Cryopreservation of Mulberry Germplasm Core Collection and Assessment of Genetic Stability through ISSR Markers

Abstract

A simple and reliable cryo technique using desiccation and slow freezing of winter dormant buds was employed for 238 core collection of mulberry germ-plasm collected from diverse geographical regions and maintained under tropical conditions in the ex situ field gene bank to develop long-term biodiversity conservation for ensuring sustainable utilization of these valuable resources. Desiccation and freezing tolerance of bud grafts and excised shoot apices in the axillary buds of different Morus species under in vivo and in vitro condition indicated species-specific variation and most of the wild Morus species were found sensitive. In vitro regeneration and cryopreservation (-196°C) protocols using differentiated bud meristem like axillary winter dormant buds were worked out for a wide range of Morus species, landraces, wild and cultivated varieties. Successful cryopreservation of mulberry winter dormant buds of different accessions belonging to M. indica, M. alba, M. latfo1ia, M. cathayana, M. laevigata, M. nigra, M. australis, M. bombycis, M. sinensis, M multicaulis and M. rotundioba was achieved. Among wild species Morus tiliaefolia, and M. serrata showed moderate recovery after cryopreservation. Survival rates did not alter after three years of cryopreservation of different Morus species. ISSR markers were used to ascertain the genetic stability of cryopre-served mulberry, which showed no difference detected among the plantlets regenerated from frozen apices in comparison to the non-frozen material.