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1.
PLoS Biol ; 22(5): e3002626, 2024 May.
Artículo en Inglés | MEDLINE | ID: mdl-38728373

RESUMEN

All plant viruses were thought to encode in its genome a movement protein that acts as a "passport," allowing active movement within the host. A new study in PLOS Biology characterizes the first plant virus that can colonize its host without encoding this protein.


Asunto(s)
Enfermedades de las Plantas , Virus de Plantas , Virus de Plantas/fisiología , Virus de Plantas/genética , Virus de Plantas/patogenicidad , Enfermedades de las Plantas/virología , Plantas/virología , Proteínas de Movimiento Viral en Plantas/metabolismo , Proteínas de Movimiento Viral en Plantas/genética , Genoma Viral , Interacciones Huésped-Patógeno
2.
J Gen Virol ; 105(5)2024 May.
Artículo en Inglés | MEDLINE | ID: mdl-38695734

RESUMEN

Members of the family Fimoviridae are plant viruses with a multipartite negative-sense enveloped RNA genome (-ssRNA), composed of 4-10 segments comprising 12.3-18.5 kb in total, within quasi-spherical virions. Fimoviruses are transmitted to plants by eriophyid mites and induce characteristic cytopathologies in their host plants, including double membrane-bound bodies in the cytoplasm of virus-infected cells. Most fimoviruses infect dicotyledonous plants, and many cause serious disease epidemics. This is a summary of the ICTV Report on the family Fimoviridae, which is available at ictv.global/report/fimoviridae.


Asunto(s)
Genoma Viral , Enfermedades de las Plantas , Virus de Plantas , Enfermedades de las Plantas/virología , Animales , Virus de Plantas/genética , Virus de Plantas/clasificación , Virus de Plantas/fisiología , ARN Viral/genética , Virión/ultraestructura , Plantas/virología , Virus ARN de Sentido Negativo/genética , Virus ARN de Sentido Negativo/clasificación , Ácaros/virología , Filogenia
3.
Methods Mol Biol ; 2788: 157-169, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38656513

RESUMEN

This chapter presents a comprehensive approach to predict novel miRNAs encoded by plant viruses and identify their target plant genes, through integration of various ab initio computational approaches. The predictive process begins with the analysis of plant viral sequences using the VMir Analyzer software. VMir Viewer software is then used to extract primary hairpins from these sequences. To distinguish real miRNA precursors from pseudo miRNA precursors, MiPred web-based software is employed. Verified real pre-miRNA sequences with a minimum free energy of < -20 Kcal/mol, are further analyzed using the RNAshapes software. Validation of predictions involves comparing them with available Expressed Sequence Tags (ESTs) from the relevant plant using BlastN. Short sequences with lengths ranging from 19 to 25 nucleotides and exhibiting <5 mismatches are prioritized for miRNA prediction. The precise locations of these short sequences within pre-miRNA structures generated using RNAshapes are meticulously identified, with a focus on those situated on the 5' and 3' arms of the structures, indicating potential miRNAs. Sequences within the arms of pre-miRNA structures are used to predict target sites within the ESTs of the specific plant, facilitated by psRNA Target software, revealing genes with potential regulatory roles in the plant. To confirm the outcome of target prediction, results are individually submitted to the RNAhybrid web-based software. For practical demonstration, this approach is applied to analyze African cassava mosaic virus (ACMV) and East African cassava mosaic virus-Uganda (EACMV-UG) viruses, as well as the ESTs of Jatropha and cassava.


Asunto(s)
Biología Computacional , MicroARNs , Virus de Plantas , ARN Viral , Programas Informáticos , MicroARNs/genética , Virus de Plantas/genética , Biología Computacional/métodos , ARN Viral/genética , Genes de Plantas , Conformación de Ácido Nucleico , Plantas/virología , Plantas/genética , Etiquetas de Secuencia Expresada
4.
Int J Mol Sci ; 25(8)2024 Apr 11.
Artículo en Inglés | MEDLINE | ID: mdl-38673821

RESUMEN

Isothermal nucleic acid amplification-based lateral flow testing (INAA-LFT) has emerged as a robust technique for on-site pathogen detection, providing a visible indication of pathogen nucleic acid amplification that rivals or even surpasses the sensitivity of real-time quantitative PCR. The isothermal nature of INAA-LFT ensures consistent conditions for nucleic acid amplification, establishing it as a crucial technology for rapid on-site pathogen detection. However, despite its considerable promise, the widespread application of isothermal INAA amplification-based lateral flow testing faces several challenges. This review provides an overview of the INAA-LFT procedure, highlighting its advancements in detecting plant viruses. Moreover, the review underscores the imperative of addressing the existing limitations and emphasizes ongoing research efforts dedicated to enhancing the applicability and performance of this technology in the realm of rapid on-site testing.


