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    Biobanking of vegetable genetic resources by in vitro conservation and cryopreservation
    (Springer Science+Business Media B.V., 2020) Ruta, Claudia; Lambardi, Maurizio; Özüdoğru, Elif Aylin
    Today, application of in vitro culture by means of slow growth storage of shoot cultures and cryopreservation of organs, tissues and cells in liquid nitrogen presents a remarkable strategic tool to support medium- and long-term conservation of plant genetic resources. Over the last 30 years, considerable progresses have been made in the development of both methods that are currently considered as ex situ conservation strategies, complementary to traditional seed banks and in-field clonal collections. Efficient protocols were developed for the conservation of a large number of crops, including strategically-important vegetables, such as garlic, artichoke, asparagus, cassava, Jerusalem artichoke, mint, potato, sweet potato, chicory, taro, thyme and yam. As a consequence, more than 45,000 accessions of vegetable crops are maintained in 22 genetic resources conservation centers (biobanks), located in 16 countries and 6 continents (Europe, Asia, Africa, Oceania, North and South America). Approximately 4/5 of these accessions are maintained in vitro by means of slow growth storage of shoot cultures, but cryopreservation is also constantly growing, with almost 8300 vegetable accessions being stored in liquid nitrogen at ? 196 °C.
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    Encapsulation of in vitro-derived propagules of two genotypes of Capparis spinosa (L.) from Pantelleria Island
    (International Society for Horticultural Science, 2020) Germana, Maria Antonietta; Lesto, F.; Lo Bosco, Fabrizia; Iacuzzi, Nicolo; Özüdoğru, Elif Aylin
    Encapsulation technology, combining the advantages of zygotic or gamic seed with those of micropropagation, has recently attracted the interest of researchers as a new propagation approach, mainly due to the unsatisfying results of the traditional propagation strategies. The encapsulation of uninodal microcuttings (3-4 mm long) from two Sicilian Capparis spinosa (L.) genotypes (from Pantelleria Island: 'Tracino' and 'Scauri') was evaluated, observing the influence of the calcium alginate coating and of three different growth regulators (PGRs): 6-benzylaminopurine (BAP), meta-topolin (MT) and zeatin (ZEA), on viability, regrowth and conversion of the propagules. Caper microcuttings were dissected and placed in different Murashige and Skoog-based artificial endosperms, enriched of either MT, ZEA or BAP. The synthetic seeds obtained were sown on a medium with full strength salts concentration, enriched with 0.4 mg L-1of naphtalene acetic acid and 0.7 mg L-1of gibbelleric acid. After 60 days, the following parameters were detected: viability, regrowth, number and length of the shoots and roots, conversion. The results confirm that encapsulation did not negatively affect the viability, which showed the highest percentage with BAP (100%) in 'Tracino' and with ZEA (100%) in 'Scauri'. Similar results were obtained in regrowth, with statistically significant differences among the three PGRs tested: 'Tracino' showed the best regrowth on capsuled enriched with BAP (100%), 'Scauri' with ZEA (100%). In addition, the synseed conversion was greatly affected by the PGR, and it was higher in artificial endosperm added with BAP in 'Tracino' (56.6%) and ZEA in 'Scauri' (23.3%) genotype.
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    Establishment of direct organogenesis protocol for arachis hypogaea cv. virginia in liquid medium by temporary immersion system (TIS)
    (MDPI, 2022) Özüdoğru, Elif Aylin; Karlık, Elif; Elazab, Doaa; Lambardi, Maurizio
    Peanuts (Arachis hypogaea L.) are a rich source of herbal oil, proteins, minerals, vitamins, fibers, essential amino acids, as well as bioactive compounds, and are thus widely used for human nutrition and animal feed, and for prevention from certain diseases. However, the in vitro regeneration response of the species is generally low, and it also displays a significant variability among its varieties. Thus, the development of advanced protocols and approaches for the in vitro propagation of peanut is still of immense importance. A recently developed in vitro propagation technique, TIS; Temporary Immersion Bioreactor System, provides a new approach for the mass propagation of plants. Accordingly, the present study provides an efficient de novo regeneration protocol for Arachis hypogaea L. cv. Virginia by using a TIS. Different concentrations of cytokinins, i.e., benzyladenine (BA) or thidiazuron (TDZ), were tested with several combinations of dry and medium immersion periods of TIS, corresponding to a total of 8, 12, 16, 24, 32, 36, 48, 64, 72, and 96 min daily immersions for the induction of direct organogenesis. The study exhibited that an MS medium added to 110 µM BA or 10 µM TDZ are the most appropriate medium formulations in TIS, when applied for 16 min every 16 h. The application of optimized procedures to cv. NC7 and two valuable Turkish autochthonous varieties, 7 × 77 and Com74, is also reported. To the best of our knowledge, the present study draws attention also for being the first study in which a TIS was used for peanuts. © 2022 by the authors.
