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1.
Mol Plant Pathol ; 24(10): 1319-1329, 2023 10.
Article in English | MEDLINE | ID: mdl-37410356

ABSTRACT

In the field of plant virology, the usage of reverse genetic systems has been reported for multiple purposes. One is understanding virus-host interaction by labelling viral cDNA clones with fluorescent protein genes to allow visual virus tracking throughout a plant, albeit this visualization depends on technical devices. Here we report the first construction of an infectious cDNA full-length clone of beet mosaic virus (BtMV) that can be efficiently used for Agrobacterium-mediated leaf inoculation with high infection rate in Beta vulgaris, being indistinguishable from the natural virus isolate regarding symptom development and vector transmission. Furthermore, the BtMV clone was tagged with the genes for the monomeric red fluorescent protein or the Beta vulgaris BvMYB1 transcription factor, which activates the betalain biosynthesis pathway. The heterologous expression of BvMYB1 results in activation of betalain biosynthesis genes in planta, allowing visualization of the systemic BtMV spread with the naked eye as red pigmentation emerging throughout beet leaves. In the case of BtMV, the BvMYB1 marker system is stable over multiple mechanical host passages, allows qualitative as well as quantitative virus detection and offers an excellent opportunity to label viruses in plants of the order Caryophyllales, allowing an in-depth investigation of virus-host interactions on the whole plant level.


Subject(s)
Beta vulgaris , Potyvirus , Transcription Factors/genetics , Transcription Factors/metabolism , Betalains , Beta vulgaris/metabolism , DNA, Complementary/genetics , Potyvirus/genetics , Plant Diseases
2.
Plant Dis ; 107(1): 188-200, 2023 Jan.
Article in English | MEDLINE | ID: mdl-35581914

ABSTRACT

Disease incidence (DI) and metrics of disease severity are relevant parameters for decision making in plant protection and plant breeding. To develop automated and sensor-based routines, a sugar beet variety trial was inoculated with Cercospora beticola and monitored with a multispectral camera system mounted to an unmanned aerial vehicle (UAV) over the vegetation period. A pipeline based on machine learning methods was established for image data analysis and extraction of disease-relevant parameters. Features based on the digital surface model, vegetation indices, shadow condition, and image resolution improved classification performance in comparison with using single multispectral channels in 12 and 6% of diseased and soil regions, respectively. With a postprocessing step, area-related parameters were computed after classification. Results of this pipeline also included extraction of DI and disease severity (DS) from UAV data. The calculated area under disease progress curve of DS was 2,810.4 to 7,058.8%.days for human visual scoring and 1,400.5 to 4,343.2%.days for UAV-based scoring. Moreover, a sharper differentiation of varieties compared with visual scoring was observed in area-related parameters such as area of complete foliage (AF), area of healthy foliage (AH), and mean area of lesion by unit of foliage ([Formula: see text]). These advantages provide the option to replace the laborious work of visual disease assessments in the field with a more precise, nondestructive assessment via multispectral data acquired by UAV flights.[Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.


Subject(s)
Beta vulgaris , Cercospora , Humans , Incidence , Plant Breeding , Vegetables , Sugars
3.
Virus Genes ; 58(6): 550-559, 2022 Dec.
Article in English | MEDLINE | ID: mdl-35960462

ABSTRACT

Beet curly top Iran virus (BCTIV) is a member of the genus Becurtovirus (Family Geminiviridae) with a circular single-strand DNA genome. BCTIV causes leaf curling and vein swelling symptoms in plants. However, the potential pathogenicity factor/s in BCTIV is/are not known. This study presents characterization of complementary-sense transcripts of BCTIV and the viral factors in directing the pathogenicity and hypersensitive response (HR) in Nicotiana benthamiana plants. In both local and systemic infection, splicing of the complementary transcripts of BCTIV was observed. Notably, a small number (8.3%) of transcripts were spliced to produce Rep (C1:C2) transcripts after deletion of 155 nt (position 1892-2046 from BCTIV). Expression of BCTIV genes in N. benthamiana using tobacco rattle virus (TRV)-based vector showed that Rep together with C1 are the main pathogenicity factors which cause typical viral leaf curling symptoms. In addition, the V2 caused a mild leaf curling, thickening, and asymmetric leaves, while the V1, V3, and C2 had no clear effect on the plant phenotype. Transient expression of individual viral genes showed that both the C1 and Rep trigger a HR response in N. benthamiana. The higher expression of HR marker genes, harpin-induced 1 (Hin1) and hypersensitivity-related (Hsr203JI), supported the role of C1 and Rep in HR response in plants. It is concluded that Rep and C1 are the main pathogenicity factors that also trigger HR response in plants.


