Faculty Opinions recommendation of Spray-Induced Gene Silencing: a Powerful Innovative Strategy for Crop Protection.

2018 ◽  
Author(s):  
Jian-Min Zhou ◽  
Xiangxiu Liang
Keyword(s):  
Gene Silencing ◽  
Crop Protection ◽  
Innovative Strategy

The economics of RNAi-based innovation: from the innovation landscape to consumer acceptance.

2021 ◽  
pp. 159-166
Author(s):  
Vera Ventura ◽  
Dario G. Frisio
Keyword(s):  
Rna Interference ◽  
Gene Silencing ◽  
Crop Protection ◽  
Consumer Perceptions ◽  
Market Potential ◽  
Consumer Acceptance ◽  
Innovative Technology ◽  
Sustainable Solutions

Abstract RNA interference (RNAi) is an innovative technology of gene silencing which offers great opportunities for the development of sustainable solutions for crop protection. This chapter discusses the market potential of RNAi innovation, the application of RNAi for biocontrol, and stakeholder and consumer perceptions of RNAi technologies.

Host-induced gene silencing and spray-induced gene silencing for crop protection against viruses.

2021 ◽  
pp. 72-85
Author(s):  
Angela Ricci ◽  
Silvia Sabbadini ◽  
Laura Miozzi ◽  
Bruno Mezzetti ◽  
Emanuela Noris
Keyword(s):  
Gene Silencing ◽  
Plant Pathogens ◽  
Crop Protection ◽  
Plant Defence ◽  
Transcriptional Gene Silencing ◽  
Target Sequence ◽  
Sequence Specificity ◽  
Rna Molecules ◽  
Political Concern ◽  
Post Transcriptional Gene Silencing

Abstract Since the beginning of agriculture, plant virus diseases have been a strong challenge for farming. Following its discovery at the very beginning of the 1990s, the RNA interference (RNAi) mechanism has been widely studied and exploited as an integrative tool to obtain resistance to viruses in several plant species, with high target-sequence specificity. In this chapter, we describe and review the major aspects of host-induced gene silencing (HIGS), as one of the possible plant defence methods, using genetic engineering techniques. In particular, we focus our attention on the use of RNAi-based gene constructs to introduce stable resistance in host plants against viral diseases, by triggering post-transcriptional gene silencing (PTGS). Recently, spray-induced gene silencing (SIGS), consisting of the topical application of small RNA molecules to plants, has been explored as an alternative tool to the stable integration of RNAi-based gene constructs in plants. SIGS has great and innovative potential for crop defence against different plant pathogens and pests and is expected to raise less public and political concern, as it does not alter the genetic structure of the plant.

scholarly journals High transgene expression is associated with systemic GFP silencing in Nicotiana benthamiana

2021 ◽  
Author(s):  
Bill Hendrix ◽  
Paul Hoffer ◽  
Rick Sanders ◽  
Steve Schwartz ◽  
Wei Zheng ◽  
...  
Keyword(s):  
Gene Silencing ◽  
Nicotiana Benthamiana ◽  
Transgene Expression ◽  
Fluorescent Protein ◽  
Integration Site ◽  
Crop Protection ◽  
Transgene Silencing ◽  
Expression Level ◽  
Transgenic Lines ◽  
Transgene Expression Level

AbstractGene silencing in plants using topical dsRNA is a new approach that has the potential to be a sustainable component of the agricultural production systems of the future. However, more research is needed to enable this technology as an economical and efficacious supplement to current crop protection practices. Systemic gene silencing is one key enabling aspect. The objective of this research was to better understand systemic transgene silencing in Nicotiana benthamiana. Previous reports details sequencing of the integration site of the Green Fluorescent Protein (GFP) transgene in the well-known N. benthamiana GFP16C event revealed inadvertent co-integration of part of a bacterial transposase. To determine the effect of this transgene configuration on systemic silencing, new GFP transgenic lines with or without the transposase sequences were produced. GFP expression levels in the 19 single-copy events and three hemizygous 16C lines produced for this study ranged from 50-72% of the homozygous 16C line. GFP expression was equivalent to 16C in a two-copy event. Local GFP silencing was observed in all transgenic and 16C hemizygous lines after topical application of delivery formulations with a GFP targeting dsRNA. The 16C-like systemic silencing phenotype was only observed in the two-copy line. The partial transposase had no impact on transgene expression level, local GFP silencing, small RNA abundance and distribution, or systemic GFP silencing in the transgenic lines. We conclude that high transgene expression level is a key enabler of systemic transgene silencing in N. benthamiana.

scholarly journals Isolation and Characterization of Barley (Hordeum vulgare) Extracellular Vesicles to Assess Their Role in RNA Spray-Based Crop Protection