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    Long-term preservation of Cicer arietinum L. germplasm by in vitro propagation and cryopreservation
    (Springer, 2020) Ruta, Claudia; De Mastro, Giuseppe; Tarraf, Waed; Ancona, Simona; Tagarelli, Anna; Özüdoğru, Elif Aylin; Lambardi, Maurizio
    Chickpea (Cicer arietinum L.) is one of the most important pulse worldwide. This crop plays a significant role to maintain soil fertility through symbiotic N fixation, as well as in the Mediterranean diet for its important content of noble protein. In South Italy, particularly in the Puglia region, many traditional landraces are still cultivated in marginal areas, becoming therefore at strong risk of genetic erosion or even extinction. In vitro culture is a useful and innovative approach for the collection and the long-term preservation of threatened germplasm by means of the cryopreservation technology, as ex situ conservation strategy complementary to traditional ones. The aim of this study was to develop an efficient protocol for the multiplication and cryopreservation of two accessions of an Apulian black chickpea threatened landrace. Seeds of Apulian black chickpea were inoculated on agarized sucrose-free nutrient medium. The cotyledonary node and axillary buds were excised from the seedlings and then cultured on the same medium, supplemented with 6-benzylaminopurine and sucrose. After three subculture cycles, shoot tips from in vitro proliferated shoots were induced to rooting on IBA or used for germplasm conservation by comparing three cryopreservation techniques, i.e. PVS2-vitrification, droplet-vitrification and V-cryoplate. The results are very promising in terms of explant survival. However, the study should now progress to optimize a recovery medium, able to further improve shoot regrowth rates and plantlet formation in post-cryopreservation. © 2019, Springer Nature B.V.
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    Türkiye'de Doğal Olarak Yayılış Gösteren ve Endemik Bazı Colchicum Türlerinde Somatik Embriyogenezis, Kriyoprezervasyon ve Sentetik Tohum Üretim Protokollerinin Geliştirilmesi
    (TÜBİTAK, 01.04.2021) Mendi, Neslihan Yeşim Yalçın; Uncuoğlu, Ahu Altınkut; Buzkan, Nihal; Özüdoğru, Elif Aylin; Şimşek, Özhan; Çürük, Pembe Evci; Kaya, Ergun; Koyuncu, Onur; Batmaz, Murat; Özdemir, Muhammet Şamil; Sevindik, Başar; Uğur, Senem; Alan, Semih
    Bu proje kapsamında, ülkemizde doğal yayılış gösteren ve bazıları endemik olan 13 Colchicum taksonunda [C. baytopiorum, C. sanguicolle, C. cilicicum, C. speciosum, C. triphyllum, C. szovitsii, C. burtii, C. atticum, C. sobolifera, C. variegatum, C. imperatoris, C. serpentinum, C. figlalii] somatik embriyogenezis protokolünün geliştirilmesi, in vitro koruma olanaklarının araştırılması (kriyoprezervasyon) ve sentetik tohum üretimi çalışmaları gerçekleştirilmiştir. Colchicum türlerinin in vitro koşullarda çoğaltım olanaklarının araştırılması amacıyla, soğan eksplantları kullanılarak in vitro rejenerasyon denemeleri kurulmuş, denemelerde farklı bitki büyüme düzenleyicilerinden BA, 2iP, TDZ?nin 2,4-D ve NAA ile kombinasyonları kullanılmış, eksplantların in vitro?da gelişimleri incelenmiş, şişkinleşme, kallus, embriyo ve çimlenme oranları saptanmıştır. Toplam sekiz Colchicum türüne, C. baytopiorum, C. sanguicolle, C. cilicicum, C. triphyllum, C. speciosum, C. szovitsii, C. imperatoris, C. figlalii ait kalluslar PVS2 vitrifikasyon yöntemiyle kriyoprezerve edilmiştir. Kriyo-saklama sonrası, kalluslarda hücre canlılığını belirlemek için tetrazolüm testi kullanılmış ve sıvı azot ile muamele edilen kalluslardan sadece C. baytopiorum, C. triphyllum, C. speciosum, ve C. figlalii türlerinde canlılık belirlenmiştir. Ayrıca, kriyoprezervasyon sonrası C. baytopiorum kallus hücrelerinde somatik embriyogenez görülmezken, C. figlalii, somatik embriyo ve sağlıklı bitkicik oluşturmuştur ve en iyi rejenerasyon %66.7 olarak hesaplanmıştır.