Defective-interfering RNAs and elevated temperatures inhibit replication of tomato bushy stunt virus in inoculated protoplasts

Jones, Richard W.; Jackson, A.O.; Morris, T. Jack

doi:10.1016/0042-6822(90)90024-l

Cited by 88 publications

(60 citation statements)

References 11 publications

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“…Infection of N. benthamiana plants with TBSV normally results in the development of severe symptoms on the leaves followed by necrosis and death of the plant, unless virus replication is inhibited by elevated temperatures or by the presence of DI (Jones et al, 1990) or satellite (Célix et al, 1999) RNAs. To test the ability of satRNA L to protect TBSV-infected plants from apical necrosis, purified virus containing satRNA L was inoculated onto N. benthamiana plants, which were grown under the following conditions: 14 h light (at 24 or 21 u C)/10 h dark (at 15 u C), or at a constant temperature of 24, 21 or 15 u C.…”

Section: Effect Of Satrna L Replication On Symptom Expressionmentioning

confidence: 99%

Properties of a novel satellite RNA associated with tomato bushy stunt virus infections

Rubino¹,

Russo²

2010

Journal of General Virology

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The biological and molecular properties of a novel satellite RNA (satRNA L) associated with tomato bushy stunt virus (TBSV) are described. satRNA L consisted of a linear single-stranded RNA of 615 nt, lacked significant open reading frames (ORFs) and had no sequence identity with the helper genome other than in the 59-proximal 7 nt and in a central region that is also conserved in all tombusvirus genomic, defective interfering and satellite RNAs. Secondary-structure analysis showed the presence of high-order domains similar to those described for other tombusvirus RNAs. Shorter-than-unit-length molecules were shown not to be related to a silencing mechanism. satRNA L did not modify the symptoms induced by TBSV under any of the temperature conditions tested. A full-length cDNA clone was constructed and used in co-inoculations with transcripts of carnation Italian ringspot virus (CIRV) and cymbidium ringspot virus (CymRSV). CIRV, but not CymRSV, supported the replication of satRNA L. Using CIRVCymRSV hybrid infectious clones, two regions were identified as possible determinants of the different ability to support satRNA L replication. The first region was in the 59-untranslated region, which folds differently in CymRSV in comparison with CIRV and TBSV; the second region was in the ORF1-encoded protein where a more efficient satRNA L-binding domain is suggested to be present in CIRV.

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Section: Effect Of Satrna L Replication On Symptom Expressionmentioning

confidence: 99%

Properties of a novel satellite RNA associated with tomato bushy stunt virus infections

Rubino¹,

Russo²

2010

Journal of General Virology

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“…The protocol of Jones et al (1990) was used for protoplasts and transfection, except that the PEG CaCI2 solution was replaced with PEG-Mg/CMSsolution (1 mL of 50% [w/v] PEG 1500 in 75 mM Hepes, pH 8.01 [Boehringer Mannheim], to which 15 pL of 1 M MgC12 and 100 pL of 1 M Ca[NO&, pH 7.9, had been added). After transfection with 2 pg of viral RNA, protoplasts were transferred to incubation media (Luciano et al, 1987).…”

Section: Protoplast Transfectionmentioning

confidence: 99%

Induction of a Highly Specific Antiviral State in Transgenic Plants: Implications for Regulation of Gene Expression and Virus Resistance.

et al. 1993

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Transgenic tobacco plants expressing either a full-length form of the tobacco etch virus (TEV) coat protein or a form truncated at the N terminus of the TEV coat protein were initially susceptible to TEV infection, and typlcal systemic symptoms developed. However, 3 to 5 weeks after a TEV infection was established, transgenlc plants "recovered" from the TEV infection, and new stem and leaf tissue emerged symptom and virus free. A TEV-resistant state was induced ln the recovered tissue. The resistance was virus speclflc. Recovered plant tissue could not be lnfected wlth TEV, but was susceptlble to the closely related virus, potato virus Y. The resistance phenotype was functional at the slngle-cell leve1 because protoplasts from recovered transgenic tissue did not support TEV repllcation. Surprlslngly, steady state transgene mRNA levels in recovered tissue were 12-to 22-fold less than transgene mRNA levels in uninoculated transgenic tissue of the same developmental stage. However, nuclear run-off studles suggested that transgene transcription rates in recovered and uninoculated plants were similar. We propose that the resistant state and reduced steady state levels of transgene transcript accumulation are mediated at the cellular leve1 by a cytoplasmic actlvity that targets speclfic RNA sequences for inactlvation.

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“…Protoplasts were prepared from B. campestris cotyledons. One microgram of helper virus RNA transcripts (plus sat C transcripts, as indicated) was used to transfect 2 x 105 protoplasts (20). Protoplasts were incubated in 0.75 ml of medium containing 30 pCi of [5,6-3H]uridine for 24 hr at room temperature under fluorescent light illumination.…”

Section: Methodsmentioning

confidence: 99%

Single amino acid change in the helicase domain of the putative RNA replicase of turnip crinkle virus alters symptom intensification by virulent satellites.

Collmer¹,

Stenzler²,

Chen³

et al. 1992

Proc. Natl. Acad. Sci. U.S.A.

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The virulent satellite [satellite C (sat C)] of turnip crinkle virus (TCV) is a small pathogenic RNA that intensifies symptoms in TCV-infected turnip plants (Brassica campesnis). The virulence of sat C is determined by properties of the satellite itself and is influenced by the helper virus. Symptoms produced in infections with sat C differ in severity depending on the helper virus. The TCV-JI helper virusproduces more severe symptoms than the TCV-B helper virus when inoculated with sat C. To rind determinants in the TCV helper virus genome that affect satellite virulence, the TCV-JI genome was doned and the sequence was compared to the TCV-B genome. The genomes were found to differ by only five base changes, and only one of the base changes, at nucleetide position 1025, produced an amino acid change, an aspartic acid glycine in the putative viral replicase. A chimeric TCV genome (TCV-B/JI) containing four of the five base changes (including the base change at position 1025) and a mutant

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Defective-interfering RNAs and elevated temperatures inhibit replication of tomato bushy stunt virus in inoculated protoplasts

Cited by 88 publications

References 11 publications

Properties of a novel satellite RNA associated with tomato bushy stunt virus infections

Properties of a novel satellite RNA associated with tomato bushy stunt virus infections

Induction of a Highly Specific Antiviral State in Transgenic Plants: Implications for Regulation of Gene Expression and Virus Resistance.

Single amino acid change in the helicase domain of the putative RNA replicase of turnip crinkle virus alters symptom intensification by virulent satellites.

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