2021 ◽  
Vol 22 (13) ◽  
pp. 7212
Author(s):  
Timo Schlemmer ◽  
Patrick Barth ◽  
Lisa Weipert ◽  
Christian Preußer ◽  
Martin Hardt ◽  
...  
Keyword(s):  
Hordeum Vulgare ◽  
Gene Silencing ◽  
Extracellular Vesicles ◽  
Fungal Pathogens ◽  
Target Genes ◽  
Crop Protection ◽  
Transgenic Expression ◽  
Isolation And Characterization ◽  
Sequence Complementarity

The demonstration that spray-induced gene silencing (SIGS) can confer strong disease resistance, bypassing the laborious and time-consuming transgenic expression of double-stranded (ds)RNA to induce the gene silencing of pathogenic targets, was ground-breaking. However, future field applications will require fundamental mechanistic knowledge of dsRNA uptake, processing, and transfer. There is increasing evidence that extracellular vesicles (EVs) mediate the transfer of transgene-derived small interfering (si)RNAs in host-induced gene silencing (HIGS) applications. In this study, we establish a protocol for barley EV isolation and assess the possibilities for EVs regarding the translocation of sprayed dsRNA from barley (Hordeum vulgare) to its interacting fungal pathogens. We found barley EVs that were 156 nm in size, containing predominantly 21 and 19 nucleotide (nts) siRNAs, starting with a 5′-terminal Adenine. Although a direct comparison of the RNA cargo between HIGS and SIGS EV isolates is improper given their underlying mechanistic differences, we identified sequence-identical siRNAs in both systems. Overall, the number of siRNAs isolated from the EVs of dsRNA-sprayed barley plants with sequence complementarity to the sprayed dsRNA precursor was low. However, whether these few siRNAs are sufficient to induce the SIGS of pathogenic target genes requires further research. Taken together, our results raise the possibility that EVs may not be mandatory for the spray-delivered siRNA uptake and induction of SIGS.

scholarly journals RNAi Crop Protection Advances

2021 ◽  
Vol 22 (22) ◽  
pp. 12148
Author(s):  
Alejandro Hernández-Soto ◽  
Randall Chacón-Cerdas
Keyword(s):  
Gene Silencing ◽  
Oral Delivery ◽  
Large Scale ◽  
Crop Protection ◽  
Scale Production ◽  
Recombinant Production ◽  
Rnai Technology ◽  
Large Scale Production ◽  
Induction Of Resistance ◽  
Multiple Chemical

RNAi technology is a versatile, effective, safe, and eco-friendly alternative for crop protection. There is plenty of evidence of its use through host-induced gene silencing (HIGS) and emerging evidence that spray-induced gene silencing (SIGS) techniques can work as well to control viruses, bacteria, fungi, insects, and nematodes. For SIGS, its most significant challenge is achieving stability and avoiding premature degradation of RNAi in the environment or during its absorption by the target organism. One alternative is encapsulation in liposomes, virus-like particles, polyplex nanoparticles, and bioclay, which can be obtained through the recombinant production of RNAi in vectors, transgenesis, and micro/nanoencapsulation. The materials must be safe, biodegradable, and stable in multiple chemical environments, favoring the controlled release of RNAi. Most of the current research on encapsulated RNAi focuses primarily on oral delivery to control insects by silencing essential genes. The regulation of RNAi technology focuses on risk assessment using different approaches; however, this technology has positive economic, environmental, and human health implications for its use in agriculture. The emergence of alternatives combining RNAi gene silencing with the induction of resistance in crops by elicitation and metabolic control is expected, as well as multiple silencing and biotechnological optimization of its large-scale production.

scholarly journals Spray-Induced Silencing of Pathogenicity Gene MoDES1 via Exogenous Double-Stranded RNA Can Confer Partial Resistance Against Fungal Blast in Rice

2021 ◽  
Vol 12 ◽  
Author(s):  
Atrayee Sarkar ◽  
Subhankar Roy-Barman
Keyword(s):  
Gene Silencing ◽  
Host Defense ◽  
Crop Protection ◽  
Defense Responses ◽  
Blast Disease ◽  
Ros Scavenging ◽  
Pathogenic Potential ◽  
Double Stranded Rna ◽  
Wide Range ◽  
Pathogenicity Gene