Asunto(s)
Técnicas de Amplificación de Ácido Nucleico , Enfermedades de las Plantas , Virus de Plantas , Técnicas de Amplificación de Ácido Nucleico/métodos , Virus de Plantas/genética , Virus de Plantas/aislamiento & purificación , Enfermedades de las Plantas/virología , Técnicas de Diagnóstico Molecular/métodos , Plantas/virología , Plantas/genética
5.
Viruses ; 16(4)2024 Apr 15.
Artículo en Inglés | MEDLINE | ID: mdl-38675953

RESUMEN

There has been substantial progress in the Mediterranean countries regarding research on viroids. Twenty-nine viroid species, all belonging to Pospiviroidae and Avsunviroidae genera, have been detected in the Mediterranean Basin. Not only have detection methods, such as reverse transcription-quantitative polymerase chain reaction and next-generation sequencing, been used for viroid detection, along with molecular hybridization techniques allowing for rapid detection, identification, and characterization of known and novel viroids in these countries, but eradication measures have also been taken that allowed for the efficient elimination of certain viroids in a number of Mediterranean countries. The eradication measures were followed as recommended by the European and Mediterranean Plant Protection Organization, which is known by its abbreviation, EPPO. The Mediterranean Region has been a niche for viroids since ancient times due to the warm climate and the socio-cultural conditions that facilitate viroid transmission among different host plant species.


Asunto(s)
Enfermedades de las Plantas , Viroides , Viroides/genética , Viroides/aislamiento & purificación , Viroides/clasificación , Región Mediterránea , Enfermedades de las Plantas/virología , Plantas/virología , Secuenciación de Nucleótidos de Alto Rendimiento
6.
FEMS Microbiol Ecol ; 100(5)2024 Apr 10.
Artículo en Inglés | MEDLINE | ID: mdl-38678007

RESUMEN

While the One Health framework has emphasized the importance of soil microbiomes for plant and human health, one of the most diverse and abundant groups-bacterial viruses, i.e. phages-has been mostly neglected. This perspective reviews the significance of phages for plant health in rhizosphere and explores their ecological and evolutionary impacts on soil ecosystems. We first summarize our current understanding of the diversity and ecological roles of phages in soil microbiomes in terms of nutrient cycling, top-down density regulation, and pathogen suppression. We then consider how phages drive bacterial evolution in soils by promoting horizontal gene transfer, encoding auxiliary metabolic genes that increase host bacterial fitness, and selecting for phage-resistant mutants with altered ecology due to trade-offs with pathogen competitiveness and virulence. Finally, we consider challenges and avenues for phage research in soil ecosystems and how to elucidate the significance of phages for microbial ecology and evolution and soil ecosystem functioning in the future. We conclude that similar to bacteria, phages likely play important roles in connecting different One Health compartments, affecting microbiome diversity and functions in soils. From the applied perspective, phages could offer novel approaches to modulate and optimize microbial and microbe-plant interactions to enhance soil health.


Asunto(s)
Bacterias , Bacteriófagos , Microbiota , Rizosfera , Microbiología del Suelo , Bacteriófagos/genética , Bacterias/virología , Bacterias/genética , Transferencia de Gen Horizontal , Plantas/microbiología , Plantas/virología , Ecosistema
8.
Nature ; 622(7981): 139-148, 2023 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-37704724

RESUMEN

Aphids transmit viruses and are destructive crop pests1. Plants that have been attacked by aphids release volatile compounds to elicit airborne defence (AD) in neighbouring plants2-5. However, the mechanism underlying AD is unclear. Here we reveal that methyl-salicylate (MeSA), salicylic acid-binding protein-2 (SABP2), the transcription factor NAC2 and salicylic acid-carboxylmethyltransferase-1 (SAMT1) form a signalling circuit to mediate AD against aphids and viruses. Airborne MeSA is perceived and converted into salicylic acid by SABP2 in neighbouring plants. Salicylic acid then causes a signal transduction cascade to activate the NAC2-SAMT1 module for MeSA biosynthesis to induce plant anti-aphid immunity and reduce virus transmission. To counteract this, some aphid-transmitted viruses encode helicase-containing proteins to suppress AD by interacting with NAC2 to subcellularly relocalize and destabilize NAC2. As a consequence, plants become less repellent to aphids, and more suitable for aphid survival, infestation and viral transmission. Our findings uncover the mechanistic basis of AD and an aphid-virus co-evolutionary mutualism, demonstrating AD as a potential bioinspired strategy to control aphids and viruses.


