Defence reactions in two potato cultivars following infection with two races ofPhytophthora infestans

Summary The kinetics of accumulation of chitinases, glucanases and phytoalexins were studied in two potato cultivars, differing in their degrees of vertical and horizontal resistance, when infected with two races ofPhytophthora infestans. Tuber disks of cvs Kennebec (susceptible, low horizontal resistance) and Huinkul (tolerant, high horizontal resistance) were infected with either race “0” (avirulent) or “C” (complex, race 1.3.5.7.11) ofPhytophthora infestans. Extracts of tuber tissue (0–7 days) from cv. Kennebec infected with race “0” showed a strong increase in phytoalexin production and in chitinase and glucanase activities when compared with those infected with race “C”. These results indicate that race “C” is able to block defensive reactions. No significant differences were observed in cv. Huinkul infected with both races. The contribution of these reactions to horizontal resistance is unknown, and our results would not support a conclusive role for them in the interaction.     

Glycoalkaloid content inSolanum species and hybrids from a breeding program for resistance to late blight (Phytophthora infestans)

As part of an effort to study the relationship between the “glycoalkaloid trait” and genetic resistance to late blight (Phytophthora infestans), glycoalkaloid content in tuber and foliar tissues from a wide genetic background withinSolanum, includingS. demissum, S. iopetalum and 15 hybrids selected from a backcrossing breeding scheme was determined. Analysis of variance indicated significant genotypic effect on total glycoalkaloid, solanidine, α-solanine, and α-chaconine content in both tubers and leaves. Tubers from wild potato species commonly contain glycoalkaloids in concentrations that exceed international health regulations for human consumption (20 mg/100 g fresh weight). In this study,S. demissum andS. iopetalum were highest in total tuber glycoalkaloids among all materials tested, with 70.4 and 76.2 mg/100 g fresh weight, respectively. In contrast, both commercial cultivars had the lowest concentration, both below the safety limit. Solanine was more abundant than chaconine in all but one genotype. All hybrids were intermediate to low in total glycoalkaloids. Except for the two wild species, glycoalkaloid concentration in leaves of all genotypes studied was at least twice that in tubers, with glycosylated forms accounting for more than 80% total glycoalkaloid content. Correlation between tuber and foliage alkaloids was poor. In view of the observed field resistance to late blight, it was concluded that tuber glycoalkaloid content may not be responsible for such resistance.     

Biochemical and morphological characterization of potato clones differing in their resistance to late blight

Summary Morphological and biochemical parameters that could be involved in resistance to late blight were studied in non-infected and in infected potato hybrids resulting from a cross betweenSolanum phureja, resistant toPhytophthora infestans, and a susceptibleSolanum tuberosum. Some morphological differences between resistant and susceptible hybrids, indicating a positive correlation between stem diameter and phloem thickness in the stem and resistance toP. infestans, were observed. The lignin content in the leaves of the resistant hybrid rose upon infection byP. infestans. In the leaves of the susceptible hybrid, a diminution of the lignin content could be observed upon infection byP. infestans. In the same context, peroxidasic activity raised upon infection byP. infestans in both resistant and susceptible hybrids. Further characterization of the hybrid clones based on the polymorphism of peroxidases was attempted using isoelectric focusing.     

Wild tuber-bearing species ofSolanum and incidence ofPhytophthora infestans (Mont.) de Bary on the Western slopes of the volcano Nevado de Toluca. 5. Type of resistance toP. infestans

Summary Plants of wildSolanum species (S. demissum (dms), S. verrucosum (ver), S. iopetalum (iop), S. brachycarpum (bra), and an unidentified species (unsp), from the natural habitat were inoculated in the laboratory withPhytophthora infestans race 0, to see if major genes for resistance were present, and with the most complex and aggressive race available in an attempt to reveal the level of general resistance. No plant with a susceptible reaction to race 0 was found in 120 plants ofdms, and 34 plants ofunsp. Plants with a hypersensitive reaction to race 0 and susceptible plants were found inver. When infected with a complex aggressive isolate of the fungus 73.5% of the WSS plants were susceptible. It was concluded that WSS in the area possess both complex race specific resistance and a high level of general resistance, whose effects cannot yet be separated.     

