Am. J. Pot Res (2009) 86:335–346 DOI 10.

1007/s12230-009-9088-9

Aggressiveness of Streptomyces on Four Potato Cultivars and Implications for Common Scab Resistance Breeding
Leslie A. Wanner & Kathleen G. Haynes

Published online: 19 May 2009 # Potato Association of America 2009

Abstract Common scab (CS), caused by several species of Streptomyces, is a serious problem for potato growers. Although the mechanism of pathogenicity, based on the phytotoxin thaxtomin, is presumably conserved in all pathogenic species, Streptomyces isolates vary in aggressiveness, and regional patterns in Streptomyces species distribution have recently emerged. We demonstrate that Streptomyces isolates differ significantly in aggressiveness, and there are specific plant genotype-pathogen isolate interactions in four contrasting potato cultivars treated with Streptomyces isolates belonging to different species and molecular types. There were significant differences between experiments, among isolates, among cultivars, and some of the two-way interactions were significant. The sum of all main and interaction effects among plant, pathogen, and experiment accounted for only 50–55% of total variation in CS disease symptoms. More information on specific plantpathogen interactions combined with knowledge of the distribution of CS-causing species could form the basis for successful recommendations of suitable potato cultivars, and will contribute to more accurate and reproducible phenotyping required for genetic studies of CS resistance in potato. Resumen La roña común (CS), causada por varias especies de Streptomyces, es un problema grave para los cultivadores de papa. Aunque el mecanismo de patogeniL. A. Wanner (*) : K. G. Haynes United States Dept. of Agriculture, Agricultural Research Service, Genetic Improvement of Fruits and Vegetables Laboratory, 10300 Baltimore Ave., Beltsville, MD 20705, USA e-mail: Leslie.Wanner@ars.usda.gov

cidad, basado en la fitotoxina taxtomina, se conserva presumiblemente en todas las especies patógenas, los aislamientos de Streptomyces varían en agresividad, y han surgido recientemente patrones regionales en la distribución de las especies de Streptomyces. Demostramos que los aislamientos de Streptomyces difieren en agresividad de forma significativa, y hay interacciones especificas entre genotipo de planta-aislamiento del patógeno en cuatro diferentes cultivares de papa tratadas con aislamientos de Streptomyces pertenecientes a diferentes especies y tipos moleculares. Hubo diferencias significativas entre experimentos, entre aislamientos, entre cultivares, y algunas de las interacciones de dos vías fueron significativas. La suma de todos los efectos principales y de interacción entre plantas, patógenos, y experimentos representaron sólo el 50–55% del total de la variación en los síntomas de la enfermedad CS. Más información sobre las interacciones planta-patógeno especificas combinado con el conocimiento de la distribución de las especies que causan CS podría constituir la base para recomendaciones exitosas de cultivares adecuados de papa, y contribuirá a la más exacta y reproducible fenotipificación necesaria para los estudios genéticos de resistencia a CS en la papa. Keywords Plant-pathogen-disease triangle . Resistance breeding Abbreviations CS common scab MA mean area tAI transformed mean area ML mean lesion PAI pathogenicity island

