A Methodological Advance of Tobacco Rattle Virus-Induced Gene Silencing for Functional Genomics in Plants

Shi, Guiyang; Hao, Mengyuan; Tian, Bin; Cao, Gangqiang; Wei, Fang; Xie, Zhengqing

doi:10.3389/fpls.2021.671091

Cited by 40 publications

(31 citation statements)

References 101 publications

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“…The target fragment was inserted into the vector using the In-Fusion ® HD Cloning Kit. VIGS was carried out according to the method of Schachtsiek et al (2019) and Shi et al (2021) . The successfully transformed GV3101 strain containing TRV-VIGS vector was placed in a YEP medium containing 10 mM MES, 20 mM acetosyringone, 25 μg/ml rifampicin, and 50 μg/ml kanamycin and incubated at 28°C for 24 h. Agrobacterium cells were collected by centrifugation and added to an infiltration buffer (OD600 = 1-2) containing 200 mM acetosyringone, 10 mM MgCl 2 , and 10 mM MES (pH 5.6).…”

Section: Methodsmentioning

confidence: 99%

LcNAC13 Is Involved in the Reactive Oxygen Species-Dependent Senescence of the Rudimentary Leaves in Litchi chinensis

et al. 2022

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Litchi is an important evergreen fruit tree. Floral formation in litchi is induced by low temperatures (LTs). However, unstable flowering is a challenge for litchi production in times of global warming and climate change. Previous studies have shown that the methyl viologen dichloride hydrate-generated reactive oxygen species (ROS) could promote flowering. Leaves in the panicles may affect the development of the inflorescence in litchi under high-temperature condition. In this study, potted litchi trees were transferred to growth chambers at LT and high temperature (HT). From a previous dataset of the RNA sequencing of the ROS-treated rudimentary leaves, a NAC transcription factor-encoding gene LcNAC13 was identified. By genetic transformation of LcNAC13 to Arabidopsis thaliana and tobacco, it was found that the ROS-induced senescence of the leaves was accelerated. Silencing LcNAC13 by virus-induced gene silencing (VIGS) delayed ROS-dependent senescence. Our results suggested that LcNAC13 regulates rudimentary leaf senescence. Our study provided a new target gene for the future molecular breeding of new cultivars that could flower under global warming conditions.

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Section: Methodsmentioning

confidence: 99%

LcNAC13 Is Involved in the Reactive Oxygen Species-Dependent Senescence of the Rudimentary Leaves in Litchi chinensis

et al. 2022

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“…The transient nature of these three distinct molecular biology tools has greatly facilitated plant gene function studies by obviating the need for the time-consuming process of producing transgenic plants (Lee et al 2012). Additionally, the development of these high-throughput tools has shed light on the limitations and capacities of the viruses themselves (Shi et al 2021;Wang et al 2020). Their emerging role as VIGE vectors adds yet another function to their repertoire for CRISPR/Cas9 mediated modification of both model and non-model plants for gene function studies and crop improvement (Wang et al 2020).…”

Section: Advantages Of Viral Delivery Of Crispr/cas9 Constructsmentioning

confidence: 99%

“…Recent work with traditional non-viral CRISPR/ Cas9 assays have targeted a number of genes impacting plant responses to drought, heat stress, salinity, and disease (Das et al 2018;Sun et al 2021). Additionally, the traditional VIGS systems have been successfully used to study plant responses to both biotic and abiotic stresses (Dommes et al 2019;Dulermo et al 2009;Shi et al 2021). Therefore, it is only a matter of time before VIGE systems will be employed to similarly study gene function during pathogen infection or under abiotic stress.…”

Section: Available Virus-mediated Crispr/cas9 Plant Genome Editing Toolsmentioning

confidence: 99%

VIGE: virus-induced genome editing for improving abiotic and biotic stress traits in plants

et al. 2022

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Agricultural production is hampered by disease, pests, and environmental stresses. To minimize yield loss, it is important to develop crop cultivars with resistance or tolerance to their respective biotic and abiotic constraints. Transformation techniques are not optimized for many species and desirable cultivars may not be amenable to genetic transformation, necessitating inferior cultivar usage and time-consuming introgression through backcrossing to the preferred variety. Overcoming these limitations will greatly facilitate the development of disease, insect, and abiotic stress tolerant crops. One such avenue for rapid crop improvement is the development of viral systems to deliver CRISPR/Cas-based genome editing technology to plants to generate targeted beneficial mutations. Viral delivery of genomic editing constructs can theoretically be applied to span the entire host range of the virus utilized, circumventing the challenges associated with traditional transformation and breeding techniques. Here we explore the types of viruses that have been optimized for CRISPR/Cas9 delivery, the phenotypic outcomes achieved in recent studies, and discuss the future potential of this rapidly advancing technology.

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“…Since VIGS can rapidly reduce the expression of target genes, it facilitates molecular function research in plants, including forest trees. Therefore, it is necessary to determine the optimal conditions for VIGS to silence target genes in forest trees, including the viral vector, ambient temperature, plant age or development stages, and inoculation method ( Shi et al, 2021 ). Overall, VIGS enables gene function analysis of trees.…”

Section: Advancements In Transgenic Technologies For Forest Treesmentioning

confidence: 99%

Advances and Perspectives of Transgenic Technology and Biotechnological Application in Forest Trees

et al. 2021

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Transgenic technology is increasingly used in forest-tree breeding to overcome the disadvantages of traditional breeding methods, such as a long breeding cycle, complex cultivation environment, and complicated procedures. By introducing exogenous DNA, genes tightly related or contributed to ideal traits—including insect, disease, and herbicide resistance—were transferred into diverse forest trees, and genetically modified (GM) trees including poplars were cultivated. It is beneficial to develop new varieties of GM trees of high quality and promote the genetic improvement of forests. However, the low transformation efficiency has hampered the cultivation of GM trees and the identification of the molecular genetic mechanism in forest trees compared to annual herbaceous plants such as Oryza sativa. In this study, we reviewed advances in transgenic technology of forest trees, including the principles, advantages and disadvantages of diverse genetic transformation methods, and their application for trait improvement. The review provides insight into the establishment and improvement of genetic transformation systems for forest tree species. Challenges and perspectives pertaining to the genetic transformation of forest trees are also discussed.

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A Methodological Advance of Tobacco Rattle Virus-Induced Gene Silencing for Functional Genomics in Plants

Cited by 40 publications

References 101 publications

LcNAC13 Is Involved in the Reactive Oxygen Species-Dependent Senescence of the Rudimentary Leaves in Litchi chinensis

LcNAC13 Is Involved in the Reactive Oxygen Species-Dependent Senescence of the Rudimentary Leaves in Litchi chinensis

VIGE: virus-induced genome editing for improving abiotic and biotic stress traits in plants

Advances and Perspectives of Transgenic Technology and Biotechnological Application in Forest Trees

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