Detección de marcadores microsatélites asociados con la resistencia al añublo bacterial de la yuca (Manihot esculenta Crantz) en Colombia

Una de las principales estrategias para el manejo del Añublo Bacterial de la Yuca (CBB), causado por Xanthomonas axonopodis pv. manihotis es el uso de resistencia varietal, que implica desarrollar variedades de yuca con resistencia genética duradera. Para tal fin, es necesario conocer los genes que dominan la resistencia a la enfermedad, detectando inicialmente marcadores moleculares asociados con la respuesta fenotipica de la planta, siendo este el principal objetivo del presente estudio. Inicialmente, se evaluó la reacción a CBB de 4 familias de yuca BCI (retrocruce 1), se seleccionó la más segregante bajo presión natural de inóculo en Villavicencio (Meta, Colombia) y se confirmó la respuesta a CBB en condiciones de invernadero en CIAT (Palmira, Valle). La familia GM 315 presentó la mejor segregación, siendo la más adecuada para buscar asociación entre su reacción fenotípica y la presencia de un marcador molecular. Para esto, se evaluaron 486 cebadores microsatélites mediante análisis de grupos segregantes (BSA), encontrándose que 17 de ellos mostraron polimorfismo entre los grupos contrastantes y solo uno de ellos, el cebador SSRY 65, mostró diferencias significativas entre individuos resistentes y susceptibles. Al evaluar este cebador en toda la familia segregante se encontró asociación entre su presencia y los individuos evaluados fenotípicamente como resistentes en campo e invernadero, con una probabilidad mínima de P=0,OOl5 y P=0,OO7 respectivamente, en una prueba de Chicuadrado de independencia. Adicionalmente, a partir de los resultados obtenidos en el análisis estadístico, se calcularon los valores predictivos, especificidad y sensibilidad del marcador SSRY 65. Con base en los valores predictivos positivos generados, es posible sugerir la utilización de este marcador en pruebas diagnósticas para detectar la presencia de una banda específica en individuos resistentes de familias genéticamente relacionadas con la familia GM 315. = A major strategy for managing Cassava Bacterial blight (CBB), caused by Xanthomonas axonopodis pv. manihotis, is to use varietal resistance, that is, to develop cassava varieties with lasting genetic resistance. A search for the genes that dominate resistance to the disease was initially conducted by seeking the molecular markers associated with the plant s phenotypic response to CBB. The response in four BC1 (backcross 1) cassava families was accordingly evaluated under natural disease pressure at Villavicencio (Meta, Colombia). The most segregating family was then selected, and its response to CBB was verified under greenhouse conditions at CIAT (Palmira, Valle). Family GM 315 presented the best segregation, so, it was the most suitable for seeking association between its phenotypic reaction and the presence of a molecular marker. Of 486 microsatellite primers evaluated by bulked segregant analysis (BSA), 17 showed polymorphism among contrasting groups. Only one primer, SSRY 65, showed significant differences between resistant and susceptible individuals. On evaluating this primer for the entire segregating family, an association was found between its presence and individuals evaluated phenotypically as resistant in the field and greenhouse (minimum P = 0.0015 and P = 0.007, respectively, in a chi-square test of independence). With the results of the statistical analysis, the predictive values, specificity, and sensibility of marker SSRY 65 were calculated. The positive predictive values generated indicate that this marker can be used in diagnostic tests to detect the presence of a specific band in resistant individuals of families genetically related to the GM 315 family

Comparison of simple sequence repeat (SSR) and diversity array technology (DArT) markers for assessing genetic diversity in cassava Manihot esculenta Crantz)

Several molecular marker systems have been developed for assessing genetic diversity in crop germplasm collections. A trade-off often exists between the number of loci that can feasibly be sampled by a marker system and the amount of information provided by each locus. We compared the usefulness of two marker systems for revealing genetic diversity and population structure in cassava (Manihot esculenta Crantz): simple sequence repeats (SSRs) and diversity array technology (DArT) markers. DArTs survey many more loci per reaction than do SSRs; however, as bi-allelic, dominant markers, DArTs provide less polymorphism information per locus. Genetic differentiation was assessed in a randomly selected set of 436 cassava accessions, consisting of 155 African and 281 Latin American accessions. A genome-wide set of 36 SSR markers and a DArT array of approximately 1000 polymorphic clones were used to assess genetic diversity and differentiation. Cluster analyses were performed using principal coordinate analysis (PCoA). Results were compared with a priori expectations of genetic differentiation based on previous genetic analyses. Analyses of the two datasets generated broadly similar clustering patterns. However, SSRs revealed greater differentiation than DArTs, and more effectively recovered patterns of genetic differentiation observed in previous analyses (differentiation between Latin American and African accessions, and some geographical differentiation within each of these groups). These results suggest that SSR markers, while low throughput in comparison with DArTs, are relatively better at detecting genetic differentiation in cassava germplasm collections. Nonetheless, DArTs will likely prove useful in ‘orphan crop’ species, where alternative molecular markers have not been developed