Effect of temperature and photoperiod on symptoms associated with resistance toPhytophthora infestans after leaf penetration in susceptible and resistant potato cultivars

The effect of temperature and photoperiod on the expression of resistance against Phytophthora infestans in five potato cultivars with and without resistance (R) genes was investigated. Four experiments were carried out under controlled conditions in growth chambers. Two cultivars (393295.236 and 391046.22) without known R genes from the International Potato Center (CIP) in Lima, Peru, two Mexican cultivars with major R genes (Tollocan and Malinche), and a susceptible cultivar (Atlantic) were used in this study. Plants were grown for 32 days in growth chambers at two temperatures (16 and 24 C) and two photoperiods (12 and 16 h day length), then inoculated with a compatible isolate of P. infestans and incubated in a mist chamber at 18 C. The inoculation efficiency, the percentage of lesions that did not grow beyond the inoculation spot, the sporangia density, and the AUDPC were recorded. The percentage of arrested lesions decreased with temperature in the two most resistant cultivars (393295.236 and Malinche), and the AUDPC was lower at 16 than at 24 C in four of the five cultivars. The inoculation efficiency and the sporangia density were not affected by change in temperature. Sporangia density increased at 16 h photoperiod; however, the final infected leaf area was not affected. Our results demonstrate that the expression of horizontal and vertical resistance was affected by temperature; however, the relative resistance ranking among cultivars was the same in the four experiments with different temperatures and photoperiods. It is assumed that the resistance in the Mexican cultivars may be conferred by minor resistance genes and by the residual effect of defeated R genes. These results emphasize the difficulty in differentiating between horizontal and vertical resistance.     

Foliar reaction toPhytophthora infestans in inoculated potato field trials in Michigan

Late blight, caused byPhytophthora infestans, is the most important disease of potato worldwide and foliar resistance is an important component of managing late blight in the field. The objective of this research was to identify germplasm for use in breeding cultivars with foliar resistance toP. infestans. More than 500 clones were tested from 1997 to 2002 in inoculated (US8 genotype) field experiments conducted at the Michigan State University Muck Soils Research Farm in Bath, Michigan. All of the current commercial cultivars tested were classified as susceptible toP. infestans. The most resistant clones were A90586-11, AWN86514-2, B0718-3, Jacqueline Lee (MSG274-3), MSI152-A, MSJ307-2, MSJ317-1, MSJ453-4Y, MSJ456-2, MSJ456-4, MSJ461-1, MSK101-2, MSK128-1, NY121, LBR8, LBR9, Tollocan, and Torridon. Some of these resistant selections were from crosses with B0718-3, Jacqueline Lee, and Tollocan suggesting that the resistance toP. infestans was transmissible. These resistant clones will provide the opportunity to breed late-blight-resistant cultivars from a diverse pool of cultivated germplasm. Consistent foliar reaction toP. infestans over years suggested that the Michigan State University Muck Soils Research Farm is a valuable location for North American breeders to assess the reaction of potato germplasm to the US8 genotype of late blight.     

Production of phytoalexins, glycoalkaloids and phenolics in leaves and tubers of potato cultivars with different degrees of field resistance after infection withPhytophthora infestans

Summary The kinetics of accumulation of phytoalexins, glycoalkaloids and phenolics was studied in two potato cultivars differing in their degrees of field resistance when infected withPhytophthora infestans. Tuber slices and leaves of cvs Pampeana INTA (high degree of field resistance, free of R genes) and Bintje (susceptible) were infected with race C (complex race 1, 3, 5, 7, 11) ofPhytophthora infestans. Phytoalexins and phenolics accumulated in tuber and leaf tissues which had been inoculated. The levels of these compounds in the susceptible cv. Bintje were relatively low and similar to those found before inoculation. Leaves of cv. Pampeana INTA had a very high glycoalkaloid content, suggesting that glycoalkaloids may play a role in protection of leaves against the fungus. However, we could find no correlation between resistance and glycoalkaloid content of tubers. Our results suggest a major role of phytoalexins, phenolics and glycoalkaloids in the complex mechanisms of field resistance.     