In vitro evaluation of resistance of potato cultivars toPhytophthora infestans

Summary The resistance of ten potato cultivars (Agria, Ajax, Désirée, Liseta, Kennebec, Majestic. Monalisa, Prima, Spunta and Tonda di Berlino) toPhytophthora infestans was analyzed in vitro using 8 fungal strains. An assay based on electrolyte leakage was used for screening leaves and tuber tissues with fungal culture filtrates. With almost all cultivars the resistance of leaves did not correlate with the resistance of tubers. Cv. Ajax appeared the least susceptible in both leaf and tuber tests, while the cv. Prima was the most susceptible in tuber tests.     

Systemic induced resistance to late blight in potato by treatment with salicylic acid andPhytophthora cryptogea

Summary Tests for systemic induced resistance (SIR) to late blight were performed with different potato cultivars in field trials and in growth chamber experiments. The non-pathogenPhytophthora cryptogea (Pc) and salicylic acid (Sa) were used as inducer agents. In the field, plants were naturally infected byPhytophthora infestans while the indoor plants were inoculated. The degree of SIR obtained varied with the cultivar used and also with the type of inducer. For example in the field the susceptible cv. St. Cecilia had enhanced resistance with Pc whereas Sa caused increased susceptibility. Inbred resistant cvs acquired increased resistance after treatment with Sa. In a separate pilot experiment in a growth chamber, Sa at two different concentrations was injected into potato seed tubers before planting. The higher Sa concentration increased resistance in genetically resistant cultivars while the lower Sa concentration brought about increased susceptibility.     

Steroid glycoalkaloids and related compounds as potato quality factors

The steroid glycoalkaloids are triterpenoid derivatives which are found in all tissues of the potato plant including the tubers. The compounds are largely localized in the peel of tubers, but tissue beneath the peel rapidly accumulates the steroid glycoalkaloids to levels equal to or greater than those in the peel as a result of injury or environmental stress. The accumulation is restricted to the outer 1–2 mm of injured or stressed tuber. Potatoes containing over 0.02% steroid glycoalkaloids are considered toxic to man, and at this concentration they would impart a distinctly bitter flavor. The accumulation of steroid glycoalkaloids is suppressed and the accumulation of sesquiterpenoids is elicited in tubers infected by various pathogens and nonpathogens including the late blight pathogen,Phytophthora infestans. Arachidonic acid and eicosapentaenoic acids, two polyunsaturated fatty acids isolated fromP. infestans, are potent inhibitors of steroid glycoalkaloid accumulation. Both acids elicit the localized accumulation of sesquiterpenoids including rishitin, lubimin, phytuberin, phytuberol and solavetivone. Rishitin and lubimin generally comprise 85–90% of the total sesquiterpenoids which accumulate. The steroid glycoalkaloids and sesquiterpenoids appear to have a role in disease resistance to some fungal pathogens. Both groups of compounds are synthesized via the acetate-mevalonate pathway. Arachidonic and eicosapentaenoic acids appear to inhibit steroid glycoalkaloid accumulation at the level of the conversion of farnesyl pyrophosphate to squalene and they activate the biosynthesis of sesquiterpenoids. The reduction of steroid glycoalkaloids in potato foliage and tubers for health and flavor considerations should be considered relative to the ability of tubers and foliage to accumulate sesquiterpenoids in response to infection and its influence on disease and insect resistance.     

Transgenic potato plants expressing oxalate oxidase have increased resistance to oomycete and bacterial pathogens

Summary Potato (cv. Bintje) was transformed with a gene encoding an oxalate oxidase from wheat under the control of the CaMV35S promoter. Transgenic potato plants produced high constitutive levels of H₂O₂ as visualized by 4-chloro-l-naphtol staining. The resistance of these plants was tested againstPhytophthora infestans. An increased level of resistance to the disease was marked by a reduced number of lesions as well as by a decreased number of sporangia formed per lesion. In addition, oxalate oxidase overexpressing plants also exhibited improved resistance toStreptomyces reticuliscabiei, the causal agent of netted scab. Increased expression of oxalate oxidase had no effect on the interaction withErwinia carotovora. These experiments show that overexpression of oxalate oxidase represents a potentially interesting approach for protection of potato to pathogens.     