1999). Pasco et al. The taxonomy of plant pathogenic streptomycetes has become more complicated by the recognition that the species Streptomyces scabies is actually a small group of different species (Bouchek-Mechiche et al. depending on soil temperature (BouchekMechiche et al. 1999. with more susceptible cultivars typically displaying warty or pitted lesions. 2000a. stelliscabiei and temporary species/strain IdahoX (Wanner 2007). scabies was apparently most common throughout much of the region. caused by soil-inhabiting Streptomyces species. but two other pathogenic species or strains. Harrison 1962). Wanner (2006) reported finding four Streptomyces species or strains that cause CS in the northern USA S. wart-like lesions or sunken pits. 2006). S. 82% of growers reported CS. Pasco et al. CS is caused by soil-inhabiting bacteria in the genus Streptomyces. J. scabies and S. Kreuze et al. In a third variation. The lineages of these four potato cultivars are quite distinct. Wanner et al. Bouchek-Mechiche et al. 1993. A netted scab-like disease referred to as russet scab has also been described in North America (Faucher et al. Symptom variation is partly determined by the potato cultivar. Wanner 2006. Losses to common scab (CS) in Canada are probably typical. Two of the potato genotypes chosen. 2005. acidiscabies. 1999. and is caused primarily by S. Loria et al. some strains of S. 2000b. 1998. Superior and Pike. Wanner 2009). 1998. Netted scab is limited to very few potato cultivars. 2000a. Streptomyces pathogenicity groups specific to potato cultivars have not been recognized in North America. 2006. Streptomycetes are abundant in soils. CS-causing Streptomyces isolates vary in aggressiveness (Bramwell et al. reticuliscabiei is genetically very closely related to another species causing CS. Mygind 1962. There seem to be two partially overlapping potato scab diseases caused by plant pathogenic Streptomyces species (Bouchek-Mechiche et al. Wanner 2006). while Chippewa and Green Mountain are highly susceptible. has also been described from the northeastern US and Canada (Faucher et al.336 Am. but most commercially successful cultivars are highly CS-susceptible. cause CS (Loria et al. S. Bukhalid et al. There is a relatively wide variation in potato cultivar susceptibility. 1997. Our objective in this study was to compare CS disease phenotypes on four potato cultivars in response to genetically different Streptomyces isolates. affects tubers only superficially. CS affects tubers (and underground stems) on all potato cultivars to differing degrees. netted scab. Kinkel et al. 2006). the biological basis for variation in aggressiveness is assumed to be differences in the production of thaxtomin. A second disease. The netted scab pathogen S. europaeiscabiei can cause either common or netted scab. Pasco et al. 1995). Lehtonen et al. is a serious problem for potato growers worldwide. All of these species or strains cause a similar spectrum of CS symptoms on potatoes. turgidiscabies in Scandinavia (Kreuze et al. 2008. Though a growing body of evidence indicates that different Streptomyces species or strains are characteristic of geographic areas (St-Onge et al. Wanner 2004). 2005). but damages roots. The only way of determining CS resistance has been years of field trials. effective controls are still lacking. S. and perhaps as many as 10. The CS situation in Europe is apparently more complex. At least three. Loria et al. thereby affecting yields. Regional and season-to-season differences in both disease severity (disease pressure) and rankings of potato cultivar susceptibility to CS have been reported in North America (Haynes et al. were characteristic of specific regions or field locations. Costs of common scab to the potato industry in Tasmania are estimated at 4% of total crop value (Wilson 2004). Pasco et al. McKee 1958. 2006). the most desirable method of control would be disease-resistant plant material. Flores-Gonzalez et al. 2005). showing a range of symptoms from superficial dark-colored scaly lesions to discrete or coalescing raised. though only a small percentage of these. a species tolerant of acidic soil conditions. europaeiscabiei in northern Europe (Bouchek-Mechiche et al. Pot Res (2009) 86:335–346 Introduction Common scab. Genes for biosynthesis of thaxtomin are carried on a distinct chromosomal region with the hallmarks of a pathogenicity island (PAI) that can be transferred horizontally among streptomycetes (Loria et al. turgidiscabies (Bouchek-Mechiche et al. Lambert and Loria 1989. 2000b. CS-causing Streptomyces species or strains have now been delineated in North America (Goyer et al. carrying genes for biosynthesis of a phytotoxin thaxtomin. 2000b. 2008. 2008) or S. europaeiscabiei (Bouchek-Mechiche et al. Since the mechanism of pathogenicity based on thaxtomin seems to be conserved. St-Onge et al. and average losses were estimated at $90 to $102 (CDN) per hectare (Hill and Lazarovits 2005). 1996. Information on the responses of individual potato cultivars to Streptomyces species and strains combined with an understanding of prevalent pathogen populations in North America is required to analyze the genetics of plant resistance to Streptomyces. reticuliscabiei or S. are considered to be relatively resistant to CS. 2005). Although CS has been described in the literature for more than one hundred years. Streptomyces isolates were chosen to represent several plant pathogenic species found in North America. 1992. 2006. with variable results from year to year and place to place. 2002. 2004. 2006). 2006). and genetically distinct Streptomyces isolates differing in thaxtomin production in vitro have been described (Beausejour et al. Like many plant diseases. and is caused by S. Lambert et al. Wanner 2009). . Progress in breeding for CS resistance is hampered by limitations in resistance screening.

