Combined Omics Approaches Reveal Distinct Mechanisms of Resistance and/or Susceptibility in Sugar Beet Double Haploid Genotypes at Early Stages of Beet Curly Top Virus Infection.

Sugar beet is susceptible to Beet curly top virus (BCTV), which significantly reduces yield and sugar production in the semi-arid growing regions worldwide. Sources of genetic resistance to BCTV is limited and control depends upon insecticide seed treatments with neonicotinoids. Through double haploid production and genetic selection, BCTV resistant breeding lines have been developed. Using BCTV resistant (R) [KDH13; Line 13 and KDH4-9; Line 4] and susceptible (S) [KDH19-17; Line 19] lines, beet leafhopper mediated natural infection, mRNA/sRNA sequencing, and metabolite analyses, potential mechanisms of resistance against the virus and vector were identified. At early infection stages (2- and 6-days post inoculation), examples of differentially expressed genes highly up-regulated in the 'R' lines (vs. 'S') included EL10Ac5g10437 (inhibitor of trypsin and hageman factor), EL10Ac6g14635 (jasmonate-induced protein), EL10Ac3g06016 (ribosome related), EL10Ac2g02812 (probable prolyl 4-hydroxylase 10), etc. Pathway enrichment analysis showed differentially expressed genes were predominantly involved with peroxisome, amino acids metabolism, fatty acid degradation, amino/nucleotide sugar metabolism, etc. Metabolite analysis revealed significantly higher amounts of specific isoflavonoid O-glycosides, flavonoid 8-C glycosides, triterpenoid, and iridoid-O-glycosides in the leaves of the 'R' lines (vs. 'S'). These data suggest that a combination of transcriptional regulation and production of putative antiviral metabolites might contribute to BCTV resistance. In addition, genome divergence among BCTV strains differentially affects the production of small non-coding RNAs (sncRNAs) and small peptides which may potentially affect pathogenicity and disease symptom development.

A robust SNP-haplotype assay for Bct gene region conferring resistance to beet curly top virus in common bean (Phaseolus vulgaris L.).

Beet curly top virus (BCTV), which is synonymous with curly top virus (CTV), causes significant yield loss in common bean (snap and dry beans) cultivars and several other important crops. Common bean cultivars have been found to be resistant to CTV, but screening for resistance is challenging due to the cyclical nature of epidemics and spotty feeding by the leafhopper that vectors the virus. We used an SNP dataset for the Snap Bean Association Panel (SnAP) agro-inoculated with CTV-Logan (CA/Logan) strain to locate the Bct gene region to a 1.7-Mb interval on chromosome Pv07 using genome-wide association study (GWAS) analysis. Recombinant lines from the SnAP were used to further narrow the Bct region to a 58.0-kb interval. A missense SNP (S07_2970381) in candidate gene Phvul.007G036300 Exonuclease V (EXO5) was identified as the most likely causal mutation, and it was the most significant SNP detected by GWAS in a dry bean population (DBP) naturally infected by the CTV-Worland (Wor) strain. Tm-shift assay markers developed for SNP S07_2970381 and two linked SNPs, S07_2970276 and S07_2966197, were useful for tracking different origins of the Bct EXO5 candidate gene resistance to CTV in common bean. The three SNPs identified four haplotypes, with haplotype 3-1 (Haplo3-1) of Middle American origin associated with the highest levels of CTV resistance. This SNP-haplotype assay will enable breeders to track resistance sources and to develop cultivars with better CTV resistance.

Bacterial Endosymbionts Identified From Leafhopper (Hemiptera: Cicadellidae) Vectors of Phytoplasmas.