Variation in tuber pathogenicity ofPhytophthora infestans in the Netherlands

Summary Variation in aggressiveness to tubers among isolates ofPhytophthora infestans sampled from three potato growing regions in the Netherlands was compared. Variation in the ability to infect tubers of cv. Bintje was found between isolates of each of the three regional populations. The most aggressive isolate of the old population matched the average level of the new population in its ability to infect tubers. As a consequence, the commonly used reference isolate VK 6C can no longer be considered to be representative of the present population ofP. infestans. Therefore it is recommended that testing tuber resistance for the official list of potato cultivars with this isolate should be discontinued. Tuber infection and subsequent spread of the fungus in the tuber tissues were not found to be correlated. The components of tuber pathogenicity studied were not correlated to pathogenicity factors in the foliage, as measured under growth chamber conditions.     

Der Einflu\ variierter Düngung, einer Pflanzenschutzmittel-Anwendung und der Lagerung auf den Glykoalkaloid-Gehalt in Kartoffelknollen zweier Sorten

Zusammenfassung Es wurden Feldversuche in drei Jahren mit zwei Speisekartoffelsorten, verschiedenen Mengen an Mineraldüngern und der Anwendungen eines Pflanzenschutzmittelns gegenPhytophthora infestans angelegt. Die Knollen wurden direkt nach der Ernte sowie nach einer sechsmonatigen Lagerung auf Glykoalkaloidgehalt untersucht. Zwischen den Sorten treten statistisch gesicherte Unterschiede im Gehalt an Glykoalkaloiden auf. Die Anwendung von Pflanzenschutzmitteln w?hrend der Vegetation erh?ht den Gesamtgehalt an Glykoalkaloiden (TGA) in den Kartoffelknollen kurz nach der Ernte. Eine variierte Mineraldüngung hat keinen statistisch signifikanten Einfluss auf den TGA-Gehalt in den Proben kurz nach der Ernte, N- und K-Düngung ver?ndern diesen jedoch im Laufe der Lagerung. Unabh?ngig von den übrigen Bedingungen erh?ht sich der TGA-Gehalt in diesem Versuch w?hrend der Lagerung im Mittel von 5,8 auf 7,7 mg/100g Frischsubstanz.

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Summary Field experiments were carried out for three years to study the effect of various magnesium, potassium and nitrogen fertilization rates on two medium-early edible potato cultivars. Additionally, one of the experimental areas was sprayed againstPhytophthora infestans to ascertain the effect of fungicides on the content of total glycoalkaloids (TGA) in potato tubers after harvest and after six months of storage. The applied fungicide treatments increased total glycoalkaloids (TGA) of potato tubers (Fig. 4). Fertilization with mineral nitrogen caused different reactions in both cultivars. There was a tendency for cv. Beryl to accumulate more glycoalkaloids with increasing nitrogen fertilization rates, while the opposite was observed for cv. Mila (Fig. 7). Fertilization with potassium and magnesium did not affect the results. On the basis of these results there was no conclusive evidence that the TGA content in the tubers was dependent on cultivar or on the weather during the growing season (Figs 1,2 and 3). The TGA content increased during storage, independently of previous treatments, (Figs 4–7).

     

Induction of systemic resistance in potato (Solanum tuberosum L.) plants to late blight by local treatment withPhytophthora infestans (Mont.) de Bary,Phytophthora cryptogea Pethyb. & laff. or dipotassium phosphate

Summary Induced systemic resistance to late blight was found in the potato cultivar Bintje which has no major or minor resistance genes to late blight. The prerequisite was a local inoculation with 5×10³ zoospores/mo, ofPhytophthora infestans race 1, 3, 4, 7, 8, a local inoculation with 2×10³ zoospores/ml ofPhytophthora cryptogea isolated from wheat or local treatment with 50–100 mM solution of K₂HPO₄. Potato plants used in the experiments were propagated in vitro from nodal cuttings. Protection caused by the different inductions was 30 to 70% (assessed as a reduced number of lesions/plant and reduced necrotic tissue/leaf or plant) and tended to positively influence yield and dry matter of the haulm. The induced protection in the potato plant was better in young developing leaves than in fully expanded leaves.     