Idaho X 3 4 5 S. Ranger Russet Aberdeen. located adjacent to the nec1 gene in S. 5 ng DNA template and MilliQ water. scabies and S. ME) and ‘Pike’ was obtained as certified seed from Dan Table 1 Characteristics of Streptomyces isolates compared in this study Species and isolate #a S. as determined using the methods of Wanner (2009. of origin mel.25 U GoTaq DNA polymerase (Promega Corp. a final extension at 72°C for 7 min and ending with a 4°C hold.Am. not noted eastern WA. and maintained in the greenhouse at USDA-ARS in Beltsville. 4. presence or absence of the nec1 gene. Anchored PCR was carried out using the primer TRR-s (5’-ACTGTCGGATGCGGTCAT-3’). europaeiscabiei 1 2 S. 1 place. production of melanin when grown on peptone-yeast extract-iron agar presence (upper case) or absence (lower case) of genes characteristic of the Streptomyces scabies pathogenicity island (PAI). Madison. WI). Pot Res (2009) 86:335–346 337 and could be useful in understanding the CS disease potential in different regions. 2001. third letter. stelliscabiei 6 a b c d Isolate name originb Melc PAI Genotype d ID-01-16c ID-02-12 ID-01-6. approx. Wanner 2007) and maintained at −80°C as spore suspensions in 20% glycerol or/and plugs cut from YME agar plates of confluent sporulating cultures (Wanner 2006). annealing at 46°C for 30 s and extension at 72°C for 2 min. potato cv. Materials and Methods Origins. BOX-PCR was carried out using the BOX1AR primer (Clark et al. 1998) in a 50 µl PCR reaction containing 1X Green GoTaq Flexi buffer (Promega Corp. 25 pmol each primer.. ID 2001 cv. followed by 40 cycles of denaturing at 95°C for 20 s. second letter. Subculture. followed by 40 cycles of denaturing at 95°C for 30 s.CGCCGAG CTCTGCG-3’) and Sph15A (5’-CACCGCATGCGCAC-3’). 2003 cv. J. not noted – + + + + + TNT Tnt TnT Tnt TnT TNT Isolate numbers are the same as those used on gel lanes in Fig. Shepody Aberdeen. approximately 5 ng DNA template (1 µl of a 1:9 dilution) and MilliQ water. 2001.2a ID-03-3A ID-05-11E NY-02-3a ID. 1. respectively). ID. Storage of Streptomyces Isolates Streptomyces isolates were cultured directly from scabby lesions on potato tissue as described (Wanner 2006.. presence or absence of the tomA gene . Amplification was carried out in a Perkin-Elmer Thermal Cycler 480 programmed for initial denaturation at 95°C for 5 min. 1.0 mM MgCl2. cv. Details on the origins of isolates are given in Table 1. 25 pmol primer BOX1AR. Desiree American Falls. a derivative of a primer designed to an open reading frame with homology to transposases. ‘Green Mountain’ and ‘Superior ’ were obtained as certified seed from Bradstreet Family Farm (Bridgewater. ID. Madison. MD. Amplification was carried out in a Perkin-Elmer Thermal Cycler 480 programmed for one initial denaturation step at 95°C for 5 min. All PCR amplification experiments were repeated at least twice for each DNA template. 2003. Isolates are named according to the state or province from which they came and the harvest year of the scabby potatoes (first two letters and two numbers. Russet Burbank Freeville. 300 µM each dNTP. coupled with random primers Sac15A (5’. annealing at 53°C for 40 s and extension at 72°C for 3 min. in press) First letter. cv.). presence of the txtAB operon. Fifty microliter PCR reactions contained 1X Green GoTaq Flexi buffer (Promega Corp.25 U GoTaq DNA polymerase (Promega Corp. cv. WI). 1998).). sp. Potato Cultivars and Sources Potato cultivar ‘Chippewa’ was originally obtained as certified seed from Jewel Brothers (Monticello. turgidiscabies (Bukhalid et al. 2.0 mM MgCl2. ME). 200 µM each dNTP. year. and ending with a 4°C hold. 2005 cv. Molecular Characterization of Isolates by PCR DNA extraction and PCR characterization of variable regions of the 16s ribosomal RNA genes (used to assign Streptomyces species) and presence or absence of genes in the Streptomyces pathogenicity island was carried out as described (Wanner 2009). NY.

