Production of siRNA targeted against TYLCV coat protein transcripts leads to silencing of its expression and resistance to the virus

Zrachya, Avi; Kumar, Pravin; Usha, R.; Levy, Yael; Loyter, Abraham; Arazi, Tzahi; Lapidot, Moshe; Gafni, Yedidya

doi:10.1007/s11248-006-9042-2

Cited by 77 publications

(45 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…This approach led to a high level of resistance to all three strains (Abhary et al 2006). Later, Gafni and colleagues obtained plants resistant to TYLCV by targeting the CP gene with an inverted-repeat construct (Zrachya et al 2007b). …”

Section: Genetically Engineered Resistancementioning

confidence: 99%

The viral etiology of tomato yellow leaf curl disease - a review

Glick¹,

Levy²,

Gafni³

2009

Plant Prot. Sci.

View full text Add to dashboard Cite

Glick E., Levy Y., Gafni Y. (2009): The viral etiology of tomato yellow leaf curl disease -a review. Plant Protect. Sci., 45: 81-97.Tomato yellow leaf curl disease (TYLCD) is one of the most devastating plant diseases in the world. As a result of its continuing rapid spread, it now afflicts more than 30 tomato growing countries in the Mediterranean basin, southern Asia, Africa, and South, Central and North America. The disease is caused by a group of viral species of the genus Begomovirus, family Geminiviridae (geminiviruses), referred to as Tomato yellow leaf curl virus (TYLCV). These are transmitted by an insect vector, the whitefly Bemisia tabaci, classified in the family Aleyrodidae. The genome of TYLCV generally consists of a single circular single-stranded (ss) DNA molecule, with only one exception in which two components were identified. It encodes six open reading frames, only one of which codes for the coat protein (CP) that represents a building block of the viral particle. TYLCV, like all other members of the Geminiviridae, has geminate particles, apparently consisting of two incomplete T = 1 icosahedra joined together to produce a structure with 22 pentameric capsomers and 110 identical CP subunits. Close to 50 years of intensive research into TYLCV epidemics has been conducted to find solutions to the severe problem caused by this virus. To date, breeding for resistance appears to be the best approach to controlling this disease, although only partially resistant varieties are commercially available. Since the virus consists of a ssDNA that replicates in the host-cell nucleus, the molecular mechanisms involved in its nuclear import have been the focus of our studies in recent years and results, as well as prospects, are discussed in this review. In addition, we describe our recent finding of a suppressor of gene silencing encoded by one of the TYLCV-Isr genes. This paper provides an overview of the most outstanding achievements in TYLCV research that may lead to more effective control strategies.

show abstract

Section: Genetically Engineered Resistancementioning

confidence: 99%

The viral etiology of tomato yellow leaf curl disease - a review

Glick¹,

Levy²,

Gafni³

2009

Plant Prot. Sci.

View full text Add to dashboard Cite

show abstract

“…Such a resistance strategy, where the transgene contained different genome fragments, has successfully been demonstrated for TYLCV, TYLCV-Mld and TYLCSV in N. benthamiana inoculated by agroinfiltration [40] and resistance to RNA [41] and DNA viruses [42]. In this study, we screened a relatively large number of independent transgenic lines, compared to previous reports, for TYLCV resistance [14][15][16] and we showed, for the first time, immunity of eight tomato lines associated with transgene-siRNA accumulation, as was previously shown with TYLCV with hpRNA for only one line [14]. The immunity of lines 417 and 450 was maintained against the closely related virus TYLCV-Mld, with 58, 100 and 100 % sequence identity to the transgene sequences of IR, V1-V2 and C1-C2, respectively.…”

Section: Size and Distribution Of T-sirna In Transgenic Rootstock Andmentioning

confidence: 99%

“…Several approaches to transgenic pathogen-derived resistance were examined in the case of TYLCV: a. coat-protein-mediated Diana Leibman, Shanmugam Prakash and Dalia Wolf have contributed equally to this work. [7], b. antisense RNA [8,9], c. defective-interfering (DI) of replication initiator protein (Rep) [9][10][11][12][13][14], d. hairpin-RNA (hpRNA) (dsRNA expression)-induced gene silencing [14][15][16] and e. peptide aptamers that suppressed virus replication [17]. In most of these strategies, transgenic plants provided a wide range of resistance to TYLCV; the hpRNA approach was most effective in triggering high levels of TYLCV resistance [15,18].…”

