Abstract
RNA interference (RNAi), a conserved RNA-mediated gene regulatory mechanism in eukaryotes, plays an important role in plant growth and development, and as an antiviral defence system in plants. As a counter-strategy, plant viruses encode RNAi suppressors to suppress the RNAi pathways and consequently down-regulate plant defence. In geminiviruses, the proteins AC2, AC4 and AV2 are known to act as RNAi suppressors. In this study, we have designed a gene silencing vector using the features of trans-acting small interfering RNA (tasiRNA), which is simple and can be used to target multiple genes at a time employing a single-step cloning procedure. This vector was used to target two RNAi suppressor proteins (AC2 and AC4) of the geminivirus, Tomato leaf curl New Delhi virus (ToLCNDV). The vector containing fragments of ToLCNDV AC2 and AC4 genes, on agro-infiltration, produced copious quantities of AC2 and AC4 specific siRNA in both tobacco and tomato plants. On challenge inoculation of the agro-infiltrated plants with ToLCNDV, most plants showed an absence of symptoms and low accumulation of viral DNA. Transgenic tobacco plants were raised using the AC2 and AC4 tasiRNA-generating constructs, and T1 plants, obtained from the primary transgenic plants, were tested for resistance separately against ToLCNDV and Tomato leaf curl Gujarat virus. Most plants showed an absence of symptoms and low accumulation of the corresponding viruses, the resistance being generally proportional to the amounts of siRNA produced against AC2 and AC4 genes. This is the first report of the use of artificial tasiRNA to generate resistance against an important plant virus.
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Claims 6
Claim · model-written summary
The AC2/AC4 tasiRNA vector produced AC2- and AC4-specific siRNA in tobacco and tomato plants.
- Taxon
- tobacco and tomato plants
Solanum lycopersicum
Nicotiana tabacume0 - Gene
- AC2 and AC4e0
- Outcome
- AC2 and AC4 specific siRNAe0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The vector containing fragments of ToLCNDV AC2 and AC4 genes, on agro-infiltration, produced copious quantities of AC2 and AC4 specific siRNA in both tobacco and tomato plants”
Claim · model-written summary
Most agro-infiltrated plants challenged with ToLCNDV showed an absence of symptoms.
- Taxon
- tobacco and tomato plantsSolanum lycopersicumNicotiana tabacume0
- Treatment
- agro-infiltrated plantse1
- Interactor
- ToLCNDVe1pathogen
- Outcome
- absence of symptomse1
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The vector containing fragments of ToLCNDV AC2 and AC4 genes, on agro-infiltration, produced copious quantities of AC2 and AC4 specific siRNA in both tobacco and tomato plants”
e1“On challenge inoculation of the agro-infiltrated plants with ToLCNDV, most plants showed an absence of symptoms and low accumulation of viral DNA”
Claim · model-written summary
Agro-infiltrated plants challenged with ToLCNDV showed low accumulation of viral DNA.
- Taxon
- tobacco and tomato plantsSolanum lycopersicumNicotiana tabacume0
- Treatment
- agro-infiltrated plantse1
- Interactor
- ToLCNDVe1pathogen
- Outcome
- accumulation of viral DNAe1
- Direction
- lowe1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The vector containing fragments of ToLCNDV AC2 and AC4 genes, on agro-infiltration, produced copious quantities of AC2 and AC4 specific siRNA in both tobacco and tomato plants”
e1“On challenge inoculation of the agro-infiltrated plants with ToLCNDV, most plants showed an absence of symptoms and low accumulation of viral DNA”
Claim · model-written summary
Transgenic tobacco T1 plants showed an absence of symptoms after challenge with ToLCNDV and Tomato leaf curl Gujarat virus.
- Taxon
- Transgenic tobacco plantsNicotiana tabacume2
- Treatment
- AC2 and AC4 tasiRNA-generating constructse2
- Interactor
- ToLCNDVe2pathogen
- Second interactor
- Tomato leaf curl Gujarat virusTomato leaf curl Gujarat viruse2pathogen
- Outcome
- absence of symptomse3
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e2“Transgenic tobacco plants were raised using the AC2 and AC4 tasiRNA-generating constructs, and T1 plants, obtained from the primary transgenic plants, were tested for resistance separately against ToLCNDV and Tomato leaf curl Gujarat virus”
e3“Most plants showed an absence of symptoms and low accumulation of the corresponding viruses, the resistance being generally proportional to the amounts of siRNA produced against AC2 and AC4 genes”
Claim · model-written summary
Transgenic tobacco plants showed low accumulation of the corresponding viruses.
- Taxon
- Transgenic tobacco plantsNicotiana tabacume2
- Treatment
- AC2 and AC4 tasiRNA-generating constructse2
- Interactor
- ToLCNDVe2pathogen
- Second interactor
- Tomato leaf curl Gujarat virusTomato leaf curl Gujarat viruse2pathogen
- Outcome
- accumulation of the corresponding virusese3
- Direction
- lowe3
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e2“Transgenic tobacco plants were raised using the AC2 and AC4 tasiRNA-generating constructs, and T1 plants, obtained from the primary transgenic plants, were tested for resistance separately against ToLCNDV and Tomato leaf curl Gujarat virus”
e3“Most plants showed an absence of symptoms and low accumulation of the corresponding viruses, the resistance being generally proportional to the amounts of siRNA produced against AC2 and AC4 genes”
Claim · model-written summary
Resistance in transgenic tobacco was related to the amount of siRNA produced against AC2 and AC4.
- Taxon
- tobaccoNicotiana tabacume2
- Gene
- AC2 and AC4e3
- Outcome
- resistancee3
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e2“Transgenic tobacco plants were raised using the AC2 and AC4 tasiRNA-generating constructs, and T1 plants, obtained from the primary transgenic plants, were tested for resistance separately against ToLCNDV and Tomato leaf curl Gujarat virus”
e3“Most plants showed an absence of symptoms and low accumulation of the corresponding viruses, the resistance being generally proportional to the amounts of siRNA produced against AC2 and AC4 genes”