Early detection of Verticillium wilt resistance in a potato breeding program

Nine potato clones/cultivars selected for varying degrees of Verticillium wilt (VW) resistance based on degree of vascular colonization, were intercrossed and outcrossed to 13 uncharacterized clones. Based on growth characteristics of 3,535 seedling transplants and an assay quantifyingV. dahliae in potato vascular tissue, 404 clones were selected for further tests. The average number ofVerticillium dahliae colony forming units (CFU) expressed as Log₁₀(CFU + l)/0.1 ml of plant sap for the 404 clones was 2.3, and 1.3 and 3.7 for resistant Reddale and susceptible Kennebec checks, respectively. Thirty-five clones (8.66%) had CFU values equal to or less than Reddale suggesting that VW resistance is a readily transmitted trait.     

The effect of wound severity, type of tissue, and use of the tuber disk model system on arachidonic acid-induced rishitin accumulation in potato tuber

The wound response within damaged potato tubers (Solanum tuberosum L.) must be a coordination of suberization with other resistance responses if infection is to be avoided. Previously, we showed that wound healing was affected by wound severity and consequently the type of tissue damaged within the tuber. Using arachidonic acid-induced accumulation of rishitin as a model for phytoalexin accumulation, we now demonstrate that this tuber resistance response is also influenced by wound severity and the type of tissue exposed to the elicitor. Kennebec and Reddale tubers that were superficially wounded by removing a thin tissue slice (0.75 mm thick) from the surface and then treated with the elicitor arachidonic acid produced significantly less rishitin than more severely wounded tubers (e.g., tubers cut in half) and excised tuber tissue disks (17 mm × 4 mm). Excised, elicitortreated tuber tissues accumulated significantly more rishitin in cortical cells than in perimedullary and pith cells. Rishitin accumulation was routinely measured 96 h after wounding and was found to be declining by 144 h regardless of wound severity. Induction of rishitin accumulation was very localized with more than 80 % of the rishitin found within 0.75 mm of the treated wound. These results indicate that excised tuber tissue disks, often used as models in wound research, are not fully representative of intact tubers. Perhaps more important, the results show that superficial wounds of intact tubers, i.e., similar to shallow nick and abrasion type wounds typically incurred during harvest, accumulate very little rishitin. However, rishitin accumulation in the tuber disk model system is much greater than that found in superficially wounded tubers, but is similar to the accumulation that could be elicited in cut (seed) tubers. The complications contributed by these wound-related and tissue-specific interactions must be factored into the model system(s) used in describing the role(s) of phytoalexins in the broad framework of disease resistance for stored potatoes and cut seed.     

Inoculum potential of verticillium—infested potato cultivars

Viable propagules ofVerticillium albo-atrum andV. dahliae were recovered from potato stems before and after burial in the field during 1968–1971. Of 14 cultivars tested: Red Pontiac and Superior ranked highest in numbers of propagules ofV. albo-atrum. Russet Burbank, Kennebec, and Irish Cobbler ranked highest withV. dahliae, respectively. Survival ofV. albo-atrum in infected stems was evident (5 ? 34 × 10³ ppg) even after burial in field soil for 17 months. In general, survival ofV. albo-atrum (47–69%) was higher thanV. dahliae (23 – 61%) following field burial of infected stems for 7 months.     

Induction of soil suppressiveness to verticillium wilt of potato by successive croppings

Nine soils collected in New Jersey and Maryland were infested with microsclerotia ofVerticillium dahliae and planted with potato cv. Superior for five successive crops in the greenhouse to induce suppressiveness to Verticillium wilt. Potatoes were harvested 9–12 wk after planting and disease incidence was determined by recovery ofV. dahliae from stem segments on selective medium. Six of the nine soils had high disease incidence during the first cropping and remained conducive to Verticillium wilt throughout the experiment. Disease incidence increased in soils 3 and 9 over time, while disease incidence decreased nonlinearly in soil 1. Viability of microsclerotia buried in soil 9 for 4 wks was reduced compared to the viability of microsclerotia buried in soils 1, 3, and 6. Addition of 50 or 100 g hydrated lime/10 kg soil to soil 9 eliminated its suppressiveness to Verticillium wilt. Repeated cropping of the same soil can induce suppressiveness to Verticillium wilt. Soils exhibiting induced suppressiveness may be a source of new antagonists againstV. dahliae.     

