cryopreservation

 Cryopreservation

Why Preservation is important?

• Until tow decades ago the genetic resources were getting depleted owing to the continuous depredation by man.
• It was imperative therefore that many of the elite Economically important and endangered species are preserved to make them available when needed.
• The conventional methods of storage of storage failed to prevent losses caused to various reason.
• A new methodology had to be devised for long term preservation of material.

There are various methods of storage

1. Cryopreservation:- Generally involves storage in liquid Nitrogen.
2. Cold storage:- It involve storage in low and non freezing temperature.
3. Low pressure:- It involves partially reducing the atmospheric pressure of surrounding.
4. Low oxygen storage:- It involves reducing the oxygen level but maintaining the pressure.

Cryopreservation;-

• Cryo is greek word, (Kroyes -frost)
• It literally means preservation in "frozen state"

The principle:- to bring plant cells or tissue to a zero metabolism and non dividing state by reducing the temperature in the presence of cryoprotectant.

It can be done:-

• Over solid carbon dioxide (at-79°C)
• Low temperature deep freezer (at-80°C)
• In vapor phase nitrogen (at-150°C)
• In liquid nitrogen (at-196°C)

Major advantages are:-

• Once the material is successfully conserved. Particular temperature it can be preserved indefinitely.
• Once in storage no chance of new contamination of fungus or Bacteria.
•  Minimal space required.
• Minimal labor required.

Mechanism of Cryopreservation:-

 

                             There cryopreservation technique followed by the regeneration of plants involves following steps:-

1. Selection of of material.
2. Addition of Cryoprotectant.
3. Freezing
4. Storage in liquid nitrogen.
5. Thawing.
6. Washing and reculturing.
7. Measurement of viability
8. Regeneration of plants.

1. Selection of plant material:-

             Two important factors depends on it such as 

                          a. Nature and

                          b. Density

      Any tissue can be selected for this purpose e.g., meristem, embryo, ovules, seeds, etc.

The density should be high.

2.Addition of Cryoprotectant:-

• They are chemical which prevent cryodestruction.
• These are sucrose, alcohol's, glycols, Some amino acid (proline), DMSO (dimethyl sulfoxide)
• Generally tow cryoprotectant should be used together instead of single one as they are more effective.

3. Freezing:-

      

               The sensitivity of cells to low temperature depends on the plant species.

  There are four different types of Freezing:-

• Slow freezing method:- The tissue or plant material is slowly frozen at slow cooling rate. The advantage is the plant cells are partially dehydrated and survive better.
• Rapid freezing method:- It involves pluming the vials in liquid nitrogen. The temperature decreases from -300°C to -1000°C  rapidly.
• Rapid freezing method:- This is combination of both slow and rapid freezing method. The process is carried out in step wise like manner.
• Dry freezing method:- In this method dehydrated cells and seeds are stored.

4.Storage:-

• The maintenance of the frozen cells or material at specific temperature is kept -70°C to -196°C
• Prolong storage is done at temperature of -196°C in liquid nitrogen.
• To prevent damage, continuous supply of Nitrogen is done.

 

5.Thawing:-

• Usually carried out by plunging the vials. into warm water bath with vigorous swirling.
• As thawing occurs the vials are transferred to another bath at 0°C degree.

6.Washing and reculturing:-

• The preserved material is washed few times to remove the cryoprotectant.
• This material is then recultured in a fresh medium.

7.Measurement of viability

• There is possibility of death of cells due to storage stress.
• Thus viability can be found at any stage.
• It is calculated by formula:-

  No. of cells growing/no of cells thawed ✖100

8.Plant regeneration:-

• The viable seeds are cultured on non specific growth medium.
• Suitable Environment Conditions are maintained. 

                                                        Application

• It is ideal method for long term conservation of material.
• DISEASE FREE PLANT CAN BE CONSERVED AND PROPAGATE.
• Peculcitrant seeds can be maintained for long time.
• Endangered species can be maintained.
• Pollens can be maintained to increase longitivity.
• Rare germplasm and other genetic manipulations can be stored.

What is Technology

Technology:-   What is technology every person have this question come in mind. If you don't know about technology. Don't worry. I will tell You about technology. We use technology everywhere in life. But You don't know It is technology. Yes, That is technology. We use Smart phone. There are many function work in Your phone by technology. We can talk any person in just minute. You can send money in just minute anywhere. And more that We use Technology in daily life. just like Mobile recharge, Communication, Current News in just minute, Play music, Television(we watch at home without any problem), Game, Online class, Money transfer, Digital India And more. That is technology.  

  

Some image of technology:-

Mobile communication

 

Money Transfer 

Television communication 

Online shopping & social media  

Computer is also biggest examples of technology Everywhere use computer in school, office, Train ticket counter, In railway, Flight communication & Booking ticket And more Uses of computer.

Some image of computer in technology:-

Movie making & Online marketing  

Study & software making

Technology uses in Medical And science :- There are many machine uses in medical science For Improve health of patient. And some machine uses in production of medicine and check their  quality. And more. Technology are uses in medical science.

