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1.
PLoS One ; 17(8): e0270918, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35914219

RESUMO

Potato mop-top virus (PMTV) is considered an emerging threat to potato production in the United States. PMTV is transmitted by a soil-borne protist, Spongospora subterranean. Rapid, accurate, and sensitive detection of PMTV in leaves and tubers is an essential component in PMTV management program. A rapid test that can be adapted to in-field, on-site testing with minimal sample manipulation could help in ensuring the sanitary status of the produce in situations such as certification programs and shipping point inspections. Toward that goal, a rapid and highly sensitive recombinase polymerase amplification (RPA)-based test was developed for PMTV detection in potato tubers. The test combines the convenience of RPA assay with a simple sample extraction procedure, making it amenable to rapid on-site diagnosis of PMTV. Furthermore, the assay was duplexed with a plant internal control to monitor sample extraction and RPA reaction performance. The method described could detect as little as 10 fg of PMTV RNA transcript in various potato tissues, the diagnostic limit of detection (LOQ) similar to that of traditional molecular methods.


Assuntos
Vírus de Plantas , Solanum tuberosum , Doenças das Plantas , Vírus de Plantas/genética , Solo
2.
Biochemistry (Mosc) ; 87(6): 548-558, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35790410

RESUMO

Structurally modified virus particles can be obtained from the rod-shaped or filamentous virions of plant viruses and bacteriophages by thermal or chemical treatment. They have recently attracted attention of the researchers as promising biogenic platforms for the development of new biotechnologies. This review presents data on preparation, structure, and properties of the structurally modified virus particles. In addition, their biosafety for animals is considered, as well as the areas of application of such particles in biomedicine. A separate section is devoted to one of the most relevant and promising areas for the use of structurally modified plant viruses - design of vaccine candidates based on them.


Assuntos
Bacteriófagos , Vírus de Plantas , Animais , Vírion
3.
J Agric Food Chem ; 70(29): 8892-8900, 2022 Jul 27.
Artigo em Inglês | MEDLINE | ID: mdl-35830295

RESUMO

Coat proteins (CPs) of RNA plant viruses play a pivotal role in virus particle assembly, vector transmission, host identification, RNA replication, and intracellular and intercellular movement. Numerous compounds targeting CPs have been designed, synthesized, and screened for their antiviral activities. This review is intended to fill a knowledge gap where a comprehensive summary is needed for antiviral agent discovery based on plant viral CPs. In this review, major achievements are summarized with emphasis on plant viral CPs as biochemical targets and action mechanisms of antiviral agents. This review hopefully provides new insights and references for the further development of new safe and effective antiviral pesticides.


Assuntos
Proteínas do Capsídeo , Vírus de Plantas , Antivirais/metabolismo , Antivirais/farmacologia , Proteínas do Capsídeo/genética , Proteínas do Capsídeo/metabolismo , Vírus de Plantas/genética , Plantas/metabolismo , RNA , Replicação Viral
4.
Biomolecules ; 12(7)2022 Jun 21.
Artigo em Inglês | MEDLINE | ID: mdl-35883420

RESUMO

Previous studies have shown that the RNA genomes of some plant viruses encode two related genetic modules required for virus movement over the host body, containing two or three genes and named the binary movement block (BMB) and triple gene block (TGB), respectively. In this paper, we predict a novel putative-related movement gene module, called the tetra-cistron movement block (TCMB), in the virus-like transcriptome assemblies of the moss Dicranum scoparium and the Antarctic flowering plant Colobanthus quitensis. These TCMBs are encoded by smaller RNA components of putative two-component viruses related to plant benyviruses. Similar to the RNA2 of benyviruses, TCMB-containing RNAs have the 5'-terminal coat protein gene and include the RNA helicase gene which is followed by two small overlapping cistrons encoding hydrophobic proteins with a distant sequence similarity to the TGB2 and TGB3 proteins. Unlike TGB, TCMB also includes a fourth 5'-terminal gene preceding the helicase gene and coding for a protein showing a similarity to the double-stranded RNA-binding proteins of the DSRM AtDRB-like superfamily. Additionally, based on phylogenetic analysis, we suggest the involvement of replicative beny-like helicases in the evolution of the BMB and TCMB movement genetic modules.


