Genetic transformation of <Emphasis Type="Italic">Agave salmiana</Emphasis> by <Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis> and particle bombardment |
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Authors: | Silvia Flores-Benítez Juan F Jiménez-Bremont Sergio Rosales-Mendoza Gerardo R Argüello-Astorga Rosalba Castillo-Collazo Ángel Gabriel Alpuche-Solís |
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Institution: | (1) División de Biología Molecular, Instituto Potosino de Investigación Científica y Tecnológica, AC, Camino a la Presa San José 2055, Lomas 4a. Sección, San Luis Potosi, San Luis Potosi, C.P. 78216, Mexico |
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Abstract: | Agave salmiana was transformed using two different protocols: co-cultivation with Agrobacterium tumefaciens and particle bombardment. The uidA (β-glucuronidase) gene was used as a reporter gene for both methods whereas the nptII and bar genes were used as selectable markers for A. tumefaciens and biolistic transformation respectively. Previous reports for in vitro regeneration of A. salmiana have not been published; therefore the conditions for both shoot regeneration and rooting were optimized using leaves and
embryogenic calli of Agave salmiana. The transgenes were detected by Polymerase Chain Reaction (PCR) in 11 month old plants. The transgenic nature of the plants
was also confirmed using GUS histochemical assays. Transformation via co-cultivation of explants with Agrobacterium harbouring the pBI121 binary vector was the most effective method of transformation, producing 32 transgenic plants and giving
a transformation efficiency of 2.7%. On the other hand, the biolistic method produced transgenic calli that tested positive
with the GUS assay after 14 months on selective medium while still undergoing regeneration. |
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Keywords: | Acetosyringone Biolistic Bar Gus Transgenic monocots |
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