Insects often harbor bacterial endosymbionts that provide them with nutritional benefit or with protection against natural enemies, plant defenses, insecticides, and abiotic stresses. Certain endosymbionts may also alter acquisition and transmission of plant pathogens by insect vectors. We identified bacterial endosymbionts from four leafhopper vectors (Hemiptera: Cicadellidae) of 'Candidatus Phytoplasma' species by direct sequencing 16S rDNA and confirmed endosymbiont presence and identity by species-specific conventional PCR. We examined three vectors of Ca. Phytoplasma pruni, causal agent of cherry X-disease [Colladonus geminatus (Van Duzee), Colladonus montanus reductus (Van Duzee), Euscelidius variegatus (Kirschbaum)] - and a vector of Ca. Phytoplasma trifolii, the causal agent of potato purple top disease [Circulifer tenellus (Baker)]. Direct sequencing of 16S identified the two obligate endosymbionts of leafhoppers, 'Ca. Sulcia' and 'Ca. Nasuia', which are known to produce essential amino acids lacking in the leafhoppers' phloem sap diet. About 57% of C. geminatus also harbored endosymbiotic Rickettsia. We identified 'Ca. Yamatotoia cicadellidicola' in Euscelidius variegatus, providing just the second host record for this endosymbiont. Circulifer tenellus harbored the facultative endosymbiont Wolbachia, although the average infection rate was only 13% and all males were Wolbachia-uninfected. A significantly greater percentage of Wolbachia-infected Ci. tenellus adults than uninfected adults carried Ca. P. trifolii, suggesting that Wolbachia may increase this insect's ability to tolerate or acquire this pathogen. Results of our study provide a foundation for continued work on interactions between leafhoppers, bacterial endosymbionts, and phytoplasma.

Leaf Bacteriome in Sugar Beet Shows Differential Response against Beet curly top virus during Resistant and Susceptible Interactions.

Beet curly top virus (BCTV) significantly reduces sugar beet yield in semi-arid production areas. Genetic resistance to BCTV is limited; therefore, identification of additional resistance-associated factors is highly desired. Using 16S rRNA sequencing and BCTV resistant (R) genotypes (KDH13, KDH4-9) along with a susceptible (S) genotype (KDH19-17), we investigated leaf bacteriome changes during BCTV post inoculation (pi). At day 6 (~6-week-old plants), Cyanobacteria were predominant (~90%); whereas, at week 4 (~10-week-old plants) Firmicutes (11-66%), Bacteroidetes (17-26%), and Verrucomicrobia (12-29%) were predominant phyla and genotype dependent. Both Bacteroidetes and Verrucomicrobia, increased post infection only in the R lines. The bacterial genera Brevibacillus increased at 6 dpi, and Akkermansia and Bacteroides at 4 wkpi in the R lines. Linear discriminant analysis effect size (LEfSe) identified potential biomarkers in the R vs. S lines. Functional profiling revealed bacterial enrichment associated with the TCA cycle, polyisoprenoid, and L-methionine biosynthesis pathways only in KDH4-9 at 6 dpi. At 4 wkpi, bacteria associated with tryptophan and palmitate biosynthesis in the R lines, and uridine monophosphate, phosphatidyl glycerol, and phospholipid biosynthesis in the S line, were enriched. Future characterization of bacterial genera with antiviral properties will help establish their use as biocontrol agents/biomarkers against BCTV.

Cell-Wall-Degrading Enzymes-Related Genes Originating from Rhizoctonia solani Increase Sugar Beet Root Damage in the Presence of Leuconostoc mesenteroides.

Sugar beet crown and root rot caused by Rhizoctonia solani is a major yield constraint. Root rot is highly increased when R. solani and Leuconostoc mesenteroides co-infect roots. We hypothesized that the absence of plant cell-wall-degrading enzymes in L. mesenteroides and their supply by R. solani during close contact, causes increased damage. In planta root inoculation with or without cell-wall-degrading enzymes showed greater rot when L. mesenteroides was combined with cellulase (22 mm rot), polygalacturonase (47 mm), and pectin lyase (57 mm) versus these enzymes (0-26 mm), R. solani (20 mm), and L. mesenteroides (13 mm) individually. Carbohydrate analysis revealed increased simpler carbohydrates (namely glucose + galactose, and fructose) in the infected roots versus mock control, possibly due to the degradation of complex cell wall carbohydrates. Expression of R. solani cellulase, polygalacturonase, and pectin lyase genes during root infection corroborated well with the enzyme data. Global mRNAseq analysis identified candidate genes and highly co-expressed gene modules in all three organisms that might be critical in host plant defense and pathogenesis. Targeting R. solani cell-wall-degrading enzymes in the future could be an effective strategy to mitigate root damage during its interaction with L. mesenteroides.

