SummaryDespite long-standing plant breeding investments and early successes in genetic engineering, plant viral pathogens still cause major losses in agriculture worldwide.Early transgenic approaches involved the expression of pathogen-derived sequences that provided limited protection against relatively narrow ranges of viral pathotypes. In contrast, this study demonstrates that the ectopic expression of pvr1 , a recessive gene from Capsicum chinense , results in dominant broad-spectrum potyvirus resistance in transgenic tomato plants ( Solanum lycopersicum ). The pvr1 locus in pepper encodes the eukaryotic translation initiation factor eIF4E. Naturally occurring point mutations at this locus result in monogenic recessive broad-spectrum potyvirus resistance that has been globally deployed via plant breeding programmes for more than 50 years. Transgenic tomato progenies that over-expressed the Capsicum pvr1 allele showed dominant resistance to several tobacco etch virus strains and other potyviruses, including pepper mottle virus, a range of protection similar to that observed in pepper homozygous for the pvr1 allele.
Summary Natural mutations in translation initiation factor eIF4E confer resistance to potyviruses in many plant species. Potato is a staple food crop plagued by several potyviruses, yet to date no known eIF4E‐mediated resistance genes have been identified. In this study, we demonstrate that transgenic expression of the pvr12 gene from pepper confers resistance to Potato virus Y (PVY) in potato. We then use this information to convert the susceptible potato ortholog of this allele into a de novo allele for resistance to PVY using site‐directed mutagenesis. Potato plants overexpressing the mutated potato allele are resistant to virus infection. Resistant lines expressed high levels of eIF4E mRNA and protein. The resistant plants showed growth similar to untransformed controls and produced phenotypically similar tubers. This technique disrupts a key step in the viral infection process and may potentially be used to engineer virus resistance in a number of economically important plant–viral pathosystems. Furthermore, the general public may be more amenable to the ‘intragenic’ nature of this approach because the transferred coding region is modified from a gene in the target crop rather than from a distant species.
Background: Environmental heterogeneity, emerging pathogens and limited access to financial and agricultural inputs prevent farmers from producing consistent crop yields in many developing countries. Yield instability impedes establishment of processing and export industries, thereby inhibiting economic development. Modern varieties offer significant increases in yield stability. Results:A deeply collaborative, multi-national germplasm trialing network was established in West Africa to identify tomato varieties well adapted to each country and mobilize those varieties into local seed distribution networks alongside an integrated pest management program. Research partners in seven West African countries evaluated over 100 tomato varieties for resistance to tomato leaf curl disease (ToLCD). Using biotechnology, the identity and distribution of the key viruses (begomoviruses) causing ToLCD in these countries were identified, and a vector-independent inoculation method (agroinoculation) was developed. The trials identified a set of high-performing varieties with resistance. Agroinoculation with the three prevalent begomoviruses confirmed resistance. Conclusions:These trial results fulfill the new Economic Community of West African States harmonized seed regulation policy, which requires at least 2 years of national performance trials prior to commercialization of a tomato variety. To compete with a rapidly expanding canned tomato import industry, West African growers need to increase productivity and processing capacity; therefore, we also assessed processing and export trade data for fresh and processed tomatoes from each of the seven countries.
Potyvirus resistance in Capsicum spp. has been attributed to amino acid substitutions at the pvr1 locus that cause conformational shifts in eukaryotic translation initiation factor eIF4E. The viral genome-linked protein (VPg) sequence was isolated and compared from three Tobacco etch virus (TEV) strains, highly aphid-transmissible (HAT), Mex21, and N, which differentially infect Capsicum genotypes encoding Pvr1(+), pvr1, and pvr1(2). Viral chimeras were synthesized using the TEV-HAT genome, replacing HAT VPg with Mex21 or N VPg. TEV HAT did not infect pepper plants homozygous for either the pvr1 or pvr1(2) allele. However, the novel chimeric TEV strains, TEVHAT(Mex21-VPg) and TEV-HAT(N-VPg), infected pvr1 and pvr1(2) pepper plants, respectively, demonstrating that VPg is the virulence determinant in this pathosystem. Three dimensional structural models predicted interaction between VPg and the susceptible eIF4E genotype in every case, while resistant genotypes were never predicted to interact. To determine whether there is a correlation between physical interaction of VPg with eIF4E and infectivity, the effects of amino acid variation within VPg were assessed. Interaction between pvr1(2) eIF4E and N VPg was detected in planta, implying that the six amino acid differences in N VPg relative to HAT VPg are responsible for restoring the physical interaction and infectivity.
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