Resistencia a Phytophthora infestans linaje clonal EC-1 en Solanum tuberosum mediante la introducción del gen RB

Abstract

Resistencia de Solanum tuberoSum a PhytoPhthora infeStanS por la introducción del gen RB Los autores no incurren en conflictos de intereses. ResumenUna de las opciones para el control de la enfermedad del tizón tardío de la papa es el desarrollo de variedades resistentes a Phytophthora infestans mediante la transferencia directa de genes de resistencia (R) por ingeniería genética. En el siguiente trabajo de investigación, se usó el gen RB de Solanum bulbocastanum, el cual otorga un amplio espectro de resistencia a razas de P. infestans. Para dicho fin, se transformó genéticamente vía Agrobacterium la variedad susceptible de papa Desiree (Solanum tuberosum) con el vector binario pCIP68 que contiene el gen RB. Como resultado, se obtuvieron 19 plantas transformadas con el gen RB, confirmadas por la prueba de resistencia a kanamicina y por la prueba de reacción en cadena de la polimerasa (PCR). Las 19 plantas transgénicas fueron sometidas a infección en invernadero bajo condiciones de bioseguridad con el aislamiento POX067 de P. infestans perteneciente al linaje clonal EC-1 que es dominante en el Perú. Tres AbstractOne of the most efficient options for the control of late blight disease in potato (Solanum tuberosum) is the development of resistant varieties to Phytophthora infestans mediated by the direct transfer of resistance (R) genes through genetic engineering. In the present work, we used Solanum bulbocastanum RB gene to confers broad spectrum resistance to P. infestans races. To that end, Agrobacterium -mediated genetic transformation was used to transform a susceptible potato variety, Desiree, with the binary vector pCIP68 harboring the RB gene. As a result, 19 transformed plants containing the RB gene were obtained. kanamycin resistance test and polymerase chain reaction (PCR) assays confirmed the integration of the T-DNA in the potato genome. The 19 transformed plants, also called transgenic events were subjected to infection under biosafety greenhouse conditions. Phytophthora infestans isolate POX067 of the EC-1 clonal lineage, commonly find in Peru, was used for the infection. Three of the 19 plants ( [RB]

Stacking three late blight resistance genes from wild species directly into African highland potato varieties confers complete field resistance to local blight races

“…Transformation efficiency of ‘Desiree’ (3.4%) was not affected by the relatively large size of the T‐DNA (18 585 bp). Indeed, transformation efficiency of ‘Desiree’ with single R gene construct was previously reported as 3.7% for RB gene, 6% for Rpi‐blb2 gene and 7.5% for Rpi‐vnt1.1 gene (Orbegozo et al ., ; Román et al ., , ). The variety ‘Victoria’ had lower transformation efficiency (1.5%) and took longer to produce regenerants.…”

“…This observation comes from the whole‐plant bioassay using the POX067 isolate which is able to infect the ‘Desiree’ and ‘Victoria’ varieties; it was used previously to select transgenic events with one of the three R genes, and has been used for selecting the transgenic events with the 3 R ‐gene stack. Indeed, the frequency of extreme resistant transgenic events is rather low when using single R genes, as we have reported previously: two out of 64 transgenic events with the RB gene alone, three out of 97 with the Rpi‐blb2 gene, and five out of 52 with the Rpi‐vnt1.1 gene (Orbegozo et al ., ; Román et al ., , ). However, out of 97 transgenic events we observed 75 with extreme resistance with the 3 R ‐gene stack from the variety ‘Desiree’ (Figure ).…”

Comparison of defense responses of transgenic potato lines expressing three different Rpi genes to specific Phytophthora infestans races based on transcriptome profiling

“…Desiree is a tetraploid potato cultivar with a high degree of susceptibility to P. infestans (Yang et al, 2018) and is a good material for genetic transformation of late blight resistance genes due to its high transformation efficiency (Armstrong et al, 2019;Ghislain et al, 2019). The transformation efficiency of Desiree with a single R gene construct was 3.7% for the RB gene, 6% for the Rpi-blb2 gene, 7.5% for the Rpi-vnt1.1 gene (Orbegozo et al, 2016;Roman et al, 2015;Roman et al, 2017), and up to 75% for gene stacking of these three R genes (Ghislain et al, 2019). Single R gene overexpression transgenic lines with the shared genetic background of variety Desiree (RPi-gene free cultivar) are ideal material for exploring the resistance and defense mechanisms mediated by R genes in potato.…”

Expresión de efectores RXLR en el linaje clonal EC-1 de Phytophthora infestans en Perú

“…Este resultado sugiere que estas cepas podrían ser avirulentes a las plantas de papa con el gen Rpi-blb1 / RB. Román et al (2015) confirma que el efector Avr-blb1 en la cepa POX067 es reconocido por el transgén RB.…”