In vitro propagation and cytogenetic stability of Tripleurospermum insularum (Asteraceae) – a critically endangered insular endemic species from Turkey

Abstract

Tripleurospermum insularum (Asteraceae) is a critically endangered (CR) insular endemic species in Turkey and is facing high risk of extinction. Here, a rapid and efficient in vitro propagation protocol using nodal segments obtained from seedling shoots cultured on Murashige and Skoog (MS) basal medium supplemented with different plant growth regulators (PRGs) was developed to conserve T. insularum. Besides, the cytogenetic fidelity of propagated plants was tested with DNA ploidy level using flow cytometry (FCM) as well as chromosome counting. The highest shoot number and length of shoot per explant were achieved in MS medium containing 4.6 µM zeatin (ZEA) and 0.5 µM indole-3-acetic acid (IAA). No variation in DNA ploidy level (2x) and somatic chromosome number (2n = 18) of all propagated plants were observed. In vitro rooting of shoots was achieved at 100% efficiency in the medium supplemented with 2.9 µM IAA. The rooted plantlets were transferred ex vitro with 74% survival. This is the first report of a successfully developed micropropagation protocol for ex situ conservation of T. insularum.

Keywords:

• Cytogenetic stability
• ex situ conservation
• insular endemic
• micropropagation
• Tripleurospermum insularum

Acknowledgements

We would like to thank Betul Ergin for technical assistance; greenhouse personnel for ex vitro studies in Faculty of Forestry at the Karadeniz Technical University. The authors also express their thanks to anonymous reviewers and subject editor for their comments and suggestions which helped to improve the manuscript considerably.

Disclosure statement

The authors declare that no conflict interests.

Additional information

Funding

This work was supported by the Scientific and Technological Research Council of Turkey [TUBITAK project no (grant numbers) 117Z588 and 106T162].