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Cercospora leaf spot (CLS) is the most destructive foliar disease in sugar beet ( Beta vulgaris ). It is caused by Cercospora beticola Sacc., a fungal pathogen that produces toxins and enzymes which affect membrane permeability and cause cell death during infection. In spite of its importance, little is known about the initial stages of leaf infection by C. beticola . Therefore, we investigated the progression of C. beticola on leaf tissues of susceptible and resistant sugar beet varieties at 12-h intervals during the first 5 days after inoculation using confocal microscopy. Inoculated leaf samples were collected and stored in DAB (3,3'-diaminobenzidine) solution until processed. Samples were stained with Alexa Fluor-488-WGA dye to visualize fungal structures. Fungal biomass accumulation, reactive oxygen species (ROS) production, and the area under the disease progress curve were evaluated and compared. ROS production was not detected on any variety before 36 h postinoculation (hpi). C. beticola biomass accumulation, percentage leaf cell death, and disease severity were all significantly greater in the susceptible variety compared with the resistant variety ( P < 0.05). Conidia penetrated directly through stomata between 48 to 60 hpi and produced appressoria on stomatal guard cells at 60 to 72 hpi in susceptible and resistant varieties, respectively. Penetration of hyphae inside the parenchymatous tissues varied in accordance with time postinoculation and varietal genotypes. Overall, this study provides a detailed account to date of events leading to CLS disease development in two contrasting varieties., Competing Interests: The author(s) declare no conflict of interest.

Cercospora leaf spot (CLS) is a destructive disease limiting sugar beet production and is managed using resistant cultivars, crop rotation, and timely applications of effective fungicides. Since 2016, its causal agent, Cercospora beticola , has been reported to be resistant to quinone outside inhibitors (QoIs) and to have reduced sensitive to demethylation inhibitors (DMIs) in sugar beet growing areas in North Dakota and Minnesota. Isolates of C. beticola resistant to QoIs, DMIs, and both QoIs and DMIs were collected from fields in Foxhome, Minnesota, in 2017. Fitness of these resistant isolates was compared with that of QoI- and DMI-sensitive isolates in laboratory and greenhouse studies. In the lab, mycelial growth, spore production, and spore germination were measured. The results showed that resistant isolates had significantly less mycelial growth and spore production than sensitive isolates, while no significant difference in spore germination was detected. In the greenhouse, six leaf-stage sugar beets were inoculated with a spore suspension made from each resistant group and incubated in separate humidity chambers. CLS disease severity was evaluated visually at 7, 14, and 21 days after inoculation (DAI), and the areas under disease progress curve (AUDPC) were calculated. Resistant isolates had significantly smaller AUDPC but still caused as high disease severity as the sensitive ones at 21 DAI. Although QoI- and/or DMI-resistant isolates had a relatively slower disease development, they still caused high disease severity and need to be factored in disease management practices., Competing Interests: The author(s) declare no conflict of interest.

Cercospora leaf spot (CLS; causal agent Cercospora beticola Sacc.) is endemic in many sugar beet production regions due to the widespread distribution of C. beticola and the inability of current management practices to provide complete control of the disease. Roots harvested from plants with CLS, therefore, are inevitably incorporated into sugar beet root storage piles, even though the effects of CLS on root storage properties are largely unknown. Research was conducted to determine the effects of CLS on storage properties including root respiration rate, sucrose loss, invert sugar accumulation, loss in recoverable sucrose yield, and changes in sucrose loss to molasses with respect to CLS disease severity and storage duration. Roots were obtained from plants with four levels of CLS severity in each of three production years, stored at 5°C and 95% relative humidity for up to 120 days, and evaluated for storage characteristics after 30, 90, and 120 days storage. No significant or repeatable effects of CLS on root respiration rate, sucrose loss, invert sugar accumulation, loss in recoverable sucrose yield, or change in sucrose loss to molasses were detected after 30, 90, or 120 days storage regardless of the severity of CLS disease symptoms. Therefore, no evidence was found that CLS accelerates sugar beet storage losses, and it is concluded that roots harvested from plants with CLS can be stored without additional or specialized precaution, regardless of CLS symptom severity., Competing Interests: The author(s) declare no conflict of interest.

