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**21**

*India* 

**Plant Cryopreservation** 

*Palode, Thiruvananthapuram, Kerala,* 

*Plant Biotechnology and Bioinformatics Division Tropical Botanic Garden and Research Institute* 

R.K. Radha, William S. Decruse and P.N. Krishnan

Two basic approaches to conservation of plant genetic resources are *ex situ* and *in situ* conservation. *Ex situ* conservation includes seed storage, *in vitro* storage, DNA storage, pollen storage, field genebanks and botanical gardens while the *in situ* approach

Cryopreservation is a part of biotechnology. Biotechnology plays an important role in international plant conservation programs and in preservation of the world's genetic resources (Bajaj, 1995; Benson, 1999). Advances in biotechnology provide new methods for plant genetic resources and evaluation (Paunesca, 2009). Cryopreservation, developed during the last 25 years, is an important and the most valuable method for long-term conservation of biological materials. The main advantages in cryopreservation are simplicity and the applicability to a wide range of genotypes (Engelmann, 2004). This can be achieved using different procedures, such as pre-growth, desiccation, pregrowth-desiccation, vitrification, encapsulationvitrification and droplet-freezing (Engelmann, 2004). Cryopreservation involves storage of plant material (such as seed, shoot tip, zygotic and somatic embryos and pollen) at ultra-low temperatures in LN (-196°C) or its vapor phase (-150°C). To avoid the genetic alterations that may occur in long tissue cultures storage, cryopreservation has been developed (Martin *et al*., 1998). At this temperature, cell division, metabolic, and biochemical activities remain suspended and the material can be stored without changes and deterioration for long time. Walters *et al*. (2009) proposed that this assumption, based on extrapolations of temperaturereaction kinetic relationships, is not completely supported by accumulating evidence that dried seeds can deteriorate during cryogenic storage. After 30 years of cryogenic storage, seeds of some species exhibited quantitatively lower viability and vigor. In cryopreservation method, subcultures are not required and somaclonal variation is reduced. Advantages of cryopreservation are that germplasm can be kept for theoretically indefinite time with low costs and little space. Besides its use for the conservation of genetic resources, cryopreservation can also be applied for the safe storage of plant tissues with specific characteristics. Different types of plant cell, tissues and organs can be cryopreserved. Cryopreservation is the most suitable long-term storage method for genetic resources of vegetatively maintained crops (Kaczmarczyk *et al*., 2008). For vegetatively propagated species, the best organs are shoot apices excised from *in vitro* plants. Shoot apices or meristems cultures are suitable because of virus-free plant production, clonal propagation, improving health status, easier recovery and

encompasses genetic reserves, on farm and home garden conservation.

**1. Introduction** 