The germplasm release of F87084, a fertile, adapted clone with multiple disease resistances

F87084 is a germplasm release that has been developed by conventional breeding methods and can be traced back to very diverse germplasm sources. This clone has excellent female fertility, round-oval tubers and is well adapted to Eastern Canada. The vine maturity is slightly later than that of Kennebec, and the mean marketable yield is 78% of Kennebec. The specific gravity, boil, bake and chip scores are somewhat lower than Kennebec. F87084 is resistant to late blight,Verticillium dahliae, early blight, and the pathotypes of wart occurring in Newfoundland. F87084 has extreme resistance to PVY^o and PVX. It is also resistant to the potato cyst nematode pathotype Ro1. Progeny analyses indicate that F87084 is duplex for the genes controlling extreme resistance to PVX as well as resistance to the potato cyst nematode (Ro1).     

Fertility and management practices to control verticillium wilt of the russet burbank potato

Management practices for the suppression of Verticillium wilt of Russet Burbank potato include sanitation, use of optimum sprinkler-irrigation practices, soil solarization, and an adequate soil fertility program. Among all cultural factors considered, nitrogen (N) deficiency in potato tissue was most commonly associated with the severity of Verticiilium wilt in Russet Burbank potato. Field studies have shown that increased N availability suppresses Verticiilium incidence on cv Russet Burbank while having no effect on cv Norgold Russet. Studies from both greenhouse and field show that the colonization ofV dahliae in potato stem tissue is suppressed in cv Russet Burbank when the availability of Phosphorous (P) is increased to the optimum. Following one season of cropping with Russet Burbank potato, the treatment providing the optimal N availability also suppressed the increases ofV dahliae populations in the soil during the following year of consecutive cropping. Similarly, after two seasons of continuous potato cropping, treatments with optimal P had lower soil populations ofV. dahliae in soil. Results show the suppression of Verticiilium wilt with optimal fertility. Verticiilium wilt [caused by eitherVerticiilium dahliae Kleb, (microsclerotial form) orVerticiilium albo- atrum Reinke and Berthold (dark mycelial form)] is one of the most severe diseases of potato in the United States. Potato yields, tuber size, and specific gravity may be substantially reduced by this disease, depending on severity, time of occurrence, and growing season. In Idaho and other arid growing regions of the United States, Verticiilium wilt is caused byVerticillium dahliae Kleb. Idaho field studies involving cropping practices, soil fumigation treatments, solarization, and Verticillium-resistant potato clones all support the importance ofV dahliae upon potato yield. Data from these studies show that yield losses due toV. dahliae commonly approximate 5 to 12 metric T/ha (5, 6). Table 1 illustrates the effect of several cropping practices upon relativeV dahliae populations in soil with the effects upon both disease severity and potato yield. With continuous cropping of Russet Burbank,V dahliae populations in soil increased, and yields were reduced by 18 to 19 metric Tlha when compared to locations that had been allowed to remain either weed-free and fallow or where corn had been previously cropped. It was estimated thatV dahliae was influencing yield by about 10 to 12 metric Tlha based on a regression analysis of yield as a function ofV dahliae populations in soil. Similarly, whenV dahliae was suppressed by fumigation treatments involving mixtures of dichloropropene and picfume, yields more significantly increased by 6.5 to 12 metric T/ha (5). Although many factors may influence yields, clones with higher Verticillium resistance than Russet Burbank generally out-yield Russet Burbank. Figure 1 demonstrates the effect of solarization (a technique involving the laying of clear plastic on soil for the purpose of elevating soil temperatures to killV dahliae). When Verticillium was controlled in this manner (lo), there was no significant yield difference between the Russet Burbank potato and the highly resistant A68113-4 clone. The A68113-4 clone grown in non-solarized soil out-yielded the Russet Burbank in non-solarized soil by 31 percent while the solarization treatment significantly increased yield for the Russet Burbank and A68113-4 clones by 46 and 18 percent, respectively. These yield responses were observed in a field where inoculum levels ofV dahliae were relatively low (2.10 cfulgrn of soil). With these losses, the need to develop control procedures is great and to achieve this control the need also exists to accurately evaluate the effect of treatments uponV dahliae. Our Idaho studies have routinely utilized such a procedure (9) in combination with comparisons of both disease severity and yield.     