Disease Scoring CS was rated as two components.5% commercial bleach (0. Quebec) was placed in the bottom of 15 cm diameter pots. and then 1. Green Mountain is a late maturing white. Harvested spores were counted using a microscope and hemocytometer slide. Since the error variances were homogeneous. lightly netted cultivar introduced from Wisconsin in 1961. Chippewa is not widely grown presently. rarely developing pits. Pot Res (2009) 86:335–346 Corey. Plants were grown under controlled conditions of 24°C constant temperature.5 cm in diameter) 4 = Coalescing raised (warty-looking) lesions (>0. Tubers were rated individually for the amount of surface area covered on a percentage basis (visual estimate). Pike is widely cultivated in Idaho. netted skinned cultivar. Pots were distributed into three growth chambers (one pot of each treatment per chamber). For each pot. and individual cultivarisolate pairs were compared using the pdiff option.5 cm−3 Vigoro All Purpose Plant Food 12-5-7 (Spectrum Group Division of United Industries Corporation. Washington. and is relatively CS-resistant. but a second lesion type was also found.5 cm in diameter) 5 = Pitted lesions of any size or depth. Minnesota. Potato cultivars utilized represent a range of genetic heritage in US cultivars.75 cm in diameter. Data Analysis MA and ML from the two experiments were analyzed separately by the general linear models procedure in SAS version 9. when plants were beginning to naturally senesce. It is still valued as a specialty cultivar in Maine. Katahdin was a great-grandparent of Pike. Least squares means for isolate. relatively early in maturity. Superior is a medium maturity. and New York. typically developing corky pits. Michigan. . Sums of squares from the sources of variation were used to determine the contribution made by each component to the host plant-diseaseexperiment disease triangle. Intermediate values were assigned when most lesions were of one type.000 cm−3 of sand: potting mix (1:1) containing 1010 CFU of Streptomyces vermiculite culture was added (final pathogen concentration. cultivar. Cary. It is highly susceptible to CS. For each pot. typically developing reddish-brown or blue-black inky deep pits. The scoring scheme used was: 0. smooth-skinned chipping cultivar. J. A single uniform mixture of sand: soil:inoculum was made for each Streptomyces isolate and distributed into pots. 500 cm−3 un-infested 1:1 sand:soil-less potting mix (ProMix PGX. 6. and grown at 28°C for 2– 3 weeks until uniformly sporulating.6×106 CFU/ cm−3 soil mixture).5 cm in diameter) 3 = Discrete raised (warty-looking) lesions (<0. New York. Chippewa is a round. but is related to Katahdin. Correlations between tMA and ML were calculated over experiments and for clones over experiments using the correlations procedure in SAS. netted skinned cultivar from New York. Dorval. Residuals from ML and the square root transformation of MA (tMA) were normally distributed between and across experiments. The experiment was conducted twice. It is still grown widely in Maine. The second measurement was the type of lesion present (lesion severity). St.09% sodium hypochlorite) containing a few drops of commercial dishwashing liquid for 2 minutes and rinsed thoroughly in reverse-osmosis purified tap water before cutting into ca. Maine. Pots were harvested individually after 3 ½ months. produced by Premier Horticulture.5 = Rough scaly or cracked surface. Potato tubers were surface-disinfested in 1. a cultivar grown widely in the northeastern US.75 cm in diameter. a mean lesion severity (ML) was calculated from all tubers larger than 0. Louis. white. Spores were harvested by gently scraping water-flooded plates. Plants were watered when the soil became dry. Pike is a recently introduced CS-resistant white. and Wisconsin as a chipping cultivar. Pathogenicity/Aggressiveness Assays on Potato Cultivars For assays of Streptomyces pathogenicity and aggressiveness on potato. Culture of Streptomyces Isolates for Soil Inoculation Original −80°C stored cultures of Streptomyces isolates were plated on YME agar.338 Am. MO). 28 g seed pieces and planting in pots containing prepared mixture of Streptomyces isolate. the mean percentage area covered (MA) was calculated from all tubers larger than 0. dark in color and distinct from russeting 1 = Discrete superficial lesions of small size (<0. Three seed pieces were planted for each cultivar for each isolate in a complete randomized design. Nebraska. Wisconsin. and fertilized every three weeks with1. and equal numbers were used to directly inoculate sterile vermiculite in plastic bags. but is highly susceptible to CS. Monticello ME.25–0. Ltd.1 (SAS Institute. 14 h photoperiod with 200–220 µmol-m2-s−1 light intensity from Phillips Natural Daylight fluorescent bulbs. Vermiculite cultures were grown for 14–18 days and then sampled to estimate CFU per cm−3 vermiculite. introduced in 1885.5 cm in diameter) 2 = Coalescing superficial lesions (>0. the data were combined over experiments and similarly analyzed. NC).

In experiment A 15 pots (out of 72) failed to produce any tubers.2 were the most aggressive isolates as measured by lesion severity (ML).27 < r <0.05.198. 750. correlations between these two measures of CS severity were highly significant (Chippewa. from top.645. and an IdX isolate ID05-11E were the least aggressive isolates. ID01-16c. ID03-3A: lane 5. and 300 bp bands are visible Aggressiveness of isolates on potato cultivars was measured as two components. Variance in symptoms within and between pots in a treatment was also high (Figs. The most aggressive isolate in terms of area covered (tMA) was NY02-3A. S. 676. Pot Res (2009) 86:335–346 339 Results Streptomyces Isolates Used for Comparison We compared six Streptomyces isolates representing three Streptomyces species (all common in North America) and at least five genotypes. Virulence and aggressiveness of all of the isolates except ID05-11E had previously been examined in radish. J. However. where all were pathogenic (Wanner 2004) and highly aggressive. which was a simple percentage of surface area showing CS symptoms at potato tuber maturity. 2a. lane 2. ID05-11E. ID01-16c. The three Idaho X (IdX) isolates also fell into at least two groups based on repetitive sequence (BOX and transposonanchored) PCR patterns (Table 1 and Fig. 500. the correlations between the two measures differed greatly depending on the level of susceptibility in the cultivars. r =0.0001. Fig. P =0. lanes 1-2.02. Both tMA and ML were significantly greater in experiment A than in experiment B. The second component was the mean lesion rating (ML). lane 6. there was no correlation between these two measures (Pike. 1 Repetitive element PCR profiles differ in Streptomyces isolates used in this study. 517. 1a) and banding patterns obtained by a RAPD method using one primer anchored in a transposase sequence (Fig. on all potato . 460.29). there were significant differences between the two experiments for tMA and ML (Table 2). Green Mountain. Promega pGEM marker.53. and in each experiment (0. r =0. 396. europaeiscabiei are different genetic types or strains. r = −0. P =0. P =0. ID01-6. lanes 3-5. from top: 2. Lane M2.78). as measured by either tMA or ML. Promega PCR markers. 1. stelliscabiei isolate NY02-3A. r =0.78. c TRR/Sac15A primer pair.90. Superior. lane 4. Lane M1.2a.30. 1. Origins and molecular characteristics for the six Streptomyces isolates are presented in Table 1 and Fig. whereas in the second replicate (experiment B) tubers were produced in all the pots. a BOX1AR primer. europaeiscabiei isolates: lane1. 1. S. All six isolates had genes encoding biosynthesis of the pathogenicity determinant thaxtomin. There were significant differences in mean aggressiveness among isolates (Tables 2. ID0212. 1). 1. 3. S. Disease Severity Measurements BOX1AR M1 1 2 3 4 europ Id 5 6 M2 ste TRR-Sph TRR-Sac Fig.0005). as they differ in PAI markers (Table 1). europaeiscabiei isolates ID02-12. repetitive sequence element (BOX) PCR patterns (Fig. while NY02-3A and ID01-6. This variable disease phenotype is reminiscent of the variability seen from field to field and year to year in field testing. 350 bp bands visible. The first component was the mean area (MA). These two measures of CS severity were positively correlated overall (r =0.000. but three isolates were missing the PAI marker gene nec1 and one was missing both the nec1 and tomA PAI marker genes.605.004). 3a). P =0. P < 0. b TRR/Sph15A primer pair. In the resistant cultivars. In the two highly susceptible cultivars. The two isolates of S. 3). species Idaho X isolates: lane 3.Am. The two S. raised lesions as more serious and pitted lesions as most serious. Despite standardization of the initial Streptomyces inoculum density and growth conditions within and between experiments. Small superficial lesions were ranked as least serious. 1b and c).

