In vitro propagation of Silene bolanthoides Quézel, Contandr. & Pamukç. and assessment of genetic stability by flow cytometry

Abstract

Silene bolanthoides Quézel, Contandr. & Pamukç. is an endemic species from Kazdagi (Mt. Ida), Canakkale-Balikesir, Turkey. In order to develop an efficient shoot regeneration protocol, the leaf, nodal and internodal explants of S. bolanthoides were cultured on Murashige and Skoog (MS) medium containing benzyladenine (BA) alone or in combination with α-naphthaleneacetic acid (NAA). The highest number of regenerated shoots (5.75±0.1) was obtained from nodal explants that were cultured on MS medium with 8.8 µM BA+0.54 µM NAA. Regenerated shoots were rooted on MS medium without plant growth regulators (PGRs). Rooted plants (2-3 cm) were separately transferred to pots containing a mixture of peat and perlite (3:1 v/v) and acclimatized successfully in a growth chamber. Genetic stability of the propagated plants was assessed by flow cytometry and cytological analysis. Flow cytometry analysis demonstrated that regenerated plants had 2.61±0.01 pg nuclear DNA (2C) and seed-derived plants had on average 2.58±0.02 pg/2C. Cytological analysis showed that the regenerated plants had the same chromosome number as seed-derived plants of S. bolanthoides (2n=24). It was determined that regenerated plants were uniform in chromosome number and had a similar DNA content to the seed-derived ones, indicating that the described efficient shoot regeneration protocol can be applied for ex situ conservation of this species.

In vitro propagation and DNA barcode analysis of the endangered Silene schimperiana in Saint Katherine protectorate

“…According to the results of the present study, when the two explant types were compared, it was found that the stem nodal segment had a higher shoot induction capacity than the shoot tip. Similarly, Çördük et al [ 9 ] found that nodal explant was the most effective explant for shoot regeneration of Silene bolanthoides .…”

“…The highest shoot multiplication ability of BA has also been reported for other Silene species; where MS medium supplemented with 4.48 μM BA was optimum for the multiplication of non-vitreous shoots of Silene fabaria with sufficient length [ 10 ]. Additionally, BA has been the most widely used cytokinin for improving the number and length of shoots in various Silene species; such as for regenerated shoots from callus of Silene vulgaris [ 32 , 33 ] and axillary shoots from direct organogenesis in Silene bolanthoides [ 9 ].…”

“…domokina , 100% rooting was obtained by incorporating 0.1 mg/L (0.49 μM) IBA into the MS medium, followed by Kin and auxins combination [ 10 ]. On the other hand, in Silene bolanthoides , regenerated shoots were rooted on PGRs-free MS medium [ 9 ].…”

“…In vitro propagation of other species of Silene have been achieved previously, such as S. leucophylla [ 6 ], S. fetissovii , S. obovata , S. sussamyrica and S. ladyginae [ 7 ], Silene cretacea [ 8 ], Silene bolanthoides [ 9 ] and Silene fabaria subsp. domokina [ 10 ].…”