Assuntos
Biologia Computacional , Vírus de Plantas , Redes Reguladoras de Genes , Filogenia , Vírus de Plantas/genética , Vírus de Plantas/metabolismo , RNA/metabolismo , Proteínas Virais/genética , Proteínas Virais/metabolismo
5.
ACS Synth Biol ; 11(7): 2384-2393, 2022 07 15.
Artigo em Inglês | MEDLINE | ID: mdl-35793201

RESUMO

Viral infections in plants threaten food security. Thus, simple and effective methods for virus detection are required to adopt early measures that can prevent virus spread. However, current methods based on the amplification of the viral genome by polymerase chain reaction (PCR) require laboratory conditions. Here, we exploited the CRISPR-Cas12a and CRISPR-Cas13a/d systems to detect three RNA viruses, namely, Tobacco mosaic virus, Tobacco etch virus, and Potato virus X, in Nicotiana benthamiana plants. We applied the CRISPR-Cas12a system to detect viral DNA amplicons generated by PCR or isothermal amplification, and we also performed a multiplexed detection in plants with mixed infections. In addition, we adapted the detection system to bypass the costly RNA purification step and to get a visible readout with lateral flow strips. Finally, we applied the CRISPR-Cas13a/d system to directly detect viral RNA, thereby avoiding the necessity of a preamplification step and obtaining a readout that scales with the viral load. These approaches allow for the performance of viral diagnostics within half an hour of leaf harvest and are hence potentially relevant for field-deployable applications.


Assuntos
Sistemas CRISPR-Cas , Vírus de Plantas , Sistemas CRISPR-Cas/genética , Genoma Viral , Vírus de Plantas/genética , Plantas/genética , RNA Viral/genética , Tabaco/genética
6.
Methods Mol Biol ; 2536: 381-394, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35819615

RESUMO

More than 80 viral species, many of which are not associated with a clear disease or symptomatology, can infect grapevine. The study of grapevine-virus interactions in recent years is playing an increasingly important role and these studies have shown that the molecular and physiological responses to a virus greatly vary depending on the viral strains, the presence of multiple viral infections, the grapevine genotype, and the environment. Moreover, due to the characteristics of the grapevine cultivation and its vegetative propagation, it is very difficult to find healthy plants in vineyards to use them as control in the experiments. Starting from these considerations, in order to investigate the plant-virus interaction in an unbiased way, it is important to set up an experimental system able to control as much of these variables as possible. The protocol here proposed provides the overcome some of these factors by: (i) the production of healthy plants by somatic embryogenesis; (ii) the virus transmission using in vitro micrografting, and (iii) the transfer of in vitro plants to ex-vitro conditions for the analysis of interest.


Assuntos
Doenças das Plantas , Técnicas de Embriogênese Somática de Plantas , Vírus de Plantas , Vitis , Interações entre Hospedeiro e Microrganismos , Doenças das Plantas/virologia , Vírus de Plantas/genética , Vitis/virologia
7.
Viruses ; 14(7)2022 07 05.
Artigo em Inglês | MEDLINE | ID: mdl-35891459

