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A rapid, simple, and highly efficient method for VIGS and in vitro-inoculation of plant virus by INABS applied to crops that develop axillary buds and can survive from cuttings

BACKGROUND: Virus-induced gene silencing (VIGS) is one of the most convenient and powerful methods of reverse genetics. In vitro-inoculation of plant virus is an important method for studying the interactions between viruses and plants. Agrobacterium-based infiltration has been widely adopted as a t...

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Autores principales: Liu, Qili, Xu, Kedong, Yi, Lun, Hou, Yalin, Li, Dongxiao, Hu, Haiyan, Zhou, Feng, Song, Puwen, Yu, Yongang, Wei, Qichao, Guan, Yuanyuan, Hu, Ping, Bu, Ruifang, Chen, Eryong, Su, Xiaojia, Li, Honglian, Li, Chengwei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8605592/
https://www.ncbi.nlm.nih.gov/pubmed/34800968
http://dx.doi.org/10.1186/s12870-021-03331-9
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author Liu, Qili
Xu, Kedong
Yi, Lun
Hou, Yalin
Li, Dongxiao
Hu, Haiyan
Zhou, Feng
Song, Puwen
Yu, Yongang
Wei, Qichao
Guan, Yuanyuan
Hu, Ping
Bu, Ruifang
Chen, Eryong
Su, Xiaojia
Li, Honglian
Li, Chengwei
author_facet Liu, Qili
Xu, Kedong
Yi, Lun
Hou, Yalin
Li, Dongxiao
Hu, Haiyan
Zhou, Feng
Song, Puwen
Yu, Yongang
Wei, Qichao
Guan, Yuanyuan
Hu, Ping
Bu, Ruifang
Chen, Eryong
Su, Xiaojia
Li, Honglian
Li, Chengwei
author_sort Liu, Qili
collection PubMed
description BACKGROUND: Virus-induced gene silencing (VIGS) is one of the most convenient and powerful methods of reverse genetics. In vitro-inoculation of plant virus is an important method for studying the interactions between viruses and plants. Agrobacterium-based infiltration has been widely adopted as a tool for VIGS and in vitro-inoculation of plant virus. Most agrobacterium-based infiltration methods applied to VIGS and virus inoculation have the characteristics of low transformation efficiencies, long plant growth time, large amounts of plant tissue, large test spaces, and complex preparation procedures. Therefore, a rapid, simple, economical, and highly efficient VIGS and virus inoculation method is in need. Previous studies have shown that the selection of suitable plant tissues and inoculation sites is the key to successful infection. RESULTS: In this study, Tobacco rattle virus (TRV) mediated VIGS and Tomato yellow leaf curl virus (TYLCV) for virus inoculation were developed in tomato plants based on the agrobacterium tumefaciens-based infiltration by injection of the no-apical-bud stem section (INABS). The no-apical-bud stem section had a “Y- type” asymmetric structure and contained an axillary bud that was about 1–3 cm in length. This protocol provides high transformation (56.7%) and inoculation efficiency (68.3%), which generates VIGS transformants or diseased plants in a very short period (8 dpi). Moreover, it greatly reduces the required experimental space. This method will facilitate functional genomic studies and large-scale disease resistance screening. CONCLUSIONS: Overall, a rapid, simple, and highly efficient method for VIGS and virus inoculation by INABS was developed in tomato. It was reasonable to believe that it can be used as a reference for the other virus inoculation methods and for the application of VIGS to other crops (such as sweet potato, potato, cassava and tobacco) that develop axillary buds and can survive from cuttings. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12870-021-03331-9.
