RESUMEN
In the current study, a novel strain of Fusarium oxysporum alternavirus 1 (FoAV1) was identified from the Fusarium oxysporum f. sp. melonis (FOM) strain T-BJ17 and was designated as Fusarium oxysporum alternavirus 1-FOM (FoAV1-FOM). Its genome consists of four dsRNA segments of 3515 bp (dsRNA1), 2663 bp (dsRNA2), 2368 bp (dsRNA3), and 1776 bp (dsRNA4) in length. Open reading frame 1 (ORF1) in dsRNA1 was found to encode a putative RNA-dependent RNA polymerase (RdRp), whose amino acid sequence was 99.02% identical to that of its counterpart in FoAV1; while ORF2 in dsRNA2, ORF3 in dsRNA3, and ORF4 in dsRNA4 were all found to encode hypothetical proteins. Strain T-BJ17-VF, which was verified to FoAV1-FOM-free, was obtained using single-hyphal-tip culture combined with high-temperature treatment to eliminate FoAV1-FOM from strain T-BJ17. The colony growth rate, ability to produce spores, and virulence of strain T-BJ17 were significantly lower than those of T-BJ17-VF, while the dry weight of the mycelial biomass and the sensitivity to difenoconazole and pydiflumetofen of strain T-BJ17 were greater than those of T-BJ17-VF. FoAV1-FOM was capable of 100% vertical transmission via spores. To our knowledge, this is the first time that an alternavirus has infected FOM, and this is the first report of hypovirulence and increased sensitivity to difenoconazole and pydiflumetofen induced by FoAV1-FOM infection in FOM.
Asunto(s)
Virus Fúngicos , Fusarium , Genoma Viral , Enfermedades de las Plantas , Triazoles , Fusarium/efectos de los fármacos , Fusarium/genética , Fusarium/virología , Fusarium/patogenicidad , Virus Fúngicos/genética , Virus Fúngicos/aislamiento & purificación , Virus Fúngicos/clasificación , Virus Fúngicos/fisiología , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/virología , Triazoles/farmacología , Dioxolanos/farmacología , Virulencia , Virus ARN/genética , Virus ARN/aislamiento & purificación , Virus ARN/efectos de los fármacos , Virus ARN/clasificación , Filogenia , Sistemas de Lectura Abierta , Triticum/microbiología , Triticum/virologíaRESUMEN
The phosphorylation of eukaryotic translational initiation factors has been shown to play a significant role in controlling the synthesis of protein. Viral infection, environmental stress, and growth circumstances cause phosphorylation or dephosphorylation of plant initiation factors. Our findings indicate that casein kinase 2 can phosphorylate recombinant wheat eIFiso4E and eIFiso4G generated from E. coli in vitro. For wheat eIFiso4E, Ser-207 was found to be the in vitro phosphorylation site. eIFiso4E lacks an amino acid that can be phosphorylated at the position corresponding to Ser-209, the phosphorylation site in mammalian eIF4E, yet phosphorylation of eIFiso4E has effects on VPg binding affinity that are similar to those of phosphorylation of mammalian eIF4E. The addition of VPg and phosphorylated eIFiso4F to depleted wheat germ extract (WGE) leads to enhancement of translation of both uncapped and capped viral mRNA. The addition of PABP together with eIFiso4Fp and eIF4B to depleted WGE increases both uncapped and capped mRNA translation. However, it exhibits a translational advantage specifically for uncapped mRNA, implying that the phosphorylation of eIFiso4F hinders cap binding while promoting VPg binding, thereby facilitating uncapped translation. These findings indicate TEV virus mediates VPg-dependent translation by engaging a mechanism entailing phosphorylated eIFiso4Fp and PABP. To elucidate the molecular mechanisms underlying these observed effects, we studied the impact of PABP and/or eIF4B on the binding of VPg with eIFiso4Fp. The inclusion of PABP and eIF4B with eIFiso4Fp resulted in about 2-fold increase in affinity for VPg (Kd = 24 ± 1.7 nM), as compared to the affinity of eIFiso4Fp alone (Kd = 41.0 ± 3.1 nM). The interactions between VPg and eIFiso4Fp were determined to be both enthalpically and entropically favorable, with the enthalpic contribution accounting for 76-97% of the ΔG at 25°C, indicating a substantial role of hydrogen bonding in enhancing the stability of the complex. The binding of PABP to eIFiso4Fp·4B resulted in a conformational alteration, leading to a significant enhancement in the binding affinity to VPg. These observations suggest PABP enhances the affinity between eIFiso4Fp and VPg, leading to an overall conformational change that provides a stable platform for efficient viral translation.