Section: Introductionmentioning

confidence: 99%

Immunity to tomato yellow leaf curl virus in transgenic tomato is associated with accumulation of transgene small RNA

et al. 2015

Self Cite

View full text Add to dashboard Cite

Gene silencing is a natural defense response of plants against invading RNA and DNA viruses. The RNA post-transcriptional silencing system has been commonly utilized to generate transgenic crop plants that are "immune" to plant virus infection. Here, we applied this approach against the devastating DNA virus tomato yellow leaf curl virus (TYLCV) in its host tomato (Solanum lycopersicum L.). To generate broad resistance to a number of different TYLCV viruses, three conserved sequences (the intergenic region [NCR], V1-V2 and C1-C2 genes) from the genome of the severe virus (TYLCV) were synthesized as a single insert and cloned into a hairpin configuration in a binary vector, which was used to transform TYLCV-susceptible tomato plants. Eight of 28 independent transgenic tomato lines exhibited immunity to TYLCV-Is and to TYLCV-Mld, but not to tomato yellow leaf curl Sardinia virus, which shares relatively low sequence homology with the transgene. In addition, a marker-free (nptII-deleted) transgenic tomato line was generated for the first time by Agrobacterium-mediated transformation without antibiotic selection, followed by screening of 1180 regenerated shoots by whitefly-mediated TYLCV inoculation. Resistant lines showed a high level of transgene-siRNA (t-siRNA) accumulation (22% of total small RNA) with dominant sizes of 21 nt (73%) and 22 nt (22%). The t-siRNA displayed hot-spot distribution ("peaks") along the transgene, with different distribution patterns than the viral-siRNA peaks observed in TYLCV-infected tomato. A grafting experiment demonstrated the mobility of 0.04% of the t-siRNA from transgenic rootstock to non-transformed scion, even though scion resistance against TYLCV was not achieved.

show abstract

“…Apart from these approaches, usage of non-coding viral RNAs [i.e. smallRNAs, antisense RNA and double-stranded (ds) RNAs homologous] have also shown to be a potential method for providing virus resistance in bean [5], blackgram [23] cassava [38], tobacco [4] and tomato [9,25,40]. In spite of all these approaches, very few promising examples of crops are available till date which have been released for cultivation and have shown resistance against broad spectrum viruses.…”

Section: Introductionmentioning

confidence: 99%

Application of molecular antiviral compounds: novel approach for durable resistance against geminiviruses

Sahu

Prasad

2015

Mol Biol Rep

View full text Add to dashboard Cite

Both transgenic as well as traditional breeding approaches have not been completely successful in inducting resistance against geminiviruses in crop plants. This demands the utilization of non-viral, non-plant compounds possessing antiviral characteristics as an alternate and effective strategy for developing durable resistance against geminiviruses. In recent years, several antiviral molecules have been developed for the treatment of plant virus infections. These molecular antiviral compounds target various geminiviral-DNA and -protein via interacting with them or by cleaving viral RNA fragments. Applications of these proteins such as GroEL, g5g and VirE2 have also provided a convincing evidence of resistance against geminiviruses. Taking advantage of this information, we can generate robust resistance against geminiviruses in diverse crop plants. In this context, the present review provides epigrammatic information on these antiviral compounds and their mode of action in modulating virus infection.

show abstract

Production of siRNA targeted against TYLCV coat protein transcripts leads to silencing of its expression and resistance to the virus

Cited by 77 publications

References 42 publications

The viral etiology of tomato yellow leaf curl disease - a review

The viral etiology of tomato yellow leaf curl disease - a review

Immunity to tomato yellow leaf curl virus in transgenic tomato is associated with accumulation of transgene small RNA

Application of molecular antiviral compounds: novel approach for durable resistance against geminiviruses

Contact Info

Product

Resources

About