RESUMO

High-throughput sequencing (HTS) of host plant small RNA (sRNA) is a popular approach for plant virus and viroid detection. The major bottlenecks for implementing this approach in routine virus screening of plants in quarantine include lack of computational resources and/or expertise in command-line environments and limited availability of curated plant virus and viroid databases. We developed: (1) virus and viroid report web-based bioinformatics workflows on Galaxy Australia called GA-VirReport and GA-VirReport-Stats for detecting viruses and viroids from host plant sRNA extracts and (2) a curated higher plant virus and viroid database (PVirDB). We implemented sRNA sequencing with unique dual indexing on a set of plants with known viruses. Sequencing data were analyzed using GA-VirReport and PVirDB to validate these resources. We detected all known viruses in this pilot study with no cross-sample contamination. We then conducted a large-scale diagnosis of 105 imported plants processed at the post-entry quarantine facility (PEQ), Australia. We detected various pathogens in 14 imported plants and discovered that de novo assembly using 21-22 nt sRNA fraction and the megablast algorithm yielded better sensitivity and specificity. This study reports the successful, large-scale implementation of HTS and a user-friendly bioinformatics workflow for virus and viroid screening of imported plants at the PEQ.


Assuntos
Vírus de Plantas , Pequeno RNA não Traduzido , Viroides , Biologia Computacional , Internet , Projetos Piloto , Doenças das Plantas , Vírus de Plantas/genética , Plantas , Quarentena , RNA de Plantas , Viroides/genética
8.
Viruses ; 14(7)2022 07 12.
Artigo em Inglês | MEDLINE | ID: mdl-35891498

RESUMO

Alfalfa (Medicago sativa L.) is one of the most important quality forages worldwide and is cultivated throughout China. Alfalfa is susceptible to a variety of viral diseases during its growth, which has caused huge amounts of commercial losses. However, the profile of the alfalfa virus in China remains ambiguous and the viruses transmitted by Odontothrips loti (Haliday), dominant insect pests in alfalfa, are also poorly understood. In the present study, virus diversity was investigated in the primary alfalfa-growing areas in China. A total of 18 alfalfa viruses were identified through RNA-sequencing (RNA-seq) and reverse transcription-polymerase chain reaction (RT-PCR). Two new plant viruses, Medicago sativa virus 1 (MsV1) and Medicago sativa luteovirus 1 (MsLV1), were detected for the first time. Another four viruses, including the Alfalfa ringspot-associated virus (ARaV), Alfalfa virus F (AVF), Alfalfa enamovirus 1 (AEV1), and Alfalfa deltaparitivirus (ADPV), were reported in China for the first time as well. Both Alfalfa mosaic virus (AMV) and Medicago sativa alphapartitivirus 2 (MsAPV2) are the dominant pathogens, with an infection incidence of 91.7-100%, and 74.4-97.2%, respectively. Additionally, O. loti with first- and second-instar nymphs were shown to acquire the AMV within 0.25 h of feeding on a virus-infected alfalfa. Transmission by thrips to healthy alfalfa plants was also demonstrated. Additionally, we clarified the dynamic changes in the AMV in pre-adult stages of O. loti, which indicated that the AMV is propagated in the nymph stage of O. loti. These findings provide valuable information for understanding the alfalfa virome, confirm the role thrips O. loti plays in alfalfa virus transmission, and improve our fundamental knowledge and management of diseases in China.


Assuntos
Luteoviridae , Vírus de Plantas , Vírus de RNA , Tisanópteros , Tymoviridae , Animais , Medicago sativa , Vírus de Plantas/genética
9.
EMBO J ; 41(13): e110060, 2022 Jul 04.
Artigo em Inglês | MEDLINE | ID: mdl-35642376

RESUMO

Viral replication and movement are intimately linked; however, the molecular mechanisms regulating the transition between replication and subsequent movement remain largely unknown. We previously demonstrated that the Barley stripe mosaic virus (BSMV) γb protein promotes viral replication and movement by interacting with the αa replicase and TGB1 movement proteins. Here, we found that γb is palmitoylated at Cys-10, Cys-19, and Cys-60 in Nicotiana benthamiana, which supports BSMV infection. Intriguingly, non-palmitoylated γb is anchored to chloroplast replication sites and enhances BSMV replication, whereas palmitoylated γb protein recruits TGB1 to the chloroplasts and forms viral replication-movement intermediate complexes. At the late stages of replication, γb interacts with NbPAT15 and NbPAT21 and is palmitoylated at the chloroplast periphery, thereby shifting viral replication to intracellular and intercellular movement. We also show that palmitoylated γb promotes virus cell-to-cell movement by interacting with NbREM1 to inhibit callose deposition at the plasmodesmata. Altogether, our experiments reveal a model whereby palmitoylation of γb directs a dynamic switch between BSMV replication and movement events during infection.