Contamination withErwinia Carotovora andVerticillium Albo-Atrum during multiplication of pathogen-tested seed potato crops,cultivar Kennebec

Contamination withErwinia carotovora andVerticillium alboatrum (dark mycelial form) occurred rapidly in certain seasons and districts in cv. Kennebec seed potato crops which were derived from pathogen-tested stem cuttings and propagated with attention to practical procedures of crop hygiene. Only varietycarotovora ofE. carotovora was found.     

Prevalence of Verticillium wilt in potatoes in the Red River Valley area of Minnesota

Verticilium spp. were present in over 60% of potato fields in Clay, Marshall, Norman, and Polk Counties in the Red River Valley in Minnesota. ThreeVerticillium spp. were isolated from 27% of the potato stems collected in commercial fields during 1968–1972.Verticillium alboatrum andV. dahliae were recovered from infected stems in approximately equal amounts (51% to 47%), butV. nigrescens was recovered from less than 2% of all host-infected plants. Of 12 cultivars sampled, Superior, Kennebec, Red Pontiac, Sebago, Irish Cobbler, and Norland were most frequently infected.     

Agronomic performance of potato germplasm selected for high resistance to verticillium wilt

A three-year study was conducted from 1991 through 1993 to evaluate the field performance of potato clones that had been selected for resistance to wilt symptom development and stem colonization byVerticillium dahliae. The total yield, size distribution, and specific-gravity of these highly resistant clones were compared with standard cultivars and two parent clones with high Verticillium resistance and high yield, A66107-51 and A68113-4. Two groups of Verticillium resistant germplasm were selected in 1991. One group was the highly resistant progeny from a cross between A66107-51 and A68113-4. The second group consisted of hybrids between wild species accessions with high Verticillium resistance and cultivated diploid and tetraploid germplasm. Twenty-one out of 125 progeny clones from the A66107-51 × A68113-4 cross were highly resistant to Verticillium infection and were tested in yield trials for two years. Another eight selections had high Verticillium resistance but such low yield of seed tubers that they were tested only one year for yield. Eleven of the original 29 highly resistant selections were significantly lower in total yield than Russet Burbank. Only one clone was significantly higher in yield than RB, and none outyielded either of the resistant parents. Reddale and Century Russet had moderate to high Verticillium resistance, respectively. Both outyielded Russet Burbank and were comparable with the high yielding resistant parents. Of 15 interspecific hybrids tested in 1991, only four had sufficient yield to produce seed for yield trials and good wilt resistance after three successive seasons of evaluation. We concluded from this study that selecting at early stages of variety development strictly for Verticillium resistance is likely to be inefficient. Rather, selection should be for yield and other agronomic criteria in Verticillium infested fields.     

