Cryopreservation of Smirnovia iranica (Sabeti) Seeds and Evaluation of Cryopreserved Seeds under Laboratory, Greenhouse and Natural Habitat Conditions

Document Type : Research and Full Length Article


Research Institute of Forests and Rangelands, AgriculturalResearch Education and Extension Organization (AREEO), Tehran, Iran


Smirnovia iranica (Sabeti) synonym: Smirnovia turkestana (Bunge) is a deciduous perennial bushy species of the Fabaceae (Papilionaceae) family. The species grow on sand dunes of desert areas, having a deep vertical root and very long horizontal or lateral roots. They have an important role in natural vegetation of desert and sand dune stabilization. Limited growing areas, grazing due to good palatability, and foliage quality of the plant have put this species under threat. Seed preservation under cryogenic conditions at -196ºC is an important approach to conserve seeds for a long period. In this study, seeds of the S. iranica were collected from natural habitats of the plants and three pre-cryopreservation treatments including PVS2, Desiccation, and 30%Glycerol were applied before transferring the seeds into liquid nitrogen (LN) at -196ºC for 1 week, 1 month, and 1 year (in 2015). Subsequently, the seeds were removed from the liquid nitrogen, imposed to heat shock and evaluated under laboratory, greenhouse and natural habitat conditions. The cryopreserved seeds of various cryopreservation periods germinated normally under laboratory and greenhouse conditions. In laboratory conditions, there were no significant differences between periods of seed storage in cryogenic conditions for seed germination trait. The highest seed germination percent (84%) was observed in the desiccation pre-cryopreservation treatment. In pre-cryopreservation treatments as well as cryogenic storage periods under greenhouse conditions, seed germination and seedling establishment were significantly different. In natural habitat, the cryopreserved seeds germinate and grow to normal seedlings and plants. The results showed that S. iranica seeds can be successfully stored in cryogenic conditions for a long period.


