Muammer CEYLAN, Ergun KAYA, Emel GÖKDOĞAN YILMAZ, Fernanda Vidigal Duerte SOUZA, Maria M. JENDEREK

Cryopreservation of citrus seed via dehydration followed by immersion in liquid nitrogen

An important method for plant germplasm conservation is offered by a biotechnology-based approach of cryopreservation. Cryopreservation refers to the storage of plant material at ultralow temperatures in liquid nitrogen. A procedure for cryopreservation of polyembryonic seeds was improved for selected citrus cultivars from Turkey. Seed dehydration was performed at different exposure times, in sterile conditions of a laminar flow-hood. The tested cultivars showed the highest tolerance to low temperature storage when the seeds were first dehydrated to a moisture content of 21.8% for Poncirus trifoliata Raf. × Citrus sinensis Osb. and to 17.6% for Citrus limonia Osbeck. The postcryopreservation germinability ranged from 73.3% (Poncirus trifoliata Raf. × C. sinensis Osb. and Fortunella margarita (Lour.) Swingle) to 93.3% (C. jambhiri Lush.). Dehydration was beneficial for germination of seeds from all of the tested citrus species after the liquid nitrogen exposure. Seedlings derived from cryopreserved seeds had well-formed shoots and roots and were easily acclimated to greenhouse conditions.