Regulatory Roles of Small Non-coding RNAs in Sugar Beet Resistance Against Beet curly top virus.

Beet curly top virus (BCTV) mediated yield loss in sugar beets is a major problem worldwide. The circular single-stranded DNA virus is transmitted by the beet leafhopper. Genetic sources of BCTV resistance in sugar beet are limited and commercial cultivars rely on chemical treatments versus durable genetic resistance. Phenotypic selection and double haploid production have resulted in sugar beet germplasm (KDH13; 13 and KDH4-9; 4) that are highly resistant to BCTV. The molecular mechanism of resistance to the virus is unknown, especially the role of small non-coding RNAs (sncRNAs) during early plant-viral interaction. Using the resistant lines along with a susceptible line (KDH19-17; 19), we demonstrate the role of sugar beet microRNAs (miRNAs) in BCTV resistance during early infection stages when symptoms are not yet visible. The differentially expressed miRNAs altered the expression of their corresponding target genes such as pyruvate dehydrogenase (EL10Ac1g02046), carboxylesterase (EL10Ac1g01087), serine/threonine protein phosphatase (EL10Ac1g01374), and leucine-rich repeats (LRR) receptor-like (EL10Ac7g17778), that were highly expressed in the resistant lines versus susceptible lines. Pathway enrichment analysis of the miRNA target genes showed an enrichment of genes involved in glycolysis/gluconeogenesis, galactose metabolism, starch, and sucrose metabolism to name a few. Carbohydrate analysis revealed altered glucose, galactose, fructose, and sucrose concentrations in the infected leaves of resistant versus susceptible lines. We also demonstrate differential regulation of BCTV derived sncRNAs in the resistant versus susceptible lines that target sugar beet genes such as LRR (EL10Ac1g01206), 7-deoxyloganetic acid glucosyltransferase (EL10Ac5g12605), and transmembrane emp24 domain containing (EL10Ac6g14074) and altered their expression. In response to viral infection, we found that plant derived miRNAs targeted BCTV capsid protein/replication related genes and showed differences in expression among resistant and susceptible lines. The data presented here demonstrate the contribution of miRNA mediated regulation of metabolic pathways and cross-kingdom RNA interference (RNAi) in sugar beet BCTV resistance.

Survey of selected antibiotic resistance genes in agricultural and non-agricultural soils in south-central Idaho.

Improving our understanding of antibiotic resistance in soils is important for the protection of human, animal and ecological health. In south-central Idaho, antibiotic resistance genes (ARGs) [blaCTX-M-1, erm(B), sul1, tet(B), tet(M) and tet(X)] and a class 1 integron-integrase gene (intI1) were quantified in agricultural and non-agricultural soils (96 total sites) under various land use practices (cropland, forestland, inactive cropland, pastureland, rangeland, recreational, residential). We hypothesized that gene occurrence and abundance would be greater in intensively managed agricultural soils. The ARGs (except blaCTX-M-1) and intI1 gene were detected in many of the soils (15 to 58 out of 96 samples), with sul1 and intI1 being detected the most frequently (60% of samples). All of the genes were detected more frequently in the cropland soils (46 sites) and at statistically greater relative abundances (per 16S rRNA gene) than in soils from the other land use categories. When the cropland gene data was separated by sites that had received dairy manure, dairy wastewater, and/or biosolids (27 sites), it was revealed that the genes [except tet(B)] were found at statistically greater abundances (7- to 22-fold higher on average) than in soils that were not treated. The results from this study provide convincing evidence that manure/biosolids use in Idaho cropland soils increases the expansion of antibiotic resistance-related determinants.