69 1. is presumably conserved in all plant pathogenic Streptomyces. Notable exceptions to this were that no pitted lesions were observed on cultivars Pike and Superior in response to isolates ID01-16c. isolate.16 2. It has long been recognized that potato .92*a 7. 2006. 4).21** 4. Isolates ID01-16c (on Superior vs. As expected. comparisons of least squares means revealed some significant specific differences in cultivar response to individual isolates (Fig.39 1.54** 3. but not for tMA (Table 2). As expected. Aggressiveness of each isolate on each cultivar is shown in Fig. 3a. 2c).21 a tMA.34* 1. and year-to-year variation in CS severity is also regularly noted. Responses to nearly all individual isolates were distinct on the most susceptible cultivar Chippewa (Fig. 1997. 3b). and cultivar was not significant for either tMA or ML. on cultivars Green Mountain.95 6. 2. Although the initial soil inoculum density was adjusted to be the same in both experiments for all isolates.f. Superior) were the only isolate-cultivar pairs showing significant differences in tMA (Fig. For example.73** 5. there was a wide spread in types of CS lesions (ML) observed in response to different isolates. 2b).f. there were significant differences among cultivars (Tables 2.30 2.49 1. J.2A was most aggressive and NY02-3A was least aggressive (Wanner 2004).63 48 3.63** 31 2. with mean surface area affected by CS symptoms ranging between 5% and 20% (Fig.27 1. Hill and Lazarovits 2005).76 ML 8. though. the range in tMA between treatments was narrow. Table 4). Discussion The mechanism of pathogenicity. However. we were also interested in whether the response of potato cultivars to individual isolates varied. 3).59 1.00 0. 3b).01 5 3 15 4. Differences in Isolate Aggressiveness on the Four Potato Cultivars Streptomyces isolates varied in mean aggressiveness on potato cultivars (Table 3). ** P <0. In general.2A were most severe on Chippewa and ID03-3A was least severe.26 5 3 15 10. Only in cultivar Superior was there no difference in response to any isolates significant at P <0. or on cultivars Green Mountain and Superior in response to isolate ID03-3A. In contrast. Notably.69 0.97** 4.96 2. and isolate NY02-3A was one of the least severe isolates on this cultivar. unexpectedly.f Experiment Pathogen isolate Potato cultivar Isolate x cultivar Experiment x isolate Experiment x cultivar Expt x isolate x cultivar Error tMAa MLa Experiment B d. The isolate x cultivar interaction was significant for ML. 2c). mean common scab lesion type as described in Materials and Methods * P <0. 2c).73 5.73 1.93 cultivars. square-root transformed mean percentage area affected by common scab.35** 13. the experiment x isolate interaction was significant for tMA. isolate ID03-3A was most severe on cultivar Pike. Pike and Superior in response to isolate ID02-12. Even with the wide variation in symptoms indicated in large interquartile ranges (Fig. significant differences in ML were seen between cultivars Pike and Chippewa in response to isolate ID01-6. ML.77* 6. with most isolates capable of producing all types of CS lesions on most potato cultivars (Fig.17* 2. in terms of area covered. Differences in Cultivar Response to Individual Isolates Mean lesion types seen on potato cultivars in response to individual Streptomyces isolates are displayed in Fig.340 Table 2 Analysis of variance on the square root of the mean area (tMA) affected by common scab and the mean common scab lesion type (ML) in replicated experiments and over both experiments Source Mean squares Experiment A d. Green Mountain) and isolate ID05-11E (on Chippewa vs.52** 2.05 (Fig. while isolates NY02-3A and ID016. tMA Am. while pitted lesions were the typical response of cultivar Chippewa to isolate NY02-3A (Fig. 1 5 3 15 5 3 15 79 tMA 11. 2a).2A and between Chippewa and Green Mountain vs. but there are well-documented and consistent regional differences in CS severity in North America (Haynes et al. Pike and Superior in response to isolate NY02-3A (Fig. lesion severity (ML) was greater in the susceptible cultivars than in resistant cultivars (Fig. These results contrast to results in radish where ID01-6. 2).05. Variability in response to different isolates is apparent among cultivars. these significant differences are a reflection of some isolatespecific difference in disease pressure between experiments among some cultivars (Figs. Pot Res (2009) 86:335–346 Both experiments ML d. Haynes et al. 2b).28 3.75* 2. 2a (tMA) and b (ML). The three way interaction involving experiment. based on the phytotoxin thaxtomin. tMA was greater in the resistant cultivars than in the susceptible cultivars. 3. but not for ML. However.15* 9.

