Abstract
Adventitious shoot organogenesis and somatic embryogenesis are the basis for implementing new genetic variability and biotechnological approaches in woody species, particularly if mature tissues from valuable cultivars are used. To date, these technologies are only applied to few tree species due to the absence of efficient regeneration protocols. In hazelnut, adventitious shoot organogenesis and somatic embryogenesis have only been carried out with zygotic embryo tissues up to now. Here we report plant regeneration from explants of somatic origin by using in vitro rejuvenated mature tissues (leaves, petioles and stipules). A histological analysis carried out on calli grown on various media, showed significant evidence of shoot regeneration, as proved by the presence of vascular elements such as tracheids with annular or helical secondary wall thickening. Subsequently, the optimization of the regeneration protocol performed by pre-treating the explants with antibiotics (carbenicillin, vancomycin, and cefotaxime) as molecules with auxin-like effects enabled us to achieve shoot organogenesis in hazelnut. The organogenic competence strictly depended on the explants and antibiotics used in the experiments. Following an antibiotic pre-treatment of cv Tonda Gentile Romana explants in the proliferation stage, organogenic responses (frequencies of 40 %) were obtained. According to the results obtained, the best protocol for inducing shoot organogenesis in hazelnut should include the use of explants (leaves and petioles) conditioned in a double-liquid layer of cefotaxime 1000 mg L−1 added to the proliferation medium, cultured on the induction medium, consisting in solid MS medium supplemented with 3 % sucrose, 6-benzylaminopurine 1 mg L−1, indole-3-butyric acid 1 mg L−1 and kinetin 2 mg L−1, and then by sub-culturing the newly-formed calli to regeneration medium, consisting of half-strength solid MS medium with sucrose (30 g L−1) and 6-benzylaminopurine (0.5 mg L−1).
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References
Ayun A, San B, Erdogan V (2009) Induction of somatic embryogenesis from mature cotyledons in Tombul hazelnut. Tarim Bilimleri Dergisi 15(2):113–118
Bacchetta L, Aramini M, Bernardini C, Rugini E (2008) In vitro propagation of traditional Italian hazelnut cultivars as a tool for the valorisation and conservation of local genetic resources. HortScience 43(2):562–566
Bosela MJ (2009) Effect of β-lactam antibiotics, auxins, and cytokinins on shoot regeneration from callus cultures of two hybrid aspens, Populus tremuloides x P-tremula and P. xcanescens x P. gradidentata. Plant Cell Tissue Organ 98:249–261
Cristofori V, Ferramondo S, Bertazza G, Bignami C (2008) Nut and kernel traits and chemical composition of hazelnut (Corylus avellana L.) cultivars. J Sci Food Agric 88(6):1091–1098
Crouch NR, van Staden LF, Van Staden J, Drewes FE, Drewes SE, Meyer HJ (1993) Accumulation of cyanidin-3-glucoside in callus and cell cultures of Oxalis reclinata. J Plant Physiol 142(1):109–111
Gatz A, Kowalski T (2011) Tracheary element differentiation and morphogenetic changes in callus derived from embryos of pepper (Capsicum annuum L.). Acta Sci Pol 10(1):131–146
Hand C, Reed BM (2014) Minor nutrients are critical for the improved growth of Corylus avellana shoot cultures. Plant Cell Tissue Organ 119(2):427–439. doi:10.1007/s11240-014-0545-x
Hand C, Maki S, Reed BM (2014) Modeling optimal mineral nutrition for hazelnut micropropagation. Plant Cell Tissue Organ 119(2):411–425. doi:10.1007/s11240-014-0544-y
Kaur A, Gill MS, Ruma D, Gosal SS (2008) Enhanced in vitro shoot multiplication and elongation in sugarcane using cefotaxime. Sugar Tech 10(1):60–64
Manchanda P, Kaur A, Gosal SS (2011) Impact of cefotaxime on in vitro shoot elongation and regeneration in banana (Musa acuminata). J Appl Hortic 13(1):52–55
Mehlenbacher SA (1997) Testing compatibility of hazelnut crosses using fluorescence microscopy. Acta Hortic 445:167–171
Mehlenbacher SA (2009) Genetic resources for hazelnut: state of the art and future perspectives. Acta Hortic 845:33–38
Mulinacci N, Giaccherini C, Santamaria AR, Caniato R, Ferrari F, Valletta A, Pasqua G (2008) Anthocyanins and xanthones in the calli and regenerated shoots of Hypericum perforatum var. angustifolium (sin. fröhlich) borkh. Plant Physiol Biochem 46(4):414–420
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497
Nas MN, Read PE, Miller V, Rutter PA (2003) In vitro “rejuvenation” of woody species is temporary. Acta Hortic 625:211–215
O’Brien TP, McCully ME (1981) The study of plant structure: principles and selected methods. Termarcarphi Pty. Ltd., Melbourne
Paulraj S, Lopez-Villalobos A, Yeung EC (2014) Abscisic acid promotes shoot regeneration in Arabidopsis zygotic embryo explants. In Vitro Cell Dev Plant 50(5):627–637
Radojevic LJ, Vujicic R, Neskovic M (1975) Embryogenesis in tissue culture of Corylus avellana L. Z Pflanzenphysiol 77(1):33–41
Rodriguez A, Vazquez A, Rodriguez R, Sanchez Tames R (1984) Cambios macromorfologicos inducidos por 6-Bencilaminopurina durante la gemrinacion de semillas de Corylus avellana L. Rev Biol Univ Oviedo 2:67–78
Rugini E, Caricato G (1995) Somatic embryogenesis and plant recovery from mature tissues 354 of olive cultivars (Olea europaea L.) “Canino” and “Moraiolo”. Plant Cell Rep 14(4):257–260
Tanprasert P, Reed BM (1997) Determination of minimal bactericidal and effective antibiotic treatment concentrations for bacterial contaminants from micropropagated strawberries. In Vitro Cell Dev Plant 33:227–230
Yepes LM, Aldwinckle HS (1994) Micropropagation of thirteen Malus cultivars and rootstocks 373 and effect of antibiotics on proliferation. Plant Growth Regul 15:55–67
Acknowledgments
This study was supported by the Italian Ministry of Agricultural and Forestry Policy (MIPAAF 2011-D.D. 17744 “VI.VA.CO.—Sviluppo del vivaismo e della piattaforma varietale corilicola”).
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Cristian Silvestri was responsible for conception and design of experiments, data analysis, drafting of the manuscript and edited the paper. Valerio Cristofori took care of study conception and design and drafting the manuscript. Marilena Ceccarelli conducted the histological analysis. Maria Eugenia Caceres conducted the histological analysis. Josep Escribà Lacuesta took care of study conception, design of experiments and data analysis. Eddo Rugini took care of study conception and design and edited the manuscript. All authors read and approved the manuscript.
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Silvestri, C., Cristofori, V., Ceccarelli, M. et al. Adventitious shoot organogenesis from leaf and petiole explants of European hazelnut. Plant Cell Tiss Organ Cult 126, 59–65 (2016). https://doi.org/10.1007/s11240-016-0976-7
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DOI: https://doi.org/10.1007/s11240-016-0976-7