or not with two concentrations ofErwinia carotovora subsp.atroseptica (Eca), were subjected to the same fertilization treatment. The incidence of blackleg and the level of seed tuber decay increased from the uninoculated treatment to the higher inoculum level, but the degree varied considerably from one year to another. Cv. Bintje was more susceptible to blackleg and yield loss caused by Eca than cv. Record. In 1990, when disease incidence was high, the proportion of blackleg stems increased with increasing doses of nitrogen fertilizer which had been applied to the seed crop, but was not significantly affected in 1988 and 1992. Seed tuber decay was retarded by the lowest nitrogen dose. The yield of the subsequent crop was not affected by the nitrogen dose applied to the seed crop.     

Preliminary selection of antagonists ofErwinia carotovora subsp.atroseptica (Van Hall) Dye for application during green crop lifting of seed potato tubers

Summary Green crop lifting (GCL) for haulm killing was developed in The Netherlands and offers ideal conditions for controlling the blackleg pathogenErwinia carotovora subsp.atroseptica (Eca) by antagonists. Based on the use of mini-tubers or young tubers from field crops, two bioassays for wound protection were developed. GCL was simulated by artificially skinning or wounding tubers, inoculating the damaged skin with Eca, treating with antagonists and incubating in either potting compost or outside in field soil. Mainly fluorescent pseudomonads, pre-screened for in vitro antagonistic activity on agar and high soft rot reducing ability on tuber slices, were tested in the mini-tuber bioassay. Strains giving the highest degree of wound protection were further tested individually and in combination under field conditions in the young tuber bioassay. One individual strain and two combinations, resulting in reduction of contamination levels on skinned surfaces of 85% and between 60% and 70%, respectively, show good potential for biological control of blackleg.     

Comparison of the interactions ofErwinia carotovora ssp.atroseptica withPhytophthora infestans, Phoma foveata andFusarium coeruleum in rotting potato tubers

Summary An interaction betweenP. infestans andE. carotovora subsp.atroseptica (Eca) in rotting tubers was confirmed and the biochemical basis for its occurrence investigated. The pH of tissue infected withP. foveata became alkaline whereas tissue infected withP. infestans orF. coeruleum did not rise above neutrality, which could promote pathogenicity of Eca by maintaining pH closer to the optimum for polygalacturonase activity. Polygalacturonase, pectate lyase and galactanase were detected in cultures of all three fungi grown on media containing cell wall material or pectin from tubers. AsP. infestans produced more polygalacturonase than the other two fungal tuber rot pathogens the possibility was investigated that oligogalacturonide products of enzymatic degradation of pectin byP. infestans stimulates pathogenicity of Eca. However, while tubers soft-rotted after infiltration with supernatant from fungal cultures grown on tuber cell wall material, controls showed that rotting resulted from infiltration rather than the products contained in the infiltrated water.     

Lightning damage to potatoes

Summary Two distinct round patches were observed in a potato field where the haulm showed symptoms like those of blackleg. The remainder of the field was in vigorous growth. Affected stems had long cracks andErwinia carotovora var.atroseptica was isolated from some. This had probably developed subsequently on damaged tissue and no rotting was apparent below soil level. Most of the tubers were decayed but noErwinia and only green pseudomonads were detected. Local meteorological records showed that thunderstorms occurred on only one day (1 August) in the six weeks before observation of the field symptoms. It was concluded from all evidence that the condition described was caused by a lightning strike on that day.     

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.     

Die Effizienz der Nassfäuleabwehr bei der Wundheilung von Kartoffelknollen. 1. Bestimmung der befallsauslösenden Erregerdichte und ihrer Umweltbeeinflussung

Zusammenfassung Nach Inokulation mit abgestuften Bakteriendichten wurde über eine Vitalf?rbung von Knollengewebe der Schwellenwert zur F?uleausl?sung bestimmt. Seine Erh?hung nach 24 h Wundheilung charakterisiert die Effizienz der biochemischen F?uleabwehr. Die Abwehrreaktionen des Knollengewebes und der Prozess der F?uleausl?sung sind in unterschiedlichem Masse von der Temperatur, der Feuchtigkeit und dem O₂- sowie dem CO₂-Gehalt der Atmosph?re abh?ngig.