Pike. are caused by Streptomyces species. C ID01-6. . McKee 1958). Pot Res (2009) 86:335–346 341 a 10 8 6 tMA 4 2 0 ID01. we obtained evidence for significant differences in susceptibility to CS among four potato cultivars in growth chamber experiments. Numbers beneath cultivars in A are total numbers of tubers scored.12 G P S 13 . C ID02. Combinations with P <0. 2 Differences in aggressiveness of 6 Streptomyces isolates on four potato cultivars. median. 2006. C 33 NY02-3A G P S 21 17 12 16 17 17 13 min 12 17 max 21 18 17 16 median mean b 5 4 ML (lesion type) 3 2 1 0 ID01-16c C G P S . Green Mountain.3A G P S .16c C 29 G P S 22 . it was recognized that Streptomyces causing CS in North America belong to several species.Am. a Box plots showing squareroot transformed tuber surface area (tMA) affected by common scab. 1997.05 are highlighted. C 26 ID02. common scab and netted scab. C 36 ID01-6. Wanner 2009) Although the existence and significance of physiological specialization between Streptomyces species/isolates and potato cultivars has been controversial (Bjor and Roer 1980. In all six tables. . J.2A G P S 7 . . 2008. C NY02-3A G P S min median mean max Fig. Superior. Haynes et al. Recently. P.12 G P S . 2005). Wanner 2006. . arranged by isolate. mean. We therefore thought that closer scrutiny of specific plant-pathogen interactions was warranted. b Box plots showing differences in type of scab lesion (ML) rated as described in the Material and Methods. C 38 ID03. Using six isolates representing some of the Streptomyces species and molecular genetic groups found in the USA. St-Onge et al. and maxima and minima are shown. C 36 ID05-11E G P S 17 .3A G P S 17 . and some evidence suggests that certain species and/or molecular types are characteristic of particular regions (Faucher et al. . G. Chippewa. C ID03. Potato cultivars are indicated as letters: C. C ID05-11E G P S . and that potato cultivars respond differentially to individual pathogenic species/ strains (Bouchek-Mechiche et al. McKee 1958). in Europe it has clearly been shown that two different diseases. differences in lesion type (ML) are displayed above the diagonal and differences in surface area affected (tMA) are displayed below the diagonal cultivars differ in CS susceptibility (Caligari and Wastie 1985. 1992. Specific interactions between individual Streptomyces isolates and individual potato cultivars were found for the tuber surface area affected by CS lesions (tMA) and for . c Matrix showing the probability of significant differences in mean response of potato cultivars to each of the Streptomyces isolates studied. Inter-quartile range. 2000b. Pasco et al.2a G P S . S. Lambert et al.

