Virus-induced gene silencing: empowering genetics in non-model organisms

Dommes, Anna Barbara; Gross, Thomas P.; Herbert, Denise; Kivivirta, Kimmo I; Becker, Annette

doi:10.1093/jxb/ery411

Cited by 87 publications

(88 citation statements)

References 151 publications

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“…Agrobacterium-based expression has proven the most significant progress in the manufacturing of plant-made proteins and compounds (Marillonnet et al, 2005;Peyret and Lomonossoff, 2015). Apart from their use for protein expression, viral vectors are also widely applied in functional genomic studies for targeted down-regulation of endogenous transcripts via RNA silencing (Dommes et al, 2018;Tang et al, 2010). Stable genome alterations in plant cells have been reported by viral expression of sequence-specific nucleases or use of virus-delivered sequences as template for targeted DNA replacement (Ali et al, 2015;Baltes et al, 2014;Gil-Humanes et al, 2017;Honig et al, 2015;Marton et al, 2010;Wang et al, 2017).…”

Section: Cutting-edge Applications Of Plant Virusesmentioning

confidence: 99%

Harnessed viruses in the age of metagenomics and synthetic biology: an update on infectious clone assembly and biotechnologies of plant viruses

Pasin

Menzel

Daròs

2019

Plant Biotechnology Journal

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Summary Recent metagenomic studies have provided an unprecedented wealth of data, which are revolutionizing our understanding of virus diversity. A redrawn landscape highlights viruses as active players in the phytobiome, and surveys have uncovered their positive roles in environmental stress tolerance of plants. Viral infectious clones are key tools for functional characterization of known and newly identified viruses. Knowledge of viruses and their components has been instrumental for the development of modern plant molecular biology and biotechnology. In this review, we provide extensive guidelines built on current synthetic biology advances that streamline infectious clone assembly, thus lessening a major technical constraint of plant virology. The focus is on generation of infectious clones in binary T‐ DNA vectors, which are delivered efficiently to plants by Agrobacterium . We then summarize recent applications of plant viruses and explore emerging trends in microbiology, bacterial and human virology that, once translated to plant virology, could lead to the development of virus‐based gene therapies for ad hoc engineering of plant traits. The systematic characterization of plant virus roles in the phytobiome and next‐generation virus‐based tools will be indispensable landmarks in the synthetic biology roadmap to better crops.

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Section: Cutting-edge Applications Of Plant Virusesmentioning

confidence: 99%

Harnessed viruses in the age of metagenomics and synthetic biology: an update on infectious clone assembly and biotechnologies of plant viruses

Pasin

Menzel

Daròs

2019

Plant Biotechnology Journal

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“…Third, they are synthesized from simple components by soluble enzymes, facilitating in vitro analysis, including pathway reconstruction in a test tube. These, combined with functional genomics tools of transformation, CRISPR-Cas9 [18,19] and viral induced gene silencing (VIGS) [20] available in the Solanaceae, permit direct and detailed analysis of how genotypic variation affects phenotypic diversity.…”

Section: Introductionmentioning

confidence: 99%

Tip of the trichome: evolution of acylsugar metabolic diversity in Solanaceae

Fan

Leong

2019

Current Opinion in Plant Biology

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Acylsugars are insecticidal plant specialized metabolites produced in the Solanaceae (nightshade family). Despite having simple constituents, these compounds are unusually structurally diverse. Their structural variations in phylogenetically closely related species enable comparative biochemical approaches to understand acylsugar biosynthesis and pathway diversification. Thus far, varied enzyme classes contributing to their synthesis were characterized in cultivated and wild tomatoes, including from core metabolism-isopropylmalate synthase (Leu) and invertase (carbon)-and a group of evolutionarily related BAHD acyltransferases known as acylsucrose acyltransferases. Gene duplication and neofunctionalization of these enzymes drove acylsugar diversification both within and beyond tomato. The broad set of evolutionary mechanisms underlying acylsugar diversity in Solanaceae make this metabolic network an exemplar for detailed understanding of the evolution of metabolic form and function.

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“…Despite the potential of HIGS for disease control, off-target effects, difficulties in plant transformation, and low public acceptance of transgene technology have restricted its application in the field [7,15,19,20]. Virus-induced gene silencing (VIGS), which exploits recombinant viruses as vectors to deliver RNAi molecules to plants, is an alternative to HIGS but to date it has mostly been utilized as a functional tool to study genes of interest in different crops, rather than for disease control [21][22][23]. The relatively recent discovery that externally applied dsRNAs and siRNAs can be taken-up by fungal cells from their environment and induce silencing in the transfected cells, has attracted much attention for its potential use as an alternative to HIGS for delivering RNAi to target pathogens [24].…”

Section: Introductionmentioning

confidence: 99%

Targeted Delivery of Gene Silencing in Fungi Using Genetically Engineered Bacteria

Niño‐Sánchez

Chen

Souza

et al. 2021

JoF

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Exploiting RNA interference (RNAi) in disease control through non-transformative methods that overcome the hurdle of producing transgenic plants has attracted much attention over the last years. Here, we explored such a method and used non-pathogenic bacteria as a versatile system for delivering RNAi to fungi. Specifically, the RNaseIII-null mutant strain of Escherichia coli HT115(DE3) was transformed with two plasmid vectors that enabled the constitutive or IPTG-inducible production of double-stranded RNAs (dsRNAs) against genes involved in aflatoxins production in Aspergillus flavus (AflC) or virulence of Botrytis cinerea (BcSAS1). To facilitate the release of the dsRNAs, the bacterial cells were further genetically engineered to undergo a bacteriophage endolysin R-mediated autolysis, following a freeze-thaw cycle. Exposure under in vitro conditions of A. flavus or B. cinerea to living bacteria or their whole-cell autolysates induced silencing of AflC and BcSAS1 in a bacteria concentration-dependent manner, and instigated a reduction in aflatoxins production and mycelial growth, respectively. In planta applications of the living bacteria or their crude whole-cell autolysates produced similar results, thus creating a basis for translational research. These results demonstrate that bacteria can produce biologically active dsRNA against target genes in fungi and that bacteria-mediated RNAi can be used to control fungal pathogens.

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Virus-induced gene silencing: empowering genetics in non-model organisms

Cited by 87 publications

References 151 publications

Harnessed viruses in the age of metagenomics and synthetic biology: an update on infectious clone assembly and biotechnologies of plant viruses

Harnessed viruses in the age of metagenomics and synthetic biology: an update on infectious clone assembly and biotechnologies of plant viruses

Tip of the trichome: evolution of acylsugar metabolic diversity in Solanaceae

Targeted Delivery of Gene Silencing in Fungi Using Genetically Engineered Bacteria

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