Polycistronic artificial <scp>miRNA</scp>‐mediated resistance to <scp><i>W</i></scp><i>heat dwarf virus</i> in barley is highly efficient at low temperature

Kis, András; Tholt, Gergely; Ivanics, M.; Várallyay, Éva; Jenes, Barnabás; Havelda, Zoltán

doi:10.1111/mpp.12291

Cited by 88 publications

(51 citation statements)

References 47 publications

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“…The multi‐targeting of plant viruses through artificial sRNAs was first achieved with transgenes expressing multiple amiRNAs from different precursors in cis (Ai et al ., ; Kung et al ., ) or in trans (Lafforgue et al ., ), or from a single polycistronic precursor (Fahim et al ., ; Kis et al ., ), and more recently with transgenes expressing multiple syn‐tasiRNAs from TAS1c (Carbonell and Daros, ; Carbonell et al ., ) or TAS3a precursors (Chen et al ., ), although neither the durability of the resistance nor the comparison with single targeting was systematically studied. Here, we analyzed the anti‐TSWV effects of two artificial sRNA constructs in parallel, one expressing an amiRNA and another expressing four syn‐tasiRNAs, stably expressed in S. lycopersicum .…”

Section: Discussionmentioning

confidence: 99%

Multi‐targeting of viral RNAs with synthetic trans‐acting small interfering RNAs enhances plant antiviral resistance

2019

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Summary RNA interference (RNAi)‐based tools are used in multiple organisms to induce antiviral resistance through the sequence‐specific degradation of target RNAs by complementary small RNAs. In plants, highly specific antiviral RNAi‐based tools include artificial microRNAs (amiRNAs) and synthetic trans‐acting small interfering RNAs (syn‐tasiRNAs). syn‐tasiRNAs have emerged as a promising antiviral tool allowing for the multi‐targeting of viral RNAs through the simultaneous expression of several syn‐tasiRNAs from a single precursor. Here, we compared in tomato plants the effects of an amiRNA construct expressing a single amiRNA and a syn‐tasiRNA construct expressing four different syn‐tasiRNAs against Tomato spotted wilt virus (TSWV), an economically important pathogen affecting tomato crops worldwide. Most of the syn‐tasiRNA lines were resistant to TSWV, whereas the majority of the amiRNA lines were susceptible and accumulated viral progenies with mutations in the amiRNA target site. Only the two amiRNA lines with higher amiRNA accumulation were resistant, whereas resistance in syn‐tasiRNA lines was not exclusive of lines with high syn‐tasiRNA accumulation. Collectively, these results suggest that syn‐tasiRNAs induce enhanced antiviral resistance because of the combined silencing effect of each individual syn‐tasiRNA, which minimizes the possibility that the virus simultaneously mutates all different target sites to fully escape each syn‐tasiRNA.

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Section: Discussionmentioning

confidence: 99%

Multi‐targeting of viral RNAs with synthetic trans‐acting small interfering RNAs enhances plant antiviral resistance

2019

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“…Indeed, a multiplex amiRNA in which five miR395s in the polycistronic miRNA backbone were replaced with amiRNAs from different Wheat streak mosaic tritimovirus (WSMV) genome regions conferred immunity to WSMV in transgenic plants (Fahim et al, 2012). Similarly, the polycistronic precursor of miR171 was used to produce three amiRNAs that targeted conserved segments in the Wheat dwarf mastrevirus (WDV) genome (Kis et al, 2016). Even though amiRNAs-derived resistance has not been reported in begomoviruses, it is likely that this approach would generate resistance in these viruses, especially if the amiRNA is designed to target the Rep gene, which is the only gene indispensable for viral replication.…”

Section: New Genomic Approaches In the Control Of Cmgsmentioning

confidence: 99%

The Search for Resistance to Cassava Mosaic Geminiviruses: How Much We Have Accomplished, and What Lies Ahead

Fondong

2017

Front. Plant Sci.