Identification of late blight, Colorado potato beetle, and blackleg resistance in three Mexican and two South American wild 2x (1EBN)Solanum species

Wild potatoes are important sources of genes for resistance to disease and insect pests. A collection of wild Mexican and South AmericanSolarium species from the US potato Genebank was evaluated under laboratory and/or field conditions for their reaction to late blight (Phytophthora infestans), Colorado potato beetle (CPB,Leptinotarsa decemlineata Say), and blackleg (Erwinia carotovora subsp.atroseptica (van Hall) Dye) in order to identify individual genotypes with multiple resistance genes. Late blight inoculations using aggressive isolates (US-8/A2 and US-11/A1 mating types) of P.infestans revealed a wide range of variation for resistance between and within the accessions of the wild species tested. For late blight, susceptible as well as moderately to highly resistant genotypes were observed in all the species tested. However, at least one accession from the three Mexican and one South American wild diploid species tested showed a relatively uniform high level of resistance toP. infestans. These includedS. bulbocastanum, S. pinnatisectum, S. cardiophyllum, andS. circaeifolium. Two accessions from South American speciesS. commersonii were highly susceptible to late blight. For the Colorado potato beetle test, only one species,S. pinnatisectum appeared uniformly resistant to CPB under field conditions. Results of screening for blackleg resistance showed that there were major differences between genotypes in the wild species. Accessions ofS. circaeifolium PI 498119 andS. bulbocastanum PI 243504 were identified as having significantly higher blackleg resistance than cultivated potato and the other wild species tested. However, genotypes from these two accessions were more susceptible to late blight and CPB. Characterization of theP. infestans isolate P1801C.16 used for late blight evaluation and multi-locus isolate tests using US-8/A2 and US-11/A1 races revealed that the resistance inS. pinnatisectum genotypes tested corresponded to a race-non-specific genetic system, which was different from any existing R genes.Solanum pin-natisectum genotypes with both high levels of late blight and CPB resistance as well as blackleg resistance genotypes identified in the present study represent a diverse gene pool that may be useful for development of new potato cultivars with multiple disease and insect resistance. The potential utilization of these valuable sources for improvement of cultivated potato is discussed.     

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.     

Effect of inoculum concentration ofPhytophthora infestans on potato late blight

Zoospore suspensions ofPhytophthora infestans applied to potato cultivars in the field with an exponential inoculum sprayer resulted in defoliation within a range of severity that was log-normal with respect to inoculum concentration. Regression lines for different cultivars differed in position and, depending upon moisture conditions, slope, indicating that the cultivars differed in resistance to penetration and to invasion of tissue by the pathogen.     

Combining Ability for Foliar Resistance to Late Blight [<Emphasis Type="Italic">Phytophthora infestans</Emphasis> (Mont.) de Bary] of Potato Cultivars with Different Levels of Resistance

Combining ability for resistance of potato to late blight (LB) was analyzed by testing segregating seedling progenies obtained from a full diallel crossing design between 3 potato parent cultivars with different levels of LB resistance and a North Carolina Mating Design II with the same resistant parents as females crossed to 2 LB susceptible male parents, against a complex race of P. infestans. Artificial inoculation was conducted under greenhouse conditions in Dalat, Vietnam at 1500 m above sea level. Percentage of foliage infection was recorded visually at 5 days intervals until 25th day after inoculation when the susceptible check was totally destroyed. General combining ability effect was significant and predominant, indicating that in the inheritance of resistance to LB, additive gene actions are more important. Specific combining ability was significant only for crosses between resistant parents, but insignificant for those between resistant and susceptible parents. In almost all cases, no significant reciprocal effect was found. Cultivar C88 was the best general combiner for resistance to LB. There were evidences that 1) the major genes R1, R3a and Rpi-blb1 alone are not enough to condition a good level of resistance to the isolate used; 2) the resistance of C88 was plausibly mostly due to presence of the gene R2, which is probably the main genetic factor of the durable resistance of this cultivar and, 3) the resistance to LB in potato seems background dependent, the background being the set of nuclear factors (major or minor genes) whose expression collectively affects the resistance level.     

