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

Abstract 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 adopte...

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Autores principales:	Qili Liu, Kedong Xu, Lun Yi, Yalin Hou, Dongxiao Li, Haiyan Hu, Feng Zhou, Puwen Song, Yongang Yu, Qichao Wei, Yuanyuan Guan, Ping Hu, Ruifang Bu, Eryong Chen, Xiaojia Su, Honglian Li, Chengwei Li
Formato:	article
Lenguaje:	EN
Publicado:	BMC 2021
Materias:	Virus-induced gene silencing (VIGS) In vitro-inoculation of plant virus Agrobacterium-based infiltration Injection of no-apical-bud stem sections (INABS) Botany QK1-989
Acceso en línea:	https://doaj.org/article/868a924851f047b2903e628575463dda
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spelling	oai:doaj.org-article:868a924851f047b2903e628575463dda2021-11-21T12:05:42ZA 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 cuttings10.1186/s12870-021-03331-91471-2229https://doaj.org/article/868a924851f047b2903e628575463dda2021-11-01T00:00:00Zhttps://doi.org/10.1186/s12870-021-03331-9https://doaj.org/toc/1471-2229Abstract 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.Qili LiuKedong XuLun YiYalin HouDongxiao LiHaiyan HuFeng ZhouPuwen SongYongang YuQichao WeiYuanyuan GuanPing HuRuifang BuEryong ChenXiaojia SuHonglian LiChengwei LiBMCarticleVirus-induced gene silencing (VIGS)In vitro-inoculation of plant virusAgrobacterium-based infiltrationInjection of no-apical-bud stem sections (INABS)BotanyQK1-989ENBMC Plant Biology, Vol 21, Iss 1, Pp 1-10 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Virus-induced gene silencing (VIGS) In vitro-inoculation of plant virus Agrobacterium-based infiltration Injection of no-apical-bud stem sections (INABS) Botany QK1-989
spellingShingle	Virus-induced gene silencing (VIGS) In vitro-inoculation of plant virus Agrobacterium-based infiltration Injection of no-apical-bud stem sections (INABS) Botany QK1-989 Qili Liu Kedong Xu Lun Yi Yalin Hou Dongxiao Li Haiyan Hu Feng Zhou Puwen Song Yongang Yu Qichao Wei Yuanyuan Guan Ping Hu Ruifang Bu Eryong Chen Xiaojia Su Honglian Li Chengwei Li 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
description	Abstract 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.
format	article
author	Qili Liu Kedong Xu Lun Yi Yalin Hou Dongxiao Li Haiyan Hu Feng Zhou Puwen Song Yongang Yu Qichao Wei Yuanyuan Guan Ping Hu Ruifang Bu Eryong Chen Xiaojia Su Honglian Li Chengwei Li
author_facet	Qili Liu Kedong Xu Lun Yi Yalin Hou Dongxiao Li Haiyan Hu Feng Zhou Puwen Song Yongang Yu Qichao Wei Yuanyuan Guan Ping Hu Ruifang Bu Eryong Chen Xiaojia Su Honglian Li Chengwei Li
author_sort	Qili Liu
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_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_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_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
publisher	BMC
publishDate	2021
url	https://doaj.org/article/868a924851f047b2903e628575463dda
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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

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