Sugar Beet Rhizomania Research Articles

Control of rhizomania in sugar beet—A success story made possible by resistance breeding

AbstractThe economic success of sugar beet production depends largely on the control of various pathogens and pests. Beet necrotic yellow vein virus (BNYVV) has historically been a major threat to sugar beet growers because of the high yield losses and the fact that the virus is transmitted by a soilborne protist vector that cannot be controlled. Resistance breeding began after the disease was first described in 1959 and led to the identification of two monogenic resistance genes. Rz1 was the first resistance gene to be introduced into all varieties, enabling sugar beet production under high disease pressure. However, the widespread use of a single resistance gene resulted in high selection pressure on the virus population. As a result, resistance‐breaking of Rz1 was first reported in the early 2000s in the United States and later in many other countries. The mechanism of resistance‐breaking is based on mutations in the viral pathogenicity factor P25 and the presence of an additional RNA5 species. The second resistance gene being used in breeding is Rz2, which is based on a different resistance mechanism and is highly effective at sites where Rz1 has been overcome by the virus. In addition, Rz2 was the first rhizomania resistance gene to be cloned, and the triple gene block I of BNYVV was identified as the corresponding avirulence gene. This article highlights that resistance breeding has been key to the control of rhizomania, but history also shows that re‐emergence of the disease due to resistance‐breaking remains a permanent threat.

Decision Strategies for Absorbance Readings from an Enzyme-Linked Immunosorbent Assay—A Case Study about Testing Genotypes of Sugar Beet (Beta vulgaris L.) for Resistance against Beet Necrotic Yellow Vein Virus (BNYVV)

The Beet necrotic yellow vein virus (BNYVV) causes rhizomania in sugar beet (Beta vulgaris L.), which is one of the most destructive diseases in sugar beet worldwide. In breeding projects towards resistance against BNYVV, the enzyme-linked immunosorbent assay (ELISA) is used to determine the virus concentration in plant roots and, thus, the resistance levels of genotypes. Here, we present a simulation study to generate 10,000 small samples from the estimated density functions of ELISA values from susceptible and resistant sugar beet genotypes. We apply receiver operating characteristic (ROC) analysis to these samples to optimise the cutoff values for sample sizes from two to eight and determine the false positive rates (FPR), true positive rates (TPR), and area under the curve (AUC). We present, furthermore, an alternative approach based upon Bayes factors to improve the decision procedure. The Bayesian approach has proven to be superior to the simple cutoff approach. The presented results could help evaluate or improve existing breeding programs and help design future selection procedures based upon ELISA. An R-script for the classification of sample data based upon Bayes factors is provided.

Pest categorisation of beet necrotic yellow vein virus.

Following a request from the EU Commission, the Panel on Plant Health performed a categorisation of beet necrotic yellow vein virus (BNYVV), the causal agent of the sugar beet rhizomania disease. The virus is currently listed in Annex III as a protected zone (PZ) quarantine pest of the Commission Implementing Regulation (EU) 2019/2072. The identity of the BNYVV is well established. BNYVV is a soil‐borne virus transmitted by the obligate root plasmodiophorid endoparasite Polymyxa betae. BNYVV is widely distributed in the EU, but is not reported in the following EU PZs: Ireland, France (Brittany), Portugal (Azores), Finland and Northern Ireland. The virus may enter, become established and spread in the PZs via P. betae resting spores with soil and growing media as such or attached to machinery and with roots and tubercles of species other than B. vulgaris and with plants for planting. Introduction of BNYVV would have a negative impact on sugar beet and other beet crops in PZs, because of yield and sugar content reduction. Phytosanitary measures are available to reduce the likelihood of entry and spread in the PZs. Once the virus and its plasmodiophorid vector have entered a PZ, their eradication would be difficult due to the persistence of viruliferous resting spores in the soil. The main knowledge gaps or uncertainties identified concerning the presence of BNYVV in the PZs and the incidence and distribution of BNYVV in Switzerland, a country to which a range of specific requirements do not apply. BNYVV meets all the criteria that are within the remit of EFSA to qualify as a potential protected zone union quarantine pest. Plants for planting are not considered as a main means of spread, and therefore BNYVV does not satisfy all the criteria evaluated by EFSA to qualify as potential Union regulated non‐quarantine pest.