PDF

___

Baümler E, Cuniberti A, Nolasco SM, Riccobene IC (2006). Moisture dependent physical and compression properties of safflower seed. J Food Eng 72: 134-140.
Benson EE (1999). Cryopreservation. In: Benson EE, editor. Plant Conservation Biotechnology. London, UK: Taylor & Francis, pp. 83-96.
Bonner FT (1990). Storage of seeds: potential and limitation for germplasm conservation. Forest Ecol Manag 35: 35-43.
Cho EG, Hor YL, Kim HH, Rao VR, Engelmann F (2001). Cryopreservation of Citrus madurensis zygotic embryonic axes by vitrification: importance of pregrowth and preculture conditions. Cryo-Lett 22: 391-396.
Cho EG, Hor YL, Kim HH, Rao VR, Engelmann F (2002a). Cryopreservation of Citrus madurensis embryonic axes by vitrification: importance of loading and treatment with vitrification solution. Cryo-Lett 23: 317-324.
Cho EG, Hor YL, Kim HH, Rao VR, Engelmann F (2002b). Cryopreservation of Citrus madurensis embryonic axes by encapsulation-dehydration. Cryo-Lett 23: 325-332.
Cho EG, Hor YL, Kim HH, Rao VR, Engelmann F (2002c). Cryopreservation of Citrus aurantifolia seeds and embyonic axes using a desiccation protocol. Cryo-Lett 23: 309-316.
Duran-Vila N (1995). Cryoconservation of germplasm of Citrus. In: Bajaj YPS, editor. Biotechnology in Agriculture and Forestry 32. Cryopreservation of Plant Germplasm. Berlin, Germany: Springer-Verlag, pp. 70-85.
Dussert S, Chabrillange N, Rocquelin G, Engelmann F, Lopez M, Hamon S (2001). Tolerance of coffee (Coffea sp.) seeds to ultralow temperature exposure in relation to calorimetric properties of tissue water, lipid composition, and cooling procedure. Physiol Plantarum 112: 495-504.
Ellis RH, Hong TD, Roberts EH (1990). An intermediate category of seed storage behaviour? I. Coffee. J Exp Bot 41: 1167-1174.
Gonzales-Arnao MT, Ortega JC, Navarro L, Duran-Vila N (2003). Cryopreservation of ovules and somatic embryos of Citrus using the encapsulation-dehydration technique. Cryo-Lett 24: 85-94.
Graiver N, Califano A, Zaritzky N (2011). Partial dehydration and cryopreservation of Citrus seeds. J Sci Food Agr 91: 2544-2550.
Hamilton KN, Ashmore SE, Pritchard HW (2009). Thermal analysis and cryopreservation of seeds of Australian wild Citrus species (Rutaceae): Citrus australasica, C. inodora and C. garrawayi. Cryo-Lett 30: 268-279.
Hor YL, Kim Y, Ugap JA, Charbrillange N, Sinniah UR, Engelmann F, Dussert S (2005). Optimal hydration status for cryopreservation of intermediate oily seeds: citrus as a case study. Ann Bot 95: 1153-1161.
Kaya E, Alves A, Rodrigues L, Jenderek M, Hernandez-Ellis M, Ozudogru A, Ellis D (2013). Cryopreservation of Eucalyptus genetic resources. Cryo-Lett 34: 608-618.
Kholina AB, Voronkova NM (2012). Seed cryopreservation of some medicinal legumes. Journal of Botany 2012: 186891.
King MW, Soetisna U, Roberts EH (1981). The dry storage of citrus seeds. Ann Bot 48: 865-872.
Koltunow AM (1993). Apomixes: embryo sac and embryos formed without meiosis or fertilization in ovules. Plant Cell 5: 1425- 1437.
Koltunow AM, Soltys K, Nito N, McClure S (1995). Anther, ovule, seed and nucellar embryo development in Citrus sinensis L. cv Valencia. Can J Bot 73: 1567-1582.
Lambardi M, De Carlo A, Biricolti S, Puglia AM, Lombardo G, Siragusam DE, Pasaquale F (2004). Zygotic and nucellar embryo survival following dehydration and cryopreservation of citrus intact seeds. Cryo-Lett 25: 81-90.
Lambardi M, Halmagyi A, Benelli C, De Carlo Vettori C (2007). Seed cryopreservation for conservation of ancient Citrus germplasm. Adv Hort Sci 21: 198-202.
Marascuilo LA, McSweeney M (1977). Post-hoc multiple comparisons in sample preparations for test of homogeneity. In: McSweeney M, Marascuilo LA, editors. Non-Parametric and Distribution-Free Methods for the Social Sciences. Pacific Grove, CA, USA: Brooks/Cole Publications, pp. 141-147.
Marin ML, Duran-Vila N (1988). Survival of somatic embryos and recovery of plants of sweet orange [Citrus sinensis (L.) Osb.] after immersion in liquid nitrogen. Plant Cell Tiss Org Cult 14: 51-57.
Marzalina M, Krishnapillay B (1999). Recalcitrant seed biotechnology application to rain forest conservation. In: Benson EE, editor. Plant Conservation Biotechnology. London, UK: Taylor & Francis, pp. 265-276.
Michalak M, Plitta BP, Chmielarz P (2013). Desiccation sensitivity and successful cryopreservation of oil seeds of European hazelnut (Corylus avellana). Ann Appl Biol 163: 351-358.
Michalak M, Plitta-Michalak BP, Chmielarz P (2015a). A new insight in desiccation tolerance and cryopreservation of mazzard cherry (Prunus avium L.) seeds. Open Life Science 10: 354-364.
Michalak M, Plitta-Michalak BP, Chmielarz P (2015b). Desiccation tolerance and cryopreservation of wild apple (Malus sylvestris) seeds. Seed Sci Technol 43: 1-12.
Mumford PM, Grout BWW (1979). Desiccation and low temperature (-196°C) tolerance of Citrus limon seed. Seed Sci Technol 7: 407-410.
Murashige T, Skoog F (1962). A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473-497.
Normah MN, Serimala SD (1995). Cryopreservation of seeds and embryonic axes of several Citrus species. In: Ellis RH, Black M, Murdoch AJ, Hong TD, editors. Basic and Applied Aspects of Seed Biology. Dordrecht, the Netherlands: Kluwer, pp. 817- 823.
Olivares-Fuster O, Asins MJ, Duran-Vila N, Navarro L (2000). Cryopreservation of callus, a source of protoplasts for Citrus improvement. J Hortic Sci Biotech 75: 635-640.
Ozudogru EA, Kaya E, Kirdok E, Issever-Ozturk S (2011). In vitro propagation from young and mature explants of thyme (Thymus vulgaris and T. longicaulis) resulting in genetically stable shoots. In Vitro Cell Dev Biol-Plant 47: 309-320.
Pixton SW (1966). Moisture contentits significance and measurement in stored products. Stored Prod Res 3: 35-47.
Roberts EH (1973). Predicting the storage life of seeds. Seed Sci Technol 1: 499-514.
Ruiz C, Breto MP, Asisns MJ (2000). A quick methodology to identify sexual seedlings in citrus breeding programs using SSR markers. Euphytica 112: 89-94.
Sakai A, Kobayashi S, Oiyama I (1991). Cryopreservation of nucellar cells of navel orange (Citrus sinensis Osb.) by a simple freezing method. Plant Sci 74: 243-248.
Stanwood PC (1985). Cryopreservation of seed germplasm for genetic conservation. In: Kartha KK, editor. Cryopreservation of Plant Cells and Organs. Boca Raton, FL, USA: CRC Press, pp 199-226.
Van Waes JM, Debergh PC (1986). Adaptation of the tetrazolium method for testing the seed viability, and scanning electron microscopy study of some Western European orchids. Physiol Plantarum 66: 435-442.
Wang ZC, Deng XX (2004). Cryopreservation of shoot tips of Citrus using vitrification: effect of reduced form of glutathione. CryoLett 25: 43-50.