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    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, Maurizio
    This 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.
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    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, Maurizio
    ElecTIS 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.