Asunto(s)
Aire , Áfidos , Enfermedades de las Plantas , Plantas , Ácido Salicílico , Transducción de Señal , Áfidos/fisiología , Áfidos/virología , Interacciones Microbiota-Huesped , Enfermedades de las Plantas/inmunología , Enfermedades de las Plantas/parasitología , Enfermedades de las Plantas/prevención & control , Enfermedades de las Plantas/virología , Proteínas de Plantas/metabolismo , Plantas/metabolismo , Plantas/parasitología , Plantas/virología , Ácido Salicílico/metabolismo , Simbiosis , Nicotiana/inmunología , Nicotiana/metabolismo , Nicotiana/parasitología , Nicotiana/virología , Proteínas Virales/metabolismo , Animales
9.
Virologie (Montrouge) ; 27(4): 225-337, 2023 08 01.
Artículo en Francés | MEDLINE | ID: mdl-37565678

RESUMEN

Resistance to viruses is an important aspect of plant breeding. One way to achieve it is to select genetic resistances based on the susceptibility factors hijacked by the virus to infect the plants. Here, we recount work done on genes encoding translation initiation factors eIF4E, some of the most successful targets for obtaining resistance to potyviruses, starting from their characterization 20 years ago. With examples from different plant species, pepper, tomato, tobacco and arabidopsis, we present the basis of this type of resistances and their characteristics, highlighting the role of gene redundancy among 4E factors, their specificity for the virus and the need for the plant of a trade-off between resistance and development. Finally, we show how the new genome editing techniques could be used in plant breeding to develop eIF4E-based resistances in crops, mimicking the functional alleles that have been selected during evolution in many crops.


Asunto(s)
Resistencia a la Enfermedad , Factor 4E Eucariótico de Iniciación , Enfermedades de las Plantas , Plantas , Potyvirus , Alelos , Factor 4E Eucariótico de Iniciación/genética , Fitomejoramiento , Potyvirus/genética , Plantas/virología , Enfermedades de las Plantas/virología
12.
J Virol ; 97(3): e0128422, 2023 03 30.
Artículo en Inglés | MEDLINE | ID: mdl-36786601

RESUMEN

The three genomic and a single subgenomic RNA of Cowpea chlorotic mottle virus (CCMV), which is pathogenic to plants, is packaged into three morphologically indistinguishable icosahedral virions with T=3 symmetry. The two virion types, C1V and C2V, package genomic RNAs 1 (C1) and 2 (C2), respectively. The third virion type, C3+4V, copackages genomic RNA3 and its subgenomic RNA (RNA4). In this study, we sought to evaluate how the alteration of native capsid dynamics by the host and viral replicase modulate the general biology of the virus. The application of a series of biochemical, molecular, and biological assays revealed the following. (i) Proteolytic analysis of the three virion types of CCMV assembled individually in planta revealed that, while retaining the structural integrity, C1V and C2V virions released peptide regions encompassing the N-terminal arginine-rich RNA binding motif. In contrast, a minor population of the C3+4V virion type was sensitive to trypsin-releasing peptides encompassing the entire capsid protein region. (ii) The wild-type CCMV virions purified from cowpea are highly susceptible to trypsin digestion, while those from Nicotiana benthamiana remained resistant, and (iii) finally, the matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) analysis evaluated the relative dynamics of C3+4V and B3+4V virions assembled under the control of the homologous versus heterologous replicase. The role of viral replicase in modulating the capsid dynamics was evident by the differential sensitivity to protease exhibited by B3+4V and C3+4V virions assembled under the homologous versus heterologous replicase. Our results collectively conclude that constant modulation of capsid dynamics by the host and viral replicase is obligatory for successful infection. IMPORTANCE Infectious virus particles or virions are considered static structures and undergo various conformational transitions to replicate and infect many eukaryotic cells. In viruses, conformational changes are essential for establishing infection and evolution. Although viral capsid fluctuations, referred to as dynamics or breathing, have been well studied in RNA viruses pathogenic to animals, such information is limited among plant viruses. The primary focus of this study is to address how capsid dynamics of plant-pathogenic RNA viruses, namely, Cowpea chlorotic mottle (CCMV) and Brome mosaic virus (BMV), are modulated by the host and viral replicase. The results presented have improved and transformed our understanding of the functional relationship between capsid dynamics and the general biology of the virus. They are likely to provide stimulus to extend similar studies to viruses pathogenic to eukaryotic organisms.


