Germplasm Storage, National Bureau of Plant Genetic Resources, International Board of Plant Genetic Resources, Cryopreservation

Scholarships Dictionary Visa Details Study Abroad Exam Results Online Test Jobs

	Home
Tissue Culture, Micropropagation and Somaclonal Variation Tissue Culture, Micropropagation and Somaclonal Variation Introduction Cell and Organ Differentiation Cytodifferentiation Organgenic Differentiation Shoot bud Differentiation Somatic Embryogenesis Clonal Propagation or Micropropagation Bioreactors for Micropropagation and for bioactive Compounds List of Plant Materials,Which can be Propagated by Tissue Culture Synthetic Seeds for Micropropagation Production of Virus free Plants ELISA for Detection of Viruses Fungi Bacteria, MLOs, Mycotoxins and Hormones DAC-ELISA PAC-ELISA Maintenance of Male Sterile Parents for Hybrids through Micropropagation Micropropagation of Hybrid Plants Somaclonal Variation

Home >> Plant Biotechnology and Genomics >>Tissue Culture, Micropropagation and Somaclonal Variation >>Germplasm Storage

Germplasm Storage
Conservation of plant genetic resources or germplasm by several national (National Bureau of Plant Genetic Resources = NBPGR) and international organizations (‘International Board of Plant Genetic Resources = IBPGR’, later renamed as ‘International Plant Genetic Resources Institute = IPGRT') has become a thrust area of biotechnology in recent years. For this purpose, various strategies for storage of germplasm without losing its viability have been developed and are being utilized. Germplasm can be stored in a variety of forms including seeds, buds, protoplasts, cells, tissues, etc. Well organized plant parts like shoot tips and plantelets in culture can also be used for long term storage.

The technique involving storage of tissues in culture may use the germplasm in the growing stage and their growth may be suspended, by any one of the following methods: (i) lowering the temperature, (ii) adding chemical retardants or hormones and (iii) reduction in oxygen concentration. Such methods, where limited growth of cultures is allowed, remains effective for about a year, so that periodic renewal of these cultures is required. However, the most popular and effective method for long term storage of cell culture, involves storage at a very low temperature using liquid nitrogen. This is described as Cryopreservation, which allows storage for virtually indefinite periods.

Cryopreservation of isolated plant protoplasts has been recommended in many cases, since survival level of up t 75% has often realized in some cases (e.g. Daucus carota). DMSO (0.5 M or 10%) or some other medium (DMSO + glycerol + sucrose) is used for step-wise freezing leading to storage in liquid nitrogen (N2). The germplasm can be used whenever required by rapid thawing.

A Typical Cryopreservation Procedure for Cell Culture

1. Culture under standard conditions	2. Pregrow for 5 days in medium containing 6% mannitol
3. Cryoprotect with 0.5% DMSO + 0.5M glycerol + 1M sucrose in culture medium	4. Pransfer cells and cryoprotectant solution in ampule; freeze at 1°C for 1min to -35°C; hold for 40 min, transfer to liquid N₂
5. Store in liquid N₂	6. Thaw in warm water
7. Pour onto semi solid medium	8. Transfer recovered culture to liquid medium

Standard Procedure for Cryopreservation of Shoot Tip

1. Culture under standard conditions	2. Dissect shoot tip
3. Pregrow for 2 days on medium containing 5% DMSO	4. Cryoprotect with 10% DMSO in culture medium
5a. Collect on a hypodermic needle; plunge into liquid N₂	5b. Transfer to an ampute containing cryprotectant solution; freeze at 0.5°C min^-1 liquid N₂
6. Store in liquid N₂	7a. Plunge into medium at +25°C
7b. Thaw in warm water	8. Place on semi-solid medium

Pollen required for hybridization can also be cryopreserved. For Cryopreservation, cells in early exponential phase have been found to be the most highly freeze-tolerant, while those in lag phase or stationary phase are susceptible to some freezing injury. This susceptibility to freezing injury actually depends on the size of individual cells and also on the size of aggregates or clumps used for Cryopreservation.

	Home
List of Some Crop Species where Desirable and Heritable Somaclonal Variation Overcoming Crossing Barriers Using Culture Technques Pre-Fertilization Crossing Barriers on Culture Techniques Post-Fertilization Crossing Barriers Embryo Rescue Embryo Culture Ovule Culture Ovary Culture Other Uses of Tissue Culture Endosperm Culture Nucellus Culture Germplasm Storage Examples of Storage of Plant Tissue Culture Status of Tissue Culture Technology in India Micropropagation Research in India Status of Tissue-Culture Technology in India-Semi Commercial Status of Tissue-Culture Technology in India Commercial Scale Production Somaclonal and Gametoclonal Work in India Secondary Metabolities Research in India

Language >> English | Français | Español | Deutsch | Italiano | Portugues | Japanese | Korean | Sim-Chinese