Abstract
MicroRNA-based gene silencing is a functional genomics tool for a wide range of eukaryotes. As a basis for broader application of virus-induced gene silencing (VIGS) to photosynthesis research, we employed a tobacco rattle virus (TRV) vector to silence expression of the nuclear psbS gene in Nicotiana benthamiana. The 22-kiloDalton psbS protein is essential for xanthophyll-and H + -dependent thermal dissipation of excitation in higher plants widely known as nonphotochemical quenching (NPQ). Controls treated with the TRV-VIGS vector containing a bacterial chloramphenicol resistance gene as the silencing target were included to test for non-silencing effects of the viral vector system. PsbS protein was undetectable and both psbS mRNA transcript levels and NPQ capacity were dramatically reduced in new leaf tissue of VIGS-psbS plants only. Photosynthetic performance in TRV-VIGS-treated and uninfiltrated plants was assessed by application of CO 2 exchange, chlorophyll fluorescence, and in vivo absorbance changes at 810 nm. TRV-VIGS caused a mild stress based on pigment content and light absorption characteristics in some cases. To assess transient complementation of NPQ, the endogenous psbS gene was silenced using only the transit sequence in the TRV vector followed by Agrobacterium-mediated transient expression of a modified gene consisting of an altered transit sequence fused to the native mature protein sequence. Nevertheless, NPQ in infused fully expanded leaves that expressed this re-introduced form was not fully restored indicating the possible importance of psbS incorporation prior to formation of grana stacks.
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Claims 5
Claim · model-written summary
VIGS-psbS rendered PsbS protein undetectable in new leaf tissue.
- Taxon
- Nicotiana benthamiana
Nicotiana benthamianat0 - Organ / stage
- new leaf tissuee0
- Treatment
- VIGS-psbSe0
- Gene
- PsbS proteine0
- Outcome
- PsbS proteine0
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“PsbS protein was undetectable and both psbS mRNA transcript levels and NPQ capacity were dramatically reduced in new leaf tissue of VIGS-psbS plants only.”
t0“Nicotiana benthamiana”
Claim · model-written summary
VIGS-psbS dramatically reduced psbS mRNA transcript levels in new leaf tissue.
- Taxon
- Nicotiana benthamianaNicotiana benthamianat0
- Organ / stage
- new leaf tissuee0
- Treatment
- VIGS-psbSe0
- Gene
- psbSe0
- Outcome
- psbS mRNA transcript levelse0
- Direction
- dramatically reducede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“PsbS protein was undetectable and both psbS mRNA transcript levels and NPQ capacity were dramatically reduced in new leaf tissue of VIGS-psbS plants only.”
t0“Nicotiana benthamiana”
Claim · model-written summary
VIGS-psbS dramatically reduced NPQ capacity in new leaf tissue.
- Taxon
- Nicotiana benthamianaNicotiana benthamianat0
- Organ / stage
- new leaf tissuee0
- Treatment
- VIGS-psbSe0
- Gene
- psbSe0
- Outcome
- NPQ capacitye0
- Direction
- dramatically reducede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“PsbS protein was undetectable and both psbS mRNA transcript levels and NPQ capacity were dramatically reduced in new leaf tissue of VIGS-psbS plants only.”
t0“Nicotiana benthamiana”
Claim · model-written summary
TRV-VIGS caused mild stress in some cases based on pigment content and light absorption.
- Taxon
- Nicotiana benthamianaNicotiana benthamianat0
- Treatment
- TRV-VIGSe0
- Outcome
- mild stresse0
- Qualifier
- in some casese0
Not reportedComparatorDirection
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“TRV-VIGS caused a mild stress based on pigment content and light absorption characteristics in some cases.”
t0“Nicotiana benthamiana”
Claim · model-written summary
NPQ was not fully restored after transient expression of the reintroduced psbS form.
- Taxon
- Nicotiana benthamianaNicotiana benthamianat0
- Organ / stage
- fully expanded leavese0
- Treatment
- re-introduced forme0
- Gene
- psbSe0
- Outcome
- NPQe0
- Direction
- not fully restorede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Nevertheless, NPQ in infused fully expanded leaves that expressed this re-introduced form was not fully restored indicating the possible importance of psbS incorporation prior to formation of grana stacks.”
t0“Nicotiana benthamiana”
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