Strategies for Fast Multiplication and Conservation of Forest Trees by Somatic Embryogenesis and Cryopreservation: a Case Study with Cypress (Cupressus sempervirens L.)

  • Maurizio LAMBARDI IVALSA/Trees and Timber Institute, National Research Council (CNR), 50019 Florence
  • Elif Aylin OZUDOGRU IVALSA/Trees and Timber Institute, National Research Council (CNR), 50019 Florence
  • Sara BARBERINI IPSP/Institute for Sustainable Plant Protection, National Research Council (CNR), 50019 Florence
  • Roberto DANTI IPSP/Institute for Sustainable Plant Protection, National Research Council (CNR), 50019 Florence

Abstract

Common cypress (Cupressus sempervirens L.) is one of the most widespread species in the Mediterranean area. It has been traditionally cultivated for its ornamental value, becoming a typical feature of urban and rural landscapes, and high timber quality. In the last 30 years, cypress has been subjected to important breeding programmes, aimed to select clones tolerant to the widespread canker, caused by the pathogenic fungus Seiridium cardinale, leading to various patented varieties today available on the market, as well as for genotypes producing null or low amount of allergenic pollen. Somatic embryogenesis is a suitable in vitro regeneration method for fast cloning of conifer trees, and the cryopreservation of embryogenic callus is a significant tool for the safe long-term conservation of valuable cell lines. Recently, a complete protocol for the production of cypress plants from somatic embryogenesis was developed for the patented clone ‘Mediterraneo’. Here, the coupling of somatic embryogenesis and cryopreservation may offer a superior tool to propagate and maintain selected genotypes of cypress by overcoming repetitive subculturing of selected embryogenic callus lines. For the above, this study aimed to compare different cryopreservation techniques (PVS2-based vitrification and slow cooling) with the ‘Mediterraneo’ embryogenic callus line. Best results were obtained after the optimization of a slow cooling procedure, based on the 30-min treatment of embryogenic masses with a cryoprotective solution containing 180 g l-1 sucrose and 7.5% DMSO, followed by the reduction of the temperature at a rate of -1 °C min-1 up to -40 °C and the subsequent immersion in liquid nitrogen  (“two-step freezing”).

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Published
2018-01-01
How to Cite
LAMBARDI, M., OZUDOGRU, E. A., BARBERINI, S., & DANTI, R. (2018). Strategies for Fast Multiplication and Conservation of Forest Trees by Somatic Embryogenesis and Cryopreservation: a Case Study with Cypress (Cupressus sempervirens L.). Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 46(1), 32-38. https://doi.org/10.15835/nbha46111011
Section
Research Articles