Incidence, Distribution, and Pathogenicity of Fungi Causing Root Rot in Idaho Long-Term Sugar Beet Storage Piles.

Fungal rots in sugar beet roots held in long-term storage can lead to considerable sucrose loss but the incidence and distribution of fungal rots inside sugar beet piles and pathogenicity for some species is poorly understood. Thus, Idaho sugar beet held in five outdoor and two indoor piles in 2014 and 2015 were investigated. The root surface area covered by fungal growth and discolored and healthy tissue were assessed in nine 1-m2 areas per pile using a stratified random sampling design. Pathogenicity was evaluated indoors via plug inoculation in 2015 and 2016. Botrytis cinerea covered more root surface area inside indoor piles (6 to 22%) than outdoor piles (0 to 3%) (P < 0.0001). No trends were evident for the Athelia-like sp. (0 to 15%) and Penicillium-type spp. (0 to 8%). Penicillium-type isolates comprised the following species: 60% Penicillium expansum, 34% P. cellarum, 3% P. polonicum, and 3% Talaromyces rugulosus. Trace levels (<1% of root surface) of other fungi, including Cladosporium and Fusarium spp., were evident on roots and in isolations. Based on sample location in a pile, there were no trends or differences; however, two outdoor piles (OVP1 and OVP2) had more healthy tissue (90 to 96%) than other piles (28 to 80%) (P < 0.0001). When the pathogenicity tests were analyzed by species, all were significantly different from each other (P < 0.0001), except for P. polonicum and P. expansum: B. cinerea (61 mm of rot), P. polonicum (36 mm), P. expansum (35 mm), P. cellarum (28 mm), Athelia-like sp. (21 mm), T. rugulosus (0 mm; not different from check), and noninoculated check (0 mm). The OVP1 and OVP2 piles had negligible fungal growth on roots after more than 120 days of storage under ambient conditions, which indicates that acceptable storage can be achieved over this time period through covering piles with tarps and cooling with ventilation pipe.

Influence of Beet necrotic yellow vein virus and Freezing Temperatures on Sugar Beet Roots in Storage.

Rhizomania caused by Beet necrotic yellow vein virus (BNYVV) is a yield-limiting sugar beet disease that was observed to influence root resistance to freezing in storage. Thus, studies were conducted to gain a better understanding of the influence of BNYVV and freezing on sugar beet roots to improve pile management decisions. Roots from five commercial sugar beet cultivars (one susceptible and four resistant to BNYVV) were produced in fields under high and trace levels of rhizomania pressure and subjected to storage using five temperature regimes ranging from 0 to -4.4°C for 24 h. After cold treatment, eight-root samples were stored in a commercial indoor storage building (set point 1.1°C) for 50 days in 2014 and 57 days in 2015. Internal root temperature, frozen and discolored tissue, and moisture and sucrose loss were evaluated. The air temperature at 0, -1.1, and -2.2°C matched internal root temperature but internal root remained near -2.2°C when air temperature was dropped to -3.3 and -4.4°C. In a susceptible cultivar produced under high rhizomania pressure, the percentage of frozen tissue increased (P < 0.0001) from an average of 0 to 7% at 0, -1.1, and -2.2°C up to 16 to 63% at -3.3°C and 63 to 90% at -4.4°C, depending on year. Roots from the susceptible cultivar produced under low rhizomania pressure and those from the resistant cultivars from both fields only had elevated (P ≤ 0.05) frozen tissue at -4.4°C in 15 of 18 cultivar-year combinations. Frozen tissue was related to discolored tissue (r2 = 0.91), weight loss (r2 = 0.12 to 0.28), and sucrose reduction (r2 = 0.69 to 0.74). Consequently, BNYVV will not only lead to yield and sucrose loss in susceptible sugar beet cultivars but also to more frozen root tissue as temperatures drop below -2.2°C. Based on these observations, the air used to cool roots in nonfrozen sugar beet piles throughout the winter should not drop below -2.2°C to maximize sucrose retention.