A detached-leaf method to evaluate late blight resistance in potato and tomato

We developed a detached leaf method for evaluating potato (Solanum tuberosum L.) and tomato (Lycopersicon esculentum Mill.) germplasm for reactions toPhytophthora infestans, the causal organism of late blight. Primary leaves from the third to the sixth node of potato plants, and the fourth to sixth node of tomato plants were excised at the stem. Their petioles were inserted into 14 mm × 100 mm floral aqua tubes containing 9 ml of sterilized distilled water. The leaves in the aqua tubes were placed with abaxial sides down on galvanized metal hardware cloth (12.5 × 12.5 mm mesh). A 12.5 mm sensi-disc containing 50μl of 2 × 10⁴ zoospores was placed in the centers of the terminal leaflet and the second leaflet pair of the potato leaf. A single disc was placed on the center of the tomato leaf. The supporting hardware cloth was placed 2.5 cm above distilled water (2.5 cm deep) in 31 cm × 17 cm × 8 cm clear plastic boxes with tight fitting lids. Leaves on intact tomato and potato plants were inoculated in a similar manner and placed in a mist chamber. Lesion growth was determined 4, 5, and 6 days following inoculation. There were no significant differences in reactions to isolates ofP. infestans on detached and intact leaves of potato cultivars Green Mountain and Kennebec and the tomato cultivar Bonnie Best.     

Assessing resistance types and levels to epidemic diseases from the analysis of disease progress curves: Principles and application to potato late blight

Both race-specific (RS) and race-non-specific (RNS) resistances exist in potato against the late blight pathogenPhytophthora infestans. Because these resistance types do not have the same epidemiological effects, their presence, alone or combined, in potato genotypes can be deduced from the analysis of disease progress curves from field experiments, a type of data commonly available to potato breeders, and their comparison with those of standard reference cultivars. The identification of RS resistance is based on the presence of a delay in epidemic onset compared to a susceptible cultivar, whereas the identification of RNS resistance is translated into a reduction of apparent infection rates. These parameters can be easily computed after linearization of the disease progress curves. This paper assesses the reliability of this identification using sets of experimental data, discusses its limitations, and highlights potential applications for breeding and cultivar assessment purposes.     

Investigation of Host Responses of Different Potato Genotypes at Tissue, Cellular and Subcellular Levels After Infection with Phytophthora infestans

In histological and cytological investigations, the infection process of Phytophthora infestans, the late blight pathogen, was comparatively studied in several potato cultivars and somatic hybrid genotypes and their parents using fluorescence microscopy and electron microscopy methods. The results showed that germination of zoospores of P. infestans and frequency of invading by infection hyphae did not differ among the cultivar-pathogen interactions, but, extension of hyphae in host cells markedly differed among these genotypes. In the susceptible genotypes the pathogen grew rapidly inter- and intracellularly, 12 h after inoculation (hai), and some digital like haustoria were formed and the cytoplasm of the host cells became disorganized. In the resistance genotypes, the pathogen was restricted to the site of initial penetration, although some hyphae could penetrate the epidermal cell, however, the host cells produced resistance responses, such as formed wall appositions when in contact with hyphae, and no haustoria like structures were found. In the somatic hybride genotypes, the host response was different according to their parents as shown by transmission electron microscopy. In the hybrid genotype 1508/2, like in the wild species S. bulbocastanum, no hyphae were found in host cells. In the other genotypes, hyphae of P. infestans spread intercellularly and formed haustoria, but the cytoplasm of hyphae and haustoria was disorganized and host cell resistant responses often appeared, such as, host cells were disorganized and necrotic and cell wall apposition were observed.     

The Relationship Between Active Oxygen Metabolism and Resistance to Late Blight in Potato

The relationship between active oxygen metabolism and resistance to late blight (Phytophthora infestans) in potato (Solanum tuberosum L.) was studied for 72 h post-inoculation by comparing three resistant cultivars (low disease index) with three susceptible ones (high disease index). Activities of superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX), and the content of ascorbic acid (ASA), were higher in the resistant cultivars than in the susceptible ones. The production rate of the superoxide anion radical (O₂^?) was lower in the resistant cultivars than in the susceptible ones. These changes, which were associated with the potato plant’s response to infection with P. infestans, provide some insight into the physiological basis of resistance and may also provide a screening tool for resistance to late blight.     

