ICAR-NBPGR Cryo Database
Toggle navigation
  • Dashboard
  • AboutUs
  • Publication
  • Books
  • Useful Links
  • Team
    • Management Team
    • Development Team
  • Report
    • Search Database
  • Login
    • Cryo Login
  1. Dashboard

Cryogenebank : the pioneering work at national level at NBPGR from 1987

CRYOGENE BANK (NBPGR):


The word ‘cryo’ is a derivative of cryogenic which is defined as very low temperatures. Cryopreservation refers to stabilizing cells at cryogenic temperatures, usually below -100°C. Cryopreservation reduces cell and tissue deterioration in storage by virtually halting metabolism and hence is an important tool for non-lethal storage of biological materials. It ensures theoretically ‘infinite’ periods of storage (long-term conservation), and reportedly causes no change in viability, vigour and genetic makeup of the conserved materials. The low temperatures are created by use of cryogen like liquid nitrogen in liquid (-196°C) or in vapour (-150°C to -180°C) phase in double walled Cryotanks without depending on mechanical system and electricity.

Cryo Bank

Cryo Tank:


Each of the six Extra large capacity (1000 liters) Cryotanks are equipped with electronically controlled filling system and alarm. They have a static holding time, which is the maximum time for which a tank can hold a particular quantity of LN, after which more LN has to be replenished. This is dependent on the rate of evaporation and the capacity of the tank. Evaporation rate of LN for most of the tanks is 0.5 to 1.5% of its capacity per day and accordingly, replenishment of LN is required about two times per week to maintain the temperature of the cryotanks between –160 to –180o C.

Cryo Tank

Liquid Nitrogen Station:


A vertical LN station / horizontal reservoir of extra large capacity (6000 lits) is maintained near the cryobank which is constantly filled to ensure continuous supply of LN to storage tanks. For this, a pipeline from the LN station directly in the Cryobank has been laid to avoid movement of heavy tanks for replenishment.

Liquid Nitrogen Station

Canisters with Cryo vials:


Storage tanks have access from the top, therefore a vertical inventory scheme is used. Inventories are metallic (aluminum) canisters in which canes are held which hold the cryovials.

Canisters with Cryo vials

Cryo vials stands:


Metallic (aluminum) canes are used on which the cryovials are mounted before dipping in liquid nitrogen. Aluminum canisters are used for holding the germplasm since 1ml, 2ml or 5ml cryovials mounted on aluminum canes or else 50ml vials and also polyolefin tubings can be effectively stored in them

Cryo vials stands

Cryo vials:


Small seeds, excised embryonic axes, pollen and buds are stored in large numbers in 1 ml/2ml polypropylene cryovials. Each Cryotank can hold up to 50,000 of 1 ml cryovials. If germplasm of a large seeded species or germplasm as twigs/ nodal sections are to be handled, then a flexible inventory system like polyolefin tubing is used.

Cryo vials

Safety Gadgets:


Safety gadgets like gloves, aprons, face masks, face shields or goggles, etc are essentially used during freezing and thawing of samples.

Safety Gadgets
Previous Next


The word ‘cryo’ is a derivative of cryogenic which is defined as very low temperatures. Cryopreservation refers to stabilizing cells at cryogenic temperatures, usually below -100°C. Cryopreservation reduces cell and tissue deterioration in storage by virtually halting metabolism and hence is an important tool for non-lethal storage of biological materials. It ensures theoretically ‘infinite’ periods of storage (long-term conservation), and reportedly causes no change in viability, vigour and genetic makeup of the conserved materials. The low temperatures are created by use of cryogen like liquid nitrogen in liquid (-196°C) or in vapour (-150°C to -180°C) phase in double walled Cryotanks without depending on mechanical system and electricity.

Seeds
Based on the seed storage characteristics, seeds have been classified as orthodox, intermediate and recalcitrant. Orthodox/desiccation tolerant are those seeds which are capable of retaining viability after being dried to less that about 5% moisture content. Seeds of most common agricultural species belong to this category and can be easily cryopreserved.