An assay to quantify vascular colonization of potato byVerticillium dahliae

A procedure was developed and tested to evaluate potato germ plasm for relative rates of vascular colonization byVerticillium dahliae. The number of colony forming units (CFU) per ml of plant sap, extracted from stems and plated on a medium, was used to assess vascular colonization from plants grown for 100 days in a field plot infested withV. dahliae. Sap extracted from main stem tissue at the ground line gave more CFU ofV. dahliae than tissue at the mid and apical region of the main stem. Aliquots of 0.05, 0.1 and 0.2 ml of plant sap were equally efficient in measuring vascular colonization. Thirty-three clones were evaluated for amount of vascular colonization in field and greenhouse tests with the former giving the best results. Cultivars and clones resistant to Verticillium wilt in Idaho, Maine, Minnesota, North Dakota and New York each had a low index of wilt and a low number of CFU/ml ofV. dahliae in plant sap in these tests. Clones intermediate in both wilt index and vascular colonization as well as selections with a high wilt index and extensive vascular colonization were found using the fresh sap assay method. Correlation values of, r = 0.92, r = 0.91 and r = 0.86 were observed between CFU/ml and percentage of foliar wilt for 11 clones/cultivars in 1986, 1987 and 1988, respectively. The correlation between CFU/ml and both moisture and temperature for the cultivars Kennebec and Russet Burbank, during a 6-yr period was r = 0.83.     

Management of nitrogen and phosphorus rates does not suppress verticillium wilt in yukon gold

Effect of nitrogen and phosphorus fertilization rates on the incidence of Verticillium wilt caused byVerticillium albo-atrum orV. dahliae and tuber yield in potato cv Yukon Gold was evaluated in field plots. In three years of study,V. albo-atrum caused a higher incidence of wilt symptoms (74%–100%) thanV. dahliae (5%–81%) resulting in lower marketable tuber yield. In plots infested withV. albo-atrum, tuber yield ranged from 12.3-22.8 T ha1 compared to 18.1– 31.5 T ha^-1 in plots infested withV. dahliae. Application of nitrogen above (280 kg ha^-1) or below (70 kg ha^-1) the recommended rate of 140 kg ha^-1 at planting did not suppress foliar symptoms or protect against yield loss caused by eitherV. albo-atrum orV. dahliae. Similarly, phosphorus rate adjustments did not consistently affect disease incidence regardless of the pathogen. Management of nitrogen or phosphorus fertility rates at planting does not appear to be a viable approach for suppressing Verticillium wilt in the determinant, wiltsusceptible cultivar Yukon Gold.     

Recovery ofVerticillium dahliae from North American certified seed potatoes and characterization of strains by vegetative compatibility and aggressiveness

Samples of tubers collected from commercial potato seed lots produced across North America were assayed forVerticillium dahliae. V. dahliae was successfully isolated from 65 of 224 seed lots tested, a successful isolation rate of nearly 30%. Vegetative compatibility of the isolates was assessed through complementation tests using nitrate non-utilizing mutants. AllV. dahliae isolates belonged to vegetative compatibility group (VCG) 4. Of the 162 isolates recovered, 64% belonged to VCG 4A, 33% to VCG 4B, and 3% to VCG 4AB. All 39 of the isolates tested in the greenhouse on potato cv. Superior were pathogenic to potato. Disease symptoms developed earlier, were more severe, and plants died earlier when inoculated with VCG 4A compared to 4B isolates. As a group, AUSPC values were significantly higher (p=0.05) for VCG 4A than for 4B isolates. These data suggest that (1) commercial certified seed tubers from diverse locations are commonly infected withV. dahliae and thus may serve as primary sources of the pathogen; (2) potato isolates ofV. dahliae in North America belong to VCG 4A and 4B and these strains are widely distributed via seed tubers; and (3) VCG 4A and 4B are distinct pathotypes ofV. dahliae that vary in their aggressiveness to potato.     