Subject(s)
Beta vulgaris , Geminiviridae , Nicotiana , Virulence Factors/genetics , Iran , Plant Diseases , Plants
4.
J Gen Virol ; 103(8)2022 08.
Article in English | MEDLINE | ID: mdl-35947097

ABSTRACT

The A-type of beet necrotic yellow vein virus (BNYVV) is widely distributed in Europe and is one of the major virus types causing rhizomania disease in sugar beet. The closely related P-type is mainly limited to a small region in France (Pithiviers). Both virus types possess four RNAs (RNA1-4), but the P-type harbours an additional fifth RNA species (RNA5). The P-type is associated with stronger disease symptoms and resistance-breaking of Rz1, one of the two resistance genes which are used to control BNYVV infection. These characteristics are presumably due to the presence of RNA5, but experimental evidence for this is lacking. We generated the first infectious cDNA clone of BNYVV P-type to study its pathogenicity in sugar beet in comparison to a previously developed A-type clone. Using this tool, we confirmed the pathogenicity of the P-type clone in the experimental host Nicotiana benthamiana and two Beta species, B. macrocarpa and B. vulgaris. Independent of RNA5, both the A- and the P-type accumulated in lateral roots and reduced the taproot weight of a susceptible sugar beet genotype to a similar extent. In contrast, only the P-type clone was able to accumulate a virus titre in an Rz1-resistant variety whereas the A-type clone failed to infect this variety. The efficiency of the P-type to overcome Rz1 resistance was strongly associated with the presence of RNA5. Only a double resistant variety, harbouring Rz1 and Rz2, prevented an infection with the P-type. Reassortment experiments between the P- and A-type clones demonstrated that both virus types can exchange whole RNA components without losing the ability to replicate and to move systemically in sugar beet. Although our study highlights the close evolutionary relationship between the two virus types, we were able to demonstrate distinct pathogenicity properties that are attributed to the presence of RNA5 in the P-type.


Subject(s)
Beta vulgaris , Plant Viruses , Clone Cells , DNA, Complementary/genetics , Plant Diseases , Plant Viruses/genetics , RNA , Sugars , Virulence/genetics
5.
Pest Manag Sci ; 78(11): 4700-4708, 2022 Nov.
Article in English | MEDLINE | ID: mdl-35866215

ABSTRACT

BACKGROUND: The planthopper Pentastiridius leporinus (Hemiptera: Cixiidae) is the main vector of the γ-3 proteobacterium 'Candidatus Arsenophonus phytopathogenicus' which causes the syndrome 'basses richesses' (SBR) in sugar beet. SBR is a new and fast-spreading disease in Central Europe that leads to high yield losses. To date, the development of management strategies has been hampered by insufficient knowledge about general life history traits of the planthopper and, most importantly, the year-round availability of insects reared under controlled conditions. Rearing of P. leporinus has been considered challenging and to date no protocol exists. RESULTS: Here we describe a method for mass rearing P. leporinus on sugar beet from egg to adult that has produced five generations and over 20 000 individuals between June 2020 and March 2022. An alternative host such as wheat is not necessary for completing the life cycle. No-choice experiments showed that P. leporinus lays 139.1 ± 132.9 eggs on sugar beet, whereas no oviposition was observed on its nymphal host wheat. Head capsule width was identified as a trait that unequivocally distinguished the five nymphal instars. Developmental time from first instar to adult was 193.6 ± 35.8 days for males and 193.5 ± 59.2 days for females. Infection rates of adults were tested with a nested polymerase chain reaction. The results demonstrated that 70%-80% of reared planthoppers across all generations carried the SBR proteobacterium. CONCLUSION: The mass-rearing protocol and life history data will help overcome an important bottleneck in SBR research and enhance efforts in developing integrated pest management tools. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.