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Summary The inoculum densities (effective dose) ofErwinia carotovora sub sp.atroseptica (Eca) needed to initiate infection of tuber tissue before and after a 24 h wound-healing period, was determined using a vital stain. The effects of various treatments on the efficiency of the defence reaction associated with wound healing, and on rot initiation were manifested as changes in the effective dose. The defence reaction was inhibited by temperatures <10°C, reduced O₂ levels ≤5%, and CO₂ proportions ≥20% (Fig. 3a and 4a). Moisture loss in stored tubers increased their defence reaction compared to that of freshly harvested ones (Table 1). The effect of these factors on Eca development was identical to that which occurs in potato production where tuber wounds are generally contaminated with Eca. Rot initiation was best prevented by removal of water (Table 1), storage at 15°C, avoidance of reduced O₂ levels <10% and CO₂ accumulation (Fig. 3b and 4b).

     

Control of potato blackleg by tuber pasteurisation: the determination of time-temperature combinations for the inactivation of pectolytic erwinia

Summary Pasteurisation of potato tubers by immersion in hot water is being examined as a method of blackleg control. Studies on the in vitro heat resistance of representative pectolytic erwinia have enabled prediction curves for the selection of suitable times and temperatures of pasteurisation to be prepared. High levels of contamination on some tubers may require heat treatment that will reduce the erwinia population by more than 80% to ensure that only innocuous levels remain.     

Detection ofErwinia carotovora var.atroseptica in potato tubers with immunofluorescence following induction of decay

Summary The simplest method of inducing tubers to decay in order to detect contamination by the blackleg organism,Erwinia carotovora var.atroseptica, was to wound them and place them in a mist chamber for four days. Incubating the decayed tubers in air for a further two days improved detection of the blackleg organism by immunofluorescence, which was a more sensitive test than the double diffusion method.     

Effects of contamination byErwinia carotovora subsp.carotovora andE. carotovora subsp.atroseptica of potato seed tubers and of cultivar resistance on blanking or nonemergence and blackleg development in Valencia,Spain

Summary High grade potato seed tubers of three cultivars were inoculated in Scotland by vacuum infiltration in suspensions ofErwinia carotovora subsp.carotovora and subsp.atroseptica in different combinations, and transported to Valencia, Spain. They were cut into two to three pieces and planted in January 1984, 1985 and 1986 in fields furrow irrigated before and after emergence. High incidences of blanking and blackleg were associated with seed inoculated with high numbers ofE.c. atroseptica alone or together withE.c. carotovora and not with high or low numbers ofE.c. carotovora alone.E.c. atroseptica predominated at emergence in March in rotting seed pieces which failed to produce a plant, regardless of the seed treatment, and in diseased stems in April but it was replaced byE.c. carotovora in May. The levels of blanking and blackleg after inoculating 16 cultivars with both erwinias were not correlated between years nor were they significantly different within years among most of the cultivars.     

Assessing six Austrian potato cultivars for resistance toErwinia carotovora subsp.atroseptica

Summary The susceptibility of six new Austrian cultivars to soft rot, stem rot and blackleg caused byErwinia carotovora subsp.atroseptica was assessed over 2 years in two locations. Soft rot susceptibility was assessed by whole tuber and half tuber tests, stem rot by stem inoculation and blackleg by planting inoculated tubers in the glasshouse. The susceptibility of the cultivars tested with the tuber and blackleg test methods were in general agreement. The half tuber test was more precise in differentiating intermediate levels of susceptibility than the whole tuber test. The reaction of the cultivars to soft rot could be classified as susceptible (Gina and Romina), moderately susceptible (Ditta, Sonja and Bintje), or slightly susceptible (Treff, Komet and Désirée). Results from the stem rot test did not agree with this classification and also depended upon the physiological state of plants as influenced by growing season and glasshouse conditions.     

Studies on natural repellents against potato tuber moth (Phthorimaea operculella Zeller) in country stores

Summary The effects of the leaves of five plant species, one from each of the generaAmbrosia, Anemone, Eupatorium, Eucalyptus andLantana, on potato tuber moth were investigated under indigenous storage conditions at the Central Potato Research Station, Shillong (1800 m above sea level). Their action was compared with that of a biological insecticide (spores ofBacillus thuringiensis), a chemical insecticide (carbaryl), and an untreated control. The data collected after six months storage on tuber damage, sprout damage and the rotting indicated that the leaves ofLantana aculeata provided most protection to the tubers, reducing damage from over 70% in the check to below 5%, and sprout damage from over 45% to below 3%. Next best wasEucalyptus globulus followed byB. thuringiensis. They may be used on tubers stored for table use or for seed as they had no adverse effect on germination or on the yield of a subsequent crop.     

