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Öğe Cold-induced genetic instability in micropropagated Pistacia lentiscus L. plantlets(Springer Heidelberg, 2014) Koc, Ibrahim; Akdemir, Hulya; Onay, Ahmet; Ciftci, Yelda OzdenGenetic stability of plants during in vitro propagation and conservation is one of the important aspects of plant biotechnology. In the present study, micropropagated P. lentiscus L. shoot cultures, which are cultivated for the mastic resin, have been cold stored up to 12 months at 4 A degrees C in the dark for different durations (2, 4, 6, 8, 10 and 12 months) and genetic alterations in cold storage conditions were evaluated. Growth parameters such as proliferation rate, shoot numbers per explant, shoot lengths and shoot forming capacity were also calculated. Since the highest proliferation rate (100 %) was obtained in 6 month-stored shoot cultures without any severe influence of cold stress on proliferation ability, amplified fragment length polymorphism (AFLP) and inter-retrotransposon amplified polymorphism (IRAP) marker systems were used to determine genetic stability in the plantlets after this storage period. Totally, 702 scorable bands were produced by 10 AFLP primer pairs. Genetic similarity value of the non-stored (control) plant and cold-stored clones ranged from 0.66 to 0.84 with a mean of 0.74. In the case of IRAP, 159 bands were produced by 8 IRAP primers. Genetic similarity value of the non-stored plant and cold-stored clones varied from 0.65 to 0.83 and the average genetic similarity value was determined as 0.72. The genetic similarity indices revealed that genetic variability was similar in both techniques. Our results showed that tissue culture and especially cold storage of P. lentiscus L. may result transposons activation, thus could cause genetic instability.Öğe In vitro regeneration and conservation of the lentisk (Pistacia lentiscus L.)(Tubitak Scientific & Technological Research Council Turkey, 2014) Koc, Ibrahim; Onay, Ahmet; Ciftci, Yelda OzdenPistacia lentiscus L. (lentisk or mastic tree) is an economically important member of the genus Pistacia due to its valuable mastic resin. Shoot tips and nodal segments were used as explant sources from in vitro-germinated seeds of lentisk. Shoot tips were found to be suitable for multiple shoot formation. Thereafter, the influences of different growth regulators [N6-benzyladenine (BA), gibberellic acid (GA(3)), naphthalene acetic acid (NAA), or jasmonic acid (JA)] together with various elicitors [silver nitrate (AgNO3) or phloroglucinol (PG)] were assessed to develop efficient micropropagation protocol. Our results showed that the combination of all tested concentrations of GA(3) with 1.0 mg/L BA resulted in enhancement of multiple shoot formation. The maximum number of shoots per explant (3.95) was recorded on MS medium containing 1.0 mg/L BA and 0.3 mg/L GA(3). In contrast, JA had a negative influence, while AgNO3 had no significant effect on multiple shoot formation. In terms of synthetic seed production, it was possible to encapsulate shoot tips at 4 degrees C in darkness for up to 6 months with a frequency of 87.5% plant regrowth. In vitro-propagated microshoots, including plantlets derived from synthetic seeds, were transferred to MS medium containing different concentrations (1.0, 2.0, or 4.0 mg/L) of indole butyric acid for rooting and successfully acclimatized to ex vitro conditions. The presented data suggest an efficient in vitro regeneration system and conservation via synthetic seed production for lentisk.Öğe Rejuvenation of mature lentisk by micrografting and evaluation of genetic stability(Tubitak Scientific & Technological Research Council Turkey, 2016) Onay, Ahmet; Tilkat, Engin; Suzerer, Veysel; Karakas Metin, Ozge; Ozden Ciftci, Yelda; Kilinc, Fatih Mehmet; Koc, IbrahimAn in vitro propagation method was established for both male and female genotypes of lentisk using actively growing shoot tips derived from forcefully lignified shoots. The effects of growth regulators on in vitro morphogenesis were investigated. Since rooting of the regenerated shoots for both genotypes was not achieved, an in vitro micrografting method was developed for the production of plantlets. Moreover, genetic stability of 3-, 6-, and 24-times subcultured clones of both genotypes was assessed and compared with the mother plants using fluorescent-based amplified fragment length polymorphism (AFLP) analysis. The set of main plants and the different subcultured clones were divided into two clusters. In the first cluster, the original male and female plants were grouped together with the 3-times subcultured female and the 6-times subcultured male and female groups, whereas the second cluster contained the 24-times subcultured clones and the 3-times subcultured male group. To the best of our knowledge, this is the first report of successfully inducing plantlets from mature lentisk genotypes and the first analysis of clonal fidelity of regenerated mature female and male P. lentiscus L. plants by AFLP.