Minnesota and North Dakota combined contain 55% of the sugar beet production area in the United States, contributing to 49% of the nation's sugar beet production in 2018. Fusarium diseases caused by Fusarium oxysporum f. betae and F. secorum on sugar beet can cause significant reduction in both root yield and sucrose concentration and purity. The objective of this research was to identify an alternative artificial inoculation method to induce Fusarium diseases on sugar beet leaves and roots caused by both Fusarium spp. in greenhouse conditions to better aid in research efforts. We tested four inoculation methods, including barley to seed, barley to root, drenching, and cutting. and compared them with the conventional root-dipping inoculation method. The inoculation method of placing Fusarium -colonized barley seed close to sugar beet seed (barley to seed) caused levels of symptom severities on both leaves and roots similar to the root-dipping method. Because the traditional root-dipping method involves a laborious transplant process, use of infected barley seed as inoculum may serve as an alternative method in the evaluation of host resistance and pathogen virulence among Fusarium diseases by Fusarium spp. on sugar beet at the seed or seedling stage.

Rhizoctonia crown and root rot of sugar beet ( Beta vulgaris L.), caused by Rhizoctonia solani , continues to be one of the important concerns for the beet industry in Minnesota and North Dakota. Use of resistant cultivars is an important strategy in the management of R. solani in combination with seed treatment and timely fungicide application during the growing season. The objective of this greenhouse study was to determine how sugar beet plants responded to increasing age in resistance to R. solani . Each of three seed companies provided three commercial cultivars with varying R. solani resistance levels: susceptible, moderately resistant, and resistant. Seed were planted at a weekly interval to create different plant age groups from seed to 10-week-old plants, with growing degree days (GDD) ranging from 0 to 1,519 thermal time (°Cd). Seed and plants were all simultaneously inoculated with R. solani AG2-2-infested barley grains. Twenty-eight days after inoculation, plants were pulled and washed, and roots were evaluated for disease severity. All cultivars were highly susceptible to R. solani when inoculated at seed to 3 weeks old (0 to 464°Cd). At 4 and 5 weeks of plant age (617 to 766°Cd), resistant cultivars started to show significant resistance to R. solani . Proportion of the affected roots with disease score ≥ 5 followed a sigmoid response, declining with increased GDD in moderately resistant and resistant cultivars, whereas it continued to decline linearly with increased GDD in susceptible cultivars. This study demonstrated that sugar beet cultivars, regardless of their assigned level of R. solani resistance, were highly susceptible to the pathogen before they reached the six- to eight-leaf stage at 4 to 5 weeks (617 to 766°Cd) after planting. Therefore, additional protection in the form of seed treatment or fungicide application may be required to protect sensitive sugar beet seed and seedlings in fields with a history of R. solani under favorable environmental conditions.

Beet necrotic yellow vein virus (BNYVV) is the causal agent of rhizomania, a disease of global importance to the sugar beet industry. The most widely implemented resistance gene to rhizomania to date is Rz1 , but resistance has been circumvented by resistance-breaking (RB) isolates worldwide. In an effort to gain greater understanding of the distribution of BNYVV and the nature of RB isolates in Minnesota and eastern North Dakota, sugar beet plants were grown in 594 soil samples obtained from production fields and subsequently were analyzed for the presence of BNYVV as well as coding variability in the viral P25 gene, the gene previously implicated in the RB pathotype. Baiting of virus from the soil with sugar beet varieties possessing no known resistance to rhizomania resulted in a disease incidence level of 10.6% in the region examined. Parallel baiting analysis of sugar beet genotypes possessing Rz1 , the more recently introgressed Rz2 , and with the combination of Rz1 + Rz2 resulted in a disease incidence level of 4.2, 1.0, and 0.8%, respectively. Virus sequences recovered from sugar beet bait plants possessing resistance genes Rz1 and/or Rz2 exhibited reduced genetic diversity in the P25 gene relative to those recovered from the susceptible genotype while confirming the hypervariable nature of the coding for amino acids (AAs) at position 67 and 68 in the P25 protein. In contrast to previous reports, we did not find an association between any one specific AA signature at these positions and the ability to circumvent Rz1 -mediated resistance. The data document ongoing virulence development in BNYVV populations to previously resistant varieties and provide a baseline for the analysis of genetic change in the virus population that may accompany the implementation of new resistance genes to manage rhizomania.