Disappointingly. We found a positive correlation between the two measures only for the two susceptible potato cultivars. carefully controlled conditions did not reduce the symptom variation within or between experiments.19 0.51 0.98 0.57 0.77 0.39 Chipp GM Pike Sup 0.06 ML 0. and conducting experiments in controlled growth chamber environments. Lambert et al.21 0. the mean surface area affected by CS was greater in the resistant cultivars.97 tMA GM 0.62 Sup 0. Goyer et al.90 0. carefully controlling inoculum density.17 Sup 0. as pitted lesions in particular are not easily removed by peeling during processing.43 0.75 tMA GM 0.01 <0. 1993.14 Sup 0.81 Pike 0.07 Pike 0.94 0.43 0. To clarify the genetics behind CS resistance. The biological reasons for this large variation in symptoms are not clear.06 0.31 Pike 0. 1998. Phenotyping CS resistance is difficult because CS symptoms are so variable.73 0.38 tMA GM 0.03 0.82 tMA GM 0.40 ML 0. more than one of these lesion types occurs on a single tuber.09 0.10 0.17 0. 2 and 3. or on tuber surface area affected by symptoms (Caligari and Wastie 1985. Both components are important in the appearance of potato tubers for fresh market sale.97 0.73 tMA GM 0. well-characterized Streptomyces isolates. J.25 0. although the variability in CS symptoms has inspired most investigators to assess both the type of lesion and the area affected. it is important to have a robust and reproducible phenotype assay. Kinkel et al. using ML as a measure of the degree of disfigurement of potato tubers and MA to express the extent of symptoms. the within-pot symptom variation equals the between-pot and between-replicate experiment variation.81 0. typically ranging from superficial cracks and scales to raised warty lesions to shallow or deep pits. but non-superficial disfigurement in the form of warts or deep pits adds an additional economic concern.41 0.14 0. Symptoms are highly variable on tubers within and between replicate experiments.06 Sup 0. Frequently.66 0. Pot Res (2009) 86:335–346 c ID01-16c Chipp Chipp ML GM Pike Sup ID02-12 Chipp Chipp ML GM Pike Sup ID01-6.02 0.342 Am.34 0. Often. but it complicates data analysis.43 0.67 ML 0.27 Chipp GM Pike Sup NY02-3A Chipp 0.26 0.63 0.01 0.57 Sup 0. although correlation coefficients were sometimes low and variable among years (Bjor and Roer 1980). and no correlation for the resistant cultivars.48 Pike 0.52 Chipp GM Pike Sup ID05-11E Chipp 0. Some investigators have assessed CS simply based on incidence of any symptoms.01 0.66 0.53 0. 2006). 1998).39 0. Our results demonstrate that differences in Streptomyces strain or species prevalence could contribute to regional differences in CS disease severity. We also note that the use of either incidence or MA data alone would give a different assessment of CS disease severity than ML.05 0. Lesions may be discrete or they may coalesce to cover large areas of the tuber surface. We have attempted to reduce the variation in symptoms by using single.84 Sup 0.79 0.17 0.2A Chipp Chipp ML GM Pike Sup 0. Goth et al. 2 (continued) the types of CS lesions (ML). This is the reason we and others have ranked pitted lesions as more severe. We assessed CS symptoms as two separate components. Others have generally reported a significantly positive correlation between affected area and the severity of lesions (Bjor and Roer 1980.11 <. Furthermore.27 0.23 0.52 Pike 0.70 0.71 0.42 0. less . as is illustrated in Figs. we found that while ML was higher for susceptible cultivars Chippewa and Green Mountain as expected.77 Fig.27 0.35 0.01 <.13 ID03-3A Chipp tMA GM 0.19 Pike 0.20 0. Disconnection of these two measures of CS in susceptible versus resistant potato cultivars suggests there are multiple plant genetic components to CS disease response. and the proportion of the tuber surface area affected by lesions varies.60 0.

88 tMA ID01-6.66 0.NY0 16c 12 6.23 0.2A 3A 11E 2-3A Superior 17 17 21 min median mean max b Chippewa ID01-16c ID02-12 ID01-6.Am.10 0.71 0.65 <0.ID01.01 <0.14 0.97 0.28 0. b Matrix showing the probability of significant differences in response of potato cultivars to each of the Streptomyces isolates studied.85 0.45 <0.11 0.92 Fig.01 0.26 0.64 0.81 0.27 0.01 0.38 0.41 0.ID05.50 0.ID01. 12 15 13 ID01.17 0.03 0.07 0.01 0.2A 3A 11E 2-3A Pike 17 18 16 .NY0 16c 12 6. a Box plots showing differences in type of scab lesion (ML).99 0.2A 3A 11E 2-3A Chippewa 29 26 36 38 36 33 .65 0.ID03. Interquartile range.45 0.53 0.06 0.ID05.32 ID0116c ID02-12 0.26 0.76 0.75 0.64 0.99 0.56 0.94 0. 3 Differences in response of four potato cultivars to 6 Streptomyces isolates.38 0.ID02.04 0.78 0.66 0.01 0.83 0.06 0.39 0.01 0.40 0.01 <0.87 0.96 0.96 <0. Differences in lesion type (ML) are displayed above the diagonal and differences in surface area affected (tMA) are displayed below the diagonal .67 0.ID03.02 0. Numbers under the cultivar names are the number of tubers rated for the isolate-cultivar combination.ID02.69 0.2A ID03-3A ID05-11E NY02-3A 0. Combinations with P <0. maxima and minima are shown.40 0.75 0.39 0.48 0.53 0.ID03.20 0.39 0.ID02.84 0.16 0.28 0.43 ID0511E 0. Pot Res (2009) 86:335–346 343 a ML (lesion type) 5 4 3 2 1 0 ID01.38 0.03 0.2A 0.49 0.97 0.13 0.01 <0.30 0.65 0.69 0.53 0.95 0.82 0.34 0.2A ID03-3A ID05-11E NY02-3A Superior ID01-16c ID02-12 ID01-6.55 0.2A ID03-3A ID05-11E ML NY02-3A Pike ID01-16c ID02-12 ID01-6.91 0.01 0.45 0.63 0.77 0.96 0.38 0.87 0.NY0 16c 12 6.55 ID03-3A 0. J.16 0.08 0.ID05.46 0.94 0.2A ID03-3A ID05-11E NY02-3A Green Mountain ID01-16c ID02-12 ID01-6.26 0.21 0.42 0. 22 13 7 ID01.88 0.01 0.ID01.43 0.01 <0.38 0.2A 3A 11E 2-3A Green Mt 17 12 21 17 .83 0.24 0.ID05. median.15 0.20 <0.94 0.NY0 16c 12 6.45 0.ID01.05 are highlighted. 17 16 ID01.26 0. rated as described in the Material and Methods.48 0.93 NY023A <0.ID02.33 <0.10 0.96 0.54 0.07 0.63 0.01 0.06 0.ID03. mean.