Resistance toErwinia carotovora subsp.atroseptica in second backcross populations of somatic hybrids betweenSolanum tuberosum andS. brevidens

Summary Eleven clones obtained from a cross between cv. Katahdin and fusion products betweenSolanum tuberosum and the non tuber-bearing speciesS. brevidens, were backcrossed again withS. tuberosum (clone AR80-127-5). Small tubers harvested from 583 seedlings of these second backcross (BC2) populations were screened for tuber tissue resistance toErwinia carotovora subsp.atroseptica under aerobic conditions. After multiplication in the field. BC2 clones again were screened for soft rot resistance under aerobic as well as anaerobic conditions. In general, the resistance of the BC2 populations was reduced in comparison with the BC1. Variation for resistance was found within the BC2 populations, but there was no correlation between the results of the three tests performed. Four BC2 populations were also screenned for resistance to blackleg in the field. Significant differences were found between populations for mean percentage of diseased plants, but these differences could not be explained by the resistance of the parental clones.     

Storage of seed potatoes in pallet boxes. 1. The role of tuber surface moisture on the population of Erwinia bacteria

Summary Three stocks of seed potatoes contaminated with soft rot Erwinias were stored for 172 days in a commercial pallet box store. Three treatments were applied: naturally ventilated boxes, forced air ventilated boxes and single tubers on trays. Skin resistance values were 2.3, 9.4 and 19.3 megohm respectively. Rewetting events numbered 4, 7 and 15.Erwinia carotovora pv.carotovora (Ecc) andErwinia carotovora pv.atroseptica (Eca) declined in all stocks and treatments. Ecc declined significantly faster in the forced air ventilated treatment. The more frequent rewettings in the trays reverse the benefit of their drier skins. In the earliest sprouting stock, the naturally ventilated, but not the forced ventilated boxes, showed a decrease in skin resistance and corresponding increase in Ecc as spring approached. Forced ventilation in the first 14 days after harvest halved the level of silver scurf compared to the naturally ventilated treatment.     

Evaluation of diploid potato clones for resistance to tuber soft rot induced by strains ofErwinia carotovora subsp.atroseptica,E. carotovora subsp.carotovora andE. chrysanthemi

Summary Tubers of clones from a diploid hybrid population ofSolanum phureja andS. stenotomum were screened for resistance to tuber soft rot caused by strains ofErwinia carotovora subsp.atroseptica (Eca),E. carotovora subsp.carotovora (Ecc) andE. chrysanthemi (Ech). Significant, positive correlations between resistance to the different strains and species were observed, indicating that screening for resistance to tuber soft rot could be accomplished by using just one of the three pathogens. Strains of Ecc and Ech were found to be much more virulent than strains of Eca. Inoculating with Ecc and Ech resulted in more distinct differences among susceptible, moderately resistant and resistant clones than inoculating with Eca.     

Effects of soil-borneRhizoctonia solani Kühn on yield and quality of ten potato cultivars

Summary The effects of soil-borneRhizoctonia solani on yield and quality of potato were studied by an extensive individual plant sampling procedure. From 1983 to 1986, stem canker and stolon pruning were examined in 10 768 plants growing on a sandy soil. Tuber yield and quality and haulm yield per plant were also recorded. The degree of attack byR. solani depended on the cropping frequency of potato. Severe and very severe stem and stolon attacks decreased fresh yield, dry matter yield and dry matter content of tubers and increased the number of deformed and small tubers, whereas the effect on haulm yield and stem number was small. Potato cultivars differed in susceptibility toR. solani, but yield response did not differ significantly between cultivars at the same levels of stem and stolon infection.     

Soft rot and blackleg reactions in diploid potato hybrids inoculated withErwinia spp.