A Novel Penicillium sp. Causes Rot in Stored Sugar Beet Roots in Idaho.

Penicillium vulpinum along with a number of other fungi can lead to rot of stored sugar beet roots. However, Penicillium isolates associated with necrotic lesions on roots from a recent sugar beet storage study were determined to be different from P. vulpinum and other recognized Penicillium species. Phylogenies based on sequencing of the internal transcribed spacer (ITS)-5.8S, ß-tubulin (BenA), and RNA polymerase II second largest subunit (RPB2) DNA regions indicate that these isolates are novel, but most closely related to the following Penicillium spp. in the section Fasiculata: P. aurantiogriseum, P. camemberti, and P. freii. Macro- and micromorphological data also support designating these isolates as a new species for which we propose the name, Penicillium cellarum sp. nov. Inoculation studies with the P. cellarum isolates on roots of the commercial sugar beet cultivar B-7 led to the formation of necrotic lesions 23 to 25 mm in diameter after 86 days in storage. These lesions were similar to those observed on sugar beet roots in commercial storage piles. These data indicate that P. cellarum is a pathogen which can cause root rot in stored sugar beet roots.

Beet curly top virus Strains Associated with Sugar Beet in Idaho, Oregon, and a Western U.S. Collection.

Curly top of sugar beet is a serious, yield-limiting disease in semiarid production areas caused by Beet curly top virus (BCTV) and transmitted by the beet leafhopper. One of the primary means of control for BCTV in sugar beet is host resistance but effectiveness of resistance can vary among BCTV strains. Strain prevalence among BCTV populations was last investigated in Idaho and Oregon during a 2006-to-2007 collection but changes in disease severity suggested a need for reevaluation. Therefore, 406 leaf samples symptomatic for curly top were collected from sugar beet plants in commercial sugar beet fields in Idaho and Oregon from 2012 to 2015. DNA was isolated and BCTV strain composition was investigated based on polymerase chain reaction assays with strain-specific primers for the Severe (Svr) and California/Logan (CA/Logan) strains and primers that amplified a group of Worland (Wor)-like strains. The BCTV strain distribution averaged 2% Svr, 30% CA/Logan, and 87% Wor-like (16% had mixed infections), which differed from the previously published 2006-to-2007 collection (87% Svr, 7% CA/Logan, and 60% Wor-like; 59% mixed infections) based on a contingency test (P < 0.0001). Whole-genome sequencing (GenBank accessions KT276895 to KT276920 and KX867015 to KX867057) with overlapping primers found that the Wor-like strains included Wor, Colorado and a previously undescribed strain designated Kimberly1. Results confirm a shift from Svr being one of the dominant BCTV strains in commercial sugar beet fields in 2006 to 2007 to becoming undetectable at times during recent years.

Leuconostoc spp. Associated with Root Rot in Sugar Beet and Their Interaction with Rhizoctonia solani.

Rhizoctonia root and crown rot is an important disease problem in sugar beet caused by Rhizoctonia solani and also shown to be associated with Leuconostoc spp. Initial Leuconostoc studies were conducted with only a few isolates and the relationship of Leuconostoc with R. solani is poorly understood; therefore, a more thorough investigation was conducted. In total, 203 Leuconostoc isolates were collected from recently harvested sugar beet roots in southern Idaho and southeastern Oregon during 2010 and 2012: 88 and 85% Leuconostoc mesenteroides, 6 and 15% L. pseudomesenteroides, 2 and 0% L. kimchi, and 4 and 0% unrecognized Leuconostoc spp., respectively. Based on 16S ribosomal RNA sequencing, haplotype 11 (L. mesenteroides isolates) comprised 68 to 70% of the isolates in both years. In pathogenicity field studies with commercial sugar beet 'B-7', all Leuconostoc isolates caused more rot (P < 0.0001; α = 0.05) when combined with R. solani than when inoculated alone in both years. Also, 46 of the 52 combination treatments over the 2 years had significantly more rot (P < 0.0001; α = 0.05) than the fungal check. The data support the conclusion that a synergistic interaction leads to more rot when both Leuconostoc spp. and R. solani are present in sugar beet roots.

