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Öğe Biobanking of vegetable genetic resources by in vitro conservation and cryopreservation(Springer Science+Business Media B.V., 2020) Ruta, Claudia; Lambardi, Maurizio; Özüdoğru, Elif AylinToday, 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.Öğe Establishment of an Efficient Somatic Embryogenesis Protocol for Giant Reed (Arundo donax L.) and Multiplication of Obtained Shoots via Semi-Solid or Liquid Culture(Mdpi, 2023) Ozudogru, Elif Aylin; Karlik, Elif; Elazab, Doaa; Lambardi, MaurizioThis study developed an efficient protocol for the in vitro propagation of giant reed (Arundo donax L.) biomass, defining a complete cycle of the induction of somatic embryogenesis from immature inflorescences, followed by the maturation of somatic embryos and the subsequent multiplication of the derived shoots in liquid culture in a temporary immersion system (TIS). The best explants were found to be 30 cm long immature inflorescences, preferably collected in spring. Such an explant type was easy to decontaminate, and the spikelets isolated from it provided over 100 embryogenic callus lines. Among the callus induction media tested, gelled MS medium supplemented with 1.1 mg/L 2,4-D provided the highest percentage of responsive spikelets and the highest density of embryogenic callus. Maturation of the embryogenic callus was easily triggered on gelled MS medium devoid of plant growth regulators. The obtained shoots could be further multiplied on previously optimized gelled DKW medium supplemented with 30 g/L sucrose, 5 mg/L BA, 0.1 mg/L IBA, and 6.8 g/L plant agar. Subsequent high multiplication of the developed shoots was achieved in liquid culture in TIS using a Plantform & TRADE; bioreactor, with an immersion cycle of 12 min every 8 h.Öğe 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, MaurizioPeanuts (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.Öğe 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, MaurizioChickpea (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.Öğe Use of Liquid Culture with the ElecTIS Bioreactor for Faster Recovery of Blackberry (Rubus fruticosus L.) Shoots from Conservation at 4 °C(Mdpi, 2023) Elazab, Doaa; Capuana, Maurizio; Ozudogru, Elif Aylin; Anichini, Monica; Lambardi, MaurizioElecTIS is a new single container bioreactor which does not require forced air blowing, instead making the culture material mobile and the liquid medium stationary. The timed up-and-down movement of the basket containing the shoot culture ensures periodic contact with the liquid medium positioned at the base of the container. In this study we tested for the first time its use in the recovery of blackberry shoot cultures (Rubus fruticosus L., cvs Thornfree and Chester), coming from 5 months of slow growth storage (SGS), at 4 & DEG;C and in the dark. The shoot recovery at standard culture conditions was performed on two different types of ElecTIS, i.e., one with a smaller basket (ElecTIS(S), 234 cm(2) of culture area), and one with a large basket (ElecTIS(L), 336 cm(2)), comparing the culture in TIS (cycle of 8 min every 6 h, equal to 32 min/day) with the traditional one in a gelled medium in glass jars (500 cc). After each one of the three 4-week subcultures, the shoot growth parameters and the relative growth rate highlighted a clear superiority of ElecTIS in promoting the recovery of shoot cultures coming from SGS. The analyses of chlorophyll content and stoma functionality confirmed the superior quality of shoots cultured in the ElecTIS bioreactor, and these shoots were afterwards easily rooted and acclimatized ex vitro.
