Agrobacterium-mediated genetic transformation of Viola cornuta
2017
Аутори:
Trajković, MilenaJeknić, Zoran
Antonić Reljin, Dragana
Subotić, Angelina
Jevremović, Slađana
Cingel, Aleksandar
Остала ауторства
Brajušković, GoranĐorđević, Ana
Тип документа:
Конференцијски прилог (Објављена верзија)
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© 2017 by the University of Belgrade, Faculty of Biology
Метаподаци
Приказ свих података о документуАпстракт:
Introduction: Viola cornuta is a valuable perennial ornamental plant. Development of new traits, such as new flower color with classical breeding suffers from many difficulties, which can be overcome using genetic engineering. With aim to develop protocol for Agrobacterium-mediated transformation of V. cornuta, we used Agrobacterium tumefaciens strain LBA4404 harbouring the superbinary vector pTOK233 carried a GUS reporter gene and hygromycin phosphotransferase selectable marker gene.
Methods: Hypocotyl explants obtained from seedlings were grown on ½MS medium supplemented with 0.1 mg/l 2,4-D and 2.0 mg/l BA for shoot induction. After two days of pre-cultivation, hypocotyl explants were inoculated in bacterial suspension for 15 min and placed on the same culture medium with addition of acetosyringone 100 μM at pH 5.2. After two days of co-cultivation, explants were transferred on shoot induction medium supplemented with cefotaxime and hygromycin B for selection. Regenerated putative transformants were analyzed by PCR for hygromycin phsphotransferase gene presence and by histochemical assay for β-glucuronidase (GUS) activity.
Results: Shoots were obtained within 8 weeks after explants were inoculated with A. tumefaciens, with 2.0% regeneration efficiency. PCR analysis confirmed selectable marker gene presence in twelve out of sixteen (75.0%) independently derived putatively transformed lines that were tested. Additionally, all analyzed lines exhibited a notable β-glucuronidase activity that was not present in untransformed plants.
Conclusion: This is the first report about V. cornuta susceptibility to A. tumefaciens. Presented protocol for genetic transformation can be used for further introduction of desirable traits in V. cornuta cultivars.
Финансирање / пројекти:
- Развој и примена биотехнолошких поступака у добијању здравог садног материјала украсних биљака (RS-MESTD-Technological Development (TD or TR)-31019)
У:
- Brajušković G, Đorđević A, editors. CoMBoS. Book of Abstracts: 1st Congress of Molecular Biologists of Serbia: CoMBoS; 2017 Sep 20-21; Belgrade, Serbia. Belgrade: University of Belgrade, Faculty of Biology; 2017. p. 83.