ISHS Acta Horticulturae 579: II Balkan Symposium on Vegetables and Potatoes
IN VITRO REGENERATION AND GENETIC TRANSFORMATION OF PARTHENOCARPIC CUCUMBER HYBRIDS |
| Authors: | G. Sapountzakis, A.S. Tsaftaris |
| Keywords: | shoot organogenesis, cotyledon, Agrobacterium, herbicide resistance |
| Abstract:
To achieve regeneration, seeds of the cucumber hybrids Brunex and Bambina were germinated in darkness. Cotyledon and hypocotyl explants that isolated from these seedlings were cultured in different media. Shoot regeneration took place when cotyledon segments (proximal to the embryonic axis) excised 5 days after the establishment of the seeds on 0.8 % w/v agar and cultured for 6 weeks on MS + 0.1 mg/l ΒΑ. The regenerated shoots were transferred to MS where they elongated and rooted. To achieve genetic transformation, cotyledonary segments were incubated for 24 hours in liquid MS + 0.1 mg/l BA in which a transformed Agrobacterium strain was suspended. This strain contains pGKB5 plasmid in which kanamycin and Basta resistance genes were inserted. After co-cultivation, explants were rinsed and transferred to solid MS medium + 0.1 mg/l BA with progressively increasing doses of kanamycin and cefotaxime. Shoots, which were formed on the cotyledon explants, were transferred to MS + 100 mg/l kanamycin + 500 mg/l cefotaxime where they elongated and rooted. Verification tests were performed to certify genetic transformation. | |
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