Subject(s)
Beta vulgaris , Hemiptera , Life History Traits , Animals , Female , Humans , Male , Nymph/microbiology , Sugars
6.
Mol Plant Pathol ; 22(7): 829-842, 2021 07.
Article in English | MEDLINE | ID: mdl-33951264

ABSTRACT

Sugar beet cultivation is dependent on an effective control of beet necrotic yellow vein virus (BNYVV, family Benyviridae), which causes tremendous economic losses in sugar production. As the virus is transmitted by a soilborne protist, the use of resistant cultivars is currently the only way to control the disease. The Rz2 gene product belongs to a family of proteins conferring resistance towards diverse pathogens in plants. These proteins contain coiled-coil and leucine-rich repeat domains. After artificial inoculation of homozygous Rz2 resistant sugar beet lines, BNYVV and beet soilborne mosaic virus (BSBMV, family Benyviridae) were not detected. Analysis of the expression of Rz2 in naturally infected plants indicated constitutive expression in the root system. In a transient assay, coexpression of Rz2 and the individual BNYVV-encoded proteins revealed that only the combination of Rz2 and triple gene block protein 1 (TGB1) resulted in a hypersensitive reaction (HR)-like response. Furthermore, HR was also triggered by the TGB1 homologues from BSBMV as well as from the more distantly related beet soilborne virus (family Virgaviridae). This is the first report of an R gene providing resistance across different plant virus families.


Subject(s)
Beta vulgaris/genetics , Disease Resistance/genetics , Plant Diseases/immunology , Plant Proteins/metabolism , Plant Viruses/physiology , Amino Acid Sequence , Beta vulgaris/immunology , Beta vulgaris/virology , Cell Death , Gene Expression , Genes, Dominant , Genetic Variation , Organ Specificity , Plant Diseases/virology , Plant Leaves/immunology , Plant Leaves/virology , Plant Proteins/genetics , Protein Domains , Sequence Alignment , Nicotiana/genetics , Nicotiana/immunology , Nicotiana/virology , Virulence
7.
Viruses ; 12(1)2020 01 08.
Article in English | MEDLINE | ID: mdl-31936258

ABSTRACT

Beet necrotic yellow vein virus (BNYVV) and Beet soil-borne mosaic virus (BSBMV) are closely related species, but disease development induced in their host sugar beet displays striking differences. Beet necrotic yellow vein virus induces excessive lateral root (LR) formation, whereas BSBMV-infected roots appear asymptomatic. A comparative transcriptome analysis was performed to elucidate transcriptomic changes associated with disease development. Many differentially expressed genes (DEGs) were specific either to BNYVV or BSBMV, although both viruses shared a high number of DEGs. Auxin biosynthesis pathways displayed a stronger activation by BNYVV compared to BSBMV-infected plants. Several genes regulated by auxin signalling and required for LR formation were exclusively altered by BNYVV. Both viruses reprogrammed the transcriptional network, but a large number of transcription factors involved in plant defence were upregulated in BNYVV-infected plants. A strong activation of pathogenesis-related proteins by both viruses suggests a salicylic acid or jasmonic acid mediated-defence response, but the data also indicate that both viruses counteract the SA-mediated defence. The ethylene signal transduction pathway was strongly downregulated which probably increases the susceptibility of sugar beet to Benyvirus infection. Our study provides a deeper insight into the interaction of BNYVV and BSBMV with the economically important crop sugar beet.


Subject(s)
Beta vulgaris/virology , Host Microbial Interactions/genetics , Mosaic Viruses/genetics , Plant Viruses/genetics , Soil Microbiology , Biosynthetic Pathways , Gene Expression Profiling , Plant Diseases/virology , Plant Roots/physiology , Plant Roots/virology , Signal Transduction
8.
Mol Plant Pathol ; 19(10): 2333-2348, 2018 10.
Article in English | MEDLINE | ID: mdl-30011123

ABSTRACT

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.


