Molecular characterization and analysis of conserved potyviral motifs in bean common mosaic virus (BCMV) for RNAi-mediated protection

Worrall, Elizabeth A.; Hayward, Alice; Fletcher, Stephen J.; Mitter, Neena

doi:10.1007/s00705-018-4065-6

Cited by 29 publications

(16 citation statements)

References 58 publications

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“…Recently, we designed and tested RNAi-inducing dsRNA molecules using transient expression via agroinfiltration in leaves of Nicotiana benthamiana (Worrall et al., 2019). Using two Australian BCMV isolates (BCMV 1 and 2), it was found that the most effective sequences to target in the BCMV genomic RNA for effective induction of RNAi-mediated resistance lie within the sequences encoding the nuclear inclusion b (NIb) protein and the CP (Worrall et al., 2019). To target these regions of the BCMV genome using BioClay, two BCMV-specific dsRNAs were synthesized.…”

Section: Resultsmentioning

confidence: 99%

“…BCMVNIb-dsRNA was designed to target a 480 nt sequence within the BCMV NIb protein coding region and BCMVCP-dsRNA which targets 461 nt of the CP coding region. Due to the constraints of the dsRNA production, BCMVNIb-dsRNA (480 nt) was shorter than the 920 nt RNAi construct expressed in the transient assay experiments (Worrall et al., 2019). BCMVNIb-dsRNA was designed to target the sequence spanning 7,865–8,344 nt of the BCMV 1 genome.…”

Section: Resultsmentioning

confidence: 99%

“…The Australian isolate BCMV 1 (GenBank accession MH220847) was revived (from freeze-dried leaf samples) and propagated in N. benthamiana (Worrall et al., 2019). Routine passaging and challenge of dsRNA-treated N. benthamiana and cowpea plants were conducted by mechanical inoculation using 50 μl sap from systemically infected N. benthamiana plants diluted 1:1500 in 0.01 M phosphate (pH 7.0) inoculation buffer with Carborundum used as an abrasive (Hamid et al., 2013).…”

Section: Methodsmentioning

confidence: 99%

“…Chemically synthesized dsRNA (Genolution Inc., Seoul, Republic of Korea) targeted to either 480 bp of the BCMV NIb coding region (7,865–8,344 nt GenBank accession MH220847) or 461 bp of the BCMV CP coding region (9,312–9,772 nt GenBank accession MH220847). BCMVNIb-dsRNA bioinformatic analysis of conserved motifs, mean pairwise percent identity and potential for 21 and 22 nt small-interfering RNAs was conducted as per Worrall et al. (2019).…”

Section: Methodsmentioning

confidence: 99%

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Exogenous Application of RNAi-Inducing Double-Stranded RNA Inhibits Aphid-Mediated Transmission of a Plant Virus

et al. 2019

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Plant viruses are difficult to control, and they decrease both the quality and yield of crops, thus threatening global food security. A new approach that uses topical application of double-stranded RNA (dsRNA) to induce antiviral RNA-interference has been shown to be effective at preventing virus infection in a range of plants following mechanical inoculation. In this study, topical application of dsRNA was effective against mechanical inoculation and aphid-mediated inoculation with the potyvirus bean common mosaic virus (BCMV). Topical application of dsRNAs targeting either the coding region of the potyviral nuclear inclusion b (NIb) protein (BCMVNIb-dsRNA) or the coat protein (CP) coding region (BCMVCP-dsRNA) protected Nicotiana benthamiana and cowpea (Vigna unguiculata) plants against mechanical inoculation with BCMV. BCMVCP-dsRNA was selected for subsequent aphid transmission experiments. BCMVCP-dsRNA was loaded onto layered double hydroxide nanoparticles to form BCMVCP-BioClay which is a more stable formulation for delivering dsRNA than naked dsRNA. BCMVCP-BioClay was shown to be successful in protecting plants against BCMV transmission by the aphid Myzus persicae. Spraying detached N. benthamiana leaves with BCMVCP-BioClay 5 days prior to exposure to viruliferous aphids protected the leaves from infection by BCMV. Importantly, spraying of intact N. benthamiana and cowpea plants with BCMVCP-BioClay 5 days prior to exposure to viruliferous aphids protected plants of both species from BCMV infection. This study demonstrates that topical application of dsRNA using BioClay protects plants from aphid-mediated virus transmission, which is an important first step toward developing practical application of this approach in crop protection.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Exogenous Application of RNAi-Inducing Double-Stranded RNA Inhibits Aphid-Mediated Transmission of a Plant Virus

et al. 2019

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“…The 3Dpol, conserved among animal picornavirues, is also structurally arranged in a right-hand orientation, and contains seven conserved motifs (A-G)) [38]. Motifs A, B, D, and E are critical for the recognition and binding of nucleoside triphosphates; A and G for the binding of metal ions and the transfer of phosphoryl groups; D for the structural integrity of the palm; E for the binding of the priming nucleotide; and B, F, and G for the binding of the template [38][39][40] Figure 1B) [41][42][43]. The NIb of the potyvirus TEV, the poliovirus 3Dpol (a picornavirus), and the RdRp of cowpea mosaic virus (CPMV, a comovirus)) share the highest sequence homology in two clusters, (T/S) GXXXTXXXN (T/S) and GDD [44].…”

Section: Structural Characterization Of Nibmentioning

confidence: 99%

The RNA-Dependent RNA Polymerase NIb of Potyviruses Plays Multifunctional, Contrasting Roles during Viral Infection

Shen

Shi

Dai

et al. 2020

Viruses

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Potyviruses represent the largest group of known plant RNA viruses and include many agriculturally important viruses, such as Plum pox virus, Soybean mosaic virus, Turnip mosaic virus, and Potato virus Y. Potyviruses adopt polyprotein processing as their genome expression strategy. Among the 11 known viral proteins, the nuclear inclusion protein b (NIb) is the RNA-dependent RNA polymerase responsible for viral genome replication. Beyond its principal role as an RNA replicase, NIb has been shown to play key roles in diverse virus–host interactions. NIb recruits several host proteins into the viral replication complexes (VRCs), which are essential for the formation of functional VRCs for virus multiplication, and interacts with the sumoylation pathway proteins to suppress NPR1-mediated immunity response. On the other hand, NIb serves as a target of selective autophagy as well as an elicitor of effector-triggered immunity, resulting in attenuated virus infection. These contrasting roles of NIb provide an excellent example of the complex co-evolutionary arms race between plant hosts and potyviruses. This review highlights the current knowledge about the multifunctional roles of NIb in potyvirus infection, and discusses future research directions.

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Induction of RNA-Mediated Resistance Against Cucumber Mosaic Virus on Exogenous Application of dsRNA

Saikia

Routhu

Borah

2023

Technologies for Rural Development

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Molecular characterization and analysis of conserved potyviral motifs in bean common mosaic virus (BCMV) for RNAi-mediated protection

Cited by 29 publications

References 58 publications

Exogenous Application of RNAi-Inducing Double-Stranded RNA Inhibits Aphid-Mediated Transmission of a Plant Virus

Exogenous Application of RNAi-Inducing Double-Stranded RNA Inhibits Aphid-Mediated Transmission of a Plant Virus

The RNA-Dependent RNA Polymerase NIb of Potyviruses Plays Multifunctional, Contrasting Roles during Viral Infection

Induction of RNA-Mediated Resistance Against Cucumber Mosaic Virus on Exogenous Application of dsRNA

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