RNAseq Analysis of Rhizomania-Infected Sugar Beet Provides the First Genome Sequence of Beet Necrotic Yellow Vein Virus from the USA and Identifies a Novel Alphanecrovirus and Putative Satellite Viruses.

“Rhizomania” of sugar beet is a soilborne disease complex comprised of beet necrotic yellow vein virus (BNYVV) and its plasmodiophorid vector, Polymyxa betae. Although BNYVV is considered the causal agent of rhizomania, additional viruses frequently accompany BNYVV in diseased roots. In an effort to better understand the virus cohort present in sugar beet roots exhibiting rhizomania disease symptoms, five independent RNA samples prepared from diseased beet seedlings reared in a greenhouse or from field-grown adult sugar beet plants and enriched for virus particles were subjected to RNAseq. In all but a healthy control sample, the technique was successful at identifying BNYVV and provided sequence reads of sufficient quantity and overlap to assemble > 98% of the published genome of the virus. Utilizing the derived consensus sequence of BNYVV, infectious RNA was produced from cDNA clones of RNAs 1 and 2. The approach also enabled the detection of beet soilborne mosaic virus (BSBMV), beet soilborne virus (BSBV), beet black scorch virus (BBSV), and beet virus Q (BVQ), with near-complete genome assembly afforded to BSBMV and BBSV. In one field sample, a novel virus sequence of 3682 nt was assembled with significant sequence similarity and open reading frame (ORF) organization to members within the subgenus Alphanecrovirus (genus Necrovirus; family Tombusviridae). Construction of a DNA clone based on this sequence led to the production of the novel RNA genome in vitro that was capable of inducing local lesion formation on leaves of Chenopodium quinoa. Additionally, two previously unreported satellite viruses were revealed in the study; one possessing weak similarity to satellite maize white line mosaic virus and a second possessing moderate similarity to satellite tobacco necrosis virus C. Taken together, the approach provides an efficient pipeline to characterize variation in the BNYVV genome and to document the presence of other viruses potentially associated with disease severity or the ability to overcome resistance genes used for sugar beet rhizomania disease management.

Massive up-regulation of LBD transcription factors and EXPANSINs highlights the regulatory programs of rhizomania disease.

Rhizomania of sugar beet, caused by Beet necrotic yellow vein virus (BNYVV), is characterized by excessive lateral root (LR) formation leading to dramatic reduction of taproot weight and massive yield losses. LR formation represents a developmental process tightly controlled by auxin signaling through AUX/IAA-ARF responsive module and LATERAL ORGAN BOUNDARIES DOMAIN (LBD) transcriptional network. Several LBD transcription factors play central roles in auxin-regulated LR development and act upstream of EXPANSINS (EXPs), cell wall (CW)-loosening proteins involved in plant development via disruption of the extracellular matrix for CW relaxation and expansion. Here, we present evidence that BNYVV hijacks these auxin-regulated pathways resulting in formation LR and root hairs (RH). We identified an AUX/IAA protein (BvAUX28) as interacting with P25, a viral virulence factor. Mutational analysis indicated that P25 interacts with domains I and II of BvAUX28. Subcellular localization of co-expressed P25 and BvAUX28 showed that P25 inhibits BvAUX28 nuclear localization. Moreover, root-specific LBDs and EXPs were greatly upregulated during rhizomania development. Based on these data, we present a model in which BNYVV P25 protein mimics action of auxin by removing BvAUX28 transcriptional repressor, leading to activation of LBDs and EXPs. Thus, the evidence highlights two pathways operating in parallel and leading to uncontrolled formation of LRs and RHs, the main manifestation of the rhizomania syndrome.