Subject(s)
Beta vulgaris/metabolism , Beta vulgaris/virology , Plant Viruses/pathogenicity , Transcription Factors/metabolism , Plant Diseases/genetics , Plant Diseases/virology , Transcription Factors/genetics , Viral Proteins/genetics , Viral Proteins/metabolism , Virulence Factors/genetics , Virulence Factors/metabolism
9.
Virology ; 518: 25-33, 2018 05.
Article in English | MEDLINE | ID: mdl-29453056

ABSTRACT

Two members of the Benyviridae family and genus Benyvirus, Beet soil-borne mosaic virus (BSBMV) and Beet necrotic yellow vein virus (BNYVV), possess identical genome organization, host range and high sequence similarity; they infect Beta vulgaris with variable symptom expression. In the US, mixed infections are described with limited information about viral interactions. Vectors suitable for agroinoculation of all genome components of both viruses were constructed by isothermal in vitro recombination. All 35S promoter-driven cDNA clones allowed production of recombinant viruses competent for Nicotiana benthamiana and Beta macrocarpa systemic infection and Polymyxa betae transmission and were compared to available BNYVV B-type clone. BNYVV and BSBMV RNA1 + 2 reassortants were viable and spread long-distance in N. benthamiana with symptoms dependent on the BNYVV type. Small genomic RNAs were exchangeable and systemically infected B. macrocarpa. These infectious clones represent a powerful tool for the identification of specific molecular host-pathogen determinants.


Subject(s)
Beta vulgaris/virology , DNA, Complementary/genetics , Mosaic Viruses/genetics , Plant Diseases/virology , Plant Viruses/genetics , Reassortant Viruses/genetics , Cloning, Molecular , Gene Expression Regulation, Viral , Plant Leaves/virology , RNA Viruses/genetics , RNA, Viral/genetics , RNA, Viral/metabolism
10.
Phytopathology ; 106(9): 1047-54, 2016 09.
Article in English | MEDLINE | ID: mdl-27143412

ABSTRACT

A direct soil DNA extraction method from soil samples (250 g) was applied for detection of the soilborne sugar-beet-infecting pathogen Rhizoctonia solani anastomosis group (AG) 2-2IIIB using a newly developed real-time polymerase chain reaction assay that showed specificity to AG2-2IIIB when tested against various R. solani AG. The assay showed a good relation between cycle threshold and amount of AG2-2IIIB sclerotia detected in three spiked field soils and was also able to detect the pathogen in naturally infested field soil samples. A field trial was conducted to quantify R. solani AG2-2IIIB soil inoculum potential (IP) before and after growing a susceptible and a resistant sugar beet variety as well as after subsequent growth of an expected nonhost winter rye. Plants of the susceptible sugar beet variety displayed a higher disease severity. A more than sixfold increase of the R. solani AG2-2IIIB soil IP was observed in contrast to the resistant variety that resulted in a constant IP. Growing winter rye significantly reduced soil IP to the initial level at sowing. Further research is required to better understand the interaction between disease occurrence and soil IP as well as the environmental influence on IP development.


Subject(s)
Beta vulgaris/microbiology , Plant Diseases/microbiology , Plants/microbiology , Rhizoctonia/isolation & purification , DNA, Fungal/genetics , DNA, Fungal/isolation & purification , Rhizoctonia/genetics , Soil/chemistry , Soil Microbiology
11.
BMC Genomics ; 17: 245, 2016 Mar 17.
Article in English | MEDLINE | ID: mdl-26988094