Length of Efficacy for Control of Curly Top in Sugar Beet With Seed and Foliar Insecticides.

Curly top in sugar beet caused by Beet curly top virus (BCTV) is an important yield-limiting disease that can be reduced via neonicotinoid and pyrethroid insecticides. The length of efficacy of these insecticides is poorly understood; therefore, field experiments were conducted with the seed treatment Poncho Beta (clothianidin at 60 g a.i. + beta-cyfluthrin at 8 g a.i. per 100,000 seed) and foliar treatment Asana (esfenvalerate at 55.48 g a.i./ha). A series of four experiments at different locations in the same field were conducted in 2014 and repeated in a neighboring field in 2015, with four treatments (untreated check, Poncho Beta, Asana, and Poncho Beta + Asana) which were arranged in a randomized complete block design with eight replications. To evaluate efficacy, viruliferous (contain BCTV strains) beet leafhoppers were released 8, 9, 10, or 11weeks after planting for each experiment, which corresponded to 1, 2, 3, and 4 weeks after Asana application. Over both years, in 30 of 32 observation dates for treatments with Poncho Beta and 14 of 16 observation dates for Asana, visual curly top ratings decreased an average of 41 and 24%, respectively, with insecticide treatments compared with the untreated check. Over both years, in eight of eight experiments for treatments with Poncho Beta and six of eight experiments for Asana, root yields increased an average of 39 and 32%, respectively, with treatment compared with the untreated check. Over both years, the Poncho Beta treatments increased estimated recoverable sucrose (ERS) yield by 75% compared with the untreated check for weeks 8 and 9. By week 10, only the Poncho Beta + Asana treatment led to increases in ERS in both years, while the influence of increasing host resistance may have made other treatments more difficult to separate. When considering curly top symptoms, root yield, and ERS among all weeks and years, there was a tendency for the insecticides in the Poncho Beta + Asana treatment to complement each other to improve efficacy.

Influence of Harvest Timing, Fungicides, and Beet necrotic yellow vein virus on Sugar Beet Storage.

Root rots in sugar beet storage can lead to multimillion dollar losses because of reduced sucrose recovery. Thus, studies were conducted to establish additional fungicide treatments for sugar beet storage and a greater understanding of the fungi involved in the sugar beet storage rot complex in Idaho. A water control treatment and three fungicides (Mertect [product at 0.065 ml/kg of roots; 42.3% thiabendazole {vol/vol}], Propulse [product at 0.049 ml/kg of roots; 17.4% fluopyram and 17.4% prothioconazole {vol/vol}], and Stadium [product at 0.13 ml/kg of roots; 12.51% azoxystrobin, 12.51% fludioxonil, and 9.76% difenoconozole {vol/vol}]) were investigated for the ability to control fungal rots of sugar beet roots held up to 148 days in storage during the 2012 and 2013 storage seasons. At the end of September into October, roots were harvested weekly for 5 weeks from each of two sugar beet fields in Idaho, treated with the appropriate fungicide, and placed on top of a commercial indoor sugar beet storage pile until early February. Differences (P < 0.0001 to 0.0150) among fungicide treatments were evident. Propulse- and Stadium-treated roots had 84 to 100% less fungal growth versus the control roots, whereas fungal growth on Mertect-treated roots was not different from the control roots in 7 of 12 comparisons for roots harvested each of the first 3 weeks in both years of this study. The Propulse- and Stadium-treated roots also reduced (P < 0.0001 to 0.0146; based on weeks 1, 3, and 4 in 2012 and weeks 1, 3, 4, and 5 in 2013) sucrose loss by 14 to 46% versus the control roots, whereas roots treated with Mertect did not change sucrose loss compared with the control roots in 7 of 10 evaluations. The predominant fungi isolated from symptomatic roots were an Athelia-like sp., Botrytis cinerea, Penicillium spp., and Phoma betae. If Propulse and Stadium are labeled for use on sugar beet in storage, these fungicides should be considered for root rot control in commercial sugar beet storage and on roots held for vernalization for seed production of this biennial plant species.

