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Öğe Clonal micropropagation of Pistacia lentiscus L. and assessment of genetic stability using IRAP markers(Springer, 2015) Kilinc, Fatih Mehmet; Suzerer, Veysel; Ciftci, Yelda Ozden; Onay, Ahmet; Yildirim, Hakan; Uncuoglu, Ahu Altinkut; Tilkat, EnginAn efficient protocol for clonal micropropagation of selected genotypes of lentisk, Pistacia lentiscus L., which is cultivated for the masticha resin, has been developed using shoot tip explants originating from in vitro seedlings. BA was found to be optimum for shoot morphogenesis in terms of the number and length of shoots among the cytokinins tested for all cloned genotypes, while the highest shoot length was noticed in the presence of 2iP at a rate 4.92 mu M. Efficient rooting (94.15 %) was achieved in a medium containing 19.6 mu M IBA with the clone II that was superior to the rest of the clones tested. The method developed for plant acclimatization was satisfactory because a high percentage of plant survival (95 %) in the growth room in the clone II was obtained and the regenerated plantlets resumed growth after 4 months. DNAs from mother seedlings and micropropagated plantlets belonging to 6, 9 and 12 times subcultured were isolated and subjected to IRAP analysis in order to evaluate their genetic stability and detect possibly existing variations among in vitro derived plantlets. The mean percentage of similarity calculated by Jaccard's similarity coefficient ranged from 78 to 86 % in the four genotypes. Although variation was observed among mother plantlets and its regenerants for all of the clones, polymorphic information content value in the range of 0.391-0.405 indicated the presence of reasonable polymorphism within the clones. The presented data confirmed that the clonal propagation of lentisk by using shoot tips could be used for commercial exploitation of the selected genotype.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 Detection of Variation in Long-Term Micropropagated Mature Pistachio via DNA-Based Molecular Markers(Springer, 2016) Akdemir, Hulya; Suzerer, Veysel; Tilkat, Engin; Onay, Ahmet; Ciftci, Yelda OzdenDetermination of genetic stability of in vitro-grown plantlets is needed for safe and large-scale production of mature trees. In this study, genetic variation of long-term micropropagated mature pistachio developed through direct shoot bud regeneration using apical buds (protocol A) and in vitro-derived leaves (protocol B) was assessed via DNA-based molecular markers. Randomly amplified polymorphic DNA (RAPD), inter-simple sequence repeat (ISSR), and amplified fragment length polymorphism (AFLP) were employed, and the obtained PIC values from RAPD (0.226), ISSR (0.220), and AFLP (0.241) showed that micropropagation of pistachio for different periods of time resulted in reasonable polymorphism among donor plant and its 18 clones. Mantel's test showed a consistence polymorphism level between marker systems based on similarity matrices. In conclusion, this is the first study on occurrence of genetic variability in long-term micropropagated mature pistachio plantlets. The obtained results clearly indicated that different marker approaches used in this study are reliable for assessing tissue culture-induced variations in long-term cultured pistachio plantlets.Öğe In vitro conservation and cryopreservation of mature pistachio (Pistacia vera L.) germplasm(Springer India, 2013) Akdemir, Huelya; Suzerer, Veysel; Tilkat, Engin; Yildirim, Hakan; Onay, Ahmet; Ciftci, Yelda OzdenAs genetic erosion of pistachio (Pistacia vera L.) has been occurring in the Mediterranean, Central and West Asia and North Africa, experiments were conducted to conserve two cultivars ('Atli' and 'Siirt') of mature pistachio germplasm by assessing both medium-and long-term conservation techniques. In medium-term conservation, our results showed that it was feasible to conserve both cultivars in the form of either microshoots or encapsulated shoot apices up to 12 months at 4 degrees C in the dark. As regards long-term conservation, encapsulation-dehydration and droplet-vitrification techniques were assessed for cryopreservation of cold-hardened and osmoprotected shoot apices of mature 'Atli' cultivar. Among the methods tested, 13.6% of regrowth was achieved with incubation of explants in the droplets of vitrification solution for 150 min at 0 degrees C followed by direct immersion in liquid nitrogen (LN), rapidly thawed and then cultured on Murashige and Skoog's (MS) medium containing 1 mg L-1 BA and 0.5 mg L-1 GA(3). The developed droplet-vitrification technique appeared as a promising procedure for long-term preservation of shoot apices of mature pistachio germplasm. Moreover, assesment of genetic fidelity by Random Amplified Polymorphic DNA analysis (RAPD) revealed out high levels of genetic stability between donor plant and cryopreserved plants (similarity indexes between 0.959 and 0.973) after they were subcultured for at least 3 months. The detected low level of genetic instability could be due to the toxic effect of PVS2 and regeneration phase. The optimized conservation techniques, especially slow growth storage, could be applied to preserve other Pistacia species.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 Micropropagation of the pistachio and its rootstocks by temporary immersion system(Springer, 2014) Akdemir, Hulya; Suzerer, Veysel; Onay, Ahmet; Tilkat, Engin; Ersali, Yusuf; Ciftci, Yelda OzdenAlthough several studies have been reported on the micropropagation of the pistachio and its rootstocks, to date none of them had been efficient on the mass production of these plants in bioreactor systems. Thus, the micropropagation of juvenile pistachio shoot tips and nodal buds was investigated in a temporary immersion bioreactor system (RITA(A (R))) and on a conventional semi-solid medium. Among the tested immersion conditions, immersion for 24 min every 16 h reduced vitrification and improved proliferation in the pistachio. Interactions were evident in immersion time and frequency in nodal segments. Nodal buds were better than shoot tips as the highest multiple shoot formation was recorded in MS medium containing 4 mg L-1 BA and 0.1 mg L-1 GA(3) in RITA(A (R)). Although shoot tip necrosis (STN) was observed in shoots proliferated on semi-solid MS medium, such a symptom did not occur in shoots sprouted in the RITA(A (R)). Additionally, these optimized conditions were applied to nodal buds of mature male pistachio 'AtlA +/- aEuro (TM) and Pistacia rootstocks (P. khinjuk Stocks and P. atlantica Desf.), and the micropropagation in the bioreactor system, in comparison to the semi-solid medium, was also improved. Furthermore, in vitro rooting of pistachio plantlets, despite the lower range (27.5 %), was also achieved in RITA(A (R)). However, rooting was better on semi-solid medium for all tested species (ranged between 50 and 70 %). The results of this study showed that RITA(A (R)) could be used for the mass propagation of pistachio and its rootstocks, as well as for other woody plant species.Öğe Oxidative Stress of Mature Pistachio (Pistacia vera L. 'Atli') Shoot Tips During In Vitro Culture.(Springer, 2016) Akdemir, Hulya; Suzerer, Veysel; Tilkat, Engin; Onay, Ahmet; Ciftci, Yelda Ozden[Abstract Not Available]
