Authors: Silvia FloresBenítez Juan F JiménezBremont Sergio RosalesMendoza Gerardo R ArgüelloAstorga Rosalba CastilloCollazo Ángel Gabriel AlpucheSolís
Publish Date: 2007/09/09
Volume: 91, Issue: 3, Pages: 215-224
Abstract
Agave salmiana was transformed using two different protocols cocultivation with Agrobacterium tumefaciens and particle bombardment The uidA βglucuronidase 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 cocultivation 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 27 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 regenerationThe authors would like to thank Dr William Cress and Dr June Simpson Williamson for critical reading of the manuscript Fellowship No176117 to support the graduate studies of Silvia FloresBenitez from CONACYT Mexico and IPICYT divisional support are also acknowledged
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