The overexpression of RXam1, a cassava gene coding for an RLK, confers disease resistance to Xanthomonas axonopodis pv. manihotis.

MAIN CONCLUSION: The overexpression of RXam1 leads to a reduction in bacterial growth of XamCIO136, suggesting that RXam1 might be implicated in strain-specific resistance. Cassava bacterial blight (CBB) caused by Xanthomonas axonopodis pv. manihotis (Xam) is a prevalent disease in all regions, where cassava is cultivated. CBB is a foliar and vascular disease usually controlled through host resistance. Previous studies have found QTLs explaining resistance to several Xam strains. Interestingly, one QTL called XM5 that explained 13% of resistance to XamCIO136 was associated with a similar fragment of the rice Xa21-resistance gene called PCR250. In this study, we aimed to further identify and characterize this fragment and its role in resistance to CBB. Screening and hybridization of a BAC library using the molecular marker PCR250 as a probe led to the identification of a receptor-like kinase similar to Xa21 and were called RXam1 (Resistance to Xam 1). Here, we report the functional characterization of susceptible cassava plants overexpressing RXam1. Our results indicated that the overexpression of RXam1 leads to a reduction in bacterial growth of XamCIO136. This suggests that RXAM1 might be implicated in strain-specific resistance to XamCIO136.

Major Novel QTL for Resistance to Cassava Bacterial Blight Identified through a Multi-Environmental Analysis.

Cassava, Manihot esculenta Crantz, has been positioned as one of the most promising crops world-wide representing the staple security for more than one billion people mainly in poor countries. Cassava production is constantly threatened by several diseases, including cassava bacterial blight (CBB) caused by Xanthomonas axonopodis pv. manihotis (Xam), it is the most destructive disease causing heavy yield losses. Here, we report the detection and localization on the genetic map of cassava QTL (Quantitative Trait Loci) conferring resistance to CBB. An F1 mapping population of 117 full sibs was tested for resistance to two Xam strains (Xam318 and Xam681) at two locations in Colombia: La Vega, Cundinamarca and Arauca. The evaluation was conducted in rainy and dry seasons and additional tests were carried out under controlled greenhouse conditions. The phenotypic evaluation of the response to Xam revealed continuous variation. Based on composite interval mapping analysis, 5 strain-specific QTL for resistance to Xam explaining between 15.8 and 22.1% of phenotypic variance, were detected and localized on a high resolution SNP-based genetic map of cassava. Four of them show stability among the two evaluated seasons. Genotype by environment analysis detected three QTL by environment interactions and the broad sense heritability for Xam318 and Xam681 were 20 and 53%, respectively. DNA sequence analysis of the QTL intervals revealed 29 candidate defense-related genes (CDRGs), and two of them contain domains related to plant immunity proteins, such as NB-ARC-LRR and WRKY.

Expresión de do genes candidatos a resistencia contra la bacteriosis vascular en ycaa / Expression Of Two Resistance Gene Candidates Against Cassava Bacterial Blight In Cassava

La yuca (Manihot esculenta Crantz, Euphorbiaceae) es el cuarto cultivo en importancia a nivel mundial como fuente de calorías para la población humana y cuya producción se ve afectada por la bacteriosis vascular, enfermedad ocasionada por Xanthomonas axonopodis pv. manihotis (Xam). La resistencia a enfermedades en plantas depende de la presencia de genes de resistencia (R), los cuales reconocen a los patógenos y simultáneamente permiten desencadenar la respuesta de defensa. A pesar de recientes esfuerzos encaminados a la identificación de genes R en yuca, aún no se ha logrado clonar el primer gen R en este cultivo. En el presente trabajo se estudió el perfil de expresión de dos Genes Candidatos a Resistencia (RGCs) asociados a QTLs de defensa contra la bacteriosis vascular en yuca. A partir de la técnica transcripción reversa y reacción en cadena de la polimerasa (RT-PCR) se evaluó la expresión de los genes RXam1 y RXam2 en tallos y hojas de las variedades resistentes SG107-35 y MBRA685 de yuca, después de ser inoculadas con la cepa CIO151 de Xam. Se observó que RXam1 es inducido a partir de los cinco días post-inoculación tanto en tallos como hojas de las dos variedades, mientras que RXam2 es expresado de manera constitutiva en la variedad MBRA685.

