**2. Spices and germplasm conservation**

Spices and herbs are aromatic plants*–*fresh or dried plant parts like foliage, young shoots, roots, bark, buds, seeds, berries and other fruits of which are mainly used to flavour our culinary preparations, confectionary. They are also major ingredients in indigenous medicine and perfumery. Spices and herbs are grown throughout the world*–*different plant species in different regions. Peninsular India is a rich repository of spices and over 100 species of spices and herbs are grown. The other major spice growing countries are Brazil, China, Guatemala, Indonesia, Madagascar, Nigeria, West Indies, Malaysia, Sri Lanka, Spain, Turkey, Mediterranean region and the Central America. Black pepper, cardamom, ginger, turmeric, vanilla, capsicum, cinnamon, clove, nutmeg, tamarind, coriander, cumin, fennel, fenugreek, dill, caraway, anise and herbs like saffron, lavender, thyme, oregano, celery, anise, sage and basil are important as spices. India being the native home of many spices, their conservation and characterization are one of the priority programmes. Deforestation, habitat degradation and overexploitation caused considerable loss of diversity in spices.

Cryopreservation of Spices Genetic Resources 459

1979) for black pepper and its related species, cinnamon, camphor and cassia. Simultaneously these tissue-cultured plants are being evaluated for their morphological and genetic stability in culture (Luckose *et al,* 1993, Chandrappa *et al*, 1997, Nirmal babu *et al* 2003, Madhusoodanan *et al* 2005). Though micropropagation protocols were standardized using growth regulators, all the *in vitro* storage experiments were carried out using growth regulators free media to reduce

> **Concentration (mgl-1) MS**

**Concentration (mgl-1) WPM**

**Concentration (mgl-1) SH** 

the rate of multiplication which in turn will reduce the extent of variation.

**formula** 

Ammonium nitrate NH4NO3 1650.00 400.00 - Ammonium phosphate NH4H2PO4 - - 300.00 Potassium nitrate KNO3 1900.00 - 2500.00 Calcium chloride CaCl2.2H2O 440.00 - - Calcium chloride CaCl2 - 72.50 151.00 Calcium nitrate Ca(NO3)2.4H2O - 386.00 -

orthophosphate KH2PO4 170.00 170.00 - Potassium sulfate K2SO4 - 990.00 - Magnesium sulphate MgSO4.7H2O 370.00 180.70 195.40

Sodium EDTA Na2EDTA 37.30 37.30 20.00 Ferrous sulphate FeSO4.7H2O 27.80 27.800 15.00 Boric acid H3BO3 6.20 6.20 5.00 Manganese sulphate MnSO4.4H2O 22.30 22.30 10.00 Potassium iodide KI 0.83 - 1.00 Zinc sulphate ZnSO4.7H2O 8.60 8.60 1.00 Sodium molybdate Na2MoO4.2H2O 0.25 0.25 0.10 Copper sulphate CuSO4.5H2O 0.025 0.25 0.20 Cobalt chloride CoCl2.6H2O 0.025 - 0.10

Myo-inositol C6H12O6 100.00 100.00 1000 Thiamine HCl C12H17CIN4OS.HCl 0.10 0.50 100 Nicotinic acid C6H5NO2 0.50 0.025 1.00 Pyridoxine HCl C6H11NO3.HCl 0.50 0.025 1.00

Glycine C2H5NO2 2.00 1 -

Protocols are available for micropropagation and multiplication of many endangerd species like *Piper hapnium, P. silent vallyensis, P.schmidtii, P. wightii, P. barberi , Vaniilla aphylla, V. pilifera, V. walkyrie, V. wightiana, K. rotunda and Alpinia galanga* are available (Peter et al 2002,

Bertaccini *et al* (2004), Du *et al (*2004) reported micropropagation and establishment of mite-

\*Murashige and Skoog, 1962, McCown and Amos, 1979, Schenk and Hildebrandt 1972

Table 1. Composition of MS\*, WPM\* and SH\* basal media

Minoo 2002, Nirmal Babu et al 1999, 2005).

brone virus-free garlic.

**Composition Molecular** 

**Macronutrients** 

Potassium di hydrogen

**Micronutrients** 

**Vitamins** 

**Amino acid** 

In many spices, conventional seed storage can satisfy most of the conservation requirements. But in crops with recalcitrant seeds and those having conservation needs cannot be satisfied by seed storage, have to be stored *in vitro*. Most field gene banks are prone to high labour cost, vulnerable to hazards like natural disasters, pests and pathogens attack (especially viruses and systemic pathogens), to which they are continuously exposed and require large areas of space. This supports *in vitro* and cryo conservation. In addition, other resources like continuous supply of standard stock cultures for experiments to examine physiological and biochemical processes, cell and callus lines developed for *in vitro*  synthesis of valuable secondary products, flavours and other important compounds will benefit strongly from *in vitro* cultures. Most of the spice crops are either vegetatively propagated or have recalcitrant seeds. The spices germplasm is mostly conserved in field gene banks. Most of the spices are plagued by destructive and epidemic diseases caused by viruses, bacteria and fungi. This makes germplasm conservation in field gene bank risky. Thus *in vitro* and cryo storage system becomes important in the overall strategy of conserving genepool. Each technology should be chosen on the basis of utility, security and complementarily to other components of the strategy. A balance needs to be struck between seed, field gene bank, *in vitro* and cryo conservation of propagules, tissues*,* pollen, cell lines and DNA storage for overall objective of conserving gene pool.
