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Establishment of virus-induced gene silencing (VIGS) system in Luffa acutangula using Phytoene desaturase (PDS) and tendril synthesis related gene (TEN)

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 stu...

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Autores principales: Qi, Xiaoyu, Mo, Qiaoping, Li, Jing, Zi, Zhibo, Xu, Mengyun, Yue, Suju, Zhao, Hongbo, Zhu, Haisheng, Wang, Guoping
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10470258/
https://www.ncbi.nlm.nih.gov/pubmed/37653449
http://dx.doi.org/10.1186/s13007-023-01064-4
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author Qi, Xiaoyu
Mo, Qiaoping
Li, Jing
Zi, Zhibo
Xu, Mengyun
Yue, Suju
Zhao, Hongbo
Zhu, Haisheng
Wang, Guoping
author_facet Qi, Xiaoyu
Mo, Qiaoping
Li, Jing
Zi, Zhibo
Xu, Mengyun
Yue, Suju
Zhao, Hongbo
Zhu, Haisheng
Wang, Guoping
author_sort Qi, Xiaoyu
collection PubMed
description 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. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13007-023-01064-4.
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spelling pubmed-104702582023-09-01 Establishment of virus-induced gene silencing (VIGS) system in Luffa acutangula using Phytoene desaturase (PDS) and tendril synthesis related gene (TEN) Qi, Xiaoyu Mo, Qiaoping Li, Jing Zi, Zhibo Xu, Mengyun Yue, Suju Zhao, Hongbo Zhu, Haisheng Wang, Guoping Plant Methods Methodology 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. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13007-023-01064-4. BioMed Central 2023-08-31 /pmc/articles/PMC10470258/ /pubmed/37653449 http://dx.doi.org/10.1186/s13007-023-01064-4 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Methodology
Qi, Xiaoyu
Mo, Qiaoping
Li, Jing
Zi, Zhibo
Xu, Mengyun
Yue, Suju
Zhao, Hongbo
Zhu, Haisheng
Wang, Guoping
Establishment of virus-induced gene silencing (VIGS) system in Luffa acutangula using Phytoene desaturase (PDS) and tendril synthesis related gene (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 Methodology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10470258/
https://www.ncbi.nlm.nih.gov/pubmed/37653449
http://dx.doi.org/10.1186/s13007-023-01064-4
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