Relationship between resistance to late blight in potato foliage and tubers of cultivars and breeding selections with different resistance levels

Due to changes in the pathogen population, late blight of potatoes caused byPhytophthora infestans has become a more difficult disease to manage and there is a need for new potato cultivars with greater and more stable disease resistance. Field studies are required to identify and characterize host resistance to late blight in both foliage and tubers but, as epidemics vary from year to year, combining multi-year results can be difficult. Residual maximum likelihood (REML) was successfully used to analyze data which demonstrated good correlations between foliar and tuber responses to late blight (US-1 genotype). In addition, studies on post-harvest inoculated tubers provided more reliable disease responses than field studies, and the disease resistance with this method correlated well with foliar disease responses. This tuber test would be a useful method of pre-screening selections for disease response prior to testing in the field.     

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.     

Breeding potato cultivars with tubers resistant toPhytophthora infestans

Summary Available information has been reviewed on tuber resistance toP. infestans, its inheritance and breeding procedures used to obtain potato cultivars superior in this character. P. infestans is worldwide the most destructive potato pathogen. Tuber resistance is an essential component of potato resistance as this pathogen is often responsible for tuber rot in storage, and infected tubers of susceptible cultivars provide an important way for its overwintering. In Europe many cultivars and advanced breeders selections have been obtained with tuber resistance toP. infestans, but in the last decades no progress is noted in the mean level of this resistance in cultivars. The expression of tuber resistance depends to a large extent on testing conditions and therefore it can be difficult to evaluate. Consequently making progress in breeding is not easy, and also cultivar assessment data from various countries sometimes differ considerably. It is concluded from published data that it should be easier to make progress in breeding potato cultivars with resistant tubers if more attention is paid to combining the various types of resistance which are already known, and if the genetic determination of durable resistance toP. infestans is better understood. Professor Dr. K.M. Świeżyński passed away on 27th July 2000     

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     

Phenotypic stability of resistance to late blight in potato clones evaluated at eight sites in the United States

Changes in the fungal pathogenPhytophthora infestans in the United States pose a significant threat to potato production. Sources of resistance to these new genotypes of P.infestans need to be identified for potato breeders to have parental materials for crossing, and the phenotypic stability of late blight resistance in these potato clones needs to be determined. Sixteen potato clones which reportedly have some resistance to late blight were evaluated at eight locations: Florida (FL), Maine (ME), Michigan (MI), Minnesota (MN), North Dakota (ND), New York (NY), Pennsylvania (PA) and Wisconsin (WI) in 1996. Percent infected foliage was recorded at approximately weekly intervals following the onset of the disease at each location. Area under the disease progress curve (AUDPC) was calculated. Clones were ranked for mean AUDPC within location and the nonparametric stability statistics, mean absolute rank differences and variance of the ranks, were analyzed for phenotypic stability. Neither of these statistics was significant, indicating a lack of genotype x environment interaction on the rankings of these clones across locations in 1996. The four clones with lowest AUDPC scores were U.S. clones AWN86514-2, B0692-4, B0718-3 and B0767-2. These clones should be useful parental materials for breeders seeking to incorporate genes for late blight resistance into potatoes.     

Testing for resistance to metalaxyl inPhytophthora infestans isolates from northwestern Washington

Outbreaks of late blight occurred in commercial potato fields in northwestern Washington in 1989 and reached epidemic proportions in 1990. Since most blighted fields reportedly had received 2–4 late-season applications of metalaxyl, the possibility existed that pathogenic strains ofPhytophthora infestans had developed resistance to this systemic fungicide. In testing this hypothesis,P. infestans was isolated consistently from diseased leaves, stems, and tubers of potato when small tissue pieces were surface-disinfected in 0.5% NaOCl for 3 min. Virulence of isolates was assessed on detached leaflets of eight differential (R-gene) potato genotypes. Although simple races (single and double) were recovered, there was also a high frequency of complex race combinations. When tested in metalaxyl-amended media, 81% of the 73 isolates recovered from diseased tissues were highly resistant to 10 μg metalaxyl/ml and 19% yielded intermediate resistance. Sensitive isolates were not recovered. Fifty-five percent of 40 isolates sporulated on potato tuber disks in the presence of ≥10 ug metalaxyl/ml. On leaf tissues, 85% of 20 isolates sporulated in the presence of 10 μg metalaxyl/ml, and 50% sporulated in the presence of 100 μg/ml. This is the first report ofP. infestans resistance to metalaxyl in the U.S.     