Difficult-to-store seeds
Non-orthodox seed species comprising intermediate and recalcitrant seeds are large sized and are shed with high moisture levels and are desiccation- and freeze-intolerant. They lose viability after being dried below a critical limit usually between 12-30% moisture. With advancement in research it has been possible to successfully cryopreserve zygotic embryos and embryonic axes of intermediate and recalcitrant seeds using new techniques of cryopreservation. The embryonic axes excised from the large seed is the preferred explant because of its organized small structure, independent identity and the presence of appreciable proportion of meristematic tissues. The aseptically excised embryonic axes are desiccated to around 11-16% moisture level using air drying (desiccation method) or by addition of cryoprotectants (pregrowth-desiccation and vitrification method) before exposing to temperatures of liquid nitrogen (-196°C). The axes are later thawed rapidly in a water bath maintained at +38°C and cultured on a defined media to obtain complete plants.

At National Cryogenebank at NBPGR cryopreservation of intermediate and recalcitrant seed species using zygotic embryos and embryonic axes has been successfully achieved for almond (Prunus amygdalus), Citrus sp., neem (Azadirachta indica), jackfruit (Artocarpus heterophyllus), Litchi (Litchi chinensis), trifoliate orange (Poncirus trifoliata), black pepper (Piper nigrum), cardamom (Elettaria cardamomum) and Wild banana (Musa balbisiana) with high percentage of recovery. In addition, cryopreservation of pollen from various crop species of importance to plant breeders has been carried out. Cryopreservation of orthodox seed species representing diverse group of crops and having special attributes and of dormant buds of mulberry has been carried out.

The goal of a Cryogenebank is to collect, store and disseminate specimens and related data. In order to provide high quality plant germplasm/ materials with well-characterized data, the management team must insure that they follow ‘best practices’. Several factors have been considered in the design and development of a Cryobank which are published in a Manual.


List of the plant species with known or likely recalcitrant or intermediate seed storage behavior needing cryobanking

Common name Botanical name
Acer Acer platanoides
Almond Prunus amygdalus Batsch.
Aonla Emblica officinalis Gaertn.
Bael Aegle marmelos L.
Baher (myrobalan) Terminalia bellirica Roxb., T. chebula Retz.
Banana (wild) Musa spp.
Ber Ziziphus spp.
Black pepper Piper nigrum L.
Cardamon Elettaria cardamomum Maton.
Citrus spp. Citrus aurantium L., C. aurantifolia (Christm.) Swingle, C. sinensis (L.) Osbeck,C. limon (L.) Burm. F. and other spp.
Cocoa Theobroma cacao L.
Coffee Coffea arabica L. and other spp.
Eucalyptus Eucalyptus L’ Herit. Spp.
Jamun Syzygium cuminii (L.) Skeels and other spp.
Karonda Carissa congesta Wt.
Ker Capparis deciduas (Forsk.)Edgew.
Jack fruit Artocarpus heterophyllus Lamk. & other Artocarpus spp.
Lac tree Schleichera oleosa (Lour.) Oken
Lasora Cordia myxa Roxb.
Litchi Litchi chinensis (Gaertn.) Sonn.
Mahua Madhuca indica J.F. Gmel.
Neem Azadirachta indica A.Juss.
Oka Quercus suber L.
Oil plam Elaeis guineensis Jacq.
Papaya Carica papaya L.
Pecan nut Carya illinoensis (Wang.) Koch.
Pilu Salvadora oleoides Decne. and other spp.
Poplar Populus deltoides and other spp.
Sal Shorea robusta Gaertn. F.
Sapota Achras zapota L.
Trifoliate orange Poncirus trifoliata (L.) Rafin.
Tea Camellia sinensis (L.) O. Kuntze
Teak Tectona grandis L. f.
Developed in ICAR National Fellow Project
Copyright (c) 2018 All Rights Resereved, National Bureau of Plant Genetic Resources,
Indian Council of Agricultural Research, Ministry of Agriculture (Govt. of India), Pusa Campus, New Delhi-110012, INDIA