Abstract
Gynura bicolor DC. is a perennial vegetable and medicinal plant. It is an important source of anthocyanins. The effects of different growth regulators on callus induction and plant regeneration were evaluated. The best SFC index (8.6) of plant regeneration was obtained in combination of 2,4-D at 2.0 mg/l and BA at 0.5 mg/l, and the frequency of regenerating explants was 78.3%. The highest number of shoots per explant was 11. The genetic stability of the regenerants was analyzed by random ampliï¬ed polymorphic DNA (RAPD), inter-simple sequence repeat (ISSR) molecular markers and flow cytometry. The results indicated that no somaclonal variation was detected among the regenerants. To our knowledge, this is the first report of somatic clone study in G. bicolor. The high efficient and reproducible protocol will be advanced for the further studies on secondary metabolic products, transformations and breeding of this potential medicinal plant. Key words: Flow cytometry analysis, genetic stability, Gynura bicolor, inter-simple sequence repeat (ISSR), random ampliï¬ed polymorphic DNA (RAPD), regeneration.
Highlights
Gynura bicolor DC. is a perennial vegetable belonging to Asteraceae and is originated from the tropical area of East Asia
This study described an efficient system for the plant regeneration of G. bicolor
6 5 3 7 4 6 2 4 6 4 7 5 5 7 6 3 5 conditions for random amplified polymorphic DNA (RAPD) analyses were consisted of an initial denaturation at 93°C for 4 min followed by 40 cycles of 45 s denaturation at 94°C, 45 s annealing at 33°C and 80 s extension at 72°C with a final extension of 72°C for 5 min using a thermal cycler (MJ Mini, Bio-Rad, USA)
Summary
Gynura bicolor DC. is a perennial vegetable belonging to Asteraceae and is originated from the tropical area of East Asia. Some strategies are available for detecting genetic variations, including DNA analysis techniques and flow cytometry analysis (Fiuk et al, 2010). Regenerated plants which offers the possibility of fast and large scale analysis of the DNA content of cells for various purposes such as, determination of species specific DNA amount, analysis of the cell cycle activity in different tissues and measurement of endopolyploidization levels ( ali -Dragosavac et al, 2010; Guo et al, 2009). The genetic variation for the regenerated plants was analyzed by RAPD, ISSR and flow cytometry techniques. High efficient protocol for producing tissue cultural based regenerants without somatic genetic variations would be useful for further studies on secondary metabolic products, gene transformations and plant breading of this potential medicinal plant species
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