The present investigation was carried out to establish an efficient and reproducible micropropagation protocol for the production of morphologically, genetically and chemically uniform plants of Curcuma zedoaria. Axillary bud explants of C. zedoaria were inoculated into MS basal medium supplemented with various combinations and concentrations of 6-benzyladenine (2.2–22.2 µM, BA), kinetin (2.3–23.2 µM, Kin), indole-3-acetic acid (2.9–11.4 µM, IAA), α-naphthalene acetic acid (2.7–10.2 µM, NAA) and adenine sulphate (33.9–203.6 µM, Ads). Almost 95% of rhizome buds sprouted on MS medium supplemented with 13.3 μM BA, 5.7 μM IAA and 63.9 μM Ads giving rise to an average of 12.89 ± 0.02 shoots within 6 weeks. However, the maximum number of roots (25.8 ± 0.07 roots per explant) was obtained on half strength MS medium supplemented with 7.4 µM of IBA after 4 weeks of inoculation. Morphological characteristics were similar in both conventionally propagated and micropropagated plants. Additionally, genetic homogeneity of in vitro plants was further confirmed through ISSR and flow cytometry analysis. A total of 27 ISSR primers were screened, out of which 13 ISSR primers generated 58 monomorphic and reproducible bands thereby confirming the genetic uniformity of obtained plants. The mean 2C DNA content of the mother plant (2.96 pg) was similar to that of in vitro derived plants (3.07 pg). Gas chromatography-mass spectrometry (GC–MS) analysis showed similarity in the qualitative profile of chemical constituents of essential oil and high-performance liquid chromatography analysis revealed no significant differences in curcumin content in the tissue culture regenerants and mother plants of C. zedoaria. Therefore, the present micropropagation protocol could be effectively employed to generate true to type plantlets of C. zedoaria.

Rapid plant regeneration in industrially important Curcuma zedoaria revealing genetic and biochemical fidelity of the regenerants

Results provide significant comparison of leaf anatomy, pigment content, antioxidant response and phenolic profile between individuals from miscellaneous populations and describe unified cultivation protocols for further research on stress biology.
 The plant communities growing on heavy metal-polluted areas have attracted considerable attention due to their unique ability to tolerate enormous amounts of toxic ions. Three ecotypes of Silene vulgaris representing calamine (CAL), serpentine (SER) and non-metallicolous (NM) populations were evaluated to reveal specific adaptation traits to harsh environment. CAL leaves presented a distinct anatomical pattern compared to leaves of SER and NM plants, pointing to their xeromorphic adaptation. These differences were accompanied by divergent accumulation and composition of photosynthetic pigments as well as antioxidant enzyme activity. In CAL ecotype, the mechanism of reactive oxygen species scavenging is based on the joint action of superoxide dismutase and catalase, but in SER ecotype on superoxide dismutase and guaiacol-type peroxidase. On the contrary, the concentration of phenylpropanoids and flavonols in the ecotypes was unchanged, implying the existence of similar pathways of their synthesis/degradation functioning in CAL and SER populations. The tested specimens showed genetic variation (atpA/MspI marker). Based on diversification of S. vulgaris populations, we focused on the elaboration of similar in vitro conditions for synchronous cultivation of various ecotypes. The most balanced shoot culture growth was obtained on MS medium containing 0.1 mg l−1 NAA and 0.25 mg l−1 BA, while the most abundant callogenesis was observed on MS medium enriched with 0.5 mg l−1 NAA and 5.0 mg l−1 BA. For the first time, unified in vitro protocols were described for metallophytes providing the opportunity to conduct basic and applied research on stress biology and tolerance mechanisms under freely controlled conditions.

/pdf/structural-physiological-and-genetic-diversification-of-4md5k6crhv.pdf

Structural, physiological and genetic diversification of Silene vulgaris ecotypes from heavy metal-contaminated areas and their synchronous in vitro cultivation.

In this study the effects of different sucrose concentrations, and the combinations of jasmonic acid (JA) with auxins (IAA or NAA) or with cytokinin (2iP) on the bulb induction and rooting of in vitro plantlets of Hyacinthus orientalis subsp. chionophyllus Wendelbo, which is endemic in Turkey, were investigated. The effect of four different sucrose concentrations (30, 45, 60 and 90 g L-1) on bulb formation in tissue culture was investigated. These plantlets were cultured on MS medium supplemented with several concentrations and combinations of JA (0.0, 1.0, 2.0 mg L-1) and 2iP (0.0, 0.25 and 0.50 mg L-1), IAA or NAA (0.5, 1.0 mg L-1). In JA- 2iP treatment, the highest number of bulblets (13.7 number/explant) was obtained by the combinations of JA 1.0 mg L-1 + 2iP 0.25 mg L-1. Also, the largest bulblets with the mean diameter of 7.9 mm were found on MS medium supplemented with JA 2.0 mg L-1. In JA – Auxin treatment, the mean root number per bulblet was highest (17.9 number/explant) and root formation rate was maximum (81.14%) on MS medium supplemented with IAA 1.0 mg L-1 + JA 2.0 mg L-1.

/pdf/the-effect-of-different-applications-on-in-vitro-bulb-ph4p6gdiiw.pdf

The Effect of Different Applications on In vitro Bulb Development of an Endemic Hyacinth Plant (Hyacinthus orientalis L. subsp. chionophyllus Wendelbo) Grown in Turkey

https://link.springer.com/content/pdf/10.1186/s43141-020-00052-8.pdf

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

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.

/pdf/in-vitro-propagation-and-cytogenetic-stability-of-bwhhcvrm.pdf

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

Silene bolanthoides Quezel, Contandr. & Pamukc. 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. https://doi.org/10.2298/ABS170410033C Received: April 10, 2017; Revised: August 9, 2017; Accepted: August 16, 2017; Published online: September 12, 2017 How to cite this article: Corduk N, Yucel G, Akinci N, Tuna M, Esen O. In vitro propagation of Silene bolanthoides Quezel, Contandr. & Pamukc. and assessment of genetic stability by flow cytometry. Arch Biol Sci. 2018;70(1):141-8.

/pdf/in-vitro-propagation-of-silene-bolanthoides-quezel-contandr-44dq96lm7f.pdf

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

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