Asunto(s)
Bromovirus , Cápside , Interacciones Microbiota-Huesped , Plantas , Proteinas del Complejo de Replicasa Viral , Bromovirus/enzimología , Bromovirus/genética , Cápside/metabolismo , Interacciones Microbiota-Huesped/fisiología , Plantas/virología , ARN Viral/genética , ARN Viral/metabolismo , Tripsina/metabolismo , Proteinas del Complejo de Replicasa Viral/metabolismo , ARN Subgenómico
13.
J Virol ; 96(24): e0026022, 2022 12 21.
Artículo en Inglés | MEDLINE | ID: mdl-35638822

RESUMEN

Our current understanding of plant viruses stems largely from those affecting economically important plants. Yet plant species in cultivation represent a small and biased subset of the plant kingdom. Here, we describe virus diversity and abundance in 1,079 transcriptomes from species across the breadth of the plant kingdom (Archaeplastida) by analyzing open-source data from the 1000 Plant Transcriptomes Initiative (1KP). We identified 104 potentially novel viruses, of which 40% were single-stranded positive-sense RNA viruses across eight orders, including members of the Hepelivirales, Tymovirales, Cryppavirales, Martellivirales, and Picornavirales. One-third of the newly described viruses were double-stranded RNA viruses from the orders Durnavirales and Ghabrivirales. The remaining were negative-sense RNA viruses from the Rhabdoviridae, Aspiviridae, Yueviridae, and Phenuiviridae and the newly proposed Viridisbunyaviridae. Our analysis considerably expands the known host range of 13 virus families to include lower plants (e.g., Benyviridae and Secoviridae) and 4 virus families to include alga hosts (e.g., Tymoviridae and Chrysoviridae). More broadly, however, a cophylogeny analysis revealed that the evolutionary history of these families is largely driven by cross-species transmission events. The discovery of the first 30-kDa movement protein in a nonvascular plant suggests that the acquisition of plant virus movement proteins occurred prior to the emergence of the plant vascular system. Together, these data highlight that numerous RNA virus families are associated with older evolutionary plant lineages than previously thought and that the apparent scarcity of RNA viruses found in lower plants likely reflects a lack of investigation rather than their absence. IMPORTANCE Our knowledge of plant viruses is mainly limited to those infecting economically important host species. In particular, we know little about those viruses infecting basal plant lineages such as the ferns, lycophytes, bryophytes, and charophytes. To expand this understanding, we conducted a broad-scale viral survey of species across the breadth of the plant kingdom. We found that basal plants harbor a wide diversity of RNA viruses, including some that are sufficiently divergent to likely compose a new virus family. The basal plant virome revealed offers key insights into the evolutionary history of core plant virus gene modules and genome segments. More broadly, this work emphasizes that the scarcity of viruses found in these species to date most likely reflects the limited research in this area.


Asunto(s)
Virus de Plantas , Plantas , Virus ARN , Transcriptoma , Filogenia , Virus de Plantas/genética , Plantas/virología , Virus ARN/genética , ARN Viral/genética
14.
Sci Rep ; 12(1): 695, 2022 01 13.
Artículo en Inglés | MEDLINE | ID: mdl-35027584

RESUMEN

In recent decades, a legion of monopartite begomoviruses transmitted by the whitefly Bemisia tabaci has emerged as serious threats to vegetable crops in Africa. Recent studies in Burkina Faso (West Africa) reported the predominance of pepper yellow vein Mali virus (PepYVMLV) and its frequent association with a previously unknown DNA-B component. To understand the role of this DNA-B component in the emergence of PepYVMLV, we assessed biological traits related to virulence, virus accumulation, location in the tissue and transmission. We demonstrate that the DNA-B component is not required for systemic movement and symptom development of PepYVMLV (non-strict association), but that its association produces more severe symptoms including growth arrest and plant death. The increased virulence is associated with a higher viral DNA accumulation in plant tissues, an increase in the number of contaminated nuclei of the phloem parenchyma and in the transmission rate by B. tabaci. Our results suggest that the association of a DNA-B component with the otherwise monopartite PepYVMLV is a key factor of its emergence.