Comparison of the effectiveness of different methods of screening for bacterial soft rot resistance of potato tubers

Tubers from eight potato cultivars (cvs) grown at two different locations in Wisconsin were tested for bacterial soft rot resistance using different inoculation techniques. The procedures included 1) point inoculations of tubers with different inoculum levels followed by incubation in ambient or low oxygen condtions, 2) inoculation of mechanically bruised tubers followed by incubation in a mist chamber, and 3) a standard slice inoculation method. The point titration test followed by incubation in dew chamber and the mist chamber-bruise test showed similar patterns of resistance for cultivars that were used in these experiments. These two methods are considered to be effective for screening potato tubers for bacterial soft rot resistance. Point titration methods are very useful if only limited numbers of tubers are available. The mist chamber-bruise test is simpler than the other procedures; however, to obtain reproducible results large numbers of tubers are required. Because of the great variability of the results obtained in inoculation of slices, the reliability of this approach can be questioned as a standardized method for evaluation of resistance. Tubers of somatic hybrids ofS. brevidens andS. tuberosum and their sexual progeny were significantly more resistant to bacterial soft rot than tubers of moderately resistant cultivars when evaluated by each of the assay procedures.     

Evaluation of potato cultivars,clones and a true seed population for resistance toRhizoctonia solani

Field and laboratory studies were conducted in 1985–1986 to evaluate the relative severity ofRhizoctonia solani damage among 26 clones and five potato (Solanum tuberosum) cultivars. A test with true potato seeds showed that resistance toR. solani could be identified in two years. One highly resistant line was identified by this method. Moderate degrees of resistance were found in lines from the USDA Potato Breeding Program (BARC) with russet types producing a higher percentage of highly resistant lines than round whites. A laboratory test revealed that apical sprout damage (nipping) was variable within and between lines and that it was an important phase of disease development to measure when determining resistance because of its effects on the resulting plant growth and production of marketable tubers. It was also determined that to accurately define resistance toR. solani in potato, all phases of the disease must be evaluated to insure against resistance “breakdown” at some point in plant development.     

Defender: A high-yielding, processing potato cultivar with foliar and tuber resistance to late blight

The potato cultivar Defender is high-yielding, white-skinned, and notable for having foliar and tuber resistance to late blight infection caused byPhytophthora infestans (Mont.) de Bary. It was released in 2004 by the USDA-ARS and the agricultural experiment stations of Idaho, Oregon, and Washington. Defender is suitable for processing into french fries and other frozen potato products directly from the field or from storage. Defender also may be used for fresh markets in regions such as California, where cultivars with long tubers and white skin are traditionally grown. Resistances to late blight and other potato diseases make Defender an ideal candidate for organic potato production. Defender consistently produced greater total and U.S. No. 1 yields than ‘Russet Burbank’ in Idaho trials. In early harvest trials conducted in the western U.S., average total yields of Defender were 17% and 23% greater than yields for ‘Ranger Russet’ and ‘Shepody’, respectively. In full-season trials conducted in the western U.S., Defender averaged 10% and 15% higher yields than Ranger Russet and Russet Burbank, respectively. Specific gravity of Defender is consistently high, with values comparable to those of Ranger Russet; tuber ascorbic acid (Vitamin C) levels are also high. In addition to late blight, Defender also is resistant to tuber early blight (Alternaria solani, (Ellis & G. Martin), L.R. Jones & Grout), potato virus X, and net necrosis; it has moderate levels of resistance to Verticillium wilt (Verticillium dahliae, Kleb), pink rot, foliar early blight, corky ringspot, and Erwinia soft rot. Defender is susceptible to scab (common and powdery) and potato leafroll virus; it has moderate susceptibility to dry rot (Fusarium spp.) and potato virus Y. Susceptibilities to internal necrosis, tuber greening, and blackspot bruise also have been noted, but may be minimized through cultural and harvest practices.