Asunto(s)
Factores Eucarióticos de Iniciación , Proteínas de Unión a Poli(A) , Potyvirus , Unión Proteica , Biosíntesis de Proteínas , Triticum , Fosforilación , Potyvirus/metabolismo , Potyvirus/genética , Triticum/virología , Triticum/metabolismo , Triticum/genética , Factores Eucarióticos de Iniciación/metabolismo , Factores Eucarióticos de Iniciación/genética , Proteínas de Unión a Poli(A)/metabolismo , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Proteínas Virales/metabolismo , Proteínas Virales/genética , Quinasa de la Caseína II/metabolismo , Quinasa de la Caseína II/genéticaRESUMEN
Triticum mosaic virus (TriMV; genus Poacevirus; family Potyviridae) is an economically important virus in the Great Plains region of the United States. TriMV is transmitted by the wheat curl mite (Aceria tosichella) Type 2 genotype but not by Type 1. Helper component-proteinase (HC-Pro) is a vector transmission determinant for several potyvirids, but the role of HC-Pro in TriMV transmission is unknown. In this study, we examined the requirement of the HC-Pro cistron of TriMV for wheat curl mite (Type 2) transmission through deletion and point mutations and constructing TriMV chimeras with heterologous HC-Pros from other potyvirids. TriMV with complete deletion of HC-Pro failed to be transmitted by wheat curl mites at detectable levels. Furthermore, TriMV chimeras with heterologous HC-Pros from aphid-transmitted turnip mosaic virus and tobacco etch virus, or wheat curl mite-transmitted wheat streak mosaic virus, failed to be transmitted by wheat curl mites. These data suggest that heterologous HC-Pros did not complement TriMV for wheat curl mite transmission. A decreasing series of progressive nested in-frame deletions at the N-terminal region of HC-Pro comprising amino acids 3 to 125, 3 to 50, 3 to 25, 3 to 15, 3 to 8, and 3 and 4 abolished TriMV transmission by wheat curl mites. Additionally, mutation of conserved His20, Cys49, or Cys52 to Ala in HC-Pro abolished TriMV transmissibility by wheat curl mites. These data suggest that the N-terminal region of HC-Pro is crucial for TriMV transmission by wheat curl mites. Collectively, these data demonstrate that the HC-Pro cistron of TriMV is a viral determinant for wheat curl mite transmission.
Asunto(s)
Ácaros , Enfermedades de las Plantas , Potyviridae , Triticum , Proteínas Virales , Triticum/virología , Animales , Enfermedades de las Plantas/virología , Ácaros/virología , Proteínas Virales/genética , Proteínas Virales/metabolismo , Potyviridae/genética , Potyviridae/fisiología , Cisteína Endopeptidasas/genética , Cisteína Endopeptidasas/metabolismoRESUMEN
WRKY transcription factors are widely involved in plant responses to biotic and abiotic stresses. However, there is currently a limited understanding of the regulation of viral infection by WRKY transcription factors in wheat (Triticum aestivum). The WRKY transcription factor TaWRKY50 in group IIb wheat exhibited a significant response to Chinese wheat mosaic virus infection. TaWRKY50 is localized in the nucleus and is an activating transcription factor. Interestingly, we found that silencing TaWRKY50 induces cell death following inoculation with CWMV. The protein kinase TaSAPK7 is specific to plants, whereas NbSRK is a closely related kinase with high homology to TaSAPK7. The transcriptional activities of both TaSAPK7 and NbSRK can be enhanced by TaWRKY50 binding to their promoters. CRP is an RNA silencing suppressor. Furthermore, TaWRKY50 may regulate CWMV infection by regulating the expression of TaSAPK7 and NbSRK to increase CRP phosphorylation and reduce the amount of programmed cell death (PCD).