試験管育苗法のコムギ縞萎縮病ベクターPolymyxa graminis への応用

試験管育苗法のコムギ縞萎縮病ベクターPolymyxa graminis への応用

Detection and molecular characterization of Polymyxa betae, transmitting agent of sugar beet rhizomania disease, in Iran

The plasmodiophorid Polymyxa betae is considered as the only natural transmitting agent of Beet necrotic yellow vein virus (BNYVV), the most devastating agent of sugar beet fields throughout the world. To evaluate for the first time the genetic diversity of P. betae isolated from different autumn and spring sugar beet fields, and also to detect the presence of virus in these isolates, susceptible sugar beet plants (cv. Regina) were grown in soil samples collected from 10 different regions of Iran. P. betae detection was carried out using root microscopic observations, DAS-ELISA, and PCR-based methods. Results showed the presence of plasmodiophorids in all soil samples. Complementary assays also revealed the presence of viruses in soil samples collected from Khorasan Razavi, Fars, Hamadan and Kermanshah regions. The genetic diversity was evaluated through comparing glutathione-S-transferase nucleotide sequences amplified by PCR with the Internal Transcribed Spacers region. Results showed no significant differences in nucleotide sequences between virus-bearing and virus-free isolates of P. betae.

Infection of Beet necrotic yellow vein virus with RNA4-encoded P31 specifically up-regulates pathogenesis-related protein 10 in Nicotiana benthamiana.

BackgroundBeet necrotic yellow vein virus (BNYVV) is the infectious agent of sugar beet rhizomania, which consists of four or five plus-sense RNAs. RNA4 of BNYVV is not essential for virus propagation in Nicotiana benthamiana but has a major effect on symptom expression. Early reports showed that RNA4-encoded P31 was associated with severe symptoms, such as curling and dwarfing, in N. benthamiana.ResultsWe discovered that the pathogenesis-related protein 10 (PR-10) gene can be up-regulated in BNYVV-infected N. benthamiana in the presence of RNA4 and that it had a close link with symptom development. Our frame-shift, deletion and substitution analysis showed that only the entire P31 could induce PR-10 up-regulation during BNYVV infection and that all the tryptophans and six cysteines (C174, C183, C186, C190, C197 and C199) in the cysteine-rich P31 had significant effects on PR-10 expression. However, P31 could not interact directly with PR-10 in yeast.ConclusionsOur data demonstrated that only integrated P31 specifically induced PR-10 transcription, which coincided closely with the appearance of severe symptoms in BNYVV-infected N. benthamiana, although they could not interact directly with each other in yeast.

Deep sequencing-based transcriptome profiling reveals comprehensive insights into the responses of Nicotiana benthamiana to beet necrotic yellow vein virus infections containing or lacking RNA4.

Background Beet necrotic yellow vein virus (BNYVV), encodes either four or five plus-sense single stranded RNAs and is the causal agent of sugar beet rhizomania disease, which is widely distributed in most regions of the world. BNYVV can also infect Nicotiana benthamiana systemically, and causes severe curling and stunting symptoms in the presence of RNA4 or mild symptoms in the absence of RNA4.ResultsConfocal laser scanning microscopy (CLSM) analyses showed that the RNA4-encoded p31 protein fused to the red fluorescent protein (RFP) accumulated mainly in the nuclei of N. benthamiana epidermal cells. This suggested that severe RNA4-induced symptoms might result from p31-dependent modifications of the transcriptome. Therefore, we used next-generation sequencing technologies to analyze the transcriptome profile of N. benthamiana in response to infection with different isolates of BNYVV. Comparisons of the transcriptomes of mock, BN3 (RNAs 1+2+3), and BN34 (RNAs 1+2+3+4) infected plants identified 3,016 differentially expressed transcripts, which provided a list of candidate genes that potentially are elicited in response to virus infection. Our data indicate that modifications in the expression of genes involved in RNA silencing, ubiquitin-proteasome pathway, cellulose synthesis, and metabolism of the plant hormone gibberellin may contribute to the severe symptoms induced by RNA4 from BNYVV.ConclusionsThese results expand our understanding of the genetic architecture of N. benthamiana as well as provide valuable clues to identify genes potentially involved in resistance to BNYVV infection. Our global survey of gene expression changes in infected plants reveals new insights into the complicated molecular mechanisms underlying symptom development, and aids research into new strategies to protect crops against viruses.