Changes in race structure of Canadian populations ofPhytophthora infestans based on specific virulence to selected potato clones

Summary A collection of 80 isolates ofPhytophthora infestans, representing multilocus genotypes of the pathogen found in Canada from 1994 to 1996, was screened on a series of nine differential potato host cultivars to determine the physiological race of the isolates. A total of 28 pathotypes were found among the 11 genotypes present in the collection. There was a significant increase in the complexity of pathotypes from 1994 to 1996 which reflected the displacement of the US-1 genotype (mean of 2.2 host differentials infected) by the US-8 genotype (mean of 8.1 host differentials infected). Eighteen of the US-8 isolates could overcome all nine of the resistance genes tested. Virulence genes which overcame host resistance conferred by R8 were the rarest in the populations studied. The predominance of complex populations ofP. infestans in Canada implies that the introduction of cultivars with specific R genes for resistance would be an ineffective control measure. AAFC Contribution No. 901     

Effect and interaction of crop management factors on the glycoalkaloid concentration of potato tubers

Summary Potato tuber glycoalkaloid content was measured in response to nitrogen fertilizer rate, storage temperature, length of storage period and cultivar. Cvs Gemchip, Norchip and Russet Burbank were grown with applied nitrogen fertilizer rates of 0, 168 or 336 kg/ha and then stored at either 4.4 or 10°C. Total glycoalkaloid content was determined one month before harvest, at harvest, after three months of storage and after nine months of storage. Higher rates of nitrogen, higher storage temperature and a period of storage all resulted in significantly (P<0.05) higher concentration of glycoalkaloids. The cv Norchip had higher glycoalkaloids than cvs Gemchip or Russet Burbank. Only the storage period had more influence than the environment (difference between years). Significant (P<0.05) two-way interactions were detected for year x cultivar, year x nitrogen, storage period x cultivar and nitrogen x cultivar. Most interactions were due to the unique responses of cultivars.     

The expression of general resistance to late blight (Phytophthora infestans) in potato leaves

Summary Resistance toPhytophthora infestans was evaluated histologically using leaves of potato cultivars with different levels of general resistance. Host genotype did not influence the preinfectional processes of encysted zoospores, e.g. frequency of germination, length of germ tubes, or frequency of formation of appressoria. Most of the appressoria on the leaf surface were attached to epidermal cells, but they were also found on cells of the stomatal complexes, on hairs and glands, and on cells adjacent to vascular tissue. There were significant differences between cultivars in the extension of intercellular hyphal growth and branching 24 and 48 h after inoculation, the differences increasing with time. In contrast to cv. Bintje, the hyphae in the most resistant cultivar Matilda were mostly confined to the epidermal and palisade cells even 48 h after infection, the number of hyphae per section was much lower, and sporulation capacity was reduced.     

Determination of the glycoalkaloids α-solanine and α-chaconine levels in 18 varieties of potato (Solarium tuberosum L.) grown in Oman

Summary The glycoalkaloid content in 18 potato varieties grown in Oman was measured by HPLC using peel and flesh tissues. The total glycoalkaloid content of the tubers was found to be variety dependent and varied greatly between the 18 varieties. The total glycoalkaloid in the tubers of seven varieties was within the safe limit of ≤200 mg kg^{− 1} FW recommended for food safety, whereas the remaining 11 varieties exceeded this upper limit. The principal glycoalkaloids α-chaconine and σ-solanine were present in higher concentration in the peel than in the flesh of all varieties with the former being the principal alkaloid in the peel. The flesh of all of the 18 potato varieties contained total glycoalkaloid levels lower than the upper safety limit. The leaves of the 18 potato varieties were also analyzed for glycoalkaloids. The levels of glycoalkaloids in expanding leaves were higher than that of expanded leaves with the exception of six varieties. The principal glycoalkaloid in expanding and expanded leaves was α-chaconine. No correlation was observed between the glycoalkaloids levels of in the leaves and tubers.     

The involvement of phenolics and phytoalexins in resistance of potato to soft rot

Summary Phenolics, for example chlorogenic, caffeic and ferulic acid, and phytoalexins, such as rishitin and phytuberin, were identified in potato tubers cv. Kufri Chandramukhi. The tissue of healthy tubers contained no detectable phytoalexins but did contain phenolics. The levels of these compounds were correlated with soft rot development. The rotting tissue either was free of these groups of compounds or had low concentrations. The wound periderm formed as a result of recovery from injury and infection contained high levels of the compounds. Much higher concentrations were detected at lower storage temperatures when oxygen supply was adequate. Antibacterial properties of the phenolics identified were tested againstErwinia carotovora which was inhibited by chlorogenic, caffeic and ferulic acids. The three phenolics were more effective together, in proportions in which they occurred in wound periderm, than individually. It was observed that none of these phenolics could inhibit pectolytic enzymes ofE. carotovora.     