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

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

The Low Potential of Teff (<Emphasis Type="Italic">Eragrostis tef</Emphasis>) as an Inoculum Source for <Emphasis Type="Italic">Verticillium dahliae</Emphasis>

Teff (Eragrostis tef) is a fine stemmed annual grass and gluten free small grain that is of interest as a forage, cover, or a rotation crop. Little is known about the susceptibility of teff to many diseases. Teff could be grown in rotation with potato in the northwestern United States provided teff cultivation is economical and does not increase soil populations for pathogens affecting rotation crops such as Verticillium dahliae. Verticillium dahliae infects a wide range of dicotyledonous plants, making it one of the most important fungal pathogens of crop plants in North America, including potato. The objective of this study was to quantify the susceptibility of teff to eight V. dahliae isolates and compare the susceptibility of teff to eggplant. Teff was confirmed as a host for V. dahliae, as indicated by the presence of microsclerotia in teff stems and roots after artificial inoculation in two years of greenhouse studies. The number of microsclerotia produced in teff did not differ between mint and potato pathotypes of V. dahliae. No V. dahliae isolate produced significantly greater numbers of microsclerotia than any of the seven other isolates tested in a two-year study. Microsclerotia production of V. dahliae in teff was consistently less than in susceptible eggplant cv. Night shadow in both greenhouse experiments (P?<?0.02). It is unlikely that teff infected by V. dahliae will proliferate microsclerotia of mint or potato-aggressive pathotypes, especially when compared to susceptible eggplant cultivars.     

Maturity-Adjusted Resistance of Potato (<Emphasis Type="Italic">Solanum tuberosum</Emphasis> L.) Cultivars to Verticillium Wilt Caused by <Emphasis Type="Italic">Verticillium dahliae</Emphasis>

Verticillium wilt is a fungal disease of potato caused by two species of Verticillium, V. dahliae and V. albo atrum. The pathogen infects the vascular tissue of potato plants through roots, interfering with the transport of water and nutrition, and reducing both the yield and quality of tubers. We have evaluated the reaction of 283 potato clones (274 cultivars and nine breeding selections) to inoculation with V. dahliae under greenhouse conditions. A significant linear correlation (r = 0.4, p < 0.0001) was detected between plant maturity and partial resistance to the pathogen, with late maturing clones being generally more resistant. Maturity-adjusted resistance, that takes into consideration both plant maturity and resistance, was calculated from residuals of the linear regression between the two traits. Even after adjusting for maturity, the difference in the resistance of clones was still highly significant, indicating that a substantial part of resistance cannot be explained by the effect of maturity. The highest maturity-adjusted resistance was found in the cv. Navajo, while the most susceptible clone was the cv. Pungo. We hope that the present abundance of data about the resistance and maturity of 283 clones will help potato breeders to develop cultivars with improved resistance to V. dahliae.     

Trichoderma asperellum is effective for biocontrol of Verticillium wilt in olive caused by the defoliating pathotype of Verticillium dahliae

Verticillium wilt caused by a highly virulent, defoliating (D) pathotype of Verticillium dahliae is threatening olive production in Spain and other Mediterranean countries. This disease must be managed by an integrated strategy, in which biocontrol agents can play an important role. We have investigated the potential of Trichoderma asperellum strains for antagonism against V. dahliae and suppression of Verticillium wilt of olive caused by the D pathotype. First, we tested the antagonistic potential of T. asperellum strains Bt2, Bt3 and T25 against six V. dahliae isolates, four of the D and two of the nondefoliating (ND) pathotypes, in different in vitro assays. All T. asperellum strains overgrew the colonies of all V. dahliae isolates to a similar extent. However, extracellular compounds from strains Bt3 and T25 showed higher anti-V. dahliae activities than those of Bt2 in membrane assays. Also, growth of Bt2 was reduced by ND V. dahliae whereas that of Bt3 and T25 was not affected by V. dahliae-secreted compounds. In planta assays using strains Bt3 and T25, and ’Picual’ olive plants, showed that the two T. asperellum strains significantly reduced the severity of symptoms and the standardized area under the disease progress curve caused by highly virulent D V. dahliae, but not the final disease incidence. Strain T25 significantly increased growth of ‘Picual’ plants and displayed higher ability for colonizing the olive rhizosphere and establishing endophytic infection in olive roots than Bt3.     