Some image of medical science technology:-

Showing image of brain surgery

 

Medical technology 

 

Temperature detector machine digital

Microscope 

And know more about technology on this website:-https://en.wikipedia.org/wiki/Technology
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Thanks for reading....................
 

Biotechnology scope and importance

Biotechnology Scope and Importance:-

                                                               

                                       Biotechnology has rapidly emerged as an area of activity having a marked realized as well as potential impact on virtually all domains of human welfare, ranging from food processing, protecting the environment, to human health. As a result, it now plays a very important role in employment, production and productivity, trade, economics and economy, human health, and the quality of human life throughout the world. This clearly reflected in the emergence of  numerous biotechnology companies throughout the world, including India, and the movement of noted scientists, including Nobel Laureates. to some of these companies. The total volume of trade in biotechnology products is increasing sharply every year, and it is expected to soon become the major contributor to world trade. Many commentators are confident that the 21th century will be the century of biotechnology, just as the 20th century is era of electronics.

A list of some of the important area in which biotechnology is making marked contributions:-

• Human health 
• Medicines
• Animal health
• Animal husbandry
• Dairy
• Chemicals and biochemical
• Food processing and beverages
• Crimes and parentage disputes
• Agriculture
• Forestry
• Horticulture and floriculture
• Environment
• Renewable energy and fuels
• Population control
• Mining
• Fisheries and aquaculture

Some selected contributions of biotechnology to human welfare:-

1.Medical Biotechnology:-

➢Monoclonal antibodies ( use for disease diagnosis, e.g, venereal diseases, hepatitis B and other viral diseases, cancer, etc.)                

➢DNA  probe ( used for disease diagnosis, e.g, kala-azar, sleeping sickness, malaria, etc.) 

➢Recombinant vaccines ( cleaner, safer, e.g, human hepatitis B virus, E. coli vaccines for pigs, rabies virus etc.)                                  

➢Valuable drug like human insulin, human interferon, human and  bovine growth hormones, etc.                                                               

➢Gene therapy to cure genetic disease , e.g., Huntington's chorea,  cystic fibrosis.              

➢Babies of specified sex ( artificial insemination with X or Y carrying sperms prepared by sperm separation techniques.)                

➢Identification of parents or criminals using DNA or auto-antibody   finger-printing.

2.Industrial Biotechnology:-

➢Production of useful compounds, e.g., ethanol, lactic acid, glycerin, citric acid, gluconic acid, acetone etc.                               

➢Production of antibiotics, e.g., penicillin, strepto-mycin, erythromycin, mitomycin, cycloheximide etc.                                      

➢Transformation of less use full and cheaper compounds into more useful and valuable ones, e.g., steroid hormones from sterols, sorbose from sorbitol etc.                                                                    

➢Production of enzymes, e.g., alpha-amylase, proteases, lipases etc. 

➢Single cell protein (SCP) from bacteria, yeast, fungi or algae for  human food and animal feed.                                               

➢Fuel (mainly ethanol, sometimes biogas) production from cheap,  less useful and abundant substrates, e.g., sugarcane bagasse, wood etc.                                                                   

➢Mineral extraction through leaching from low grade ores, e.g., copper, uranium etc.                                                                            
➢Immobilization of enzymes for their repeated industrial application.                                                                          

➢Protein or enzyme engineering to change the primary structure of existing proteins or enzymes to make them more efficient, change their substrate specificity, e.g., successes with T4 lysozyme, trypsin, subtilisin, lactate dehydrogenases etc.              

➢Production of immunotoxins by joining a natural toxin with a specific antibody.

3.Animal Biotechnology:-                                                                           

➢Test tube babies in human; involves in vitro fertilization and embryo transfer.

➢Hormone-induced superovulation and splitting in farm animal; involves transfer and, in many cases, in vitro fertilization.

➢Production of transgenic animals for increased milk, growth rate,  resistance to disease etc. and production of some valuable proteins in milk.

4.Environmental Biotechnology:-

➢Efficient sewage treatment, deodorization of human excreta.

➢Degradation of petroleum and management of oil spills.

➢Detoxification of wastes and industrial effluents.

➢Bio control of plant disease and insect pets by using viruses, bacteria, amoebae fungi etc.    

5.Plant Biotechnology:-

➢Embryo culture to rescue otherwise invariable hybrids, to recover haploid plants from interspecific hybrids, micro propagation of orchids etc..

➢Rapid clonal multiplication through meristem culture, e.g., of many fruit and forest trees, such as, teak.

➢Recovery of virus and other pathogen free stocks of clonal crops; meristem culture is generally combined with thermotherapy.

➢Germplasm conservation through storage in liquid nitrogen (-196'C, cryopreservation) or through slow growth.

➢Rapid isolation of homozygous lines by chromosome doubling of haploid produced through anther culture, interspecific hybridization, ovary culture.

➢Isolation of stable somaclonal variants with improved yield traits  resistance to cold, herbicides, metal toxicity, salt and other abiotic stresses.

➢Gene transfer for insect resistance, protection against viruses, herbicide resistance, storage protein improvement etc.

➢Molecular marker, e.g., RFLPs and RAPDs, for linkage mapping and mapping of quantitative trait loci.