Assessment of the genetic stability of indirect shoot organogenesis-derived plantlets of Digitalis trojana Ivanina by flow cytometry and cytological analyses
In this study, flow cytometry and cytological analysis was used to evaluate the genetic stability of Digitalis trojana Ivanina plants regenerated via indirect shoot organogenesis. For in vitro propagation, leaf explants were excised from seedlings grown in sterile conditions and cultured MS medium supplemented with 3.0 mg/L BAP, 0.1 mg/L NAA. Shoots and calli were subcultured for a period of 2 weeks for shoot multiplication. For rooting, shoots were separated individually and transferred to MS medium containing 0.1% activated charcoal. Genetic stability of the regenerated plants was assessed by flow cytometry and cytological analyses. Flow cytometric analysis revealed that regenerated plantlets had as 2.82 pg nuclear DNA (2C) and seed-derived plants had on average 2.80 pg/2C. Cytological analysis showed that regenerated plantlets have the same number chromosome with seed-derived plantlets of D. trojana (2n=56). Our results have showed that the plantlets propagated in MS medium with 3 mg/L BAP, 0.1 mg/L NAA didn’t differ genetically from donor plants. Therefore, this system can be effective and suitable for clonal propagation of D. trojana. Our result also confirmed that flow cytometry is fast, easy, accurate and relatively cheap method to determine ploidy of in vitro propagated D. trojana plantlets.
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