Asunto(s)
Apoptosis , Regulación de la Expresión Génica de las Plantas , Enfermedades de las Plantas , Proteínas de Plantas , Factores de Transcripción , Triticum , Triticum/virología , Triticum/genética , Triticum/metabolismo , Enfermedades de las Plantas/virología , Fosforilación , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Factores de Transcripción/metabolismo , Factores de Transcripción/genética , Interacciones Huésped-PatógenoRESUMEN
Maize rough dwarf disease (MRDD) threatens the sustainable production of major cereal crops. Recently, Xu et al. reported a new resistance gene, ZmGLK36, which promotes MRDD resistance in maize by increasing jasmonic acid (JA)-mediated defence. This discovery provides opportunities to develop resistance to rice black-streaked dwarf virus (RBSDV) in other cereal crops such as rice and wheat.
Asunto(s)
Resistencia a la Enfermedad , Oryza , Enfermedades de las Plantas , Proteínas de Plantas , Triticum , Oryza/virología , Oryza/genética , Enfermedades de las Plantas/virología , Triticum/virología , Triticum/genética , Resistencia a la Enfermedad/genética , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Factores de Transcripción/metabolismo , Factores de Transcripción/genética , Oxilipinas/metabolismo , Ciclopentanos/metabolismo , Zea mays/virología , Zea mays/genética , Regulación de la Expresión Génica de las Plantas , Virus de Plantas/fisiologíaRESUMEN
Wheat yellow mosaic virus (WYMV) is a pathogen transmitted into its host's roots by the soil-borne vector Polymyxa graminis. Ym1 and Ym2 genes protect the host from the significant yield losses caused by the virus, but the mechanistic basis of these resistance genes remains poorly understood. Here, it has been shown that Ym1 and Ym2 act within the root either by hindering the initial movement of WYMV from the vector into the root and/or by suppressing viral multiplication. A mechanical inoculation experiment on the leaf revealed that the presence of Ym1 reduced viral infection incidence, rather than viral titer, while that of Ym2 was ineffective in the leaf. To understand the basis of the root specificity of the Ym2 product, the gene was isolated from bread wheat using a positional cloning approach. The candidate gene encodes a CC-NBS-LRR protein and it correlated allelic variation with respect to its sequence with the host's disease response. Ym2 (B37500) and its paralog (B35800) are found in the near-relatives, respectively, Aegilops sharonensis and Aegilops speltoides (a close relative of the donor of bread wheat's B genome), while both sequences, in a concatenated state, are present in several accessions of the latter species. Structural diversity in Ym2 has been generated via translocation and recombination between the two genes and enhanced by the formation of a chimeric gene resulting from an intralocus recombination event. The analysis has revealed how the Ym2 region has evolved during the polyploidization events leading to the creation of cultivated wheat.
Asunto(s)
Aegilops , Triticum , Aegilops/genética , Aegilops/metabolismo , Triticum/genética , Triticum/metabolismo , Triticum/virología , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raíces de Plantas/genética , Raíces de Plantas/metabolismo , Raíces de Plantas/virología , Clonación Molecular , Transcripción Genética , Filogenia , Enfermedades de las PlantasRESUMEN
Since 1948, pale yellow wheat spike have been reported in southern Brazil. This symptom was associated with tenuiviruses due to the observation of cytoplasmic inclusions constituted by a mass of filamentous particles (7-10 nm in diameter) with indeterminate length, identical to those found in "leaf dip" preparations. Such symptoms are still seen in wheat crops; however, there is a lack of information regarding this pathosystem. Decades after the first report, the first sequences of wheat white spike virus were characterized. Wheat plants with symptoms such as pale yellowing, chlorotic streaks, and leaf mosaic were collected in Paraná State, Southern Brazil. High-throughput sequencing was used to determine the nearly complete nucleotide sequence of the viral genome. The genome is composed of five RNAs with a total size of 18,129 nucleotides, with eight open reading frames (ORFs). The virus identified in this study can be included in a new species in the family Phenuiviridae, genus Tenuivirus, and we have tentatively named this virus "wheat white spike virus".
Asunto(s)
Enfermedades de las Plantas/virología , Tenuivirus , Triticum/virología , Brasil , Filogenia , Tenuivirus/clasificaciónRESUMEN
Maize chlorotic dwarf virus (MCDV) encodes a 3C-like protease that cleaves the N-terminal polyprotein (R78) as previously demonstrated. Here, we examined amino acid residues required for catalytic activity of the protease, including those in the predicted catalytic triad, amino acid residues H2667, D2704, and C2798, as well as H2817 hypothesized to be important in substrate binding. These and other residues were targeted for mutagenesis and tested for proteolytic cleavage activity on the N-terminal 78 kDa MCDV-S polyprotein substrate to identify mutants that abolished catalytic activity. Mutations that altered the predicted catalytic triad residues and H2817 disrupted MCDV-S protease activity, as did mutagenesis of a conserved tyrosine residue, Y2774. The protease activity and R78 cleavage of orthologs from divergent MCDV isolates MCDV-Tn and MCDV-M1, and other waikavirus species including rice tungro spherical virus (RTSV) and bellflower vein chlorosis virus (BVCV) were also examined.