Control of Curly Top in Sugar Beet with Seed and Foliar Insecticides.

Curly top in sugar beet is a serious problem that is caused by Beet curly top virus and other closely related species and transmitted by the beet leafhopper. In order to find a means of reducing curly top in sugar beet, 15 combinations of insecticide seed (Poncho, Poncho Beta, and Poncho Votivo) and foliar (Asana, Cyazypyr, Lorsban, Mustang, Scorpion, and Sivanto) treatments were evaluated versus an untreated check during the 2012 and 2013 growing seasons. An epiphytotic was created by releasing viruliferous beet leafhoppers 58 to 59 days after planting. The foliar sprays were applied 6 to 7 days before and again 6 to 8 days after leafhopper release. Seed treatments (active ingredient: clothianidin) were able to reduce symptoms by 26 to 42% and increase recoverable sucrose by 16 to 21%. The pyrethroids Asana and Mustang also performed well by reducing symptoms 22 to 56% and increasing yields 13 to 20%. The neonicotinoid seed treatments should be an effective way of supplementing host resistance for early-season (at least 59 days after planting) curly top control in sugar beet. The pyrethroid foliar applications could be used to extend curly top control during the midseason period and provide resistance management.

Selection for Resistance to the Rhizoctonia-Bacterial Root Rot Complex in Sugar Beet.

The Rhizoctonia-bacterial root rot complex continues to be a concerning problem in sugar beet production areas. To investigate resistance to this complex in 26 commercial sugar beet cultivars, field studies and greenhouse studies with mature roots from the field were conducted with Rhizoctonia solani anastomosis group 2-2 IIIB strains and Leuconostoc mesenteroides. Based on means for the 26 cultivars in the 2010 and 2011 field studies, fungal rot ranged from 0 to 8%, bacterial rot ranged from 0 to 37%, total internal rot ranged from 0 to 44%, and surface rot ranged from 0 to 52%. All four rot variables resulted in significant (P < 0.0001) cultivar differences. Based on regression analysis, strong positive relationships (r2 from 0.6628 to 0.9320; P < 0.0001) were present among the rot variables. When ranking cultivars, the most consistent rot variable was surface rot, because 12 of 13 variable-year combinations had significant (P ≤ 0.05) correlations. When cultivar ranking in greenhouse assays was compared, there was frequently a positive correlation with storage data but no relationship with field results. Thus, the greenhouse assays will identify storage rot resistance but field screening will be required to find resistance to this rot complex in the field.

Interaction of Sugar Beet Host Resistance and Rhizoctonia solani AG-2-2 IIIB Strains.

Rhizoctonia crown and root rot caused by Rhizoctonia solani can cause serious economic losses in sugar beet fields. Preliminary evidence suggests that there could be interactions between different strains and resistance sources. Thus, field studies were conducted to determine whether nine R. solani AG-2-2 IIIB strains varied for virulence when compared with a noninoculated check and interacted with five sugar beet lines (four resistant lines and a susceptible check). The studies were arranged in a randomized complete block design with six replications. Roots were evaluated for surface rot and internal fungal and bacterial rot in September. All strains were virulent on the susceptible check, FC901/C817, and had a similar ranking (r = 0.80 to 0.97; P = 0.0096 to <0.0001) regardless of disease variable. Line FC709-2 was resistant (response not different from noninoculated check, P ≥ 0.1042) to all strains, while the strain responses resulted in weak interactions with less-resistant lines in 14 of 19 variable-year combinations. Because most commercial sugar beet cultivars contain low to intermediate resistance to Rhizoctonia crown and root rot, the strain used to screen should be considered in order to maintain consistent responses between nurseries and commercial fields.