ABSTRACT

BACKGROUND: Sugar beet (Beta vulgaris) is a crop cultivated for its high content in sugar, but it is vulnerable to many soil-borne pathogens. One of them is the basidiomycete Rhizoctonia solani. This fungal species has a compatibility system regulating hyphal fusions (anastomosis). Consequently, R. solani species are categorized in anastomosis groups (AGs). AG2-2IIIB isolates are most aggressive on sugar beet. In the present study, we report on the draft genome of R. solani AG2-2IIIB using the Illumina technology. Genome analysis, interpretation and comparative genomics of five sequenced R. solani isolates were carried out. RESULTS: The draft genome of R. solani AG2-2IIIB has an estimated size of 56.02 Mb. In addition, two normalized EST libraries were sequenced. In total 20,790 of 21,980 AG2-2IIIB isotigs (transcript isoforms) were mapped on the genome with more than 95 % sequence identity. The genome of R. solani AG2-2IIIB was predicted to harbor 11,897 genes and 4908 were found to be isolate-specific. R. solani AG2-2IIIB was predicted to contain 1142 putatively secreted proteins and 473 of them were found to be unique for this isolate. The R. solani AG2-2IIIB genome encodes a high number of carbohydrate active enzymes. The highest numbers were observed for the polysaccharide lyases family 1 (PL-1), glycoside hydrolase family 43 (GH-43) and carbohydrate estarase family 12 (CE-12). Transcription analysis of selected genes representing different enzyme clades revealed a mixed pattern of up- and down-regulation six days after infection on sugar beets featuring variable levels of resistance compared to mycelia of the fungus grown in vitro. CONCLUSIONS: The established R. solani AG2-2IIIB genome and EST sequences provide important information on the gene content, gene structure and transcriptional activity for this sugar beet pathogen. The enriched genomic platform provides an important platform to enhance our understanding of R. solani biology.


Subject(s)
Beta vulgaris/microbiology , Expressed Sequence Tags , Genome, Fungal , Rhizoctonia/genetics , Chromosome Mapping , Comparative Genomic Hybridization , Plant Diseases/microbiology , Rhizoctonia/enzymology , Sequence Analysis, DNA
12.
J Biotechnol ; 222: 11-2, 2016 Mar 20.
Article in English | MEDLINE | ID: mdl-26851388

ABSTRACT

Rhizoctonia solani is a widespread plant pathogenic fungus featuring a broad host range including several economically important crops. Accordingly, genome analyses of R. solani isolates are important to uncover their pathogenic potential. Draft genome sequences for four R. solani isolates representing three of the 14 R. solani anastomosis groups (AGs) are available. Here, we present the first draft genome sequence for an R. solani AG2-2IIIB isolate that is pathogenic on sugar beet. The fungal genome was assembled in 2065 scaffolds consisting of 5826 contigs amounting to a size of about 52 Mb which is larger than any other R. solani isolate known today. Genes potentially encoding cellulolytic, lignolytic and pectinolytic enzymes were identified.


Subject(s)
Beta vulgaris/microbiology , Genome, Fungal/genetics , Rhizoctonia/genetics , Crops, Agricultural/microbiology , Sequence Analysis, DNA
13.
FEMS Microbiol Ecol ; 92(2)2016 Feb.
Article in English | MEDLINE | ID: mdl-26738557

ABSTRACT

Post-harvest colonization of sugar beets accompanied by rot development is a serious problem due to sugar losses and negative impact on processing quality. Studies on the microbial community associated with rot development and factors shaping their structure are missing. Therefore, high-throughput sequencing was applied to describe the influence of environment, plant genotype and storage temperature (8°C and 20°C) on three different communities in stored sugar beets, namely fungi (internal transcribed spacers 1 and 2), Fusarium spp. (elongation factor-1α gene fragment) and oomycetes (internal transcribed spacers 1). The composition of the fungal community changed during storage mostly influenced by the storage temperature followed by a weak environmental effect. Botrytis cinerea was the prevalent species at 8°C whereas members of the fungal genera Fusarium and Penicillium became dominant at 20°C. This shift was independent of the plant genotype. Species richness within the genus Fusarium also increased during storage at both temperatures whereas the oomycetes community did not change. Moreover, oomycetes species were absent after storage at 20°C. The results of the present study clearly show that rot development during sugar beet storage is associated with pathogens well known as causal agents of post-harvest diseases in many other crops.


