Abstract
Effects of the empty pYL156 vector (pYL156∷00), the most commonly used control vector for Tobacco rattle virus (TRV)-induced gene silencing analysis on viral symptom development and growth of VIGS-treated tomato and Nicotiana benthamiana plants were investigated. It was shown that VIGS-treatment for pYL156∷00 caused severe systemic viral symptoms and obvious growth repression in treated plants. Therefore, pYL156∷00 is not a good control vector for TRV-induced gene silencing analysis in these plant species. To set a better control for this analysis, two new constructs, pYL156∷NIRi and pYL156∷eGFP, were made. They were released by inserting a 253 bp fragment of a Phaseolus vulgaris nitrite reductase gene intron and a 400 bp fragment of the jellyfish GFP coding sequence in pYL156∷00, respectively. Effects of the two new control constructs on viral symptom development and growth of VIGS-treated N. benthamiana plants were compared with pYL156∷00. The results showed that VIGS-treatment for pYL156∷00 caused severe systemic viral symptoms and significant growth repression in treated plants, resulting in death of 26.7% plants. VIGS-treatment for pYL156∷NIRi also led to obvious viral symptoms and growth repression, but in a less severe extent than that for pYL156∷00, resulting in death of 13.3% plants. However, VIGS-treatment for pYL156∷eGFP did not cause any obvious viral symptom and growth repression. Severity of viral symptom and growth repression was well correlated with the accumulation level of TRV virus. These results demonstrate that pYL156∷eGFP is an excellent control vector for TRV-induced gene silencing analysis, and provide some insights into the direction to establish an excellent control for VIGS analysis.
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Claims 8
Claim · model-written summary
VIGS treatment with pYL156∷00 caused severe systemic viral symptoms in treated Nicotiana benthamiana plants.
- Taxon
- Nicotiana benthamiana
Nicotiana benthamianat0 - Treatment
- VIGS-treatment for pYL156∷00e1
- Outcome
- systemic viral symptomse1
- Direction
- increaseinferrede1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“in Nicotiana benthamiana”
e1“VIGS-treatment for pYL156∷00 caused severe systemic viral symptoms and obvious growth repression in treated plants”
Claim · model-written summary
VIGS treatment with pYL156∷00 caused significant growth repression in treated Nicotiana benthamiana plants.
- Taxon
- Nicotiana benthamianaNicotiana benthamianat0
- Treatment
- VIGS-treatment for pYL156∷00e2
- Outcome
- growthe2
- Direction
- repressione2
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“in Nicotiana benthamiana”
e2“VIGS-treatment for pYL156∷00 caused severe systemic viral symptoms and significant growth repression in treated plants”
Claim · model-written summary
VIGS treatment with pYL156∷00 resulted in the death of 26.7% of treated plants.
- Taxon
- Nicotiana benthamianaNicotiana benthamianat0
- Treatment
- VIGS-treatment for pYL156∷00e2
- Outcome
- deathe3
- Value
- 26.7%e3
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“in Nicotiana benthamiana”
e2“VIGS-treatment for pYL156∷00 caused severe systemic viral symptoms and significant growth repression in treated plants”
e3“resulting in death of 26.7% plants”
Claim · model-written summary
VIGS treatment with pYL156∷NIRi caused less severe viral symptoms than pYL156∷00 in Nicotiana benthamiana.
- Taxon
- Nicotiana benthamianaNicotiana benthamianat0
- Treatment
- VIGS-treatment for pYL156∷NIRie4
- Outcome
- viral symptomse4
- Direction
- less severee4
- Comparator
- pYL156∷00e4
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“in Nicotiana benthamiana”
e4“VIGS-treatment for pYL156∷NIRi also led to obvious viral symptoms and growth repression, but in a less severe extent than that for pYL156∷00, resulting in death of 13.3% plants”
Claim · model-written summary
VIGS treatment with pYL156∷NIRi resulted in the death of 13.3% of treated plants.
- Taxon
- Nicotiana benthamianaNicotiana benthamianat0
- Treatment
- VIGS-treatment for pYL156∷NIRie4
- Outcome
- deathe4
- Value
- 13.3%e4
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“in Nicotiana benthamiana”
e4“VIGS-treatment for pYL156∷NIRi also led to obvious viral symptoms and growth repression, but in a less severe extent than that for pYL156∷00, resulting in death of 13.3% plants”
Claim · model-written summary
VIGS treatment with pYL156∷eGFP did not cause obvious viral symptoms or growth repression, compared with pYL156∷00.
- Taxon
- Nicotiana benthamianaNicotiana benthamianat0
- Treatment
- VIGS-treatment for pYL156∷eGFPe5
- Outcome
- viral symptom and growth repressione5
- Comparator
- pYL156∷00e6
Not reportedDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“in Nicotiana benthamiana”
e5“However, VIGS-treatment for pYL156∷eGFP did not cause any obvious viral symptom and growth repression”
e6“Effects of the two new control constructs on viral symptom development and growth of VIGS-treated N. benthamiana plants were compared with pYL156∷00”
Claim · model-written summary
Severity of viral symptoms and growth repression was positively associated with the accumulation level of TRV virus.
- Taxon
- Nicotiana benthamianaNicotiana benthamianat0
- Interactor
- TRV viruse7pathogen
- Outcome
- Severity of viral symptom and growth repressione7
- Direction
- increaseinferrede7
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“in Nicotiana benthamiana”
e7“Severity of viral symptom and growth repression was well correlated with the accumulation level of TRV virus”
Claim · model-written summary
The pYL156∷eGFP vector is an excellent control vector for TRV-induced gene silencing analysis, compared with pYL156∷00.
- Taxon
- Nicotiana benthamianaNicotiana benthamianat0
- Outcome
- control vector for TRV-induced gene silencing analysise0
- Comparator
- pYL156∷00e1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
t0“in Nicotiana benthamiana”
e0“These results demonstrate that pYL156∷eGFP is an excellent control vector for TRV-induced gene silencing analysis”
e1“Effects of the two new control constructs on viral symptom development and growth of VIGS-treated N. benthamiana plants were compared with pYL156∷00”
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