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spelling pubmed-86055922021-11-22 A rapid, simple, and highly efficient method for VIGS and in vitro-inoculation of plant virus by INABS applied to crops that develop axillary buds and can survive from cuttings Liu, Qili Xu, Kedong Yi, Lun Hou, Yalin Li, Dongxiao Hu, Haiyan Zhou, Feng Song, Puwen Yu, Yongang Wei, Qichao Guan, Yuanyuan Hu, Ping Bu, Ruifang Chen, Eryong Su, Xiaojia Li, Honglian Li, Chengwei BMC Plant Biol Methodology Article BACKGROUND: Virus-induced gene silencing (VIGS) is one of the most convenient and powerful methods of reverse genetics. In vitro-inoculation of plant virus is an important method for studying the interactions between viruses and plants. Agrobacterium-based infiltration has been widely adopted as a tool for VIGS and in vitro-inoculation of plant virus. Most agrobacterium-based infiltration methods applied to VIGS and virus inoculation have the characteristics of low transformation efficiencies, long plant growth time, large amounts of plant tissue, large test spaces, and complex preparation procedures. Therefore, a rapid, simple, economical, and highly efficient VIGS and virus inoculation method is in need. Previous studies have shown that the selection of suitable plant tissues and inoculation sites is the key to successful infection. RESULTS: In this study, Tobacco rattle virus (TRV) mediated VIGS and Tomato yellow leaf curl virus (TYLCV) for virus inoculation were developed in tomato plants based on the agrobacterium tumefaciens-based infiltration by injection of the no-apical-bud stem section (INABS). The no-apical-bud stem section had a “Y- type” asymmetric structure and contained an axillary bud that was about 1–3 cm in length. This protocol provides high transformation (56.7%) and inoculation efficiency (68.3%), which generates VIGS transformants or diseased plants in a very short period (8 dpi). Moreover, it greatly reduces the required experimental space. This method will facilitate functional genomic studies and large-scale disease resistance screening. CONCLUSIONS: Overall, a rapid, simple, and highly efficient method for VIGS and virus inoculation by INABS was developed in tomato. It was reasonable to believe that it can be used as a reference for the other virus inoculation methods and for the application of VIGS to other crops (such as sweet potato, potato, cassava and tobacco) that develop axillary buds and can survive from cuttings. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12870-021-03331-9. BioMed Central 2021-11-20 /pmc/articles/PMC8605592/ /pubmed/34800968 http://dx.doi.org/10.1186/s12870-021-03331-9 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis 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 Article
Liu, Qili
Xu, Kedong
Yi, Lun
Hou, Yalin
Li, Dongxiao
Hu, Haiyan
Zhou, Feng
Song, Puwen
Yu, Yongang
Wei, Qichao
Guan, Yuanyuan
Hu, Ping
Bu, Ruifang
Chen, Eryong
Su, Xiaojia
Li, Honglian
Li, Chengwei
A rapid, simple, and highly efficient method for VIGS and in vitro-inoculation of plant virus by INABS applied to crops that develop axillary buds and can survive from cuttings
title A rapid, simple, and highly efficient method for VIGS and in vitro-inoculation of plant virus by INABS applied to crops that develop axillary buds and can survive from cuttings
title_full A rapid, simple, and highly efficient method for VIGS and in vitro-inoculation of plant virus by INABS applied to crops that develop axillary buds and can survive from cuttings
title_fullStr A rapid, simple, and highly efficient method for VIGS and in vitro-inoculation of plant virus by INABS applied to crops that develop axillary buds and can survive from cuttings
title_full_unstemmed A rapid, simple, and highly efficient method for VIGS and in vitro-inoculation of plant virus by INABS applied to crops that develop axillary buds and can survive from cuttings
title_short A rapid, simple, and highly efficient method for VIGS and in vitro-inoculation of plant virus by INABS applied to crops that develop axillary buds and can survive from cuttings
title_sort rapid, simple, and highly efficient method for vigs and in vitro-inoculation of plant virus by inabs applied to crops that develop axillary buds and can survive from cuttings
topic Methodology Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8605592/
https://www.ncbi.nlm.nih.gov/pubmed/34800968
http://dx.doi.org/10.1186/s12870-021-03331-9
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