Rhizoctonia damping-off and crown and root rot caused by Rhizoctonia solani are major diseases of sugar beet (Beta vulgaris L.) worldwide, and growers in the United States rely on fungicides for disease management. Sensitivity of R. solani to fungicides was evaluated in vitro using a mycelial radial growth assay and by evaluating disease severity on R. solani AG 2-2 inoculated plants treated with fungicides in the greenhouse. The mean concentration that caused 50% mycelial growth inhibition (EC 50 ) values for baseline isolates (collected before the fungicides were registered for sugar beet) were 49.7, 97.1, 0.3, 0.2, and 0.9 μg ml -1 and for nonbaseline isolates (collected after registration and use of fungicides) were 296.1, 341.7, 0.9, 0.2, and 0.6 μg ml -1 for azoxystrobin, trifloxystrobin, pyraclostrobin, penthiopyrad, and prothioconazole, respectively. The mean EC 50 values of azoxystrobin, trifloxystrobin, and pyraclostrobin significantly increased in the nonbaseline isolates compared with baseline isolates, with a resistant factor of 6.0, 3.5, and 3.0, respectively. Frequency of isolates with EC 50 values >10 μg ml -1 for azoxystrobin and trifloxystrobin increased from 25% in baseline isolates to 80% in nonbaseline isolates. Although sensitivity of nonbaseline isolates of R. solani to quinone outside inhibitors decreased, these fungicides at labeled rates were still effective at controlling the pathogen under greenhouse conditions.

Cercospora leaf spot (CLS) of sugar beet is caused by the fungus Cercospora beticola. CLS management practices include the application of the sterol demethylation inhibitor (DMI) fungicides tetraconazole, difenoconazole, and prothioconazole. Evaluating resistance to DMIs is a major focus for CLS fungicide resistance management. Isolates were collected in 1997 and 1998 (baseline sensitivity to tetraconazole, prothioconazole, or difenoconazole) and 2007 through 2010 from the major sugar-beet-growing regions of Minnesota and North Dakota and assessed for in vitro sensitivity to two or three DMI fungicides. Most (47%) isolates collected in 1997-98 exhibited 50% effective concentration (EC 50 ) values for tetraconazole of <0.01 μg ml -1 , whereas no isolates could be found in this EC 50 range in 2010. Since 2007, annual median and mean tetraconazole EC 50 values have generally been increasing, and the frequency of isolates with EC 50 values >0.11 μg ml -1 increased from 2008 to 2010. In contrast, the frequency of isolates with EC 50 values for prothioconazole of >1.0 μg ml -1 has been decreasing since 2007. Annual median difenoconazole EC 50 values appears to be stable, although annual mean EC 50 values generally have been increasing for this fungicide. Although EC 50 values are important for gauging fungicide sensitivity trends, a rigorous comparison of the relationship between in vitro EC 50 values and loss of fungicide efficacy in planta has not been conducted for C. beticola. To explore this, 12 isolates exhibiting a wide range of tetraconazole EC 50 values were inoculated to sugar beet but no tetraconazole was applied. No relationship was found between isolate EC 50 value and disease severity. To assess whether EC 50 values are related to fungicide efficacy in planta, sugar beet plants were sprayed with various dilutions of Eminent, the commercial formulation of tetraconazole, and subsequently inoculated with isolates that exhibited very low, medium, or high tetraconazole EC 50 values. The high EC 50 isolate caused significantly more disease than isolates with medium or very low EC 50 values at the field application rate and most reduced rates. Because in vitro sensitivity testing is typically carried out with the active ingredient of the commercial fungicide, we investigated whether loss of disease control was the same for tetraconazole as for the commercial product Eminent. The high EC 50 isolate caused more disease on plants treated with tetraconazole than Eminent but disease severity was not different between plants inoculated with the very low EC 50 isolate.

In all, 98 isolates of three Fusarium spp. (18 Fusarium oxysporum, 30 F. graminearum, and 50 Fusarium sp. nov.) obtained from sugar beet in Minnesota were characterized for pathogenicity and virulence on sugar beet in the greenhouse by a bare-root inoculation method. Among the 98 isolates tested, 80% of isolates were pathogenic: 83% of the F. oxysporum isolates, 57% of the F. graminearum isolates, and 92% of the Fusarium sp. nov. isolates. Symptoms varied from slight to moderate wilting of the foliage, interveinal chlorosis and necrosis, and vascular discoloration of the taproot without any external root symptoms. Among the pathogenic isolates, 14% were highly virulent and 12% were moderately virulent. Most of the highly virulent isolates (91%) and moderately virulent isolates (89%) were Fusarium sp. nov. All pathogenic isolates of F. graminearum and most pathogenic isolates (87%) of F. oxysporum were less virulent. In general, more-virulent isolates induced first foliar symptoms earlier compared with less-virulent isolates. This study indicates that both F. oxysporum and Fusarium sp. nov. should be used in greenhouse and be present in field studies used for screening and developing sugar beet cultivars resistant to Fusarium yellows complex for Minnesota and North Dakota.

