Abstract
As a defense against viral infection, plants are thought to use RNA-induced silencing complexes (RISCs) to target and cleave viral RNA. To counteract this, some viruses have evolved proteins to inhibit RISC-mediated activity, thus ensuring continued virulence. This research focused on the study and analysis of the anti-viral RNAi response to various viruses in plants to gain an understanding of how the plant defense operates on the molecular and biochemical levels. Nicotiana benthamiana plants were infected with Tomato bushy stunt virus (TBSV) and Tobacco rattle virus (TRV). These plants were subjected to column chromatography methods, and fractions contained a virus-specific ribonuclease activity, co-eluting with small interfering RNAs (siRNA), that was shown to be sensitive to inhibition with EDTA and enhanced by the addition of divalent metal cations. This ribonuclease activity co-purified with proteins that contained a domain from the hallmark RISC protein Argonaute family. To further study host responses to viral infection, monocots were infected with Panicum mosaic virus (PMV) and satellite panicum mosaic virus (SPMV) and also were subjected to column chromatography following infection. Preliminary studies show that fractions contained ribonuclease activity as well as siRNAs and proteins containing an Argonaute domain. Additionally, silencing suppressors have been directly implicated in interfering with RNAi pathways in plants. Studies involving Agrobacterium- and virus-vectored cDNA to express green fluorescent protein (GFP) were used to establish that cointroduced suppressors of RNAi can extend the production of a foreign protein for enhancement of biotechnological applications. It was found that the hordeivirus protein ?b contributes to enhancement of expression for the foreign protein GFP early in the infection, while the potyvirus protein HcPro and tombusvirus protein P19 enhance and extend protein production later in the infection.
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Claims 8
Claim · model-written summary
Fractions from Nicotiana benthamiana infected with TBSV and TRV contained a virus-specific ribonuclease activity co-eluting with siRNAs.
- Taxon
- Nicotiana benthamiana
Nicotiana benthamianae0 - Interactor
- Tomato bushy stunt virusTomato bushy stunt viruse0pathogen
- Second interactor
- Tobacco rattle virusTobacco rattle viruse0pathogen
- Outcome
- virus-specific ribonuclease activitye1
- Qualifier
- co-eluting with small interfering RNAse1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Nicotiana benthamiana plants were infected with Tomato bushy stunt virus (TBSV) and Tobacco rattle virus (TRV)”
e1“fractions contained a virus-specific ribonuclease activity, co-eluting with small interfering RNAs (siRNA)”
Claim · model-written summary
Ribonuclease activity in Nicotiana benthamiana fractions was sensitive to inhibition by EDTA.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae0
- Treatment
- EDTAe2
- Outcome
- ribonuclease activitye1
- Direction
- sensitive to inhibitione2
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Nicotiana benthamiana plants were infected with Tomato bushy stunt virus (TBSV) and Tobacco rattle virus (TRV)”
e1“fractions contained a virus-specific ribonuclease activity”
e2“that was shown to be sensitive to inhibition with EDTA”
Claim · model-written summary
Divalent metal cations enhanced the ribonuclease activity in Nicotiana benthamiana fractions.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae0
- Treatment
- addition of divalent metal cationse3
- Outcome
- ribonuclease activitye1
- Direction
- enhancede3
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Nicotiana benthamiana plants were infected with Tomato bushy stunt virus (TBSV) and Tobacco rattle virus (TRV)”
e1“fractions contained a virus-specific ribonuclease activity”
e3“enhanced by the addition of divalent metal cations”
Claim · model-written summary
The ribonuclease activity co-purified with proteins containing an Argonaute-family domain.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae0
- Gene
- RISC protein Argonautee2
- Outcome
- ribonuclease activitye2
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Nicotiana benthamiana plants were infected with Tomato bushy stunt virus (TBSV) and Tobacco rattle virus (TRV)”
e2“This ribonuclease activity co-purified with proteins that contained a domain from the hallmark RISC protein Argonaute family”
Claim · model-written summary
Preliminary results: fractions from monocots infected with Panicum mosaic virus and satellite panicum mosaic virus contained ribonuclease activity, siRNAs and Argonaute-domain proteins.
- Taxon
- monocotsLiliopsidae3
- Interactor
- Panicum mosaic virusPanicum mosaic viruse3pathogen
- Second interactor
- satellite panicum mosaic virusPanicum mosaic satellite viruse3pathogen
- Outcome
- ribonuclease activitye4
- Qualifier
- Preliminarye4
Verbatim
e3“monocots were infected with Panicum mosaic virus (PMV) and satellite panicum mosaic virus (SPMV)”
e4“Preliminary studies show that fractions contained ribonuclease activity as well as siRNAs and proteins containing an Argonaute domain”
Claim · model-written summary
The hordeivirus protein ?b enhanced expression of the foreign protein GFP early in infection.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae0
- Treatment
- hordeivirus protein ?be5
- Interactor
- hordeivirusHordeiviruse5pathogen
- Outcome
- expression for the foreign protein GFPe5
- Direction
- enhancemente5
- Qualifier
- early in the infectione5
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Nicotiana benthamiana plants were infected with Tomato bushy stunt virus (TBSV) and Tobacco rattle virus (TRV)”
e5“the hordeivirus protein ?b contributes to enhancement of expression for the foreign protein GFP early in the infection”
Claim · model-written summary
The potyvirus protein HcPro and tombusvirus protein P19 enhanced and extended protein production later in infection.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae0
- Treatment
- potyvirus protein HcPro and tombusvirus protein P19e6
- Outcome
- protein productione6
- Direction
- enhance and extende6
- Qualifier
- later in the infectione6
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Nicotiana benthamiana plants were infected with Tomato bushy stunt virus (TBSV) and Tobacco rattle virus (TRV)”
e6“potyvirus protein HcPro and tombusvirus protein P19 enhance and extend protein production later in the infection”
Claim · model-written summary
Cointroduced suppressors of RNAi can extend the production of a foreign protein.
- Taxon
- Nicotiana benthamianaNicotiana benthamianae0
- Treatment
- cointroduced suppressors of RNAie7
- Outcome
- production of a foreign proteine7
- Direction
- extende7
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Nicotiana benthamiana plants were infected with Tomato bushy stunt virus (TBSV) and Tobacco rattle virus (TRV)”
e7“cointroduced suppressors of RNAi can extend the production of a foreign protein”