Over the past years, RNA interference (RNAi) has been used as a promising combat strategy against a wide range of pests and pathogens in ensuring global food security. It involves the induction of highly specific posttranscriptional regulation of target essential genes from an organism, via the application of precursor long, non-coding double-stranded RNA (dsRNA) molecules that share sequence-complementarity with the mRNAs of the targets. Fungal blast disease caused by Magnaporthe oryzae is one of the most deadly diseases of rice and wheat incurring huge losses in global crop yield. To date, the host-induced gene silencing (HIGS) and virus-induced gene silencing (VIGS) aspects of RNAi have been successfully exploited in developing resistance against M. oryzae in rice. Spray-induced gene silencing (SIGS) is a current, potential, non-transformative, and environment-friendly pest and pathogen management strategy, where naked or nanomaterial-bound dsRNA are sprayed on leaves to cause selective knockdown of pathogenicity genes. Although it relies on the ability of fungal pathogens to uptake sprayed RNA, its efficiency varies largely across phytopathogens and their genes, targeted for silencing. Here, we report a transient dsRNA supplementation system for the targeted knockdown of MoDES1, a host-defense suppressor pathogenicity gene from M. oryzae. We validate the feasibility of in vivo SIGS and post-uptake transfer of RNA signals to distal plant parts in rice-M. oryzae pathosystem through a GFP-based reporter system. A protocol for efficient silencing via direct foliar spray of naked dsRNA was optimized. As proof-of-concept, we demonstrate the phenotypic impacts of in vitro dsDES1 treatment on growth, conidiation, ROS-scavenging ability, and pathogenic potential of M. oryzae. Furthermore, our extrapolatory dsDES1 spray experiments on wounded leaves and whole rice plants indicate resultant silencing of MoDES1 that conferred significant resistance against the fungal blast disease. The evaluation of primary and secondary host defense responses provides evidence supporting the notion that spray of sequence-specific dsRNA on wounded leaf tissue can cause systemic and sustained silencing of a M. oryzae target gene. For the first time, we establish a transgene-free SIGS approach as a promising crop protection strategy against the notorious rice-blast fungus.

scholarly journals RNAi Crop Protection Advances

2021 ◽  
Author(s):  
Alejandro Hernández-Soto ◽  
Randall Chacón-Cerdas
Keyword(s):  
Gene Silencing ◽  
Oral Delivery ◽  
Large Scale ◽  
Crop Protection ◽  
Scale Production ◽  
Recombinant Production ◽  
Rnai Technology ◽  
Large Scale Production ◽  
Induction Of Resistance ◽  
Multiple Chemical

RNAi technology is a versatile, effective, safe, and eco-friendly alternative for crop protection. There is plenty of evidence of its use through Host-induced gene silencing (HIGS) and spray-induced gene silencing (SIGS) techniques to control viruses, bacteria, fungi, insects, and nematodes. As for SIGS, its most significant challenge is achieving stability and avoiding premature degradation of RNAi in the environment or during its absorption in the target organism. One alternative is the encapsulation in liposomes, virus-like particles, polyplex nanoparticles, and bio-clay, which can be obtained through the recombinant production of RNAi in vectors, transgenesis, and micro/nanoencapsulation. The materials must be safe, biodegradable, and stable in multiple chemical environments favoring the controlled release of RNAi. Most of the current research of encapsulated RNAi focuses primarily on oral delivery to control insects by silencing essential genes. The regulation of RNAi technology focuses on risk assessment from different approaches; however, this technology has positive characteristics for its use in agriculture from the economic, environmental, and human health implications. The emergence of alternatives combining RNAi gene silencing with the induction of resistance in crops by elicitation and metabolic control is expected, as well as multiple silencing and biotechnological optimization of its large-scale production.

scholarly journals RNAi Crop Protection Advances

2021 ◽  
Author(s):  
Alejandro Hernández-Soto ◽  
Randall Chacón-Cerdas
Keyword(s):  
Gene Silencing ◽  
Oral Delivery ◽  
Large Scale ◽  
Crop Protection ◽  
Scale Production ◽  
Recombinant Production ◽  
Rnai Technology ◽  
Large Scale Production ◽  
Induction Of Resistance ◽  
Multiple Chemical

RNAi technology is a versatile, effective, safe, and eco-friendly alternative for crop protection. There is plenty of evidence of its use through Host-induced gene silencing (HIGS) and spray-induced gene silencing (SIGS) techniques to control viruses, bacteria, fungi, insects, and nematodes. As for SIGS, its most significant challenge is achieving stability and avoiding premature degradation of RNAi in the environment or during its absorption in the target organism. One alternative is the encapsulation in liposomes, virus-like particles, polyplex nanoparticles, and bio-clay, which can be obtained through the recombinant production of RNAi in vectors, transgenesis, and micro/nanoencapsulation. The materials must be safe, biodegradable, and stable in multiple chemical environments favoring the controlled release of RNAi. Most of the current research of encapsulated RNAi focuses primarily on oral delivery to control insects by silencing essential genes. The regulation of RNAi technology focuses on risk assessment from different approaches; however, this technology has positive characteristics for its use in agriculture from the economic, environmental, and human health implications. The emergence of alternatives combining RNAi gene silencing with the induction of resistance in crops by elicitation and metabolic control is expected, as well as multiple silencing and biotechnological optimization of its large-scale production.

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