Assuntos
Lipoilação , Vírus de Plantas , Tabaco/metabolismo , Proteínas não Estruturais Virais/metabolismo , Replicação Viral
10.
Viruses ; 14(6)2022 06 09.
Artigo em Inglês | MEDLINE | ID: mdl-35746722

RESUMO

Tomato is the most economically important vegetable crop worldwide and the second most important for Mexico. However, viral diseases are among the main limiting factors that affect the productivity of this crop, causing total losses in some cases. This review provides key information and findings on the symptoms, distribution, transmission, detection, and management of diseases caused by viruses of major importance in tomato crops in Mexico. Currently, about 25 viruses belonging to nine different families have been reported infecting tomato in Mexico, but not all of them cause economically significant diseases. Viruses of economic importance include tomato brown rugose fruit virus (ToBRFV), tomato spotted wilt virus (TSWV), tomato yellow leaf curl virus (TYLCV), pepino mosaic virus (PepMV), and tomato marchitez virus (ToMarV). The topics discussed here will provide updated information about the status of these plant viruses in Mexico as well as diverse management strategies that can be implemented according to the specific circumstances of each viral pathosystem. Additionally, a list of tomato-affecting viruses not present in Mexico that are continuous threats to the crop health is included.


Assuntos
Lycopersicon esculentum , Vírus de Plantas , Tospovirus , Produtos Agrícolas , Humanos , México , Doenças das Plantas
11.
Viruses ; 14(6)2022 06 18.
Artigo em Inglês | MEDLINE | ID: mdl-35746802

RESUMO

Plant viruses threaten agricultural production by reducing the yield, quality, and economical benefits. Tomato mosaic virus (ToMV) from the genus Tobamovirus causes serious losses in the quantity and quality of tomato production. The management of plant protection is very difficult, mainly due to the vector-less transmission of ToMV. Resistant breeding generally has low effectiveness. The most practical approach is the use of a rapid diagnostic assay of the virus' presence before the symptoms occur in plants, followed by the eradication of virus-infected plants. Such approaches also include serological detection methods (ELISA and Western immunoblotting), where antibodies need to be developed for an immunochemical reaction. The development and characterization of polyclonal antibodies for the detection of ToMV with appropriate parameters (sensitivity, specificity, and cross-reactivity) were the subjects of this study. A new polyclonal antibody, AB-1, was developed in immunized rabbits using the modified oligopeptides with antigenic potential (sequences are revealed) derived from the coat protein of ToMV SL-1. the developed polyclonal antibody. AB-1, showed higher sensitivity when compared with commercially available analogs. It also detected ToMV in infected pepper and eggplant plants, and detected another two tobamoviruses (TMV and PMMoV) and ToMV in soil rhizosphere samples and root residues, even two years after the cultivation of the infected tomato plant.


Assuntos
Lycopersicon esculentum , Vírus de Plantas , Tobamovirus , Animais , Humanos , Melhoramento Vegetal , Doenças das Plantas , Plantas , Coelhos , Tobamovirus/genética
12.
Acta Virol ; 66(2): 192-194, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35766477