From 1993 to 1996 three groups of potato genotypes were evaluated for resistance toErwinia spp.: (1) 31 interspecific diploid hybrids (28 resistant and three susceptible), (2) five hexaploid or pentaploid somatic hybrids ofSolanum tuberosum (tbr) ×S. brevidens (brd), and (3) eight cultivars. Two evaluation methods were applied: tuber point inoculation with eitherErwinia carotovora ssp.atroseptica (Eca) orE. chrysanthemi (Ech) to test tuber soft rot resistance and stem base inoculation with Eca to test blackleg resistance. Some resistant diploid hybrids and somatic hybrids oftbr × brd were superior to cultivars for both tuber and stem resistance. Tuber and stem resistance toErwinia spp. in the most resistant diploid hybrids were equal to the highly resistant somatic hybrids oftbr xbrd. Tuber resistance to Eca was highly correlated to tuber resistance to Ech (r=0.815***). In two years of evaluation for stem resistance, three diploid hybrids and a derivative of one of the somatic hybrids (USA M 264) failed to develop symptoms of blackleg following inoculation with Eca, Analyses of variance for tuber and stem resistance indicated significant effects of genotype, year and genotype × year interaction. A positive relationship between tuber and stem resistance toErwinia spp. has been found, however the genetic control of resistance in tuber and stem is partially independent. In the case of Eca the correlation coefficient was r = 0.725***. Therefore it should be possible to obtain resistant genotypes to both blackleg and tuber soft rot. Several resistant diploid hybrids were selected from among those tested, which also have several other characters desirable for potato breeders.     

Transgenic potatoes expressing anErwinia pectate lyase gene — results of a 4-year field experiment

Summary Four potato lines of cv. Désirée that express the pectate lyase (PL) isoenzyme 3 ofErwinia carotovora subsp.atroseptica were examined in a 4-year field experiment with respect to plant development, tuber yield and resistance of tuber tissue toErwinia soft rot. The PL3 degrades plant cell wall pectin into unsaturated oligogalacturonates eliciting plant defence responses. In one line, enzyme expression was controlled by the CaMV 35S promoter (C) and in three lines it was driven by the potato patatin B33 promoter (D). Plant development of the D-lines in field plots was not distinguishable from that of the non-transgenic counterpart. Also tuber yield was not too different. By contrast, plants of the C-line were smaller than those of the nontransformed counterpart and also showed reduced tuber yield. There were no significant differences in dry mass, starch and protein content of tuber tissue between PL transgenic and non-transgenic potatoes. But compared with the latter, field-grown tubers expressing the PL displayed an enhanced resistance toErwinia soft rot. Thus, average rotting caused byEc-bacteria was diminished in tubers of PL-transgenic lines by 34.1%. The resistance of tubers toEc soft rot was significantly correlated with the PPO activity in tuber tissue.     

The effect of temperature on the development of wilting and on progeny tuber infection of potatoes inoculated with South African strains of biovar 2 and 3 ofPseudomonas solanacearum

Summary Potato plants, after inoculation with a South African strain of biovar 2 or a strain of biovar 3 ofPseudomonas solanacearum were grown in growth chambers at temperatures ranging from 14/16°C (dark/light) up to 25/30°C. When grown at 14/16°C or higher, plants inoculated with the biovar 2 strain developed wilting symptoms and internal symptoms on progeny tubers and the disease was transmitted by the progeny tubers. When the plants were inoculated with the biovar 3 strain, wilting symptoms developed from 18/20°C, internal symptoms on progeny tubers from 20/22°C and the disease was transmitted by progeny tubers from plants grown at 16/18°C or higher.     

Evaluation of a PCR kit for the detection ofErwinia carotovora subsp.atroseptica on potato tubers

Summary A PCR-based kit, Probelia^TM, for the detection ofErwinia carotovora subsp.atroseptica (Eca) on potatoes was evaluated at five laboratories in four countries. The kit is based on DNA-specific PCR amplification followed by detection of amplicons by hybridization to a peroxidase-labelled DNA probe in a microplate. Specificity of the PCR primers for Eca, regardless of serogroups, was confirmed by testing against 246 bacterial, fungal and plant species. Detection limits of the assay varied little between six Eca strains in pure cultures (1.3×10² to 1.5×10³ cells ml⁻¹). When Eca-free tuber peel extract from four cultivars was inoculated with known numbers of 15 Eca strains, detection limits were more variable (1.0×10¹ to 6.2×10³ cells ml⁻¹ peel extract), attributed probably to inconsistency in the recovery of DNA during extraction. When the PCR assay was compared with three current commercial Eca detection methods, using naturally contaminated tubers, results matched most closely those from viable counts on a selective medium, the most sensitive method (88%), followed by enrichment ELISA (72%) and last ELISA (30%), the least sensitive method.