Tobacco rattle virus-induced PHYTOENE DESATURASE (PDS) and Mg-chelatase H subunit (ChlH) gene silencing in Solanum pseudocapsicum L.
Virus-induced gene silencing (VIGS) is an attractive tool for determining gene function in plants. The present study constitutes the first application of VIGS in S. pseudocapsicum, which has great ornamental and pharmaceutical value, using tobacco rattle virus (TRV) vectors. Two marker genes, PHYTOE...
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PeerJ Inc.
2018-03-01
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| author | Hua Xu Leifeng Xu Panpan Yang Yuwei Cao Yuchao Tang Guoren He Suxia Yuan Jun Ming |
| author_facet | Hua Xu Leifeng Xu Panpan Yang Yuwei Cao Yuchao Tang Guoren He Suxia Yuan Jun Ming |
| author_sort | Hua Xu |
| collection | DOAJ |
| description | Virus-induced gene silencing (VIGS) is an attractive tool for determining gene function in plants. The present study constitutes the first application of VIGS in S. pseudocapsicum, which has great ornamental and pharmaceutical value, using tobacco rattle virus (TRV) vectors. Two marker genes, PHYTOENE DESATURASE (PDS) and Mg-chelatase H subunit (ChlH), were used to test the VIGS system in S. pseudocapsicum. The photobleaching and yellow-leaf phenotypes of the silenced plants were shown to significantly correlate with the down-regulation of endogenous SpPDS and SpChlH, respectively (P ≤ 0.05). Moreover, the parameters potentially affecting the efficiency of VIGS in S. pseudocapsicum, including the Agrobacterium strain and the inoculation method (leaf syringe-infiltration, sprout vacuum-infiltration and seed vacuum-infiltration), were compared. The optimized VIGS parameters were the leaf syringe-infiltration method, the Agrobacterium strain GV3101 and the growth of agro-inoculated plants at 25°. With these parameters, the silencing efficiency of SpPDS and SpChlH could reach approximately 50% in S. pseudocapsicum. Additionally, the suitability of various reference genes was screened by RT-qPCR using three candidate genes, and the results demonstrated that glyceraldehyde 3-phosphate dehydrogenase (GAPDH) can serve as a suitable reference for assessing the gene expression levels of VIGS systems in S. pseudocapsicum. The proven application of VIGS in S. pseudocapsicum and the characterization of a suitable reference gene in the present work will expedite the functional characterization of novel genes in S. pseudocapsicum. |
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| last_indexed | 2024-03-09T08:20:02Z |
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| spelling | doaj.art-cccf3a51fbe142b7acfdacaf5db0790d2023-12-02T21:54:16ZengPeerJ Inc.PeerJ2167-83592018-03-016e442410.7717/peerj.4424Tobacco rattle virus-induced PHYTOENE DESATURASE (PDS) and Mg-chelatase H subunit (ChlH) gene silencing in Solanum pseudocapsicum L.Hua Xu0Leifeng Xu1Panpan Yang2Yuwei Cao3Yuchao Tang4Guoren He5Suxia Yuan6Jun Ming7Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, ChinaInstitute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, ChinaInstitute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, ChinaInstitute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, ChinaInstitute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, ChinaInstitute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, ChinaInstitute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, ChinaInstitute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, ChinaVirus-induced gene silencing (VIGS) is an attractive tool for determining gene function in plants. The present study constitutes the first application of VIGS in S. pseudocapsicum, which has great ornamental and pharmaceutical value, using tobacco rattle virus (TRV) vectors. Two marker genes, PHYTOENE DESATURASE (PDS) and Mg-chelatase H subunit (ChlH), were used to test the VIGS system in S. pseudocapsicum. The photobleaching and yellow-leaf phenotypes of the silenced plants were shown to significantly correlate with the down-regulation of endogenous SpPDS and SpChlH, respectively (P ≤ 0.05). Moreover, the parameters potentially affecting the efficiency of VIGS in S. pseudocapsicum, including the Agrobacterium strain and the inoculation method (leaf syringe-infiltration, sprout vacuum-infiltration and seed vacuum-infiltration), were compared. The optimized VIGS parameters were the leaf syringe-infiltration method, the Agrobacterium strain GV3101 and the growth of agro-inoculated plants at 25°. With these parameters, the silencing efficiency of SpPDS and SpChlH could reach approximately 50% in S. pseudocapsicum. Additionally, the suitability of various reference genes was screened by RT-qPCR using three candidate genes, and the results demonstrated that glyceraldehyde 3-phosphate dehydrogenase (GAPDH) can serve as a suitable reference for assessing the gene expression levels of VIGS systems in S. pseudocapsicum. The proven application of VIGS in S. pseudocapsicum and the characterization of a suitable reference gene in the present work will expedite the functional characterization of novel genes in S. pseudocapsicum.https://peerj.com/articles/4424.pdfPHYTOENE DESATURASE (PDS)VIGSMg-chelatase H subunit (ChlH)Solanum pseudocapsicum L.TRVReference genes |
| spellingShingle | Hua Xu Leifeng Xu Panpan Yang Yuwei Cao Yuchao Tang Guoren He Suxia Yuan Jun Ming Tobacco rattle virus-induced PHYTOENE DESATURASE (PDS) and Mg-chelatase H subunit (ChlH) gene silencing in Solanum pseudocapsicum L. PeerJ PHYTOENE DESATURASE (PDS) VIGS Mg-chelatase H subunit (ChlH) Solanum pseudocapsicum L. TRV Reference genes |
| title | Tobacco rattle virus-induced PHYTOENE DESATURASE (PDS) and Mg-chelatase H subunit (ChlH) gene silencing in Solanum pseudocapsicum L. |
| title_full | Tobacco rattle virus-induced PHYTOENE DESATURASE (PDS) and Mg-chelatase H subunit (ChlH) gene silencing in Solanum pseudocapsicum L. |
| title_fullStr | Tobacco rattle virus-induced PHYTOENE DESATURASE (PDS) and Mg-chelatase H subunit (ChlH) gene silencing in Solanum pseudocapsicum L. |
| title_full_unstemmed | Tobacco rattle virus-induced PHYTOENE DESATURASE (PDS) and Mg-chelatase H subunit (ChlH) gene silencing in Solanum pseudocapsicum L. |
| title_short | Tobacco rattle virus-induced PHYTOENE DESATURASE (PDS) and Mg-chelatase H subunit (ChlH) gene silencing in Solanum pseudocapsicum L. |
| title_sort | tobacco rattle virus induced phytoene desaturase pds and mg chelatase h subunit chlh gene silencing in solanum pseudocapsicum l |
| topic | PHYTOENE DESATURASE (PDS) VIGS Mg-chelatase H subunit (ChlH) Solanum pseudocapsicum L. TRV Reference genes |
| url | https://peerj.com/articles/4424.pdf |
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