Cross-kingdom host shifts of phytomyxid parasites

BackgroundPhytomyxids (plasmodiophorids and phagomyxids) are cosmopolitan, obligate biotrophic protist parasites of plants, diatoms, oomycetes and brown algae. Plasmodiophorids are best known as pathogens or vectors for viruses of arable crops (e.g. clubroot in brassicas, powdery potato scab, and rhizomania in sugar beet). Some phytomyxid parasites are of considerable economic and ecologic importance globally, and their hosts include important species in marine and terrestrial environments. However most phytomyxid diversity remains uncharacterised and knowledge of their relationships with host taxa is very fragmentary.ResultsOur molecular and morphological analyses of phytomyxid isolates–including for the first time oomycete and sea-grass parasites–demonstrate two cross-kingdom host shifts between closely related parasite species: between angiosperms and oomycetes, and from diatoms/brown algae to angiosperms. Switching between such phylogenetically distant hosts is generally unknown in host-dependent eukaryote parasites. We reveal novel plasmodiophorid lineages in soils, suggesting a much higher diversity than previously known, and also present the most comprehensive phytomyxid phylogeny to date.ConclusionSuch large-scale host shifts between closely related obligate biotrophic eukaryote parasites is to our knowledge unique to phytomyxids. Phytomyxids may readily adapt to a wide diversity of new hosts because they have retained the ability to covertly infect alternative hosts. A high cryptic diversity and ubiquitous distribution in agricultural and natural habitats implies that in a changing environment phytomyxids could threaten the productivity of key species in marine and terrestrial environments alike via host shift speciation.

A SIMPLE METHOD FOR DETECTION OF POLYMYXA BETAE AND BEET NECROTIC YELLOW VEIN VIRUS IN SOIL

Extraction of high quality DNA and RNA from soil is a most challenging step to identify soil organisms due to the presence of a variety of inhibitors and microbial nucleases. In this study, we have developed a simple extraction method for simultaneous purification of DNA and RNA from soil. The isolated nucleic acids were of high quality allowing for sensitive detection of the oomycete Polymyxa betae and Beet necrotic yellow vein virus (BNYVV) by nested-PCR. The vector and BNYVV were detected in all infested soils but only the vector was detected in some apparently non-infested soils or fallow fields. The method may help in the management of sugar beet rhizomania.

SELECTION OF SPECIFIC SINGLE CHAIN VARIABLE FRAGMENTS (SCFV) AGAINST POLYMYXA BETAE FROM PHAGE DISPLAY LIBRARIES

Abstract Sugar beet is one of the most important industrial crops in Iran. For the last two decades it has been mainly affected by a destructive virus, beet necrotic yellow vein virus (BNYVV). The Polymyxa betae is the only natural transmitting agent of the disease among the plants. Developing accurate diagnostic methods may have a major impact on the rising of resistant germplasms. In the present study, specific monoclonal recombinant antibodies in the form of single chain variable fragments (scFv) were obtained from naïve phage display libraries. The fungus specific glutathione-S-transferase (GST) protein was chosen as an antigen for developing antibodies and diagnostic purposes. To generate specific scFv, screening of Tomlinson phage display libraries was performed by applying both recombinant and native fungal GST. Using the recombinant GST in the panning process resulted in the isolation of an antibody only bound to recombinant GST but it failed to detect native GST in the infected plants. Alternatively, the process of panning was carried out by applying native fungal GST trapped to immunotubes through specific polyclonal antibody intermediate. The recent approach resulted in the selection of a specific scFv binding to native GST which was able to detect the presence of the fungi within infected plants. To the best of our knowledge, this is the first report on the generation of recombinant antibodies against Polymyxa betae, fungal vector of sugar beet rhizomania disease.

