Targeted Delivery of Gene Silencing in Fungi Using Genetically Engineered Bacteria

Niño‐Sánchez, Jonatan; Chen, Li-Hung; Souza, Jorge Teodoro de; Mosquera, Sandra; Stergiopoulos, Ioannis

doi:10.3390/jof7020125

Cited by 16 publications

(9 citation statements)

References 67 publications

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“…SIGS constitutes an environmentally friendly approach for the control of fungal plant diseases. However, its commercial application is still limited due to the instability of dsRNA under field conditions (Cagliari et al, 2019; Dalakouras et al, 2020; Niño‐Sánchez et al, 2021). This work investigated using LDH as an RNA carrier, extending on previous studies which have shown that LDH‐bound dsRNA is protected from environmental degradation (Mitter et al, 2017; Jain et al, 2022).…”

Section: Discussionmentioning

confidence: 99%

BioClay™ prolongs RNA interference‐mediated crop protection against Botrytis cinerea

Niño‐Sánchez

Sambasivam

Sawyer

et al. 2022

JIPB

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One of the most promising tools for the control of fungal plant diseases is spray‐induced gene silencing (SIGS). In SIGS, small interfering RNA (siRNA) or double‐stranded RNA (dsRNA) targeting essential or virulence‐related pathogen genes are exogenously applied to plants and postharvest products to trigger RNA interference (RNAi) of the targeted genes, inhibiting fungal growth and disease. However, SIGS is limited by the unstable nature of RNA under environmental conditions. The use of layered double hydroxide or clay particles as carriers to deliver biologically active dsRNA, a formulation termed BioClay™, can enhance RNA durability on plants, prolonging its activity against pathogens. Here, we demonstrate that dsRNA delivered as BioClay can prolong protection against Botrytis cinerea, a major plant fungal pathogen, on tomato leaves and fruit and on mature chickpea plants. BioClay increased the protection window from 1 to 3 weeks on tomato leaves and from 5 to 10 days on tomato fruits, when compared with naked dsRNA. In flowering chickpea plants, BioClay provided prolonged protection for up to 4 weeks, covering the critical period of poding, whereas naked dsRNA provided limited protection. This research represents a major step forward for the adoption of SIGS as an eco‐friendly alternative to traditional fungicides.

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Section: Discussionmentioning

confidence: 99%

BioClay™ prolongs RNA interference‐mediated crop protection against Botrytis cinerea

Niño‐Sánchez

Sambasivam

Sawyer

et al. 2022

JIPB

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“…In addition to inorganic and organic carriers, another delivery method being investigated is the use of vectors that have been genetically modified to produce dsRNAs. Niño-Sánchez et al first demonstrated that silencing can be induced by the application of living or lysed bacteria engineered to produce biologically active dsRNAs targeting fungal genes [ 215 ]. Further development and improvement of methods to minimize the degradation of dsRNA molecules and to improve their stability and durability are needed to allow this SIGS to be used in the field.…”

Section: Rna Interference (Rnai): the Targeted Management For Plant D...mentioning

confidence: 99%

“…Thus, large quantities of dsRNA can be produced with bacteria-based systems at a cost-effective price. Moreover, the application of living dsRNA-producing bacteria for gene silencing has been used in nematodes [ 160 ], insects [ 225 ], crustaceans [ 226 ], mammalian cells [ 227 ], and filamentous fungi [ 215 ]. However, in order to avoid bacterial release into the environment, it is necessary to purify the RNA, which can make the process more expensive and limit the amount of dsRNAs produced [ 26 , 228 , 229 , 230 , 231 , 232 ].…”

Section: Rna Interference (Rnai): the Targeted Management For Plant D...mentioning

confidence: 99%

New Insights on the Integrated Management of Plant Diseases by RNA Strategies: Mycoviruses and RNA Interference

Bocos-Asenjo

Niño‐Sánchez

Ginésy

2022

IJMS

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RNA-based strategies for plant disease management offer an attractive alternative to agrochemicals that negatively impact human and ecosystem health and lead to pathogen resistance. There has been recent interest in using mycoviruses for fungal disease control after it was discovered that some cause hypovirulence in fungal pathogens, which refers to a decline in the ability of a pathogen to cause disease. Cryphonectria parasitica, the causal agent of chestnut blight, has set an ideal model of management through the release of hypovirulent strains. However, mycovirus-based management of plant diseases is still restricted by limited approaches to search for viruses causing hypovirulence and the lack of protocols allowing effective and systemic virus infection in pathogens. RNA interference (RNAi), the eukaryotic cell system that recognizes RNA sequences and specifically degrades them, represents a promising. RNA-based disease management method. The natural occurrence of cross-kingdom RNAi provides a basis for host-induced gene silencing, while the ability of most pathogens to uptake exogenous small RNAs enables the use of spray-induced gene silencing techniques. This review describes the mechanisms behind and the potential of two RNA-based strategies, mycoviruses and RNAi, for plant disease management. Successful applications are discussed, as well as the research gaps and limitations that remain to be addressed.

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“…Foliar sprays, trunk injections, root drench, or delivery from clay granules can also be used for sustained release of dsRNA ( Ghosh et al, 2018 ). Another promising approach, exemplified in some of the studies described below, is to use transgenic bacteria as a source of sRNA ( Niño-Sánchez et al, 2021 ).…”

Section: Spray-induced Gene Silencingmentioning

confidence: 99%

Small RNA-based plant protection against diseases

Bilir¹,

Göl

Hong

et al. 2022

Front. Plant Sci.

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Plant diseases cause significant decreases in yield and quality of crops and consequently pose a very substantial threat to food security. In the continuous search for environmentally friendly crop protection, exploitation of RNA interferance machinery is showing promising results. It is well established that small RNAs (sRNAs) including microRNA (miRNA) and small interfering RNA (siRNA) are involved in the regulation of gene expression via both transcriptional and post-transcriptional RNA silencing. sRNAs from host plants can enter into pathogen cells during invasion and silence pathogen genes. This process has been exploited through Host-Induced Gene Silencing (HIGS), in which plant transgenes that produce sRNAs are engineered to silence pest and pathogen genes. Similarly, exogenously applied sRNAs can enter pest and pathogen cells, either directly or via the hosts, and silence target genes. This process has been exploited in Spray-Induced Gene Silencing (SIGS). Here, we focus on the role of sRNAs and review how they have recently been used against various plant pathogens through HIGS or SIGS-based methods and discuss advantages and drawbacks of these approaches.

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Targeted Delivery of Gene Silencing in Fungi Using Genetically Engineered Bacteria

Cited by 16 publications

References 67 publications

BioClay™ prolongs RNA interference‐mediated crop protection against Botrytis cinerea

BioClay™ prolongs RNA interference‐mediated crop protection against Botrytis cinerea

New Insights on the Integrated Management of Plant Diseases by RNA Strategies: Mycoviruses and RNA Interference

Small RNA-based plant protection against diseases

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