Subject(s)
Beta vulgaris/microbiology , Botrytis/genetics , Crops, Agricultural/microbiology , Fusarium/genetics , Oomycetes/genetics , Penicillium/genetics , Base Sequence , Botrytis/classification , Botrytis/growth & development , Carbohydrates , DNA, Intergenic/genetics , Environment , Fusarium/classification , Fusarium/growth & development , Genetic Markers/genetics , High-Throughput Nucleotide Sequencing , Oomycetes/classification , Penicillium/classification , Penicillium/growth & development , Sequence Analysis, DNA , Temperature
14.
Phytopathology ; 106(1): 76-86, 2016 Jan.
Article in English | MEDLINE | ID: mdl-26524545

ABSTRACT

Sugar beet root rot diseases that occur during the cropping season or in storage are accompanied by high yield losses and a severe reduction of processing quality. The vast diversity of microorganism species involved in rot development requires molecular tools allowing simultaneous identification of many different targets. Therefore, a new microarray technology (ArrayTube) was applied in this study to improve diagnosis of sugar beet root rot diseases. Based on three marker genes (internal transcribed spacer, translation elongation factor 1 alpha, and 16S ribosomal DNA), 42 well-performing probes enabled the identification of prevalent field pathogens (e.g., Aphanomyces cochlioides), storage pathogens (e.g., Botrytis cinerea), and ubiquitous spoilage fungi (e.g., Penicillium expansum). All probes were proven for specificity with pure cultures from 73 microorganism species as well as for in planta detection of their target species using inoculated sugar beet tissue. Microarray-based identification of root rot pathogens in diseased field beets was successfully confirmed by classical detection methods. The high discriminatory potential was proven by Fusarium species differentiation based on a single nucleotide polymorphism. The results demonstrate that the ArrayTube constitute an innovative tool allowing a rapid and reliable detection of plant pathogens particularly when multiple microorganism species are present.


Subject(s)
Bacteria/genetics , Beta vulgaris/microbiology , Fungi/genetics , Oligonucleotide Array Sequence Analysis/methods , Oomycetes/genetics , Plant Diseases/microbiology , Bacteria/isolation & purification , Fungi/isolation & purification , Oomycetes/isolation & purification , Species Specificity
15.
Phytopathology ; 106(1): 65-75, 2016 Jan.
Article in English | MEDLINE | ID: mdl-26474333

ABSTRACT

Storage rots represent an economically important factor impairing the storability of sugar beet by increasing sucrose losses and invert sugar content. Understanding the development of disease management strategies, knowledge about major storage pathogens, and factors influencing their occurrence is crucial. In comprehensive storage trials conducted under controlled conditions, the effects of environment and genotype on rot development and associated quality changes were investigated. Prevalent species involved in rot development were identified by a newly developed microarray. The strongest effect on rot development was assigned to environment factors followed by genotypic effects. Despite large variation in rot severity (sample range 0 to 84%), the spectrum of microorganisms colonizing sugar beet remained fairly constant across all treatments with dominant species belonging to the fungal genera Botrytis, Fusarium, and Penicillium. The intensity of microbial tissue necrotization was strongly correlated with sucrose losses (R² = 0.79 to 0.91) and invert sugar accumulation (R² = 0.91 to 0.95). A storage rot resistance bioassay was developed that could successfully reproduce the genotype ranking observed in storage trials. Quantification of fungal biomass indicates that genetic resistance is based on a quantitative mechanism. Further work is required to understand the large environmental influence on rot development in sugar beet.


Subject(s)
Beta vulgaris/genetics , Beta vulgaris/microbiology , Food Microbiology , Food Storage , Plant Roots/microbiology , Agriculture , Environment , Time Factors
16.
Fungal Biol ; 118(9-10): 764-75, 2014.
Article in English | MEDLINE | ID: mdl-25209635

ABSTRACT

This study characterized a novel sugar beet (Beta vulgaris L.) pathogen from the Red River Valley in north central USA, which was formally named Fusarium secorum. Molecular phylogenetic analyses of three loci (translation elongation factor1α, calmodulin, mitochondrial small subunit) and phenotypic data strongly supported the inclusion of F. secorum in the Fusarium fujikuroi species complex (FFSC). Phylogenetic analyses identified F. secorum as a sister taxon of F. acutatum and a member of the African subclade of the FFSC. Fusarium secorum produced circinate hyphae sometimes bearing microconidia and abundant corkscrew-shaped hyphae in culture. To assess mycotoxin production potential, 45 typical secondary metabolites were tested in F. secorum rice cultures, but only beauvericin was produced in detectable amounts by each isolate. Results of pathogenicity experiments revealed that F. secorum isolates are able to induce half- and full-leaf yellowing foliar symptoms and vascular necrosis in roots and petioles of sugar beet. Inoculation with F. acutatum did not result in any disease symptoms. The sugar beet disease caused by F. secorum is named Fusarium yellowing decline. Since Fusarium yellowing decline incidence has been increasing in the Red River Valley, disease management options are discussed.


