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

Rezvanolsadat Kazeroonian, Amir Mousavi, Sepideh Kalatejari, Masoud Tohidfar

Department of Horticultural Sciences, Science and Research Branch, Islamic Azad University, Tehran, Iran

National Institute of Genetic Engineering and Biotechnology, Iran

Seed and Plant Improvement Campus, Agricultural Biotechnology Research Institute of Iran, Karaj, Iran

Key words: Chrysanthemum morifolium, transformation, Agrobacterium tumefaciens, gus reporter gene, leaf explants.


DisplayImage.ashx_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 cocultivated 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.

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