Carmen Castresana

Bio: Carmen Castresana is an academic researcher. The author has contributed to research in topics: Plant virus & Genomic organization. The author has an hindex of 1, co-authored 1 publications receiving 94 citations.

Nucleotide sequence of the genomic RNA of pepper mild mottle virus, a resistance-breaking tobamovirus in pepper

Abstract: The entire genomic RNA of a Spanish isolate of pepper mild mottle virus (PMMV-S), a resistance-breaking virus in pepper, was cloned and sequenced and shown to be similar to other tobamoviruses in its genomic organization. It consisted of 6357 nucleotides (nt) and contained four open reading frames (ORFs) which encode a 126K protein and a readthrough 183K protein (nt 70 to 4908), a 28K protein (nt 4909 to 5682) and a 17.5K coat protein (nt 5685 to 6158). This is the first tobamovirus in which none of the ORFs overlap. Both its nucleic acid and predicted protein sequences were compared with the previously determined sequences of other tobamoviruses. The variations and similarities found and their relationship with the pathogenicity of this virus are discussed.

Double-Stranded RNA-Mediated Interference with Plant Virus Infection

Abstract: Double-stranded RNA (dsRNA) has been shown to play a key role as an inducer of different interference phenomena occurring in both the plant and animal kingdoms. Here, we show that dsRNA derived from viral sequences can interfere with virus infection in a sequence-specific manner by directly delivering dsRNA to leaf cells either by mechanical inoculation or via an Agrobacterium-mediated transient-expression assay. We have successfully interfered with the infection of plants by three viruses belonging to the tobamovirus, potyvirus, and alfamovirus groups, demonstrating the reliability of the approach. We suggest that the effect mediated by dsRNA in plant virus infection resembles the analogous phenomenon of RNA interference observed in animals. The interference observed is sequence specific, is dose dependent, and is triggered by dsRNA but not singlestranded RNA. Our results support the view that a dsRNA intermediate in virus replication acts as efficient initiator of posttranscriptional gene silencing (PTGS) in natural virus infections, triggering the initiation step of PTGS that targets viral RNA for degradation. Gene sequences derived from different plant viruses have been introduced into a wide variety of plant species to produce transgenic plants protected against virus infection. In a number of cases, it is known that the mechanism of resistance is a posttranscriptional, RNA-mediated process that targets both the viral RNA and the transgene mRNA for degradation in a sequence-specific manner (11, 19). RNA-mediated virus resistance is a manifestation of posttranscriptional gene silencing (PTGS), a more general phenomenon which was first described as a coordinated and reciprocal inactivation of host gene and transgenes encoding the same sense RNA (reviewed in references 33 and 41). More recently, three components in the dynamics of PTGS have been proposed: initiation, propa

Crude extracts of bacterially expressed dsRNA can be used to protect plants against virus infections

Abstract: Double-stranded RNA (dsRNA) is a potent initiator of gene silencing in a diverse group of organisms that includes plants, Caenorhabditis elegans, Drosophila and mammals. We have previously shown and patented that mechanical inoculation of in vitro-transcribed dsRNA derived from viral sequences specifically prevents virus infection in plants. The approach required the in vitro synthesis of large amounts of RNA involving high cost and considerable labour. We have developed an in vivo expression system to produce large amounts of virus-derived dsRNAs in bacteria, with a view to providing a practical control of virus diseases in plants. Partially purified bacterial dsRNAs promoted specific interference with the infection in plants by two viruses belonging to the tobamovirus and potyvirus groups. Furthermore, we have demonstrated that easy to obtain, crude extracts of bacterially expressed dsRNAs are equally effective protecting plants against virus infections when sprayed onto plant surfaces by a simple procedure. Virus infectivity was significantly abolished when plants were sprayed with French Press lysates several days before virus inoculation. Our approach provides an alternative to genetic transformation of plant species with dsRNA-expressing constructs capable to interfere with plant viruses. The main advantage of this mode of dsRNA production is its simplicity and its extremely low cost compared with the requirements for regenerating transgenic plants. This approach provides a reliable and potential tool, not only for plant protection against virus diseases, but also for the study of gene silencing mechanisms in plant virus infections.

Analysis of a tobacco mosaic virus strain capable of overcoming N gene-mediated resistance.

Abstract: The genome of Ob, a tobamovirus that overcomes the N gene-mediated hypersensitive response (HR), was cloned as a cDNA, and its nucleotide sequence was determined. The genomic organization of Ob is similar to that of other tobamoviruses, consisting of 6506 nucleotides and containing at least four open reading frames. These open reading frames encode a 126-kD polypeptide with a 183-kD readthrough product, a 30.6-kD movement protein, and an 18-kD coat protein. A bacteriophage T7 promoter sequence was fused to the full-length cDNA clone to obtain infectious RNA transcripts. These transcripts, when inoculated onto tobacco plants, induced disease symptoms indistinguishable from plants inoculated with Ob viral RNA. To determine which viral factor is responsible for the resistance-breaking character of Ob, a recombinant virus was constructed in which the movement protein gene of tobacco mosaic virus was replaced with that of Ob. Cultivar Xanthi NN tobacco plants infected with this virus responded with an HR, indicating that the Ob movement protein alone does not act to overcome the N gene-mediated response. Following mutagenesis of the infectious Ob cDNA clone with hydroxylamine, populations of transcripts from the mutagenized DNA were inoculated onto Xanthi NN tobacco, and a variant that induced the HR was identified. The mutant was analyzed and found to contain a single nucleotide change in the 126-kD gene. Recreating the mutation in the Ob cDNA clone by site-directed mutagenesis resulted in a virus that caused symptoms identical to the chemically induced mutant.

Pepper mild mottle virus as a water quality indicator

Abstract: Pepper mild mottle virus (PMMoV) was recently found to be the most abundant RNA virus in human feces, and is a plant virus belonging to the genus Tobamovirus in the family Virgoviridae. When in human feces, it is of dietary origin from peppers and their processed products, and is excreted from a large proportion of healthy human populations, but rarely found in animal feces. Over the past decade, this virus has been increasingly attracting research attention as a potential viral indicator for human fecal pollution in aquatic environments and water treatment systems. Results presented in the literature reveal that PMMoV is globally distributed and present in various water sources in greater abundance than human pathogenic viruses, without substantial seasonal fluctuations. Several studies report that increased concentrations of PMMoV tend to be correlated with increased fecal contamination in general, along with more frequent detection of pathogenic enteric viruses. PMMoV also exhibits remarkable stability in water under various environmental conditions. Here, we review recent advancements in our understanding of the occurrence and persistence of PMMoV in natural and engineered water systems and discuss its advantages and limitations as a viral indicator for improved microbial water quality management.