Subject(s)
Beta vulgaris/microbiology , Fusarium/classification , Fusarium/isolation & purification , Plant Diseases/microbiology , Calmodulin/genetics , Cluster Analysis , DNA, Fungal/chemistry , DNA, Fungal/genetics , Fusarium/cytology , Fusarium/physiology , Hyphae/cytology , Hyphae/growth & development , Molecular Sequence Data , Mycotoxins/metabolism , Peptide Elongation Factor 1/genetics , Phylogeny , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Sequence Homology , United States
17.
Mol Plant Pathol ; 14(4): 356-64, 2013 May.
Article in English | MEDLINE | ID: mdl-23282068

ABSTRACT

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.


Subject(s)
Beta vulgaris/genetics , Beta vulgaris/virology , Disease Resistance/immunology , Plant Diseases/immunology , Plant Diseases/virology , Plant Viruses/pathogenicity , Viral Proteins/metabolism , Amino Acids/genetics , Genes, Plant/genetics , Genetic Variation , Genotype , High-Throughput Nucleotide Sequencing , Phenotype , Plant Roots/virology , Soil Microbiology
18.
Mol Plant Microbe Interact ; 25(8): 1058-72, 2012 Aug.
Article in English | MEDLINE | ID: mdl-22512382

ABSTRACT

P25, a Beet necrotic yellow vein virus (BNYVV) pathogenicity factor, interacts with a sugar beet protein with high homology to Arabidopsis thaliana kelch repeat containing F-box family proteins (FBK) of unknown function in yeast. FBK are members of the Skp1-Cullin-F-box (SCF) complex that mediate protein degradation. Here, we confirm this sugar beet FBK-P25 interaction in vivo and in vitro and provide evidence for in planta interaction and similar subcellular distribution in Nicotiana tabacum leaf cells. P25 even interacts with an FBK from A. thaliana, a BNYVV nonhost. FBK functional classification was possible by demonstrating the interaction with A. thaliana orthologs of Skp1-like (ASK) genes, a member of the SCF E3 ligase. By means of a yeast two-hybrid bridging assay, a direct effect of P25 on SCF-complex formation involving ASK1 protein was demonstrated. FBK transient Agrobacterium tumefaciens-mediated expression in N. benthamiana leaves induced a hypersensitive response. The full-length F-box protein consists of one F-box domain followed by two kelch repeats, which alone were unable to interact with P25 in yeast and did not lead to cell-death induction. The results support the idea that P25 is involved in virus pathogenicity in sugar beet and suggest suppression of resistance response.


Subject(s)
Beta vulgaris/metabolism , Beta vulgaris/virology , F-Box Proteins/metabolism , Plant Proteins/metabolism , Plant Viruses/pathogenicity , Proteasome Endopeptidase Complex/metabolism , Virulence Factors/metabolism , Agrobacterium tumefaciens/genetics , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Disease Resistance , F-Box Proteins/genetics , Host-Pathogen Interactions , Molecular Sequence Data , Plant Diseases/virology , Plant Leaves/metabolism , Plant Leaves/virology , Plant Proteins/genetics , Plant Roots/genetics , Plant Roots/metabolism , Protein Interaction Maps , Protein Structure, Tertiary , SKP Cullin F-Box Protein Ligases/genetics , SKP Cullin F-Box Protein Ligases/metabolism , Sequence Homology, Amino Acid , Subcellular Fractions , Nicotiana/metabolism , Nicotiana/virology , Two-Hybrid System Techniques , Viral Proteins/genetics , Viral Proteins/metabolism , Virulence Factors/genetics
19.
Phytopathology ; 101(11): 1330-7, 2011 Nov.
Article in English | MEDLINE | ID: mdl-21770776