RESUMO

Presence of alternate hosts of plants is a great threat to the agriculture industry. Plants from several species growing in the papaya orchards affected by papaya sticky disease were examined for Papaya meleira virus (PMeV) infection causing this disease. The viral dsRNA was already detected in some plants from the family Poaceae or in watermelon. To identify new hosts of PMeV, we have collected 38 plant species belonging to 15 families of common weed species found in papaya-growing areas in México and used reverse-transcription PCR (RT-PCR) or quantitative real-time RT-PCR (RT-qPCR) for virus detection. We have detected the viral RNA in 11 species belonging to the families Acanthaceae, Fabaceae and Poaceae. Under experimental conditions, PMeV-Mx in Panicum hirsutum and Ruellia nudiflora inoculated weed species, showed that PMeV-Mx is able to replicate in plant cells of these species and spread in a systemic way. These results highlight the importance of weed species as potential virus reservoirs for PMeV-Mx Keywords: Papaya meleira virus; papaya sticky disease; Carica papaya; RT-PCR; TaqMan.


Assuntos
Carica , Vírus de Plantas , México , Doenças das Plantas , Vírus de Plantas/genética , RNA Viral
13.
Sci China Life Sci ; 65(8): 1498-1503, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-35661965

RESUMO

Geminiviruses are a group of plant viruses that cause severe diseases in many economically important crops worldwide, leading to devastating losses to agricultural production. Here we summarize the occurrence and distribution of geminiviruses in China, which provides valuable information for further epidemiological studies and supports the development of effective disease management strategies.


Assuntos
Geminiviridae , Vírus de Plantas , China , Produtos Agrícolas , Geminiviridae/genética , Doenças das Plantas
14.
Viruses ; 14(5)2022 04 21.
Artigo em Inglês | MEDLINE | ID: mdl-35632605

RESUMO

As rivals over the long history of co-evolution, viruses and host plants have each developed specialized strategies and machineries to cope with the rivalry [...].


Assuntos
Vírus de Plantas , Vírus não Classificados , Vírus de DNA , Doenças das Plantas , Vírus de Plantas/genética , Plantas
15.
Sci Rep ; 12(1): 8429, 2022 05 19.
Artigo em Inglês | MEDLINE | ID: mdl-35589977

RESUMO

There is widespread evidence of plant viruses manipulating behavior of their insect vectors as a strategy to maximize infection of plants. Often, plant viruses and their insect vectors have multiple potential host plant species, and these may not overlap entirely. Moreover, insect vectors may not prefer plant species to which plant viruses are well-adapted. In such cases, can plant viruses manipulate their insect vectors to preferentially feed and oviposit on plant species, which are suitable for viral propagation but less suitable for themselves? To address this question, we conducted dual- and no-choice feeding studies (number and duration of probing events) and oviposition studies with non-viruliferous and viruliferous [carrying beet curly top virus (BCTV)] beet leafhoppers [Circulifer tenellus (Baker)] on three plant species: barley (Hordeum vulgare L.), ribwort plantain (Plantago lanceolata L.), and tomato (Solanum lycopersicum L.). Barley is not a host of BCTV, whereas ribwort plantain and tomato are susceptible to BCTV infection and develop a symptomless infection and severe curly top symptoms, respectively. Ribwort plantain plants can be used to maintain beet leafhopper colonies for multiple generations (suitable), whereas tomato plants cannot be used to maintain beet leafhopper colonies (unsuitable). Based on dual- and no-choice experiments, we demonstrated that BCTV appears to manipulate probing preference and behavior by beet leafhoppers, whereas there was no significant difference in oviposition preference. Simulation modeling predicted that BCTV infection rates would to be higher in tomato fields with barley compared with ribwort plantain as a trap crop. Simulation model results supported the hypothesis that manipulation of probing preference and behavior may increase BCTV infection in tomato fields. Results presented were based on the BCTV-beet leafhopper pathosystem, but the approach taken (combination of experimental studies with complementary simulation modeling) is widely applicable and relevant to other insect-vectored plant pathogen systems involving multiple plant species.


