Cryopreservation has proven to be an important tool for the storage and conservation of plant genetic resources. This book is a unique resource for plant scientists, providing more than 100 ready-to-use cryopreservation protocols for plant types from algae and bryophytes to a range of flowering plants. It includes techniques for diverse plant parts such as dormant buds, pollen, and apical meristems and for cell types such as suspension and callus cultures.

Cryopreservation is the storage of cells, tissues and organs at ultra-low temperatures (-196 °C) and can be an important tool for long-term conservation of plant genetic resources. A wide range of cryogenic procedures have been developed for in vitro explants, dormant buds and non-orthodox seeds in many plant species; however, research that provides novelty, insight and advances for developing and improving new protocols in plant species is still needed. This book includes the development of new protocols, physical and chemical studies related to preservation in liquid nitrogen, and some reviews on several aspects of cryopreservation. This publication will be a useful document for the community of cryopreservation scientists.

Plant cell culture. Basic principles pf freezing injury to plant cells: natural tolerance and approaches to cryopreservation. Cryobiology of isolated protoplasts: application to plant cell cryopreservation. Biophysical and ultrastructural studies of membrane alterations in plant cells during extracellular freezing: molecular mechanisms of membrane injury. Cryoprotective compounds in the viable freezing of plant tissues. Meristem culture and germplasm preservation. Cryopreservation of shoot-tips of fruit trees and herbaceous plants. Cryopreservation of potato meristems...

The germ plasm of numerous plant species, especially those of forest trees, some agricultural crops, and medicinal plants, is endangered and threatened with extinction. This depletion of germplasm pools and the shrinkage of naturally occurring genetic resources have caused international concern. Conventionally, the germplasm of plants is conserved through seeds, tubers, roots, corms, rhizomes, bulbs, cuttings, etc. However, the germ plasm of a number of trees and plantation crops (such as coconut, cocao, coffee, oil palm, rubber, mango, horse chestnut, etc. ) cannot be preserved since their seed are short-lived (recalcitrant). Likewise, germplasm of vegetatively propagated crops (such as potato and cassava) cannot be stored on a long term basis and has to be grown and multiplied periodically in nurseries and fields. The plants are thus exposed to unpredictable weather conditions and diseases, with the result that instances are known where entire genetic stocks are lost. Therefore, unconventional methods are being developed for the storage and international exchange of germplasm. For this purpose in vitro cultures have been employed, but they can only enable short-to medium term preservation; moreover, cell cultures upon repeated subculture undergo genetic erosion. In view of the recent developments in the in vitro induction of genetic variability through somaclonal variation, somatic hybridization, recombinant DNA technology, etc. , new methods need to be employed for the storage of desirable cultures. In this regard freeze preservation of cells in liquid nitrogen (-196 0q, like that of semen, enables long-term storage, theoretically, for an indefinite period of time.

In addition to outlining the fundamental principles associated with the conservation of biological resources, freeze-drying and cryopreservation, this text is a compilation of cryptopreservation and freeze-drying methodologies applicable to different biological materiels, developed by expert laboratories.

While it is barely 50 years since the first reliable reports of the recovery of living cells frozen to cryogenic temperatures, there has been tremendous growth in the use of cryobiology in medicine, agriculture, horticulture, forestry, and the conservation of endangered or economically important species. As the first major text on cryobiolog

Ex situ preservation of germplasm for higher plant species has been accom plished using either seeds or clones, but storage of these under typical condi tions does not provide the extreme longevities that are needed to minimize risk of loss. Costs of maintenance and regeneration of stocks are also high. Systems that provide virtually indefinite storage should supplement existing methods and it is within this context that cryopreservation is presented. The use of low temperature preservation was initially more a concern of medicine and animal breeding, and was expanded to plants in the 1970s. Sur vival after cryogenic exposure has now been demonstrated for diverse plant groups including algae, bryophytes, fungi and higher plants. If survival is com monplace, then the eventual application is a cryopreservation system, whereby cells, tissues and organs are held indefinitely for use, often in the unforeseen future. The increasing interest and capabilities for application could not have occurred at a more opportune time since expanding human populations have placed unprecedented pressures on plant diversity. This book emphasizes cry opreservation of higher plants and was initially driven by the concern for loss of diversity in crops and the recognized need that this diversity would be essential for continued improvement of the many plants used by society for food, health and shelter. The interest in cryopreservation has been expanded by conservationists and their concerns for retaining, as much as possible, the diversity of natural populations. The need for cryopreservation, thus, is well established.