Development of an asymmetric PCR-ELISA typing method for citrus tristeza virus based on the coat protein gene

Nolasco, Gustavo; Santos, Cláudia Simone Silveira dos; Silva, Gonçalo; Fonseca, Filomena

doi:10.1016/j.jviromet.2008.09.030

Cited by 33 publications

(48 citation statements)

References 27 publications

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“…1). The classification obtained supports that published by Nolasco et al (2009) grouping isolates with similar biological characteristics. All virus groups were supported by high bootstrap values (Fig.…”

supporting

confidence: 88%

“…The basis of current CTV classification is mainly biological indexing on different indicator plants, although many authors support the existence of a relationship between virus phenotype and coat protein (CP) gene sequences (Pappu et al 1993;Rubio et al 2001;Zemzami et al 2002;Nolasco et al 2009). As a result of error-prone replication, recombination events, and repeated vector-mediated transmission, naturally occurring CTV isolates exist mostly as populations of genetic variants (Kong et al 2000) that can be distinguished by the analysis of singlestrand conformation polymorphism (SSCP) profiles (Rubio et al 1996).…”

mentioning

confidence: 99%

“…0 no symptoms were observed, SP stem-pitting symptoms, NT not tested a Phylogenetic groups are defined according to classification published by Nolasco et al (2009) both ends of the CP gene was performed according to the procedure described by Nolasco et al (2002). To separate different virus variants presumably present in sample populations, amplicons were cloned into pTZ57R/T vector (Fermentas) according to the manufacturer's instructions.…”

mentioning

confidence: 99%

“…The sequences from the East Adriatic clustered into six out of the seven groups. Two of these groups (2 and M) harbour mild CTV isolates, while four of them (1, 3a, 4, and 5) harbour CP sequences of isolates that cause severe CTV symptoms of quick decline (groups 1, 3a, 4 and The bar represents 0.01 nucleotide substitution per site 5), SP (groups 3a and 5), and seedling yellows (Nolasco et al 2009). Surprisingly, sequences of almost half of our isolates clustered into one of the severe groups (Table 1) with the highest occurrence of sequences in group 3a.…”

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confidence: 99%

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East Adriatic—a reservoir region of severe Citrus tristeza virus strains

et al. 2009

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Citrus tristeza virus (CTV) represents one of the major threats to citrus production worldwide. In the East Adriatic region, CTV symptoms are mostly absent due to traditional citrus grafting on trifoliate orange (Poncirus trifoliata), a CTV-tolerant rootstock. Therefore, the virus has been continuously spreading by the propagation of infected material. The genetic variability of CTV was studied on nineteen citrus samples, collected from orchards in the coastal region of Croatia, Montenegro and Albania, that previously tested positive by ELISA and immunocapture RT-PCR. Single-strand conformation polymorphism of the amplified coat protein gene demonstrated the presence of different CTV variants in each amplicon, while sequence analysis of cloned CP gene variants confirmed their clustering into six out of the seven phylogenetic groups so far delineated. Four of these groups include sequences of severe quick decline, seedling yellows and stem-pitting (SP) isolates, thought to be found only rarely in the Mediterranean region. Regardless of the lack of symptoms in the field, CTV isolates from the East Adriatic displayed high genetic variability and pathogenic potential, additionally confirmed by biological characterisation. The high percentage of mixed infections suggest the potential for further diversification and a greater risk of severe variants spreading into new areas.

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confidence: 99%

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confidence: 99%

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confidence: 99%

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East Adriatic—a reservoir region of severe Citrus tristeza virus strains

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“…The so called mild strains do not produce noticeable symptoms of any kind. A study on the genomic variability of the p25 gene (coat protein gene, CP) obtained from worldwide isolates revealed the existence of seven phylogenetic groups that could be correlated with the distinct syndromes caused by the virus (Nolasco et al 2009). Lu et al (2004) showed that the CTV genome has three genes encoding VSR activity: CP, p20 and p23 which affect the RNA silencing mechanism differently.…”

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confidence: 99%

Comparing p20’s RNA silencing suppressing activity among five phylogenetic groups of Citrus Tristeza virus

et al. 2012

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The p20 protein encoded by the Citrus Tristeza Virus (CTV) was previously identified as a RNA silencing suppressor. In this study, we analyzed the p20's suppressing activity from five phylogenetic groups of CTV, using the co-infiltration assay of Green fluorescence protein (GFP) gene and the suppressor gene in 16C line Nicotiana benthamiana plants. Green fluorescence, GFP mRNA relative levels and GFP specific siRNAS were compared showing in most cases, only slight differences. Contrary to previous studies, the p20 suppressor was not able to impede neither short range nor systemic spreading of RNA silencing. The suppressor from the phylogenetic group 4 revealed a much reduced activity when compared with the others. At present we still don't know whether this property is a characteristic of this group or an atypical feature due to a unique point mutation. The differences in the symptom type and intensity originated by isolates belonging to the phylogenetic groups assayed could not be related to differences to the p20 suppressor's activity.

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Biology and Infection Process of Viruses and Viroids

2017

Microbial Plant Pathogens

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Development of an asymmetric PCR-ELISA typing method for citrus tristeza virus based on the coat protein gene

Cited by 33 publications

References 27 publications

East Adriatic—a reservoir region of severe Citrus tristeza virus strains

East Adriatic—a reservoir region of severe Citrus tristeza virus strains

Comparing p20’s RNA silencing suppressing activity among five phylogenetic groups of Citrus Tristeza virus

Biology and Infection Process of Viruses and Viroids

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