A simple and efficient genetic transformation protocol for Eucalyptus camaldulensis using internodal explants

Venkatachalam Rathinavel, Boby Vattekkattu Unnikrishnan, Suribabu Thaliyakula, Gurumurthy Demalapura Shankaranarayana

Abstract


A simple and efficient genetic transformation protocol was developed for Eucalyptus camaldulensis. Selected line of eucalyptus was established under in vitro conditions by using nodal cuttings.Assessment of hygromycin sensitivity on explants collected from in vitro established cultures showed that hygromycin concentration of 2.5 mg/L was optimum for transformation. Transformation procedure was standardized with internodal explants through indirect organogenesis. Transformation efficiency of intermodal explants was assessed by plasmids pCAMBIA1305.1 and pRNAiLIC-Hyg carrying GA2ox gene. The transformation efficiency of 20 % was achieved with the present protocol and regeneration of transgenic shoot was obtained in two months of culture compared to three to four months in previous studies.Confirmation of gene integration in eucalyptus was confirmed by histochemical staining and PCR based methods.


Keywords


Eucalyptus, transformation, internodes, efficient, Protocol

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