Assuntos
Beta vulgaris , Geminiviridae , Hemípteros , Vírus de Plantas , Animais , Feminino , Insetos Vetores , Doenças das Plantas , Plantas
16.
Genes (Basel) ; 13(5)2022 Apr 22.
Artigo em Inglês | MEDLINE | ID: mdl-35627117

RESUMO

Potato is a major food crop that has the potential to feed the increasing global population. Potato is the fourth most important crop and a staple food for many people worldwide. The traditional breeding of potato poses many challenges because of its autotetraploid nature and its tendency toward inbreeding depression. Moreover, potato crops suffer considerable production losses because of infections caused by plant viruses. In this context, RNA silencing technology has been successfully applied in model and crop species. In this review, we describe the RNA interference (RNAi) mechanisms, including small-interfering RNA, microRNA, and artificial microRNA, which may be used to engineer resistance against potato viruses. We also explore the latest advances in the development of antiviral strategies to enhance resistance against potato virus X, potato virus Y, potato virus A, potato leafroll virus, and potato spindle tuber viroid. Furthermore, the challenges in RNAi that need to be overcome are described in this review. Altogether, this report would be insightful for the researchers attempting to understand the RNAi-mediated resistance against viruses in potato.


Assuntos
MicroRNAs , Vírus de Plantas , Solanum tuberosum , Humanos , MicroRNAs/genética , Melhoramento Vegetal , Vírus de Plantas/genética , Interferência de RNA , RNA Interferente Pequeno , Solanum tuberosum/genética
17.
Int J Mol Sci ; 23(10)2022 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-35628126

RESUMO

Plant viruses are devastating plant pathogens that severely affect crop yield and quality. Plants have developed multiple lines of defense systems to combat viral infection. Gene silencing/RNA interference is the key defense system in plants that inhibits the virulence and multiplication of pathogens. The general mechanism of RNAi involves (i) the transcription and cleavage of dsRNA into small RNA molecules, such as microRNA (miRNA), or small interfering RNA (siRNA), (ii) the loading of siRNA/miRNA into an RNA Induced Silencing Complex (RISC), (iii) complementary base pairing between siRNA/miRNA with a targeted gene, and (iv) the cleavage or repression of a target gene with an Argonaute (AGO) protein. This natural RNAi pathway could introduce transgenes targeting various viral genes to induce gene silencing. Different RNAi pathways are reported for the artificial silencing of viral genes. These include Host-Induced Gene Silencing (HIGS), Virus-Induced Gene Silencing (VIGS), and Spray-Induced Gene Silencing (SIGS). There are significant limitations in HIGS and VIGS technology, such as lengthy and time-consuming processes, off-target effects, and public concerns regarding genetically modified (GM) transgenic plants. Here, we provide in-depth knowledge regarding SIGS, which efficiently provides RNAi resistance development against targeted genes without the need for GM transgenic plants. We give an overview of the defense system of plants against viral infection, including a detailed mechanism of RNAi, small RNA molecules and their types, and various kinds of RNAi pathways. This review will describe how RNA interference provides the antiviral defense, recent improvements, and their limitations.


Assuntos
MicroRNAs , Vírus de Plantas , Proteínas Argonauta/genética , Vírus de Plantas/genética , Plantas/genética , Interferência de RNA , RNA de Cadeia Dupla/genética , RNA Interferente Pequeno/genética , Complexo de Inativação Induzido por RNA/genética
18.
BMC Genomics ; 23(1): 371, 2022 May 16.
Artigo em Inglês | MEDLINE | ID: mdl-35578183