Asunto(s)
Begomovirus/genética , Begomovirus/patogenicidad , ADN Viral/genética , ADN Viral/metabolismo , Enfermedades de las Plantas/virología , Plantas/virología , Virulencia/genética , Animales , Hemípteros/virología , Plantas/metabolismo
15.
Nat Commun ; 13(1): 523, 2022 01 26.
Artículo en Inglés | MEDLINE | ID: mdl-35082293

RESUMEN

Pollen is a unique vehicle for viral spread. Pollen-associated viruses hitchhike on or within pollen grains and are transported to other plants by pollinators. They are deposited on flowers and have a direct pathway into the plant and next generation via seeds. To discover the diversity of pollen-associated viruses and identify contributing landscape and floral features, we perform a species-level metagenomic survey of pollen from wild, visually asymptomatic plants, located in one of four regions in the United States of America varying in land use. We identify many known and novel pollen-associated viruses, half belonging to the Bromoviridae, Partitiviridae, and Secoviridae viral families, but many families are represented. Across the regions, species harbor more viruses when surrounded by less natural and more human-modified environments than the reverse, but we note that other region-level differences may also covary with this. When examining the novel connection between virus richness and floral traits, we find that species with multiple, bilaterally symmetric flowers and smaller, spikier pollen harbored more viruses than those with opposite traits. The association of viral diversity with floral traits highlights the need to incorporate plant-pollinator interactions as a driver of pollen-associated virus transport into the study of plant-viral interactions.


Asunto(s)
Fenotipo , Plantas/virología , Polen/virología , Viroma , Secuencia de Aminoácidos , Animales , Ecología , Flores , Genoma Viral , Filogenia , Polinización , Semillas , Viroma/genética , Virus/clasificación , Virus/genética
16.
Microbiol Res ; 254: 126888, 2022 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-34700185

RESUMEN

Phyllosphere or aerial surface of plants represents the globally largest and peculiar microbial habitat that inhabits diverse and rich communities of bacteria, fungi, viruses, cyanobacteria, actinobacteria, nematodes, and protozoans. These hyperdiverse microbial communities are related to the host's specific functional traits and influence the host's physiology and the ecosystem's functioning. In the last few years, significant advances have been made in unravelling several aspects of phyllosphere microbiology, including diversity and microbial community composition, dynamics, and functional interactions. This review highlights the current knowledge about the assembly, structure, and composition of phyllosphere microbial communities across spatio-temporal scales, besides functional significance of different microbial communities to the plant host and the surrounding environment. The knowledge will help develop strategies for modelling and manipulating these highly beneficial microbial consortia for furthering scientific inquiry into their interactions with the host plants and also for their useful and economic utilization.


Asunto(s)
Biodiversidad , Interacciones Microbiota-Huesped , Microbiota , Plantas , Microbiota/fisiología , Plantas/microbiología , Plantas/parasitología , Plantas/virología
17.
Plant Cell Rep ; 41(2): 281-291, 2022 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-34665312

RESUMEN

The genome of most plant viruses consists of a single positive-strand of RNA (+ ssRNA). Successful replication of these viruses is fully dependent on the endomembrane system of the infected cells, which experiences a massive proliferation and a profound reshaping that enables assembly of the macromolecular complexes where virus genome replication occurs. Assembly of these viral replicase complexes (VRCs) requires a highly orchestrated interplay of multiple virus and co-opted host cell factors to create an optimal microenvironment for efficient assembly and functioning of the virus genome replication machinery. It is now widely accepted that VRC formation involves the recruitment of high levels of sterols, but the specific role of these essential components of cell membranes and the precise molecular mechanisms underlying sterol enrichment at VRCs are still poorly known. In this review, we intend to summarize the most relevant knowledge on the role of sterols in ( +)ssRNA virus replication and discuss the potential of manipulating the plant sterol pathway to help plants fight these infectious agents.


