Abstract
Virus induced gene silencing (VIGS) is an important tool for functional genomics studies in plants. With this method, it is possible to target most endogenous genes and down-regulate the messenger RNA (mRNA) in a sequence-specific manner. Chinese wheat mosaic virus (CWMV) has a bipartite, single-strand positive RNA genome and can infect both wheat and N. benthamiana, and the optimal temperature for systemic infection in plants is 17℃. To assess the potential of the virus as a vector for gene silencing at low temperature, a fragment of the N. benthamiana or wheat phytoene desaturase (PDS) gene was expressed from a modified CWMV RNA2 clone and the resulting photo bleaching in infected plants was used as a reporter for silencing. Down-regulation of PDS mRNA was also measured by quantitative reverse-transcriptase polymerase chain reaction (RT-qPCR). In experiments using fragments of PDS ranging from 500 to 1500 nucleotides, insert length influenced the stability and the efficiency of VIGS. The CWMV induced silencing system was also used to suppress miR165/166 and miR3134a through expression of miRNA target mimics. The relative expression levels of mature miR165/166 and miR3134a decreased whereas the transcript levels of their target genes increased. Interestingly, we also found the CWMV induced silencing system was more efficient compare with the vector based on Barley stripe mosaic virus (BSMV) or Foxtail mosaic virus (FoMV) in wheat or the vector based on TRV in N. benthamiana at 17 ℃. In summary, the CWMV vector is effective in silencing endogenous genes and miRNAs at 17 ℃, thereby providing a powerful tool for gene function analysis in both N. benthamianaand and wheat at low temperature.
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Claims 8
Claim · model-written summary
PDS silencing produced photobleaching in infected plants.
Not reportedComparatorValue
Verbatim
e0“the resulting photo bleaching in infected plants was used as a reporter for silencing”
e1“N. benthamiana or wheat phytoene desaturase (PDS) gene”
Claim · model-written summary
Insert length influenced stability of CWMV-based VIGS.
Not reportedTaxon
Verbatim
e0“In experiments using fragments of PDS ranging from 500 to 1500 nucleotides, insert length influenced the stability and the efficiency of VIGS”
t0“Monocots and Dicots”
Claim · model-written summary
Insert length influenced the efficiency of CWMV-based VIGS.
Not reportedTaxon
Verbatim
e0“In experiments using fragments of PDS ranging from 500 to 1500 nucleotides, insert length influenced the stability and the efficiency of VIGS”
Claim · model-written summary
CWMV-based target mimics decreased mature miR165/166 expression.
- Taxon
- N. benthamianae1
- Treatment
- expression of miRNA target mimicse0
- Gene
- miR165/166e0
- Outcome
- relative expression levels of mature miR165/166e0
- Direction
- decreasede0
Not reportedComparator
Verbatim
e0“CWMV induced silencing system was also used to suppress miR165/166 and miR3134a through expression of miRNA target mimics. The relative expression levels of mature miR165/166 and miR3134a decreased”
e1“in wheat or the vector based on TRV in N. benthamiana at 17 ℃”
Claim · model-written summary
CWMV-based target mimics decreased mature miR3134a expression.
- Taxon
- N. benthamianae1
- Treatment
- expression of miRNA target mimicse0
- Gene
- miR3134ae0
- Outcome
- relative expression levels of mature miR165/166 and miR3134ae0
- Direction
- decreasede0
Not reportedComparator
Verbatim
e0“CWMV induced silencing system was also used to suppress miR165/166 and miR3134a through expression of miRNA target mimics. The relative expression levels of mature miR165/166 and miR3134a decreased”
e1“in wheat or the vector based on TRV in N. benthamiana at 17 ℃”
Claim · model-written summary
Transcripts of the miRNA target genes increased under CWMV-based silencing.
Not reportedTaxonValue
Verbatim
e0“the transcript levels of their target genes increased”
e1“CWMV induced silencing system was also used to suppress miR165/166 and miR3134a”
Claim · model-written summary
In wheat at 17 ℃, the CWMV vector was more efficient than BSMV and FoMV vectors.
- Taxon
- wheat
Triticum aestivume0 - Outcome
- more efficiente0
- Direction
- more efficiente0
- Comparator
- vector based on Barley stripe mosaic virus (BSMV) or Foxtail mosaic virus (FoMV)e0
- Setting
- 17 ℃e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“CWMV induced silencing system was more efficient compare with the vector based on Barley stripe mosaic virus (BSMV) or Foxtail mosaic virus (FoMV) in wheat or the vector based on TRV in N. benthamiana at 17 ℃”
Claim · model-written summary
In N. benthamiana at 17 ℃, the CWMV vector was more efficient than the TRV vector.
Verbatim
e0“CWMV induced silencing system was more efficient compare with the vector based on Barley stripe mosaic virus (BSMV) or Foxtail mosaic virus (FoMV) in wheat or the vector based on TRV in N. benthamiana at 17 ℃”
The same questions in other papers
transgenic · molecular: expression · Nicotiana benthamianaPositive and negative results
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- negative
- negative
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See all 29 papers →Claims are grouped by organism, treatment and what was measured or targeted. A paper counts as positive when any of its claims in the group is positive.