Characterization of a Basidiomycete fungus from stored sugar beet roots.

Eighteen isolates from sugar beet roots associated with an unknown etiology were characterized based on observations of morphological characters, hyphal growth at 4-28 C, production of phenol oxidases and sequence analysis of internal transcribed spacer (ITS) and large subunit (LSU) regions of the ribosomal DNA (rDNA). The isolates did not produce asexual or sexual spores, had binucleate hyphal cells with clamp connections, grew 4-22 C with estimated optimal growth at 14.5 C and formed a dark brown pigment on potato dextrose or malt extract agar amended with 0.5% tannic acid. Color changes observed when solutions of gum guiac, guiacol and syringaldzine were applied directly to mycelium grown on these media indicated that all isolates produced phenol oxidases. Sequences of ITS and LSU regions on the rDNA gene from 15 isolates were 99.2-100% identical, and analysis of sequence data with maximum likelihood and maximum parsimony suggest that the isolates from sugar beet roots are phylogenetically related to Athelia bombacina, Granulobasidium vellereum and Cyphella digitalis. High statistical support for both loci under different criteria confirmed that Athelia bombacina was consistently the closest known relative to the sugar beet isolates. Additional taxonomic investigations are needed before species can be clarified and designated for these isolates.

Management of Severe Curly Top in Sugar Beet with Insecticides.

Curly top, caused by Curtovirus spp., is a widespread disease problem vectored by the beet leafhopper in semiarid sugar beet production areas. The insecticide seed treatment Poncho Beta has proven to be effective in controlling curly top in sugar beet but was only evaluated under light to moderate disease pressure. Thus, the insecticide seed treatments Poncho Beta, NipsIt INSIDE, and Cruiser Force were evaluated under severe curly top pressure (six viruliferous beet leafhoppers per plant) in field studies during the 2010 and 2011 growing seasons on two commercial sugar beet cultivars. In addition, the foliar insecticides Movento, Provado, and Scorpion were also evaluated. The seed treatments and Scorpion reduced curly top symptoms by 33 to 41% (P < 0.0001) and increased root yield by 55 to 95% (P < 0.0001), sucrose content by 6.5 to 7.2% (P = 0.0013 to <0.0001), and estimated recoverable sucrose by 58 to 96% (P < 0.0001) when compared with the untreated check. Movento and Provado did not improve control beyond that provided by Poncho Beta. Even under severe disease pressure 50 to 55 days after planting, neonicotinoid seed treatments can effectively reduce curly top, increase yield, and help protect against early-season insect pest pressure.

Immunodetection of Two Curtoviruses Infecting Sugar Beet.

Beet leafhopper-transmitted curly top virus is a serious problem in many different crops in the semiarid western United States, including sugar beet, tomatoes, and beans. Curly top is caused by a genetically diverse complex of phloem-limited curtoviruses. Due to the phloem restriction of curtoviruses and the lack of a convenient laboratory host-vector system for curly top virus propagation and purification, no commercial immunodetection tests are available for curtoviruses. Routine diagnostics for curly top rely either on visual symptoms or on polymerase chain reaction (PCR) tests. Lack of an enzyme-linked immunosorbent assay (ELISA) system is one of the factors hampering development and screening of the curly top resistant germplasm in, for instance, sugar beet and bean breeding programs. To fill in this gap, we developed an ELISA-based detection system for curtoviruses which utilizes virus-specific antibodies generated against bacterially expressed capsid protein (CP) of Beet mild curly top virus. Bacterially expressed CP was affinity purified and used as an antigen for antibody production in two animal species. Specificity of the resulting antisera was tested in Western blots and various triple-antibody sandwich (TAS)-ELISA formats with sugar beet, bean, and Nicotiana benthamiana leaf tissue. We demonstrate reliable detection of two curtoviruses in different crops in TAS-ELISA format, suitable for large-scale screening of germplasm in breeding programs.