Effect of sugar beet genotype on the Beet necrotic yellow vein virus P25 pathogenicity factor and evidence for a fitness penalty in resistance‐breaking strains

Beet necrotic yellow vein virus (BNYVV), vectored by Polymyxa betae, causes rhizomania in sugar beet. For disease control, the cultivation of hybrids carrying Rz1 resistance is crucial, but is compromised by resistance-breaking (RB) strains with specific mutations in the P25 protein at amino acids 67-70 (tetrad). To obtain evidence for P25 variability from soil-borne populations, where the virus persists for decades, populations with wild-type (WT) and RB properties were analysed by P25 deep sequencing. The level of P25 variation in the populations analysed did not correlate with RB properties. Remarkably, one WT population contained P25 with RB mutations at a frequency of 11%. To demonstrate selection by Rz1 and the influence of RB mutations on relative fitness, competition experiments between strains were performed. Following a mixture of strains with four RNAs, a shift in tetrad variants was observed, suggesting that strains did not mix or transreplicate. The plant genotype exerted a clear influence on the frequency of RB tetrads. In Rz1 plants, the RB variants outcompeted the WT variants, and mostly vice versa in susceptible plants, demonstrating a relative fitness penalty of RB mutations. The strong genotype effect supports the hypothesized Rz1 RB strain selection with four RNAs, suggesting that a certain tetrad needs to become dominant in a population to influence its properties. Tetrad selection was not observed when an RB strain, with an additional P26 protein encoded by a fifth RNA, competed with a WT strain, supporting its role as a second BNYVV pathogenicity factor and suggesting the reassortment of both types.

Investigation of the persistence of Beet necrotic yellow vein virus in rootlets of sugar beet during biogas fermentation

Rhizomania of sugar beet caused by the Beet necrotic yellow vein virus (BNYVV) and transmitted by Polymyxa betae leads to great losses of yield in sugar beet production worldwide. The virus is multiplying in the sugar beet and its roots are finally enclosed in P. betae sporosori and released into the soil during decomposing. Virus and vector sporosori remain infectious in the soil for years. Setting free sporosori und BNYVV into the environment during sugar production and deposing of contaminated waste implies a phytosanitary risk for spreading rhizomania. Fermentation of plant residues in biogas plants could possibly lead to inactivation of P. betae and/or BNYVV. Within a research project, conducted at the Bavarian State Research Center for Agriculture (LfL), the persistence of different pathogens — including P. betae and BNYVV — during biogas fermentation was studied to evaluate the hygienisation potential of the biogas process and to assess the risk of spreading pathogens with the fermentation residues. To investigate BNYVV tenacity, infected rootlets of sugar beet plantlets were incubated at 38°C and 55°C in 36 l-continuous fermenters up to 36 days and analyzed by ELISA directly after fermentation. In addition, to check the infectivity of the vector/virus complex a bioassay was performed. When fermented at 38°C, BNYVV could be detected by ELISA in the rootlets for up to 36 days, but only during the first 4–8 days of incubation infectivity of virus and/or vector were retained. Compared with incubation at 38°C, incubation at 55°C led to an enhanced degradation of BNYVV and accelerated loss of viral and/or vector infectivity: BNYVV antigen concentration was dramatically reduced and reached a minimum after 3 weeks of incubation. Viral and/or vector infectivity were lost very fast already during the first 4 days of incubation in the fermenter. It can be assumed that biogas fermentation under thermophilic but also mesophilic conditions leads to a relatively fast inactivation of BNYVV and/or its vector P. betae. At least with respect to rhizomania, widely harmless fermentation residues should be produced if the retention time in the fermenter at temperatures between 38°C and 55°C is at least 1 week. But nevertheless, for a full assessment of the phytosanitary risk additional studies are required.