Asunto(s)
Proteasas Virales 3C/química , Regulación Viral de la Expresión Génica , Genoma Viral , Waikavirus/genética , Proteasas Virales 3C/genética , Proteasas Virales 3C/metabolismo , Secuencia de Aminoácidos , Sitios de Unión , Sistema Libre de Células/metabolismo , Modelos Moleculares , Mutación , Unión Proteica , Biosíntesis de Proteínas , Conformación Proteica en Hélice alfa , Conformación Proteica en Lámina beta , Dominios y Motivos de Interacción de Proteínas , Proteolisis , Semillas/química , Semillas/metabolismo , Alineación de Secuencia , Homología de Secuencia de Aminoácido , Relación Estructura-Actividad , Especificidad por Sustrato , Transcripción Genética , Triticum/virología , Waikavirus/enzimología , Zea mays/virologíaRESUMEN
GDSL-type esterase/lipase proteins (GELPs) characterized by a conserved GDSL motif at their N-terminus belong to the lipid hydrolysis enzyme superfamily. In plants, GELPs play an important role in plant growth, development and stress response. The studies of the identification and characterization of the GELP gene family in Triticeae have not been reported. In this study, 193 DvGELPs were identified in Dasypyrum villosum and classified into 11 groups (clade A-K) by means of phylogenetic analysis. Most DvGELPs contain only one GDSL domain, only four DvGELPs contain other domains besides the GDSL domain. Gene structure analysis indicated 35.2% DvGELP genes have four introns and five exons. In the promoter regions of the identified DvGELPs, we detected 4502 putative cis-elements, which were associated with plant hormones, plant growth, environmental stress and light responsiveness. Expression profiling revealed 36, 44 and 17 DvGELPs were highly expressed in the spike, the root and the grain, respectively. Further investigation of a root-specific expressing GELP, DvGELP53, indicated it was induced by a variety of biotic and abiotic stresses. The knockdown of DvGELP53 inhibited long-distance movement of BSMV in the tissue of D. villosum. This research provides a genome-wide glimpse of the D. villosum GELP genes and hints at the participation of DvGELP53 in the interaction between virus and plants.
Asunto(s)
Hidrolasas de Éster Carboxílico/genética , Genes de Plantas , Enfermedades de las Plantas/genética , Proteínas de Plantas/genética , Virus de Plantas/fisiología , Plantas/genética , Plantas/virología , Triticum/genética , Triticum/virología , Hidrolasas de Éster Carboxílico/química , Hidrolasas de Éster Carboxílico/clasificación , Exones , Regulación de la Expresión Génica de las Plantas , Silenciador del Gen , Interacciones Microbiota-Huesped/genética , Intrones , Filogenia , Enfermedades de las Plantas/virología , Proteínas de Plantas/química , Proteínas de Plantas/clasificación , Regiones Promotoras Genéticas/genética , Dominios Proteicos , TranscriptomaRESUMEN
Although peroxisomes play an essential role in viral pathogenesis, and viruses are known to change peroxisome morphology, the role of genotype in the peroxisomal response to viruses remains poorly understood. Here, we analyzed the impact of wheat streak mosaic virus (WSMV) on the peroxisome proliferation in the context of pathogen response, redox homeostasis, and yield in two wheat cultivars, Patras and Pamir, in the field trials. We observed greater virus content and yield losses in Pamir than in Patras. Leaf chlorophyll and protein content measured at the beginning of flowering were also more sensitive to WSMV infection in Pamir. Patras responded to the WSMV infection by transcriptional up-regulation of the peroxisome fission genes PEROXIN 11C (PEX11C), DYNAMIN RELATED PROTEIN 5B (DRP5B), and FISSION1A (FIS1A), greater peroxisome abundance, and activation of pathogenesis-related proteins chitinase, and ß-1,3-glucanase. Oppositely, in Pamir, WMSV infection suppressed transcription of peroxisome biogenesis genes and activity of chitinase and ß-1,3-glucanase, and did not affect peroxisome abundance. Activity of ROS scavenging enzymes was higher in Patras than in Pamir. Thus, the impact of WMSV on peroxisome proliferation is genotype-specific and peroxisome abundance can be used as a proxy for the magnitude of plant immune response.