Cercospora leaf spot, caused by the fungus Cercospora beticola Sacc., is the most serious and important foliar disease of sugar beet (Beta vulgaris L.) wherever it is grown worldwide. Cercospora leaf spot first caused economic damage in North Dakota and Minnesota in 1980, and the disease is now endemic. This is the largest production area for sugar beet in the United States, producing 5.5 to 6.0 million metric tons on approximately 300,000 ha, which is 56% of the sugar beet production in the United States. This Plant Disease feature article details a cooperative effort among the participants in the sugar beet industry in this growing area and represents a successful collaboration and team effort to confront and change a fungicide resistance crisis to a fungicide success program. As a case study of success for managing fungicide resistance, it will serve as an example to other pathogen-fungicide systems and provide inspiration and ideas for long-term disease management by fungicides.

Cercospora beticola survives as stromata in infected crop residue. Spores produced on these survival structures serve as primary inoculum during the next cropping season. This study was conducted to determine how long C. beticola can survive at different soil depths, the mechanism of inoculum dispersal, and the primary infection site in sugar beet. Longevity of C. beticola was studied over a 3-year period under field conditions at Fargo, ND. C. beticola-infected leaves were placed at depths of 0, 10, and 20 cm and retrieved after 10, 22, and 34 months. Survival of C. beticola inoculum declined with time and soil depth. Inoculum left on the soil surface, 0 cm in depth, survived the longest (22 months) compared with that buried at 10 cm (10 months) and 20 cm (10 months). C. beticola dispersal from the primary source of inoculum was studied in the field for three growing seasons. Sugar beet plants were surrounded with plastic cages with and without ground cover, or exposed with and without ground cover. Significantly higher disease severity was observed on exposed plants than caged plants with or without ground cover, suggesting that wind was the major dispersal factor for C. beticola inoculum. The primary infection site by C. beticola was determined in a greenhouse study. Leaves, roots, and stems of healthy sugar beet plants were inoculated with C. beticola. Cercospora leaf spot symptoms were observed only on plants that were leaf inoculated, suggesting that the leaf was the primary infection site for C. beticola.

Management of Cercospora leaf spot, caused by Cercospora beticola, is necessary for the economic production of sugar beet (Beta vulgaris). The objectives of this study were to evaluate the impact of two relative humidity thresholds (87 and 90%) on the daily infection values (DIVs) used to determine when fungicide applications were required, to determine whether current Cercospora management recommendations for northern areas of Minnesota and North Dakota could be used by growers in the southern areas of these states, and to compare the utility of calendar-based fungicide applications with the Cercospora management model. Research was conducted in Breckenridge, MN and St. Thomas, ND in 2003 and 2004. Fungicide applications significantly (P = 0.05) reduced maximum disease severity (y max ) and area under the disease progress curve (AUDPC) when compared with the nontreated control at both locations during 2003 and 2004. Fungicides applied according to DIVs calculated at RH ≥ 87% or RH > 90% gave similar results. The mandatory second fungicide application 14 days after the first application for southern areas did not significantly decrease disease severity or AUDPC, or improve root yield or recoverable sucrose compared with treatments without the mandatory application. This research illustrates that a DIV calculated at RH ≥ 87% would result in similar timing of fungicide applications compared with DIVs calculated at RH > 90%. The results further show that the recommendation of fungicide applications at initial symptom and subsequent applications based on DIV and disease severity should be used for both northern and southern growers. Finally, this research showed that fungicide applications based on the Cercospora management model provided similar, effective disease control with fewer fungicide applications compared with calendar-based applications.

Powdery mildew caused by Erysiphe polygoni was widespread on sugar beet (Beta vulgaris) in North Dakota during 2006. This disease is generally not prevalent in the state because of the application of fungicides, which also have efficacy against powdery mildew, for control of Cercospora leaf spot caused by Cercospora beticola. Because Cercospora leaf spot pressure was low in 2006, fewer fungicide applications were made in the state, thus allowing for more observations of powdery mildew. Leaf samples from four fields near Amenia, Minto, Prosper, and St. Thomas, ND were collected in mid-September to look for the perfect stage of E. polygoni, since this has recently been observed in Idaho, Colorado, Montana, and Nebraska (1-3). Only the leaves collected from the field near Amenia had visible immature (yellow and brown) globose ascomata; ascomata were not observed on the leaves collected in the other fields. Additional leaves were collected from the field near Amenia in early October; these leaves had immature and mature (black) globose ascomata that were 70 to 105 μm in diameter. Mature ascomata contained ovoid to elliptic asci with one to four hyaline-to-golden pigmented ascospores (20 to 25 × 12 to 20 μm). The color, shape, and size of ascomata, asci, and ascospores were similar to previously reported observations (1-4). The prevalence of the perfect stage in North Dakota is unknown, since no statewide surveys were conducted. To our knowledge, this is the first report of the perfect stage of E. polygoni on sugar beet in North Dakota. The occurrence of the perfect stage could lead to a means for overwintering in this area. Because of the means for genetic recombination, the risk of fungicide resistance and the development of races may increase. References: (1) J. J. Gallian and L. E. Hanson. Plant Dis. 87:200, 2003. (2) R. M. Harveson. Plant Dis. 88:1049, 2004. (3) B. Jacobsen et al. Plant Dis. 89:1362, 2005. (4) E. G. Ruppel. Powdery mildew. Pages 13-15 in: Compendium of Beet Diseases and Insects. E. D. Whitney and J. E. Duffus, eds. The American Phytopathological Society. St. Paul, MN, 1986.