Asunto(s)
Interacciones Huésped-Patógeno/fisiología , Fitosteroles/metabolismo , Virus de Plantas/fisiología , Plantas/metabolismo , Plantas/virología , Membrana Celular/metabolismo , Membrana Celular/virología , Genoma Viral , Enfermedades de las Plantas/virología , Virus de Plantas/patogenicidad , Virus ARN/patogenicidad , Virus ARN/fisiología , Replicación Viral
18.
Nucleic Acids Res ; 50(D1): D432-D438, 2022 01 07.
Artículo en Inglés | MEDLINE | ID: mdl-34751403

RESUMEN

We introduce ViroidDB, a value-added database that attempts to collect all known viroid and viroid-like circular RNA sequences into a single resource. Spanning about 10 000 unique sequences, ViroidDB includes viroids, retroviroid-like elements, small circular satellite RNAs, ribozyviruses, and retrozymes. Each sequence's secondary structure, ribozyme content, and cluster membership are predicted via a custom pipeline optimized for handling circular RNAs. The data can be explored via a purpose-built user interface that features visualizations, multiple sequence alignments, and a portal for downloading bulk data. Users can browse the data by sequence type, taxon, or typo-tolerant search of metadata fields. The database is freely accessible at https://viroids.org.


Asunto(s)
Bases de Datos de Ácidos Nucleicos , ARN Catalítico/genética , ARN Circular/genética , ARN Viral/genética , Programas Informáticos , Viroides/genética , Secuencia de Bases , Internet , Metadatos , Conformación de Ácido Nucleico , Enfermedades de las Plantas/virología , Plantas/virología , ARN Catalítico/química , ARN Catalítico/clasificación , ARN Catalítico/metabolismo , ARN Circular/química , ARN Circular/clasificación , ARN Circular/metabolismo , ARN Viral/química , ARN Viral/clasificación , ARN Viral/metabolismo , Alineación de Secuencia , Viroides/clasificación , Viroides/metabolismo
19.
Methods Mol Biol ; 2400: 125-137, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-34905197

RESUMEN

The plant phloem vasculature is crucial for plant growth and development, and is essential for the systemic movement (SM) of plant viruses. Recent transcriptomic studies of the phloem during virus infection have shown the importance of this tissue, yet transcript levels do not provide definitive answers how virus-host interactions favour successful viral SM. Proteomic analyses have been used to identify host-virus protein interactions, uncovering a variety of ways by which viruses utilize host cellular machinery for completion of the viral infection cycle. Despite this new evidence through proteomics, very few phloem centric studies during viral infection have been performed. Here, we describe a protocol for the isolation of phloem tissues and proteins and the subsequent label-free quantitation (LFQ), for identification of proteomic alterations caused by viral infection.


Asunto(s)
Virus de Plantas , Virosis , Floema , Plantas/virología , Proteómica
20.
Methods Mol Biol ; 2316: 23-28, 2022.
Artículo en Inglés | MEDLINE | ID: mdl-34845681

RESUMEN

The characterization of the elusive disease agent of the potato spindle tuber disease, potato spindle tuber viroid (PSTVd), was aided by the ability to obtain large amounts of infected tomato tissue in a simple bioassay where PSTVd was easily mechanically transmissible to an alternate herbaceous host in which it thrived and produced dramatic symptoms in a relatively short period (Diener, Viroids. Handbook of plant virus infections: comparative diagnosis. Elsevier/North-Holland, Amsterdam, pp 913-934, 1981; Diener, Virology 45:411-428, 1971; Raymer and O'Brien, Am Pot J, 39:401-408, 1962). Reactions in the primary, or secondary, herbaceous indicator host can range from asymptomatic to severe depending upon the viroid strain, host species, and environmental conditions and can provide evidence of a viroid infection, but do not permit identification of the viroid in question. Further characterization by molecular hybridization, RT-PCR, and sequence analysis is used to determine the etiology of the disease agent. In this chapter, methods are described for mechanical inoculation of viroids to herbaceous hosts to determine the viroid nature of diseases and the experimental host range of the viroid or to shorten the time required for obtaining relatively large amounts of viroid for subsequent purification and characterization.


Asunto(s)
Virus de Plantas , Viroides , Bioensayo , Solanum lycopersicum , Enfermedades de las Plantas , Virus de Plantas/genética , Plantas/virología , ARN Viral/genética , Solanum tuberosum , Viroides/genética
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