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Genetic transformation of Agave salmiana by Agrobacterium tumefaciens and particle bombardment

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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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Abbreviations

BAP:

6-Benzylaminopurine

IAA:

Indole-3-acetic acid

MS:

Murashige and Skoog medium

NAA:

Naphthaleneacetic acid

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Acknowledgements

The authors would like to thank Dr. William Cress and Dr. June Simpson Williamson for critical reading of the manuscript. Fellowship No.176117 to support the graduate studies of Silvia Flores-Benitez, from CONACYT Mexico and IPICYT divisional support are also acknowledged.

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Authors and Affiliations

  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

    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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  1. Silvia Flores-Benítez
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  2. Juan F. Jiménez-Bremont
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  3. Sergio Rosales-Mendoza
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  4. Gerardo R. Argüello-Astorga
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  5. Rosalba Castillo-Collazo
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  6. Ángel Gabriel Alpuche-Solís
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Correspondence to Ángel Gabriel Alpuche-Solís.

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Flores-Benítez, S., Jiménez-Bremont, J.F., Rosales-Mendoza, S. et al. Genetic transformation of Agave salmiana by Agrobacterium tumefaciens and particle bombardment. Plant Cell Tiss Organ Cult 91, 215–224 (2007). https://doi.org/10.1007/s11240-007-9287-3

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  • DOI: https://doi.org/10.1007/s11240-007-9287-3

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