Asunto(s)
Resistencia a la Enfermedad/inmunología , Peroxisomas/metabolismo , Enfermedades de las Plantas/inmunología , Enfermedades de las Plantas/microbiología , Potyviridae , Triticum/inmunología , Triticum/virología , Quitinasas/metabolismo , Clorofila/metabolismo , Glucano 1,3-beta-Glucosidasa/metabolismo , Oxidación-Reducción , Peroxidasas/metabolismo , Peroxisomas/genética , Peroxisomas/virología , Fenotipo , Hojas de la Planta/inmunología , Hojas de la Planta/virología , Especies Reactivas de Oxígeno/metabolismoRESUMEN
Wheat (Triticum aestivum) is one of the most important food crops around the world. China is the largest wheat production country and wheat yellow mosaic virus (WYMV) is a non-negligible threat to wheat production. This study aimed to explore miRNAs and their corresponding target genes responsive to WYMV in wheat. Linmai and Jimai were used for miRNA and degradome high-throughput sequencing. After comparison and analysis, differentially expressed miRNAs and their target genes between normal wheat and WYMV-infected wheat were identified. GO and KEGG pathway enrichment analysis were then performed on target genes. A total of 530 miRNAs were identified in all samples, including 106 known miRNAs and 424 novel miRNAs. Among them, 131 miRNAs, corresponding to 85 target genes, were differentially expressed between normal wheat and WYMV-infected wheat. 85 target genes were significantly enriched in 21 GO terms and two KEGG pathways, Plant hormone signal transduction and Monobactam biosynthesis. In conclusion, 131 differentially expressed miRNAs, corresponding to 85 target genes, were identified between normal wheat and WYMVinfected wheat. Our findings provide more evidence on the roles of miRNAs and their target genes in wheat- WYMV interactions.
Asunto(s)
Regulación de la Expresión Génica de las Plantas/inmunología , MicroARNs/metabolismo , Virus del Mosaico/fisiología , Enfermedades de las Plantas/virología , Triticum/virología , MicroARNs/genética , Virus del Mosaico/inmunología , Enfermedades de las Plantas/inmunología , ARN de PlantaRESUMEN
Genome editing provides novel strategies for improving plant traits but mostly relies on conventional plant genetic transformation and regeneration procedures, which can be inefficient. In this study, we have engineered a Barley stripe mosaic virus-based sgRNA delivery vector (BSMV-sg) that is effective in performing heritable genome editing in Cas9-transgenic wheat plants. Mutated progenies were present in the next generation at frequencies ranging from 12.9% to 100% in three different wheat varieties, and 53.8%-100% of mutants were virus free. We also achieved multiplex mutagenesis in progeny using a pool of BSMV-sg vectors harboring different sgRNAs. Furthermore, we devised a virus-induced transgene-free editing procedure to generate Cas9-free wheat mutants by crossing BSMV-infected Cas9-transgenic wheat pollen with wild-type wheat. Our study provides a robust, convenient, and tissue culture-free approach for genome editing in wheat through virus infection.
Asunto(s)
Edición Génica/métodos , Genoma de Planta , Virus de Plantas/genética , Triticum/genética , Proteína 9 Asociada a CRISPR/genética , Patrón de Herencia , Plantas Modificadas Genéticamente/genética , ARN Guía de Kinetoplastida , Triticum/virologíaRESUMEN
Wheat plants showing yellowing and mosaic in leaves and stunting were collected from wheat fields in Henan Province, China. Analysis of these plants by transmission electron microscopy showed that they contained two types of filamentous virus-like particles with a length of 200-500 nm and 1000-1300 nm, respectively. RNA-seq revealed a coinfection with wheat yellow mosaic virus (WYMV) and an unknown wheat-infecting virus. The genome of the unknown virus is 8,410 nucleotides long, excluding its 3' poly(A) tail. It has six open reading frames (ORFs). ORF1 encodes a putative viral replication-associated protein (Rep), and ORFs 2, 3, and 4 encode the triple gene block (TGB) proteins. ORFs 5 and 6 encode the capsid protein (CP) and a protein with unknown function, respectively. Phylogenetic analysis showed that this novel virus is evolutionarily related to members of the subfamily Quinvirinae, family Betaflexiviridae. It is, however, distinct from the viruses in the currently established genera. Based on the species and genus demarcation criteria set by the International Committee on Taxonomy of Viruses (ICTV), we tentatively name this novel virus "wheat yellow stunt-associated betaflexivirus" (WYSaBV), and we propose it to be a member of a new genus in the family Betaflexiviridae.