Cassava (Manihot esculenta Crantz, Euphorbiaceae) is the fourth food crop used as an important energy source for human population worldwide. Cassava Bacterial Blight (CBB) is the most important disease of this crop. CBB is caused by the pathogenic bacterium Xanthomonas axonopodis pv. manihotis (Xam). Plants have developed sophisticated mechanisms to detect and respond to infection by pathogens. These mechanisms depend on the presence of resistance (R) genes, which recognize proteins produced by pathogens. Although efforts have been conducted to identify R genes in cassava, the first R gene in this crop has not been cloned. The present work studied the expression profile of two resistance gene candidates (RGCs) which are linked to QTL associated with resistance to CBB. Gene expression of RXam1 and RXam2 was evaluated by reverse transcription-polymerase chain reaction (RT-PCR) in stems and leaves of SG107-35 and MBRA685, two cassava resistant cultivars, which were challenged with Xam CIO151. We observed that the expression of RXam1 is induced starting five days post-inoculation in the two cultivars studied and in both tissues while the gene RXam2 was constitutively expressed in MBRA685 cultivar.

CONSTRUCCIÓN DE UNA LIBRERÍA DE ADNc EN YUCA: UNA HERRAMIENTA PARA EL DESARROLLO BIOTECNOLÓGICO DEL CULTIVO / Cassava cDNA Library Construction: One Tool for Biotechnological Development of the Crop

La yuca es un cultivo de importancia en la seguridad alimentaria mundial ya que constituye la base de la alimentación de más de 600 millones de personas en el mundo. También es un alto productor de almidón con niveles que oscilan entre 73,7 y 84,9% de su peso seco total en raíces (FAO, 2007). El almidón de yuca puede utilizarse en una gama amplia de industrias (textil, cosmética, alimentaria, etc). Además, es empleado en la producción de biocombustibles. Una de las principales limitantes en la producción de yuca es la bacteriosis vascular producida por la bacteria Xanthomonas axonopodis pv. manihotis (Xam). Esta enfermedad puede comprometer no solo el suministro de almidón para las plantas industriales productoras de bioetanol sino que también puede amenazar la seguridad alimentaria. El mejoramiento genético convencional de yuca es complicado dado su largo ciclo reproductivo, su alta heterocigocidad y su naturaleza tetraploide. Por estas razones se deben buscar alternativas que involucren desarrollos en biotecnología que permitan un mejoramiento eficaz y rápido. En la actual era genómica y postgenómica muchos de los experimentos dependen de la posibilidad de contar con la secuencia de transcritos clonados. Dado que estos clones provienen de librerías de ADNc contar con este tipo de recursos de excelente calidad es un paso esencial. En este artículo reportamos la construcción de una librería de ADNc empleando el sistema Gateway® a partir de plantas de yuca que han sido inoculadas con la cepa CIO151 de Xam. La librería presentó un título de 1 x 10(7) unidades formadoras de colonias (ufc)/ml y el tamaño de los insertos osciló entre 600-1.500 pb. Los análisis de secuencia de 14 clones al azar confirmaron que se trata de genes expresados y mostraron similitud con genes previamente reportados en especies estrechamente relacionadas a yuca. Esta librería se convierte en un excelente recurso para identificar novedosos genes y para el estudio de su función a través de la identificación y la interacción entre proteínas.

Cassava is one of the most important crops for global food security and provides food and livelihood for 600 million people in the developing world. It is also good source of starch, with levels between 73.7 y 84.9% of total dry weight in roots (FAO, 2007). Cassava starch can be used in a wide range of industries (textile, cosmetic, nourishing, etc) and it has a high potential for the production of biofuel. Cassava bacterial blight, caused by Xanthomonas axonopodis pv. manihotis (Xam), is one of the most important diseases that affects cassava. This disease can compromise the starch supply not only for bioetanol production but also affect global food security. The long reproductive cycle, high heterozigosity and tetraploid character of cassava are characteristics that have complicated the genetic breeding for this crop. For these reasons new alternatives based on biotechnology are necessary to accelerate its improvement. In the postgenomic era many experiments rely on the availability of transcript sequences for cloning. As these clones usually originate from cDNA libraries, the quality of these libraries is crucial. In this article we report the construction of the first cassava cDNA library employing the Gateway® system. For this, in vitro grown plants were inoculated with the Xam strain CIO151. The expression library shows a high titer of 1 x 10(7) cfu/ml, with inserts ranging between 600 and 1500 bp. The sequence analyses from 14 random clones confirmed that these are expressed genes and showed similarity with previously cloned genes from species related to cassava. This library is an excellent resource for the identification of novel genes and for functional studies through the identification of their interactions with other proteins.