Establishment of virus-induced gene silencing (VIGS) system in Luffa acutangula using Phytoene desaturase (PDS) and tendril synthesis related gene (TEN)
Abstract Background Virus-induced gene silencing (VIGS) is a reverse genetics technology that can efficiently and rapidly identify plant gene functions. Although a variety of VIGS vectors have been successfully used in plants, only a few reports on VIGS technology in Luffa exist. Results In the pres...
| Main Authors: | , , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2023-08-01
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| Series: | Plant Methods |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s13007-023-01064-4 |
| _version_ | 1797452501292679168 |
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| author | Xiaoyu Qi Qiaoping Mo Jing Li Zhibo Zi Mengyun Xu Suju Yue Hongbo Zhao Haisheng Zhu Guoping Wang |
| author_facet | Xiaoyu Qi Qiaoping Mo Jing Li Zhibo Zi Mengyun Xu Suju Yue Hongbo Zhao Haisheng Zhu Guoping Wang |
| author_sort | Xiaoyu Qi |
| collection | DOAJ |
| description | Abstract Background Virus-induced gene silencing (VIGS) is a reverse genetics technology that can efficiently and rapidly identify plant gene functions. Although a variety of VIGS vectors have been successfully used in plants, only a few reports on VIGS technology in Luffa exist. Results In the present study, a new cucumber green mottle mosaic virus (CGMMV)-based VIGS vector, pV190, was applied to establish the CGMMV-VIGS to investigate the feasibility of the silencing system for Luffa. Phytoene desaturase (PDS) gene was initially selected as a VIGS marker gene to construct a recombinant vector. Plants infected with Agrobacterium harboring pV190-PDS successfully induced effective silencing in Luffa, and an effective gene silencing phenotype with obvious photobleaching was observed. To further validate the efficiency, we selected TEN for gene-silencing, which encodes a CYC/TB1-like transcription factor and is involved in tendril development. Luffa plants inoculated with the pV190-TEN exhibited shorter tendril length and nodal positions where tendrils appear are higher compared to those of non-inoculated plants. RT-qPCR showed that the expression levels of PDS and TEN were significantly reduced in the CGMMV-VIGS plants. Moreover, we evaluated the CGMMV-VIGS efficiency in three cucurbits, including cucumber, ridge gourd, and bottle gourd. Conclusion We successfully established a CGMMV-based VIGS system on ridge gourd and used marker genes to identify the feasibility of the silencing system in Luffa leaves and stems. |
| first_indexed | 2024-03-09T15:09:34Z |
| format | Article |
| id | doaj.art-3430ccc7747a44cc8838387588014a94 |
| institution | Directory Open Access Journal |
| issn | 1746-4811 |
| language | English |
| last_indexed | 2024-03-09T15:09:34Z |
| publishDate | 2023-08-01 |
| publisher | BMC |
| record_format | Article |
| series | Plant Methods |
| spelling | doaj.art-3430ccc7747a44cc8838387588014a942023-11-26T13:26:46ZengBMCPlant Methods1746-48112023-08-0119111010.1186/s13007-023-01064-4Establishment of virus-induced gene silencing (VIGS) system in Luffa acutangula using Phytoene desaturase (PDS) and tendril synthesis related gene (TEN)Xiaoyu Qi0Qiaoping Mo1Jing Li2Zhibo Zi3Mengyun Xu4Suju Yue5Hongbo Zhao6Haisheng Zhu7Guoping Wang8College of Horticulture, South China Agricultural UniversityCollege of Horticulture, South China Agricultural UniversityCollege of Horticulture, South China Agricultural UniversityCollege of Horticulture, South China Agricultural UniversityCollege of Horticulture, South China Agricultural UniversityCollege of Life Sciences, Zhongkai University of Agriculture and EngineeringCollege of Horticulture, South China Agricultural UniversityFujian Key Laboratory of Vegetable Genetics and Breeding/Crops Research Institute, Fujian Academy of Agricultural SciencesCollege of Horticulture, South China Agricultural UniversityAbstract Background Virus-induced gene silencing (VIGS) is a reverse genetics technology that can efficiently and rapidly identify plant gene functions. Although a variety of VIGS vectors have been successfully used in plants, only a few reports on VIGS technology in Luffa exist. Results In the present study, a new cucumber green mottle mosaic virus (CGMMV)-based VIGS vector, pV190, was applied to establish the CGMMV-VIGS to investigate the feasibility of the silencing system for Luffa. Phytoene desaturase (PDS) gene was initially selected as a VIGS marker gene to construct a recombinant vector. Plants infected with Agrobacterium harboring pV190-PDS successfully induced effective silencing in Luffa, and an effective gene silencing phenotype with obvious photobleaching was observed. To further validate the efficiency, we selected TEN for gene-silencing, which encodes a CYC/TB1-like transcription factor and is involved in tendril development. Luffa plants inoculated with the pV190-TEN exhibited shorter tendril length and nodal positions where tendrils appear are higher compared to those of non-inoculated plants. RT-qPCR showed that the expression levels of PDS and TEN were significantly reduced in the CGMMV-VIGS plants. Moreover, we evaluated the CGMMV-VIGS efficiency in three cucurbits, including cucumber, ridge gourd, and bottle gourd. Conclusion We successfully established a CGMMV-based VIGS system on ridge gourd and used marker genes to identify the feasibility of the silencing system in Luffa leaves and stems.https://doi.org/10.1186/s13007-023-01064-4LuffaVIGSCucumber green mottle mosaic virus (CGMMV)PDSTEN |
| spellingShingle | Xiaoyu Qi Qiaoping Mo Jing Li Zhibo Zi Mengyun Xu Suju Yue Hongbo Zhao Haisheng Zhu Guoping Wang Establishment of virus-induced gene silencing (VIGS) system in Luffa acutangula using Phytoene desaturase (PDS) and tendril synthesis related gene (TEN) Plant Methods Luffa VIGS Cucumber green mottle mosaic virus (CGMMV) PDS TEN |
| title | Establishment of virus-induced gene silencing (VIGS) system in Luffa acutangula using Phytoene desaturase (PDS) and tendril synthesis related gene (TEN) |
| title_full | Establishment of virus-induced gene silencing (VIGS) system in Luffa acutangula using Phytoene desaturase (PDS) and tendril synthesis related gene (TEN) |
| title_fullStr | Establishment of virus-induced gene silencing (VIGS) system in Luffa acutangula using Phytoene desaturase (PDS) and tendril synthesis related gene (TEN) |
| title_full_unstemmed | Establishment of virus-induced gene silencing (VIGS) system in Luffa acutangula using Phytoene desaturase (PDS) and tendril synthesis related gene (TEN) |
| title_short | Establishment of virus-induced gene silencing (VIGS) system in Luffa acutangula using Phytoene desaturase (PDS) and tendril synthesis related gene (TEN) |
| title_sort | establishment of virus induced gene silencing vigs system in luffa acutangula using phytoene desaturase pds and tendril synthesis related gene ten |
| topic | Luffa VIGS Cucumber green mottle mosaic virus (CGMMV) PDS TEN |
| url | https://doi.org/10.1186/s13007-023-01064-4 |
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