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The cassava mosaic disease (CMD), which occurs in all cassava growing regions of Africa and the Indian subcontinent, is caused by cassava mosaic geminiviruses (CMGs). CMGs are considered to be the most damaging vector-borne plant pathogens. So far, the most successful approach used to control these viruses has been the transfer of a polygenic recessive resistance locus, designated CMD1, from wild cassava to cassava cultivars. Further progress in harnessing natural resistance to contain CMGs has come from the discovery of the dominant monogenic resistance locus, CMD2, in some West African cassava cultivars. CMD2 has been combined with CMD1 through genetic crosses. Because of the limitations of the cassava breeding approach, especially with regard to time required to produce a variety and the loss of preferred agronomic attributes, efforts have been directed toward the deployment of genetic engineering approaches. Most of these approaches have been centered on RNA silencing strategies, developed mainly in the model plant Nicotiana benthamiana. Early RNA silencing platforms assessed for CMG resistance have been use of viral genes for co-suppression, antisense suppression or for hairpin RNAs-mediated gene silencing. Here, progress and challenges in the deployment of these approaches in the control of CMGs are discussed. Novel functional genomics approaches with potential to overcome some of the drawbacks of the current strategies are also discussed.

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“…Resistance was assessed over two generations. The same polycistronic type of vector, which was based on a barley miRNA precursor backbone and targeted different conservative sequences of Wheat dwarf virus ( WDV ), conferred barley resistance to the virus at low temperature (Kis et al, 2016). Recent research indicated that synthetic/artificial trans-acting siRNAs (syn-tasiRNAs) carrying multiple virus resistance cassettes might be more effective than amiRNA for enhanced antiviral defense (Carbonell et al, 2016; Chen et al, 2016).…”

Section: Introductionmentioning

confidence: 99%

Artificial MicroRNA-Based Specific Gene Silencing of Grain Hardness Genes in Polyploid Cereals Appeared to Be Not Stable Over Transgenic Plant Generations

et al. 2017

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Gene silencing by RNA interference is a particularly important tool in the study of gene function in polyploid cereal species for which the collections of natural or induced mutants are very limited. Previously we have been testing small interfering RNA-based approach of gene silencing in wheat and triticale. In this research, artificial microRNAs (amiRs) were studied in the same species and the same target genes to compare effectiveness of both gene silencing pathways. amiR cassettes were designed to silence Puroindoline a (Pina) and Puroindoline b (Pinb) hardness genes in wheat and their orthologues Secaloindoline a (Sina) and Secaloindoline b (Sinb) genes in triticale. Each of the two cassettes contained 21 nt microRNA (miR) precursor derived from conserved regions of Pina/Sina or Pinb/Sinb genes, respectively. Transgenic plants were obtained with high efficiency in two cultivars of wheat and one cultivar of triticale after using the Pinb-derived amiR vector for silencing of Pinb or Sinb, respectively. Lack of transgenic plants in wheat or very low transformation efficiency in triticale was observed using the Pina-derived amiR cassette, despite large numbers of embryos attempted. Silencing of Pinb in wheat and Sinb in triticale was highly efficient in the T1 generation. The transcript level of Pinb in wheat was reduced up to 92% and Sinb in triticale was reduced up to 98%. Moreover, intended silencing of Pinb/Sinb with Pinb-derived amiR cassette was highly correlated with simultaneous silencing of Pina/Sina in the same transgenic plants. High downregulation of Pinb/Pina genes in T1 plants of wheat and Sinb/Sina genes in T1 plants of triticale was associated with strong expression of Pinb-derived amiR. Silencing of the target genes correlated with increased grain hardness in both species. Total protein content in the grains of transgenic wheat was significantly lower. Although, the Pinb-derived amiR cassette was stably inherited in the T2 generation of wheat and triticale the silencing effect including strongly decreased expression of silenced genes as well as strong expression of Pinb-derived amiR was not transmitted. Advantages and disadvantages of posttranscriptional silencing of target genes by means of amiR and siRNA-based approaches in polyploid cereals are discussed.

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Polycistronic artificial miRNA‐mediated resistance to Wheat dwarf virus in barley is highly efficient at low temperature

Cited by 88 publications

References 47 publications

Multi‐targeting of viral RNAs with synthetic trans‐acting small interfering RNAs enhances plant antiviral resistance

Multi‐targeting of viral RNAs with synthetic trans‐acting small interfering RNAs enhances plant antiviral resistance

The Search for Resistance to Cassava Mosaic Geminiviruses: How Much We Have Accomplished, and What Lies Ahead

Artificial MicroRNA-Based Specific Gene Silencing of Grain Hardness Genes in Polyploid Cereals Appeared to Be Not Stable Over Transgenic Plant Generations

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