In vitro techniques are very useful for conserving plant biodiversity, including (a) genetic resources of recalcitrant seed and vegetatively propagated species, (b) rare and endangered plant species and (c) biotechnology products such as elite genotypes and genetically engineered material. Explants from recalcitrant seed and vegetatively propagated species can be efficiently collected under field conditions using in vitro techniques. In vitro culture techniques ensure the production and rapid multiplication of disease-free material. Medium-term conservation is achieved by reducing growth of plant material, thus increasing intervals between subcultures. For long-term conservation, cryopreservation (liquid nitrogen, −196°C) allows storing plant material without modification or alteration for extended periods, protected from contaminations and with limited maintenance. Slow growth storage protocols are routinely employed for a large number of species, including numerous endangered plants, from temperate and tropical origin. Cryopreservation is well advanced for vegetatively propagated species, and techniques are ready for large-scale experimentation in an increasing number of cases. Research is much less advanced for recalcitrant species due to their seed characteristics, viz., very high sensitivity to desiccation, structural complexity and heterogeneity in terms of developmental stage and water content at maturity. However, various technical approaches should be explored to develop cryopreservation techniques for a larger number of recalcitrant seed species. A range of analytical techniques are available, which allow understanding physical and biological processes taking place in explants during cryopreservation. These techniques are extremely useful to assist in the development of cryopreservation protocols. In comparison with crop species, only limited research has been performed on cryopreservation of rare and endangered species. Even though routine use of cryopreservation is still limited, an increasing number of examples where cryopreservation is used on a large scale can be found both in genebanks for crops and in botanical gardens for endangered species.
In vitro medium term conservation of tropical plant germplasm is used routinely in many laboratories. Growth reduction is achieved by modifying various parameters, such as temperature, culture medium, gaseous environment. For long term conservation, cryopreservation (i.e. storage in liquid nitrogen, -196 ° C) is the only current method available. Each successive step of the process requires precise conditions which have to be defined for each material. Cryopreservation protocols have been set up for more than 40 tropical species. Results obtained with various culture systems such as cell suspensions, protoplasts, calluses, meristems and embryos are discussed. The first example of the large scale application of cryopreservation (oil palm somatic embryos) is presented.
Advances in plant biotechnology provide new options for collection, multiplication and short-to long-term conservation of plant biodiversity, using in vitro culture techniques. Significant progress has been made for conserving endangered, rare, crop ornamental, medicinal and forest species, especially for non-orthodox seed and vegetatively propagated plants of temperate and tropical origin. Cell and tissue culture techniques ensure the rapid multiplication and production of plant material under aseptic conditions. Medium-term conservation by means of in vitro slow growth storage allows extending subcultures from several months to several years, depending on the species. Cryopreservation (liquid nitrogen, −196 °C) is the only technique ensuring the safe and cost-effective long-term conservation of a wide range of plant species. Cryopreservation of shoot tips is also being applied to eradicate systemic plant pathogens, a process termed cryotherapy. Slow growth storage is routinely used in many laboratories for medium-conservation of numerous plant species. Today, the large-scale, routine application of cryopreservation is still restricted to a limited number of cases. However, the number of plant species for which cryopreservation techniques are established and validated on a large range of genetically diverse accessions is increasing steadily.
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