Produção de antissoro policlonal utilizando a proteína capsidial recombinante do Rupestris stem pitting-associated virus

Basso, Marcos Fernando; Fajardo, T. V. M.; Eiras, Marcelo; Ayub, Ricardo Antônio; Nickel, Osmar

doi:10.1590/s0103-84782010001100022

Cited by 5 publications

(5 citation statements)

References 5 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The obtained results were similar to the average yields of recombinant coat proteins found for other viruses (Fajardo et al, 2007;Basso et al, 2010). Good quality antiserum is essential for the efficiency of serological detection, particularly for a low titer virus such as PLYV (Nascimento et al, 2010).…”

Section: Resultssupporting

confidence: 82%

Screening of papaya accessions resistant to Papaya lethal yellowing virus and capacity of Tetranychus urticae to transmit the virus

Basso

Pereira

et al. 2015

Pesq. agropec. bras.

Self Cite

View full text Add to dashboard Cite

-The objective of this work was to produce a polyclonal antiserum against the coat protein (CP) of Papaya lethal yellowing virus (PLYV) and to determine its specificity and sensibility in the diagnosis of the virus, as well as to evaluate the genetic resistance to PLYV in papaya (Carica papaya) accessions and to investigate the capacity of the two-spotted spider mite Tetranychus urticae to acquire and transmit PLYV to the plants. Sixty-five papaya accessions were evaluated. For each accession, ten plants were mechanically inoculated using PLYV-infected plant extracts, and three plants were mock inoculated with phosphate buffer alone and used as negative controls. Ninety days after inoculation, newly-emerging systemic leaves were collected from the inoculated plants, and viral infection was diagnosed by indirect Elisa, using polyclonal antiserum sensible to the in vitro-expressed PLYV CP. Viral transmission by T. urticae was evaluated in greenhouse. The experiments were repeated twice. Polyclonal antiserum recognized the recombinant PLYV CP specifically and discriminated PLYV infection from infections caused by other plant viruses. Out of the 65 papaya accessions evaluated, 15 were considered resistant, 18 moderately resistant, and 32 susceptible. The two-spotted spider mite T. urticae was capable of acquiring PLYV, but not of transmitting it to papaya.Index terms: Carica papaya, Elisa, genetic resistance, plant breeding, PLYV, two-spotted spider mite. Termos para indexação: Carica papaya, Elisa, resistência genética, melhoramento de plantas, PLYV, ácaro rajado. Identificação de acessos de mamoeiro resistentes ao

show abstract

Section: Resultssupporting

confidence: 82%

Screening of papaya accessions resistant to Papaya lethal yellowing virus and capacity of Tetranychus urticae to transmit the virus

Basso

Pereira

et al. 2015

Pesq. agropec. bras.

Self Cite

View full text Add to dashboard Cite

show abstract

“…Serological diagnostic techniques based on specific mono or polyclonal antibodies produced from pre-defined antigens search for the presence of a certain pathogen within an infected tissue sample. 22 In the 1970s, the 'enzyme-linked immuno-sorbent assay' (ELISA) was developed to detect plant viruses present in aqueous solutions. 23 ELISAs are currently the most widely used immunodiagnostic techniques for their high-throughput potential and low cost.…”

Section: Serological Methodsmentioning

confidence: 99%

Next‐generation methods for early disease detection in crops

Trippa,

Scalenghe,

Basso

et al. 2023

Pest Management Science

Self Cite

View full text Add to dashboard Cite

BackgroundPlant pathogens are commonly identified in the field by the typical disease symptoms that they can cause. The efficient early detection and identification of pathogens are essential procedures to adopt effective management practices that reduce or prevent their spread in order to mitigate the negative impacts of the disease. Herein, in this review were presented and discussed the traditional and innovative methods for early detection of the plant pathogens highlighting their major advantages and limitations.Resultstraditional techniques of diagnosis used for plant pathogen identification are focused typically on the DNA, RNA (when molecular methods), and proteins or peptides (when serological methods) of the pathogens. Serological methods based on mainly enzyme‐linked immunosorbent assay (ELISA) are the most common method used for pathogen detection due to their high‐throughput potential and low cost. This technique is not particularly reliable and sufficiently sensitive for many pathogens detection during the asymptomatic stage of infection. For non‐cultivable pathogens in the laboratory, nucleic acid‐based technology is the best choice for consistent pathogen detection or identification. Lateral flow systems are innovative tools that allow fast and accurate results even in the field conditions, but they have sensitivity issues to be overcome. PCR assays performed on last‐generation portable thermocyclers may provide rapid detection results in situ.ConclusionThe advent of portable instruments can speed pathogen detection, reduce commercial costs, and potentially can revolutionize plant pathology. This review provides information on current methodologies and procedures for the effective detection of different plant pathogens.This article is protected by copyright. All rights reserved.

show abstract

“…Serological methods for the detection of GRSPaV in field-collected material have shown limited success. Several polyclonal antibodies have been developed for the use in Western blotting and enzyme-linked immunosorbent assays (ELISAs) (Basso et al, 2010;. These methods are more rapid than biological indexing, but are limited to specific viruses and will therefore not be efficient at detecting diseases for which the associated infectious agents are not known.…”

Section: Serological Methodsmentioning

confidence: 99%

“…Expression of the capsid protein greatly varied based on the genetic make-up of cultivars, types of tissue tested, and climate (Meng et al, 2000;Minafra et al, 2015). Antiserums for GRSPaV continue to be developed and tested (Basso et al, 2010), but no serological assay has been successful enough to be commercially implemented on a global level.…”

Section: Serological Methodsmentioning

confidence: 99%

“…Specific protocols for the isolation and purification of nucleic acids from grapevines have been proposed, however, the protocols are not equally efficient at extracting high quality RNA from all tissue types (Xiao et al, 2015). A low virus titre and a suspected uneven localisation in the plant, especially during early stages of infection, can also lead to unreliable detection via RT-PCR or RT-qPCR (Basso et al, 2010;Beuve et al, 2007;Meng et al, 2013).…”

Section: Nucleic-acid Based Detection Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Genetic diversity and identification of putative recombination events in grapevine rupestris stem pitting-associated virus

2018

View full text Add to dashboard Cite

The impact of recombination on variant classification and the use of different genomic regions to identify virus variants were investigated using a diversity study performed on grapevine rupestris stem pitting-associated virus (GRSPaV). Three surveys were conducted to investigate the genetic diversity of GRSPaV and to compare the ability of the GRSPaV coat protein and replicase domains to classify virus variants. GRSPaV variants identified in the surveys clustered into five of the six currently recognised lineages, and a seventh, previously unclassified lineage was detected. A correlation was observed between the detection of recombinant GRSPaV sequences and inconsistencies in classification when using different genome regions for analysis.

show abstract

Produção de antissoro policlonal utilizando a proteína capsidial recombinante do Rupestris stem pitting-associated virus

Abstract: RESUMO O Rupestris stem pitting-associated virus (RSPaV) é o agente causal das caneluras do lenho da videira. Este trabalho teve como objetivo produzir antissoro policlonal a partir da proteína capsidial (CP) recombinante do

Cited by 5 publications

References 5 publications

Screening of papaya accessions resistant to Papaya lethal yellowing virus and capacity of Tetranychus urticae to transmit the virus

Screening of papaya accessions resistant to Papaya lethal yellowing virus and capacity of Tetranychus urticae to transmit the virus

Next‐generation methods for early disease detection in crops

Genetic diversity and identification of putative recombination events in grapevine rupestris stem pitting-associated virus

Contact Info

Product

Resources

About