Identificación de genes análogos de resistencia a enfermedades en yuca, Manihot esculenta Crantz, y su relación con la resistencia a tres especies de Phytophthora
Los genes de resistencia se buscaron mediante dos estrategias. La primera, por medio de hibridización con sondas de maíz y arroz, utilizando RFLP. La segunda consistió en la amplificación de regiones conservadas de ADN, con cebadores degenerados NBS y Pto kinasa, en tres genotipos de yuca resistentes a Phytophthora tropicalis y P. palmivora, obteniendo clones que se secuenciaron y se homologaron con genes de resistencia conocidos. Con las secuencias se diseñaron cebadores específicos que permitieron amplificar regiones de ADN de los parentales e individuos resistentes y susceptibles. Las bandas se separaron mediante electroforesis en geles de poliacrilamida denaturantes y no denaturantes (SSCP - polimorfismo en la conformación de cadenas simples). Se identificaron cinco QTLs asociados con resistencia a Phytophthora. La yuca tuvo muy baja homología con los genes de maíz y arroz. Se obtuvieron 28 clones NBS y 2 Pto kinasa, de los cuales 5 mostraron secuencia homóloga con RGAs (genes análogos de resistencia) NBS-LRR y cuatro de ellos mostraron marco abierto de lectura con motivos conservados de la región NBS, y se consideraron como RGAs. Se identificaron tres clases de RGAs aunque no hubo evidencia de su asociación con resistencia a Phytophthora. = Identification of gene analogs for resistance to cassava (Manihot esculenta Crantz) Diseases, and their relationship to resistance to three Phytophthora species. Two strategies were used to find resistance genes in cassava. The first through hybridizing probes from maize and rice, using RFLP. The second strategy consisted of amplifying conserved regions of DNA, with degenerated NBS and Pto kinase primers, in three cassava genotypes resistant to Phytophthora tropicalis and P. palmivora, obtaining clones that were sequenced and compared with known resistance genes. Specific primers were designed from the sequences, allowing DNA regions of parental material and resistant and susceptible individuals, to be amplified. Bands were separated by denaturing polyacrylamide gel electrophoresis, and non-denaturing polyacrylamide gel (SSCP - single strand conformation polymorphism-). Five QTLs associated to Phytophthora spp resistance were identified. Cassava has a very low homology with the genes of the monocotyledons tested. A total of 28 NBS and 2 Pto kinase clones were obtained; of these, 5 showed homologous sequence with NBS-LRR RGAs (resistance gene analogs). Four of them had open reading frames with conserved motifs of the NBS region, and were considered as RGAs. Three different RGAs classes were identified. It remain to be shown if there are association to resistance to Phytophthora.
| Main Authors: | , , , |
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| Format: | Journal Article biblioteca |
| Language: | Spanish / Castilian |
| Published: |
2004
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| Subjects: | manihot esculenta, phytophthora, genes, selection, disease resistance, genetic resistance, hybridization, genetic maps, cloning, nucleotide sequence, selección, resistencia a la enfermedad, resistencia genética, hibridación, mapas genéticos, clonación, secuencia nucleotídica, |
| Online Access: | https://hdl.handle.net/10568/44339 http://www.revistas.unal.edu.co/index.php/acta_agronomica/article/download/88/196 |
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