Asunto(s)
Flexiviridae/genética , Triticum/virología , China , Flexiviridae/clasificación , Flexiviridae/patogenicidad , Flexiviridae/ultraestructura , Genoma Viral/genética , Sistemas de Lectura Abierta/genética , Filogenia , Enfermedades de las Plantas/virología , Hojas de la Planta/virología , ARN Viral/genética , Especificidad de la Especie , Proteínas Virales/genética , Virión/ultraestructuraRESUMEN
Wheat yellow mosaic virus (family Potyviridae; genus Bymovirus), is an important soil-borne virus that causes serious economic losses in wheat. In this study, we constructed infectious cDNA clones of WYMV genomic RNAs under the control of 35S or SP6 promoter for versatile usage (agroinfiltration or in vitro RNA transcription). Our results showed that an Agrobacterium-mediated inoculation system enabled WYMV to infect the leaves of Nicotiana benthamiana without causing WYMV systemic infection. However, in vitro transcripts from infectious cDNA clones using the SP6 promoter promoted WYMV systemic infection of wheat plants, which was then developed for further assays. The optimal temperature for virus multiplication and systemic infection of wheat was 8 °C. Additionally, a synergistic effect between WYMV and Chinese wheat mosaic virus (CWMV) was also detected. This is the first report of the construction of a Chinese isolate of WYMV and should facilitate the investigation of viral pathogenesis.
Asunto(s)
Enfermedades de las Plantas/virología , Virus de Plantas , Triticum/virología , Virus de Plantas/genética , Virus de Plantas/aislamiento & purificación , Virus de Plantas/patogenicidadRESUMEN
Cereal aphids are vectors of at least 11 species of Barley Yellow Dwarf Viruses (BYDV) in wheat that alone and/or in combination can cause between 5%-80% grain yield losses. They establish complex virus-vector interactions, with variations in specificity and transmission efficiency that need to be considered for control purposes. In general, these viruses and vectors have a global distribution, however, BYDV-PAV is the most prevalent and abundant virus species worldwide, likely due to its vectoring efficiency and the wide distribution of its primary vector Rhopalosiphum padi. Host plant resistance (HPR) is an environmentally friendly, efficient and cost-effective tool to reduce crop losses to biotic stressors such as aphids and viruses. Finding resistance sources is paramount to breed for HPR. Currently, most of the resistance identified for aphids and BYDV derives from wheat related and wild relative species. However, breeding for HPR to BYDV and its vectors has additional challenges besides the source identification, for example, the lack of selection tools for certain aphid species, which likely prevents the development of elite wheat germplasm carrying resistance to these constraints. Nonetheless, modern technologies such as high-throughput phenotyping, genomic and advanced statistical tools can contribute to make HPR to aphids and BYDV more efficient. In the present review we describe the main sources of resistance, discuss the challenges and opportunities for incorporating the resistance in wheat breeding programs and present a workflow to breed for BYDV and its vectors in wheat.
Asunto(s)
Antibiosis , Áfidos/fisiología , Luteovirus/fisiología , Fitomejoramiento , Defensa de la Planta contra la Herbivoria , Triticum/genética , Animales , Herbivoria , Insectos Vectores/fisiología , Enfermedades de las Plantas/virología , Triticum/virologíaRESUMEN
Wheat dwarf bunt is caused by Tilletia controversa Kühn, which is one of the most destructive diseases of wheat worldwide. To explore the interaction of T. controversa and wheat, we analysed the transcriptome profile of spikes of the susceptible wheat cultivar Dongxuan 3, which was subjected to a T. controversa infection and a mock infection. The results obtained from a differential expression analysis of T. controversa-infected plants compared with mock-infected ones showed that 10,867 out of 21,354 genes were upregulated, while 10,487 genes were downregulated, and these genes were enriched in 205 different pathways. Our findings demonstrated that the genes associated with defence against diseases, such as PR-related genes, WRKY transcription factors and mitogen-activated protein kinase genes, were more highly expressed in response to T. controversa infection. Additionally, a number of genes related to physiological attributes were expressed during infection. Three pathways were differentiated based on the characteristics of gene ontology classification. KEGG enrichment analysis showed that twenty genes were expressed differentially during the infection of wheat with T. controversa. Notable changes were observed in the transcriptomes of wheat plants after infection. The results of this study may help to elucidate the mechanism governing the interactions between this pathogen and wheat plants and may facilitate the development of new methods to increase the resistance level of wheat against T. controversa, including the overexpression of defence-related genes.
