In vitro conservation (slow growth storage)

The in vitro conservation (slow growth storage) of shoot cultures is an important part of the productive cycle of commercial micropropagation laboratories, as it allows to reduce markedly the frequency of subcultures and the consequent risks of contamination, to widen the laboratory offer of species and varieties and to facilitate the productive organization. In spite of that, the research never examined thoroughly the various parameters influencing this important step of the micropropagation cycle which, in commercial laboratory, is always accomplished in simple cold chambers at 4-5°C and in darkness. The maintenance of shoot cultures is obtained through the slowing down of cell metabolism e, as a consequece, of shoot growth. Although the temperature is the most effective factor, other conservation conditions (e.g., the photoperiod, light intensity, container characteristics, medium composition and the use of osmotically active compounds, such as sucrose and mannitol) can interfere on shoot growth and influence markedly the maximum storage time. By means of the optimization of such parameters, the subculture time is greatly enhanced, the quality of the stored material is maintained and the recover of the shoot proliferative potential is assured. Recent studies, carried out at the CNR-IVALSA, show that, through the optimization of the medium composition and the choice of appropriate containers, it is possible to improve the conservation at 4°C and in darkness of fruit species shoot cultures. The storage in media with high sucrose concentrations (up to 60 g/l), for istance, markedly improves the quality of shoot cultures of kiwi, Gisela 5® and Mirabolano 29C, stored for 18 months at 4°C and in the dark. The gas-chromatographic analysis of CO2 and ethylene, accumulating in the storage jars, allows to monitor the level of cell metabolism and the sanitary condition of shoot cultures. Experiments on the in vitro slow growth storage of ornamental species (Ranunculus, Anthurium, Carex) are presently in progress.


Carla Benelli (person in charge)
Anna De Carlo
Maurizio Lambardi
Elif Aylin Ozudogru


Lambardi M., Roncasaglia R., Previati A., De Carlo A., Dradi G., Da Re F., Calamai L., 2006. In vitro slow growth storage of fruit rootstocks inside gas-tight or gas-permeable containers. Acta Hortic. 725: 483-488.

Previati A., Benelli C., Da Re F., Ozudogru E.A., Lambardi M., 2008. Micropropagation and in vitro conservation of virus-free rose germplasm. Prop. Ornam. Plants 8(2): 93-98.

Ozudogru E.A., Previati A., Lambardi M., 2010. In vitro conservation and cryopreservation of ornamental plants. In: S.M. Jain and S.J. Ochatt (eds) Protocols for In Vitro Propagation of Ornamental Plants. Methods in Molecular Biology, vol. 589. Humana Press-Springer, New York, pp. 303-324.

[Last updated on: 02-11-2015]
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