Selection and characterization of resistance to Polymyxa betae, vector of Beet necrotic yellow vein virus, derived from wild sea beet

The plasmodiophoromycete Polymyxa betae is an obligate root parasite that transmits Beet necrotic yellow vein virus (BNYVV), the cause of sugar beet rhizomania disease. Currently, control of this disease is achieved through the use of cultivars with monogenic (Rz1) partial resistance to the virus. To improve the level and durability of this resistance, sources of resistance to the virus vector, P. betae, were sought. Over 100 accessions of the wild sea beet (Beta vulgaris ssp. maritima) from European coastal regions were evaluated for resistance in controlled environment tests. Quantification of P. betae biomass in seedling roots was achieved using recombinant antibodies raised to a glutathione‐s‐transferase expressed by the parasite in vivo. Several putative sources of resistance were identified and selected plants from these were hybridized with a male‐sterile sugar beet breeding line possessing partial virus resistance (Rz1). Evaluation of F1 hybrid populations identified five in which P. betae resistance had been successfully transferred from accessions originating from Mediterranean, Adriatic and Baltic coasts. A resistant individual from one of these populations was backcrossed to the sugar beet parent to produce a BC1 population segregating for P. betae resistance. This population was also tested for resistance to BNYVV. Amplified fragment length polymorphism and single‐nucleotide polymorphism markers were used to map resistance quantitative trait loci (QTL) to linkage groups representing specific chromosomes. QTL for resistance to both P. betae and BNYVV were co‐localized on chromosome IV in the BC1 population, indicating resistance to rhizomania conditioned by vector resistance. This resistance QTL (Pb1) was shown in the F1 population to reduce P. betae levels through interaction with a second QTL (Pb2) found on chromosome IX, a relationship confirmed by general linear model analysis. In the BC1 population, vector‐derived resistance from wild sea beet combined additively with the Rz1 virus resistance gene from sugar beet to reduce BNYVV levels. With partial virus resistance already deployed in a number of high‐yielding sugar beet cultivars, the simple Pb1/Pb2 two‐gene system represents a valuable additional target for plant breeders.

Preparation of polyclonal antiserum to beet necrotic yellow vein virus and its application for immunodiagnostics

* Corresponding author. E-mail: marija.zizyte@botanika.lt Sugar beet rhizomania, caused by the beet necrotic yellow vein virus (BNYVV), was recorded in several areas of Lithuania. From symptomatic sugar beet leaves the virus was mechanically transmitted to the indicator plants for maintenance, purifi cation and storage. Th e virus was maintained in Chenopodium quinoa which served as a propagation host for BNYVV. Transmission of the virus to indicator plants was confi rmed by the DAS-ELISA, immunosorbent electron microscopy (IEM) and PCR methods. BNYVV purifi ed by PEG and density gradient centrifugation was used for the production of polyclonal antiserum suitable for immunodiagnostics by ELISA and Western blot analysis. Th e polyclonal antibody titer obtained in indirect ELISA reactions with a purifi ed virus suspension was found to be 1 : 1600. For the immunoenzyme system formation polyclonal antibodies were conjugated to horseradish peroxidase (HPR). Th e developed system allowed BNYVV detection in the range of 0.3–0.15 μg/ml.

Progress towards the understanding and control of sugar beet rhizomania disease.

Rhizomania is a soil-borne disease that occurs throughout the major sugar beet growing regions of the world, causing severe yield losses in the absence of effective control measures. It is caused by Beet necrotic yellow vein virus (BNYVV), which is transmitted by the obligate root-infecting parasite Polymyxa betae. BNYVV has a multipartite RNA genome with all natural isolates containing four RNA species, although some isolates have a fifth RNA. The larger RNA1 and RNA2 contain the housekeeping genes of the virus and are always required for infection, whereas the smaller RNAs are involved in pathogenicity and vector transmission. RNA5-containing isolates are restricted to Asia and some parts of Europe, and these isolates tend to be more aggressive. With no acceptable pesticides available to restrict the vector, the control of rhizomania is now achieved almost exclusively through the use of resistant cultivars. A single dominant resistance gene, Rz1, has been used to manage the disease worldwide in recent years, although this gene confers only partial resistance. More recently, new variants of BNYVV have evolved (both with and without RNA5) that are able to cause significant yield penalties on resistant cultivars. These isolates are not yet widespread, but their appearance has resulted in accelerated searches for new sources of resistance to both the virus and the vector. Combined virus and vector resistance, achieved either by conventional or transgenic breeding, offers the sugar beet industry a new approach in its continuing struggle against rhizomania.