Reaction toPhytophthora infestans of potato genotypes with race-specific resistance

Summary Phytophthora infestans isolates collected in Poland in 1987–1995 were evaluated on detached leaflets of a differential series consisting of potato genotypes possessing resistance genes R1–R11. On the same differentials was evaluated over three years a highly virulentP. infestans isolate MP 245. Isolate MP 245 and those from natural populations ofP. infestans were seldom or inconsistently pathogenic to R5, R8 and R9. When leaflets of the differentials were infected with MP 245 the lesions were often small and sporulation weak. A statistically significant interaction was found between years and pathogenicity of MP 245 to individual differentials. Attempts to train the isolate MP 245 for pathogenicity to R9 were unsuccessful. The variation in specific resistance of potato genotypes may complicate the evaluation of their resistance toP. infestans, as well as the evaluation of the virulence spectrum of natural fungus populations. This variation may be due to changes in specific pathogenicity of the fungus or in specific resistance of the potato. Some differentials have a possibly useful resistance toP. infestans.     

Resistance to late blight and soft rot in six potato progenies and glycoalkaloid contents in the tubers

Glycoalkaloids are anti-nutritional compounds commonly found in wildSolanum species used as resistance sources to major potato pathogens. It is therefore important for breeding purposes to know whether selecting for resistance using such species necessarily selects also for high glycoalkaloid contents in the tubers. To test this hypothesis, we used six partial progenies from crosses betweenSolanum tuberosum and accessions ofS. andigena, S. berthaultii, S. phureja, andS. vernei to investigate the possible correlation between resistance toPhytophthora infestans and/or toErwinia carotovora subsp.atroseptica and the concentration of glycoalkaloids in tubers. Concentrations of α-solanine and α-chaconine in the tubers segregated in each progeny, as did resistance to each pathogen. Some, but not all, clones from each progeny showed hypersensitive reactions to the isolate ofP. infestans used. Furthermore, clones within each progeny also differed for components of partial resistance toP. infestans, suggesting that all four wild species could be used as sources of both race-specific and partial resistance to late blight. With the exception of low, but statistically significant, correlations between concentration of α-solanine and two late blight resistance components (incubation period and spore production per unit lesion area) in progenies derived fromS. vernei, and despite a trend towards higher glycoalkaloid concentrations in the tubers of the clones most resistant to soft rot within progenies derived fromS. berthaultii andS. vernei, no consistent relationship between resistance to either disease and concentrations of α-solanine and/or α-chaconine was observed. These results indicate that neither race-specific nor partial resistance to late blight and soft rot in the accessions used as progenitors of resistance depend on high solanine or chaconine concentrations. These resistance sources could thus prove useful in breeding programs for improved behaviour againstP. infestans and/orE. carotovora.     

Indications of soil borne inoculum ofPhytophthora infestans

Summary In this paper we present observations made during three years in a potato field in Sweden that indicated the presence of soil borne inoculum ofPhytophthora infestans. In 1994, a fungicide trial against late blight was left in the field with various degrees of late blight infection. In 1995, cereals was grown in the field. In 1996, certified potato seed tubers were planted and blight appeared early in the season in a pattern corresponding to the heavily infected areas in 1994. No volunteer potato plants were found in 1995 or 1996. Both mating types A1 and A2 were isolated in 1996 and 1997 and oospores were found in leaves, stems as well as in stolons.     

Inheritance of resistance to late blight in foliage and tubers of progenies from parents differing in resistance

Summary TenSolanum tuberosum genotypes differing in resistance to late blight (Phytophthora infestans (Mont.) de Bary) in foliage and tubers were intercrossed to determine the inheritance of disease resistance in their progenies. Plots of 10–15 clones per progeny were established in each of 2 years and resistance assessed by field or laboratory tests. The parental genotypes were similarly tested each year. The parents differed in general combining ability (GCA) for both foliage blight (FB) and tuber blight (TB). The parental and GCA scores were significantly correlated for both aspects of the disease, but the correlations between foliage and tuber scores for parents and for GCAs were not significant. Three parental genotypes were highly resistant in both foliage and tubers, and the genotype with the highest GCA for resistance to both FB and TB (cv. stirling) is recommended as the best parent. There was no evidence of strong genetic correlation between both aspects of resistance, and it is suggested that both be selected for in a breeding programme.