Main Subjects

Al Zoubi, O. M., Normah, M. N., 2012. Desiccation sensitivity and cryopreservation of excised embryonic axes of Citrus suhuiensis cv.Limau Madu, Citrumelo [Citrus paradisi Macf.× Poncirus trifoliata (L.) Raf.] and Fortunella polyandra. CryoLetters, 33: 240-250.
Avrami, M., 1941. Granulation phase change and microstructure, III. Kinetics of phase change. Journal of Chemical Physics, 9: 177-184.
Blakesley, D., Kiernan, R. J., 2001. Cryopreservation of axillary buds of a Eucalyptus grandis x Eucalyptus camaldulensis hybrid. CryoLetters, 22: 13-18.
Caswell, K. L., Kartha, K. K., 2009. Recovery of plants from pea and strawberry meristems cryopreserved for 28 years. CryoLetters, 30: 41-46.
Chmielarz, P., March, G. G., Boucaud, M. T., 2005. Cryopreservation of Quercus robur L. embryogenic calli. CryoLetters, 26: 349-356.
Flachsland, E., Terada G., Scocchi, A., Rey, H., Mroginski, L., Engelmann, F., 2006. Cryopreservation of seeds and in vitro–cultured protocorms of Oncidium bifolium Sims. (Orchidaceae) by encapsulation-dehydration. CryoLetters, 27: 235-242.
Gale, S., John, A., Harding, K., Benson, E., 2008. Developing cryopreservation for Picea sitchensis (sitka spruce) somatic embryos: a comparison of vitrification protocols. CryoLetters, 29: 135-144.
Gratton, S., Grieve, C. M., Poss, J. A., Benes S. E., 2004. Evaluation of salt-tolerant forages for sequential water reuse system: I. Biomass production. Agricultural water management, 70: 109-120.
Hao, Y. J., You, C. X., Deng, X. X., 2002. Analysis of ploidy and the patterns of amplified fragment length polymorphism and methylation sensitive amplified polymorphism in strawberry plants recovered from cryopreservation. CryoLetters, 23: 37-46.
Harding, K., Benson, E. E., 2001. The use of microsatellite analysis in Solanum tuberosum L. in vitro plantlets derived from cryopreserved germplasm. CryoLetters, 22: 199 – 208.
Hay, F. R., Muir. J. S. K., 2000. Low temperature survival of slender Naiad (Najas flexilis) seeds. CryoLetters, 21: 271-278.
Jeyendran, R. S., Van der Ven, H. H., Perez-Pelaez, M., Zaneveld, L. J., 1985. Effect of glycerol and cryopreservation on oocyte penetration by human spermatozoa. Andrologia, 17 : 241-248.
Kapustina, L. A., 2001. Biodiversity, ecology and microelement composition of Kyzylkum desert shrubs (Uzbekistan) in: USDA Forest Service Proceedings RMRS-P-21. 2001.
Kuleshova, L. L., MacFarlane, D. R., Trounson, A. O., Shaw, J. M., 1999. Sugars exert a major influence on the vitrification properties of ethylene glycol–based solutions and have low toxicity to embryos and oocytes. Cryobiology, 38: 119-130.
Makeen, A. M., Noor, N. M., Dussert, S., Clyde, M. M., 2005. Cryopreservation of whole seeds and excised embryonic axes of Citrus suhuiensis cv. Limau Langkat in accordance to their desiccation sensitivity. CryoLetters, 26: 259-268.
Naderi Shahab, M., Hatami, F., Tabari, M., Jafari. A. A., 2009. Cryopreservation and evaluation of chinese arbor-vitae (Biota orientalis) Seeds. Journal of New Seeds, 10: 264-276.
Naderi Shahab, M., Jebelli, M., Shahmoradi, A. A., Jafari A. A., 2013. Seed cryopreservation and evaluation of Ferula gummosa and Kelussia odoratissima. Seed Technology Journal, 31: 103-116.
Ozden-Tokatli, Y., Ozudogru, E. A., Gumusel, F., Lambardi, M., 2007. Cryopreservation of Pistacia spp. seeds by dehydration and one-step freezing. CryoLetters, 28 (2): 83-94
Rall, W. F., 1987. Factors affecting the survival of mouse embryos cryopreserved by vitrification. Cryobiology, 24: 387–402.
Rechinger, K. H., 1984. Flora Iranica. Akademiche Druk-U. Verlagsanstalt, Graz, Austria.
Sakai, A., Kobayashi, S., Oiyama, I., 1991. Survival by vitrification of nucellar cells of navel orange (Citrus sinensis Osb. var. brasiliensis Tanaka) cooled to -196°C. Journal of Plant Physiology, 137: 465–470.
Sánchez, C., Martinez, M. T., Vidal, N., San–José, M. C., Valladares, S., Vieitez, A. M., 2008. Preservation of Quercus robur germplasm by cryostorage of embryogenic cultures derived from mature trees and RAPD analysis of genetic stability. CryoLetters, 29: 493-504.
Stanwood, P. C., 1985. Cryopreservation of seed germplasm for genetic conservation. in: Kartha, K. K., (ed) Cryopreservation of Plant Cells and Tissues. Boca Raton, FL: CRC, 199-226.
Thammasiri, K., 2000. Cryopreservation of seeds of a Thai orchid (Doritis pulcherrima Lindl.) by vitrification. CryoLetters, 21: 237-244.
Tyagi, R. K., Hymowitz, T., 2003. Pollen from Glycine species survive cryogenic exposure. CryoLetters, 24: 119-124.
Walters, C., Wheeler, L. J., Stanwood, P. C., 2004. Longevity of cryogenically stored seeds. Cryobiology, 48: 229-244.
Wesley-Smith, J., Walters, C., Berjak, P., Pammenter, N. W., 2004. The influence of water content, cooling and warming rate upon survival of embryonic axes of Poncirus trifoliata (L.). CryoLetters, 25: 129-138.
Wood, C. B., Pritchard, H. W., Lindegaard, K., 2003. Seed cryopreservation and longevity of two Salix hybrids. CryoLetters, 24: 17-26.
Zhai, Z., Wu, Y., Engelmann, F., Chen, R., Zhao, Y., 2003. Genetic stability assessments of plantlets regenerated from cryopreserved in vitro cultured grape and kiwi shoot–tips using RAPD. CryoLetters, 24: 315-322.
Volume 7, Issue 2 - Serial Number 2
April 2017
Pages 122-137
  • Receive Date: 27 May 2016
  • Revise Date: 09 August 2016
  • Accept Date: 24 August 2016
  • First Publish Date: 01 April 2017