It seems likely that further specificity in what isolates or species are most aggressive on individual potato cultivars will be discovered. respectively (Fig. and the interactions between these sources of variance. followed by the plant genotype (6. after unexplained variation.2 and 76. 2006. Area Index (tAI) Host 6.0% H x P x E 11.1 and 55% of the total variation. By doing experiments in growth chambers with carefully controlled initial pathogen pressure.3% HxPxE 11.05 accurately gauging acceptability of affected tubers by the potato processing industry.33 3.01 aa a b b ML 2.1% PxE 3. Lesion Index (ML) Host 6. showing that a large amount of variation remains unexplained by host plant genotype. Decomposition of Variation Due to Plant Genotype. The significant cultivar-isolate interaction for ML and experiment x isolate interaction for tMA we document here have implications for screening potato cultivars for CS resistance.71 a a ab a a b ML 1. different isolates were most aggressive on particular potato cultivars. The ANOVA model for tMA and ML explained only 50.0 and 2. Haynes et al. isolate(s) used to test cultivars for CS resistance should reflect Streptomyces species and types found in the target growing region.1% Experiment 4. and necessitates multiple years of field evaluation of plant cultivars to assess CS resistance in commercial potato cultivars.6%). 4). were able to explain 74. We would A.0% of variation is unexplained B. Pot Res (2009) 86:335–346 a means followed by the same letter do not differ at P ≤ 0.16 4.24 3. Variation in CS phenotype is commonly attributed to lack of control of the environmental component of the plant-pathogen-environment disease triangle. respectively (Haynes et al.6% HxP 16.1 and 12.7%) for tMA and ML.17 3. and then the experiment (4.2% HxP 10.3% Experiment 2.21 2.62 4. However. It therefore seems important to include a lesion type component when phenotyping plants for CS susceptibility. 2000b.22 4. Pathogen genotype explained the most variation (8. were the two-way interaction effects between plant genotype x pathogen genotype x experiment and plant genotype x pathogen genotype which explained 10.39 1. in field studies looking at CS area index and lesion index at two locations over 2 years.71 2. In contrast. We were only partially successful in doing this.05 . 4 Decomposition of variation in common scab disease phenotype. a Host-PathogenExperiment disease triangle for mean lesion. Bouchek-Mechiche et al.46 1.3% of the variation for tMA and ML. pathogen genotype. rather than using an evaluation scheme based only on incidence or affected area.27 1. We did not find evidence for specialized disease types with cultivar specificities such as those that characterize scab diseases caused by Streptomyces species in Europe (Bouchek-Mechiche et al. and an isolate that was most aggressive on one cultivar was least aggressive on another cultivar.2% Pathogen PxE 8.9% of variation is unexplained Fig.2 and 6.9% of the total variation.344 Table 3 Mean tMA and ML by Streptomyces isolate from both experiments Isolate ID01-16ca ID02-12 ID01-6. The two largest sources of variation.2a ID03-3A ID05-11E NY02-3A tMA 3. we attempted to reduce the contribution of experimental and extraneous variation. respectively. respectively.1%). Pasco et al.14 3. Pathogen Genotype and “Environment” The difficulty in reproducing disease severities between experiments remains an obstacle to statistical power in genetic analysis.0% 45. 1997) and computations from their data).0% Pathogen 12.31 1. Possibly we were able to discern cultivar x isolate interactions because we deliberately selected widely divergent plant genetic material and Streptomyces isolates. b Host-PathogenExperiment Disease Triangle for square root transformed mean area means followed by the same letter do not differ at P ≤ 0.39 a a b ab ab b Am.9% HxE 1. J. experiment.1% 7.0 and 16.79 3.7% HxE 4.78 1.96 1. 2005).50 b ab a a 49. in our small selection of only 6 Streptomyces isolates.7% Table 4 Mean tMA and ML by potato cultivar from both experiments Cultivar Chippewa Green Mountain Pike Superior a tMA 3. and standardize the pathogen pressure.

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