Paper Details

Using leaf explants for transformation of Chrysanthemum morifolium Ramat. mediated by Agrobacterium tumefaciens

Rezvanolsadat Kazeroonian, Amir Mousavi, Sepideh Kalatejari, Masoud Tohidfar

DOI: https://dx.doi.org/10.12692/ijb/6.4.124-132

Int. J. Biosci. 6(4), 124-132. February, 2015. (PDF)

Abstract:

An Agrobacterium-mediated transformation for chrysanthemum using leaf explants of two different cultivars was conducted. Leaves were first infected with Agrobacterium tumefaciens LBA4404 strain, harbouring a binary vector pBI121 carrying the GUS reporter gene and the nptII as the selectable marker gene. 30 seconds for infection of leaves proved to be the optimum duration. Best results were achieved when explants were co-cultivated with A. tumefaciens for two days. Shoot induction medium was supplemented with 7 mgl-1 kanamycin and 300 mgl-1 cefotaxime. Putative transgenic plants were obtained after rooting on 1/2 MS medium containing 7mgl-1 kanamycin in the absence of cefotaxime. Transformation efficiency of ‘Reagan Elite Salmon’ (29.1%) was higher than ‘Resomee Splendid’ (9.1%). Presence of the gus gene was verified with histochemical GUS assay and polymerase chain reaction (PCR) of transformed plants.