Asunto(s)
Basidiomycota/genética , Resistencia a la Enfermedad/genética , Micosis/genética , Enfermedades de las Plantas/virología , Transcriptoma , Triticum/virología , Perfilación de la Expresión Génica , Enfermedades de las Plantas/genética , Triticum/genéticaRESUMEN
BACKGROUND: Wheat yellow dwarf virus disease is infected by barley yellow dwarf virus (BYDV), which causes leaf yellowing and dwarfing symptoms in wheat, thereby posing a serious threat to China's food production. The infection of plant viruses can produce large numbers of vsiRNAs, which can target host transcripts and cause symptom development. However, few studies have been conducted to explore the role played by vsiRNAs in the interaction between BYDV-GAV and host wheat plants. METHODS: In this study, small RNA sequencing was conducted to profile vsiRNAs in BYDV-GAV-infected wheat plants. The putative targets of vsiRNAs were predicted by the bioinformatics software psRNATarget. RT-qPCR and VIGS were employed to identify the function of selected target transcripts. To confirm the interaction between vsiRNA and the target, 5' RACE was performed to analyze the specific cleavage sites. RESULTS: From the sequencing data, we obtained a total of 11,384 detected vsiRNAs. The length distribution of these vsiRNAs was mostly 21 and 22 nt, and an A/U bias was observed at the 5' terminus. We also observed that the production region of vsiRNAs had no strand polarity. The vsiRNAs were predicted to target 23,719 wheat transcripts. GO and KEGG enrichment analysis demonstrated that these targets were mostly involved in cell components, catalytic activity and plant-pathogen interactions. The results of RT-qPCR analysis showed that most chloroplast-related genes were downregulated in BYDV-GAV-infected wheat plants. Silencing of a chlorophyll synthase gene caused leaf yellowing that was similar to the symptoms exhibited by BYDV-GAV-inoculated wheat plants. A vsiRNA from an overlapping region of BYDV-GAV MP and CP was observed to target chlorophyll synthase for gene silencing. Next, 5' RACE validated that vsiRNA8856 could cleave the chlorophyll synthase transcript in a sequence-specific manner. CONCLUSIONS: This report is the first to demonstrate that BYDV-GAV-derived vsiRNAs can target wheat transcripts for symptom development, and the results of this study help to elucidate the molecular mechanisms underlying leaf yellowing after viral infection.
Asunto(s)
Ligasas de Carbono-Oxígeno/genética , Hordeum/virología , Interacciones Huésped-Patógeno , Luteovirus/genética , Enfermedades de las Plantas/virología , Hojas de la Planta/virología , ARN Interferente Pequeño/genética , Triticum/virología , Luteovirus/patogenicidad , Hojas de la Planta/enzimología , Interferencia de ARN , Triticum/enzimologíaRESUMEN
Wheat dwarf virus (WDV) is considered as one of the most common viruses on cereal crops. Recently, severe outbreaks of WDV have been observed especially on winter wheat in southwestern part of Poland. Moreover, the presence of genetically different WDV-barley-specific and WDV-wheat-specific forms (WDV-B and WDV-W, respectively) was confirmed. In this study, a loop-mediated isothermal amplification assay (LAMP) was developed for the first time for efficient and rapid detection of WDV-B and WDV-W in infected plants. The reaction was performed using a set of three primer pairs: WDVF3/WDVB3, WDVFIB/WDVBIP and WDVLoopF/WDVLoopB specific for coat protein coding sequence. The amplified products were analyzed by direct staining of DNA, gel electrophoresis and real-time monitoring of the amplification curves. The sensitivity of optimized reaction was tenfold higher in comparison with conventional PCR. LAMP assay developed here is a useful and practical method for the rapid detection of different WDV isolates and can be implemented by phytosanitary services.