Postharvest Storage Losses Associated with Rhizomania in Sugar Beet.

During storage of sugar beet, respiration and rots consume sucrose and produce invert sugar. Diseases that occur in the field can affect the magnitude of these losses. This research examines the storage of roots with rhizomania (caused by Beet necrotic yellow vein virus) and the effectiveness of rhizomania-resistant hybrids in reducing postharvest losses. Roots of susceptible hybrids from sites with rhizomania had respiration rates 30 days after harvest (DAH) that ranged from 0.68 to 2.79 mg of CO2 kg-1 h-1 higher than roots of the resistant hybrids. This difference ranged from 2.60 to 13.88 mg of CO2 kg-1 h-1 120 DAH. Roots of resistant hybrids from sites with rhizomania had 18 kg more sucrose per ton than roots from susceptible hybrids 30 DAH, with this difference increasing to 55 kg Mg-1 120 DAH. The invert sugar concentration of susceptible hybrids from sites with rhizomania ranged from 8.38 to 287 g per 100 g of sucrose higher than that for resistant hybrids 120 DAH. In contrast, differences between susceptible and resistant hybrids in respiration rate, sucrose loss, and invert sugar concentration in the absence of rhizomania were relatively small. Storage losses due to rhizomania can be minimized by planting resistant hybrids and processing roots from fields with rhizomania soon after harvest.

Etiology of the sugar beet rhizomania

Beet necrotic yellow vein virus is responsible for sugar beet rhizomania. Root proliferation is characteristic of the viral infection and lead to sugar losses. Pathogenicity is particularly linked to the expression of RNA-3-encoded p25. The extensive use of viral tolerant crops allows maintenance of sugar yields but also permits viruliferous vector to be maintained and therefore the appearance of resistance breaking isolates. The resistance breaking isolates present some amino acid variations within the p25 protein sequence, a key determinant in BNYVV pathogenicity. Here, we will review the molecular biology of BNYVV, of its vector and the antiviral strategies that may be used against rhizomania.

Distribution and Molecular Characterization of Resistance-Breaking Isolates of Beet necrotic yellow vein virus in the United States.

Beet necrotic yellow vein virus (BNYVV) is the causal agent of rhizomania in sugar beet (Beta vulgaris). The virus is transmitted by the plasmodiophorid Polymyxa betae. The disease is controlled primarily by the use of partially resistant cultivars. During 2003 and 2004 in the Imperial Valley of California, partially resistant sugar beet cultivars with Rz1 allele seemed to be compromised. Field trials at Salinas, CA have confirmed that Rz1 has been defeated by resistance-breaking isolates. Distinct BNYVV isolates have been identified from these plants. Rhizomania-infested sugar beet fields throughout the United States were surveyed in 2004-05. Soil surveys indicated that the resistance-breaking isolates not only existed in the Imperial Valley and San Joaquin Valley of California but also in Colorado, Idaho, Minnesota, Nebraska, and Oregon. Of the soil samples tested by baited plant technique, 92.5% produced infection with BNYVV in 'Beta 6600' (rz1rz1rz1), 77.5% in 'Beta 4430R' (Rz1rz1), 45.0% in 'Beta G017R' (Rz2rz2), and 15.0% in 'KWS Angelina' (Rz1rz1+Rz2rz2). Analyses of the deduced amino acid sequence of coat protein and P-25 protein of resistance-breaking BNYVV isolates revealed the high percentage of identity with non-resistance-breaking BNYVV isolates (99.9 and >98.0%, respectively). The variable amino acids in P-25 proteins were located at the residues of 67 and 68. In the United States, the two amino acids found in the non-resistance-breaking isolates were conserved (AC). The resistance-breaking isolates were variable including, AF, AL, SY, VC, VL, and AC. The change of these two amino acids cannot be depended upon to differentiate resistance-breaking and non-resistance-breaking isolates of BNYVV.