Verticillium wilt of potato: a model of key factors related to disease severity and tuber yield in Southeastern Idaho

In three years (1994, 1995, and 1996), a total of 100 commercial potato fields in southeastern Idaho were surveyed for soil variables, severity of Verticillium wilt, soil inoculum density ofVerticillium dahliae andColletotrichum coccodes, colonization of stems, root, and tubers byV. dahliae andC. coccodes, and tuber yield, size, and quality. As a generalization, factors related to soil integrity (organic matter, organic nitrogen, and increased nutrient availability) were most closely related to wilt suppression and higher tuber yields, whereas factors related to loss of soil integrity (sodium and reduced nutrient availability) were related to increased wilt and lower tuber yields. In a multiple regression analysis, three independent variables, feeder-root infections byV. dahliae, sodium content in soil, and soil organic content, were significant predictors of tuber yield. With these three factors, this model accounted for 49%, 53%, and 62% of the field variability related to total yield in 1994, 1995, and 1996, respectively. Throughout this investigation,V. dahliae root infections had the most direct effect on tuber yield, which emphasizes the importance of quantifying root infections in epidemiological studies of Verticillium wilt. Based on these results, organic matter may be one factor that can be manipulated for suppression of Verticillium wilt without reducing soil populations of the pathogen.     

Are plant cell wall hydrolysing enzymes of saprobe fungi implicated in the biological control of the Verticillium dahliae pathogenesis?

We studied the possibility that the antagonistic saprobe fungi Coriolopsis rigida, Trametes versicolor, Fusarium lateritium, Penicillium chrysogenum and Verticillium dahliae-2379 may control V. dahliae disease through the inhibition of the activity of hydrolytic enzymes produced by the pathogen. These saprobe fungi were able to decrease the growth in vitro of V. dahliae. The exudates produced by these fungi seemed to be the main factor responsible for their antagonistic effect. The exudates of all the fungi tested had endopolymethylgalacturonase, endoglucanase and endoxyloglucanase activities. Exudates of P. chrysogenum and V. dahliae-2379 had higher hydrolytic activities than those of V. dahliae. The saprobe V. dahliae-2379 did not decrease the growth of the pathogen V. dahliae but decreased its harmful effect on the growth of tomato. However, no inhibition of the activities of the hydrolytic enzyme of the pathogenic V. dahliae by the no pathogenic V. dahliae-2379 or by the other saprobe fungi was observed. Nevertheless, the protection of tomato by the five antagonistic saprobe fungi against V. dahliae disease could be carried out by the induction of plant defense by the hydrolytic enzymes produced by these fungi.     

The Effect of Alfalfa Residue Incorporation on Soil Bacterial Communities and the Quantity of <Emphasis Type="Italic">Verticillium dahliae</Emphasis> Microsclerotia in Potato Fields in the Columbia Basin of Washington State,USA

Verticillium wilt, caused by the soil-borne fungus Verticillium dahliae, is one of the most important diseases of potato in North America. Soil incorporation of alfalfa residues prior to planting potato could be a nonchemical Verticillium wilt management tactic by reducing the number of viable microsclerotia in field soil. Verticillium dahliae microsclerotia were quantified in field soils where organic material from alfalfa was incorporated, and numbers of microsclerotia were compared to fields where alfalfa residue was not incorporated. In addition, bacterial metagenomics was utilized to characterize soils where organic material from alfalfa was or was not incorporated to determine if alfalfa residue incorporation facilitates the formation of soils that suppress or kill V. dahliae microsclerotia. The number of V. dahliae microsclerotia in soil was greater (P = 0.0003) in fields where crop residue was incorporated than fields without incorporation when chloropicrin was used as a fumigant. Conversely, the number of V. dahliae microsclerotia observed in potato plants did not differ (P = 0.4020) between fields where residues were or were not incorporated if chloropicrin was used. Alfalfa residue incorporation did not significantly alter the soil bacterial metagenome compared to fields not subject to residue incorporation in both years of study. Despite these conclusions, the method can be employed to analyze the effect of grower practices with the intent of linking a field practice to increasing soil bacterial diversity and decreasing Verticillium wilt severity on potato.