Asunto(s)
Geminiviridae/genética , Hordeum/virología , Técnicas de Diagnóstico Molecular , Técnicas de Amplificación de Ácido Nucleico , Filogenia , Enfermedades de las Plantas , Triticale/virología , Triticum/virología , Enfermedades de las Plantas/genética , Enfermedades de las Plantas/virologíaRESUMEN
BACKGROUND: Plant viruses maintain intricate interactions with their vector and non-vector insects and can impact the fitness of insects. However, the details of their molecular and cellular mechanisms have not been studied well. We compared the transcriptome-level responses in vector and non-vector aphids (Schizaphis graminum and Rhopalosiphum padi, respectively) after feeding on wheat plants with viral infections (Barley Yellow Dwarf Virus (BYDV) and Wheat dwarf virus (WDV), respectively). We conducted differentially expressed gene (DEG) annotation analyses and observed DEGs related to immune pathway, growth, development, and reproduction. And we conducted cloning and bioinformatic analyses of the key DEG involved in immune. RESULTS: For all differentially expressed gene analyses, the numbers of DEGs related to immune, growth, development, reproduction and cuticle were higher in vector aphids than in non-vector aphids. STAT5B (signal transducer and activator of transcription 5B), which is involved in the JAK-STAT pathway, was upregulated in R. padi exposed to WDV. The cloning and bioinformatic results indicated that the RpSTAT5B sequence contains a 2082 bp ORF encoding 693 amino acids. The protein molecular weight is 79.1 kD and pI is 8.13. Analysis indicated that RpSTAT5B is a non-transmembrane protein and a non-secreted protein. Homology and evolutionary analysis indicated that RpSTAT5B was closely related to R. maidis. CONCLUSIONS: Unigene expression analysis showed that the total number of differentially expressed genes (DEGs) in the vector aphids was higher than that in the non-vector aphids. Functional enrichment analysis showed that the DEGs related to immunity, growth and reproduction in vector aphids were higher than those in non-vector aphids, and the differentially expressed genes related to immune were up-regulated. This study provides a basis for the evaluation of the response mechanisms of vector/non-vector insects to plant viruses.
Asunto(s)
Áfidos/genética , Insectos Vectores/genética , Transcriptoma , Animales , Áfidos/metabolismo , Áfidos/patogenicidad , Áfidos/virología , Dicistroviridae/patogenicidad , Geminiviridae/patogenicidad , Proteínas de Insectos/genética , Proteínas de Insectos/metabolismo , Insectos Vectores/metabolismo , Insectos Vectores/patogenicidad , Insectos Vectores/virología , Quinasas Janus/genética , Quinasas Janus/metabolismo , Luteovirus/patogenicidad , Factor de Transcripción STAT5/genética , Factor de Transcripción STAT5/metabolismo , Triticum/parasitología , Triticum/virologíaRESUMEN
The leaf rust pathogen, Puccinia triticina (Pt), threatens global wheat production. The deployment of leaf rust (Lr) resistance (R) genes in wheat varieties is often followed by the development of matching virulence in Pt due to presumed changes in avirulence (Avr) genes in Pt. Identifying such Avr genes is a crucial step to understand the mechanisms of wheat-rust interactions. This study is the first to develop and apply an integrated framework of gene expression, single nucleotide polymorphism (SNP), insertion/deletion (InDel), and copy number variation (CNV) analysis in a rust fungus and identify candidate avirulence genes. Using a long-read based de novo genome assembly of an isolate of Pt ('Pt104') as the reference, whole-genome resequencing data of 12 Pt pathotypes derived from three lineages Pt104, Pt53, and Pt76 were analyzed. Candidate avirulence genes were identified by correlating virulence profiles with small variants (SNP and InDel) and CNV, and RNA-seq data of an additional three Pt isolates to validate expression of genes encoding secreted proteins (SPs). Out of the annotated 29,043 genes, 2392 genes were selected as SP genes with detectable expression levels. Small variant comparisons between the isolates identified 27-40 candidates and CNV analysis identified 14-31 candidates for each Avr gene, which when combined, yielded the final 40, 64, and 69 candidates for AvrLr1, AvrLr15, and AvrLr24, respectively. Taken together, our results will facilitate future work on experimental validation and cloning of Avr genes. In addition, the integrated framework of data analysis that we have developed and reported provides a more comprehensive approach for Avr gene mining than is currently available.