ABSTRACT

Based on a 2-year field trial at two locations in Lower Saxony (Germany), 395 Fusarium isolates belonging to 13 species were collected from more than 3,000 sugar beet roots that were apparently healthy at harvest. In a comparative screen, subsamples were analyzed for Fusarium infection directly after harvest and after different storage conditions. Depending on the storage duration, a different species composition was observed. F. redolens was predominant in freshly harvested beets, while F. culmorum, F. cerealis, and F. graminearum comprised 50.0% (2006) and 84.8% (2007) of the Fusarium mycoflora of sugar beets subjected to long-term pile storage. Randomly selected isolates of all species detected were tested for pathogenicity to sugar beet, but only isolates of F. graminearum and F. sambucinum caused severe root symptoms. Overall, 34 isolates of all species detected were characterized for their mycotoxin profile in rice culture to determine potentially produced toxins for future analysis of sugar beet. A total of 26 Fusarium mycotoxins were detected by liquid chromatography-tandem mass spectrometry, including trichothecenes, zearalenone, and especially high amounts of beauvericin, enniatins, and moniliformin. Further work is required to analyze the natural occurrence of these mycotoxins in sugar beet.


Subject(s)
Beta vulgaris/microbiology , Fusarium/classification , Fusarium/pathogenicity , Mycotoxins/biosynthesis , Cyclobutanes/analysis , Depsipeptides/analysis , Depsipeptides/biosynthesis , Edible Grain/microbiology , Fusarium/isolation & purification , Fusarium/metabolism , Germany , Mycotoxins/analysis , Oryza/microbiology , Plant Leaves/microbiology , Plant Roots/microbiology , Trichothecenes/analysis , Trichothecenes/biosynthesis , Zearalenone/analysis , Zearalenone/biosynthesis
20.
Phytopathology ; 101(11): 1338-45, 2011 Nov.
Article in English | MEDLINE | ID: mdl-21635142

ABSTRACT

Crop rotations with putative non-host crops such as sugar beet are often recommended to reduce Fusarium head blight (FHB) in cereals. However, recent observations have shown pathogenic, endophytic, and saprotrophic colonization of sugar beet with various Fusarium spp. Therefore, strains of seven species frequently isolated from sugar beet were tested for pathogenicity on wheat. Species-specific symptoms on heads and kernels were evaluated and the grains were analyzed for 20 mycotoxins with liquid chromatography-tandem mass spectrometry. Fusarium graminearum, F. culmorum, and F. cerealis from sugar beet caused typical FHB symptoms and mycotoxin contamination with deoxynivalenol and nivalenol, while a high incidence of black point was observed in heads inoculated with F. tricinctum or F. equiseti. Black point kernels revealed 3.4 to 14.5 times higher mycotoxin concentrations than symptomless grains, containing enniatin B1 at 38,000 µg/kg, moniliformin at 4,900 µg/kg, and 2-amino-14,16-dimethyloctadecan-3-ol at 5,500 µg/kg, as well as monoacetoxyscirpenol at 2,600 µg/kg and nivalenol at 3,800 µg/kg. Monitoring of these latter two species in the field is hampered by the lack of typical head symptoms after infection. In further experiments, the impact of sugar beet residues on FHB severity and the correlation between mycotoxin contamination of cereal lots and the amount of black point have to be evaluated.


Subject(s)
Beta vulgaris/microbiology , Fusarium/pathogenicity , Mycotoxins/biosynthesis , Plant Diseases/microbiology , Triticum/microbiology , Cyclobutanes/analysis , Depsipeptides/analysis , Depsipeptides/biosynthesis , Edible Grain/microbiology , Fusarium/chemistry , Fusarium/metabolism , Mycotoxins/analysis , Plant Leaves/microbiology , Plant Roots/microbiology , Species Specificity , Sphingolipids/analysis , Sphingolipids/biosynthesis , Trichothecenes/analysis , Trichothecenes/biosynthesis
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