Synergizing cryopreservation, thermal analysis, and in vitro regeneration: Strategies for long-term plant germplasm preservation

Abstract

The storage of tissue under low temperature has been originated in the 18th century. During the mid-19th century, the osmotic stress has been identified as the only factor responsible for the tissues to get deteriorated during the low temperature storage. Cryopreservation should indeed be considered a vital backup method for the conservation of plant germplasm. This technique offers an effective and long-term solution for storing plant cells, tissues, seeds, and embryos at ultra-low temperatures, typically in liquid nitrogen at -196 °C. By halting all biological activity, cryopreservation ensures that the genetic material remains viable for extended periods without degradation. This approach is particularly valuable for preserving species that are rare, endangered, or have seeds that are not suitable for conventional storage methods. The cell deterioration has been overcome by the addition of penetrating and non-penetrating cryoprotective agents such as DMSO, Glycerol, PEG, Sucrose, Glucose alone or in combinations for the successful cryopreservation. In addition to this, the explants size, shape and the isolation are the critical factors in cryopreservation. The structural integrity of the tissue is the crucial factor as it determines the no changes occurred in the stored tissue during the cryopreservation. Thermal analysis using Differential Scanning Calorimetry has been employed to identify all possible factors responsible for the cell survival during the cryopreservation which provide the standardization of protocols. The DSC helps in rapid standardization of cryopreservation protocol compared to the traditional methods.