In 2002, somel sugar beet (Beta vulgaris L.) fields in the Red River Valley (RRV) of Minnesota and North Dakota had symptoms characteristic of Fusarium yellows (4). In 2004, ≈5% of fields in the RRV had symptomatic plants. Interveinal yellowing of older leaves typically began in mid-July and as the disease progressed, younger leaves turned yellow. Sometimes, one side of the leaf was yellow or necrotic while the other side remained green. As leaves died, they remained attached to the crown. Transverse sections of roots revealed a light gray-brown discoloration of the vascular tissue but no external rotting of roots. Isolations from 35 symptomatic roots collected in eight fields yielded 25 isolates identified as F. oxysporum (from single conidia grown on homemade potato dextrose agar and carnation leaf agar) (3). Pathogenicity was determined by dipping roots of 5-week-old sugar beet plants (cv. ACH 9363) in a suspension of 10 4 conidia per ml for 8 min (12 isolates, 10 to 12 plants per isolate). Plants were planted in Cone-tainers (3.8 cm diameter × 21 cm; Stuewe and Sons, Inc. Corvallis, OR) containing sterile soil. Three known cultures of F. oxysporum Schlecht. emend. Snyd. & Hans. f. sp. betae Stewart (= F. conglutinans var. betae Stewart [4]) also were included (13 and 216c from L. Hanson, USDA-ARS, Fort Collins, CO; 0-1122 from The Pennyslvania State University Fusarium Research Center). The control was sterile water. Plants were placed in a greenhouse at 24 to 27°C with natural light supplemented with illumination from high-pressure sodium-vapor lamps for 16 h daily and lightly fertilized biweekly to avoid chlorosis from nutrient deficiency. After 6 to 7 weeks, plants were rated for disease on a 0 to 4 scale: 0 = no disease; 1 = slight to extreme plant stunting, leaves may be wilted; 2 = chlorotic leaves, some with necrosis at margins; 3 = tap root dried and brown to black in color, leaves dying; and 4 = plant dead (1). The experiment was repeated. Disease severity differed between trials, but all isolates of F. oxysporum and F. oxysporum f. sp. betae resulted in disease ratings statistically (P < 0.05) greater than that of the water control. In Trial 1, isolates of F. oxysporum averaged a rating of 2.1 (range of 1.8 to 3.3) and F. oxysporum f. sp. betae averaged 2.1 (range of 2.0 to 2.2) compared with 0.1 for the water control. One isolate of F. oxysporum had a statistically higher rating than did the cultures of F. oxysporum f. sp. betae. In Trial 2, isolates of F. oxysporum averaged a rating of 3.3 (range of 2.7 to 3.7) and F. oxysporum f. sp. betae averaged 3.1 (range of 2.7 to 3.4) compared with 0.2 for the water control. Cultures of F. oxysporum (8 of 12) resulted in ratings statistically higher than that of the least pathogenic culture of F. oxysporum f. sp. betae. Cultures of F. oxysporum and F. oxysporum f. sp. betae recovered from inoculated plants were identical to those used to inoculate plants. To our knowledge, this is the first report of F. oxysporum f. sp. betae on sugar beet in the Red River Valley of Minnesota and North Dakota. The disease has been reported in California, Colorado, Montana, Nebraska, Oregon, Texas, and Wyoming (1,2). References: (1) R. A. Cramer et al. J. Phytopathol. 151:352, 2003. (2) G. A. Fisher and J. S. Gerik. Phytopathology 84:1098, 1994. (3) P. E. Nelson et al. Fusarium Species: An illustrated Manual for Identification. The Pennsylvania State University Press. University Park, 1983. (4) D. Stewart. Phytopathology 21:59, 1931.