RESUMO

BACKGROUND: Cucurbita pepo is highly susceptible to Zucchini yellow mosaic virus (ZYMV) and the resistance found in several wild species cannot be considered as complete or broad-spectrum resistance. In this study, a source of tolerance introgressed in C. pepo (381e) from C. moschata, in True French (TF) background, was investigated 12 days post-inoculation (DPI) at transcriptomic and genomic levels. RESULTS: The comparative RNA-sequencing (RNA-Seq) of TF (susceptible to ZYMV) and 381e (tolerant to ZYMV) allowed the evaluation of about 33,000 expressed transcripts and the identification of 146 differentially expressed genes (DEGs) in 381e, mainly involved in photosynthesis, transcription, cytoskeleton organization and callose synthesis. By contrast, the susceptible cultivar TF triggered oxidative processes related to response to biotic stimulus and activated key regulators of plant virus intercellular movement. In addition, the discovery of variants located in transcripts allowed the identification of two chromosome regions rich in Single Nucleotide Polymorphisms (SNPs), putatively introgressed from C. moschata, containing genes exclusively expressed in 381e. CONCLUSION: 381e transcriptome analysis confirmed a global improvement of plant fitness by reducing the virus titer and movement. Furthermore, genes implicated in ZYMV tolerance in C. moschata introgressed regions were detected. Our work provides new insight into the plant virus recovery process and a better understanding of the molecular basis of 381e tolerance.


Assuntos
Cucurbita , Vírus de Plantas , Potyvirus , Cucurbita/genética , Genômica , Doenças das Plantas/genética , Vírus de Plantas/genética , Potyvirus/genética , Transcriptoma
19.
J Agric Food Chem ; 70(19): 5838-5848, 2022 May 18.
Artigo em Inglês | MEDLINE | ID: mdl-35532753

RESUMO

Potato virus X (PVX), a species of the genus Potexvirus, is a plant pathogenic virus that causes severe symptoms such as mild mosaic, crinkling, necrosis, and mottling on leaves. The objectives of the present study were to investigate the effect of PVX virus infection on the metabolic system in nontransgenic and Arabidopsis thaliana production of anthocyanin pigment 1 (AtPAP1) transgenic tobacco using transcript expression analysis and metabolic profiling. Potato virus X inoculation increased the gene expression of phenylpropanoid and flavonoid biosynthesis and the production of chlorogenic acid, p-coumaric acid, benzoic acid, rutin, quercetin, and kaempferol in nontransgenic tobacco leaves. However, in the AtPAP1 transgenic tobacco leaves, PVX inoculation decreased the expression of AtPAP1 and phenylpropanoid and flavonoid biosynthesis genes, and the production of phenolics and anthocyanin also declined. In contrast, the levels of amino acids and tricarboxylic acid (TCA) cycle intermediates increased after infection in the AtPAP1 transgenic plant leaves. To date, these results have not been reported previously. We suggest that PVX infection decreases AtPAP1 expression, leading to the downregulation of phenylpropanoid and flavonoid biosynthesis in transgenic plants.


Assuntos
Arabidopsis , Vírus de Plantas , Potexvirus , Antocianinas , Arabidopsis/genética , Expressão Gênica , Plantas Geneticamente Modificadas/genética , Potexvirus/genética , Tabaco/genética
20.
Methods Mol Biol ; 2480: 103-111, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35616860

RESUMO

Recent discoveries in the dynamics of genome replication and packaging in the plant virus Cowpea mosaic virus (CPMV) has led to the development of a novel method for specifically packaging an RNA molecule of choice into virus-like particles (VLPs) of CPMV. Thanks to modern gene synthesis and molecular cloning methods, the DNA sequence corresponding to an RNA sequence of interest can be cloned into a suitable expression plasmid for transient expression in plants. We describe here a method for ensuring that this RNA sequence will be packaged within VLPs of CPMV in plant cells by replication-dependent RNA packaging. This requires co-expression of the CPMV replication machinery alongside the CPMV coat protein precursor. These components are co-expressed in the leaves of the Nicotiana benthamiana plant and this co-expression results in the production of large quantities of VLPs that contain the RNA sequence of choice. These VLPs are easy to extract and purify from the plant tissue, and are stable for months in refrigerated conditions. These VLPs can then be used for a variety of different applications, such as RNA delivery or control reagents in RT-qPCR.


Assuntos
Comovirus , Vírus de Plantas , Comovirus/genética , Comovirus/metabolismo , Vírus de Plantas/genética , Plasmídeos , RNA/metabolismo , Tabaco/genética
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