Factores que afectan la expresión transitoria del gen GUS en yuca (Manihot esculenta Crantz)
Factors affecting the transient expression of gene GUS in cassava (Manihot esculenta Crantz)
Fatores que afetam a expressão transitória do gene GUS na mandioca (Manihot esculenta Crantz)
DOI:
https://doi.org/10.15446/rev.colomb.biote.v20n2.77063Keywords:
Agrobacterium tumefaciens, Beta-glucuronidasa, VirG, agroinfiltración, yuca (es)Agrobacterium tumefaciens, beta-glucuronidase, VirG, agroinfiltration, cassava (en)
Agrobacterium tumefaciens, Beta-glucuronidase, VirG, agroinfiltração, mandioca (pt)
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La expresión transitoria es una métodología ampliamente utilizada para el estudio de genes. Sin embargo, hasta la fecha no existe un reporte en donde se utilice esta técnica en hojas de yuca de plantas adultas. Por esta razón este trabajo se centró en la determinación de algunos parámetros críticos para la expresión transitoria del gen GUS en yuca como son: la metodología para introducir la bacteria, la cepa de Agrobacterium, el tiempo post-inoculación, la introducción del gen VirG y la expresión del gen GUS en algunas variedades de yuca. Los resultados indicaron niveles más altos de expresión del gen GUS entre 5-7 días post-inoculación (dpi), agroinfiltrando con la cepa GV3101 y un incremento en la virulencia de esta cepa mediante la introducción del gen VirG. Por último se observaron diferentes niveles de expresión del gen GUS entre las variedades de yuca evaluadas, lo que indica que el factor genético es clave en la eficiencia de la agroinfiltración en este cultivo.
Transient expression is a common technique used for the co-expression of proteins for a wide range of experiments in molecular biology. This technique has been frequently used for gene study in plant pathogen interaction and has proved to be versatile and effective. However, there are still few reports and sparse data for transient expression in cassava leaves from adult/mature plants. For this reason, in this study we evaluated some regards of transient expression for the gene GUS in cassava: bacteria introduction methods, type of Agrobacterium strains, time post-inoculation, introduction of gene VirG and GUS and their expression into different cassava varieties. The results show that GUS expression is more efficient between 5-7 days post-inoculation with the GV3101 strain, moreover the virulence of this strain increases with the VirG gene. Finally, we observed a range of GUS expression pattern between different cassava varieties, altogether showing that the genetic factor is important for an accurate and effective agroinfiltration in cassava.
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