anther culture and haploid plant production

   

      ANTHER CULTURE AND HAPLOID PLANT PRODUCTION

All cells are having chromosome which are having genetic information . Many cells are having single set of chromosome are haploid structures , some are having two set of chromosomes , and others  may be having three or four are triploid and tetraploid respectively.                 

N=  SINGLE SET OF CHROMOSOMES I.e HAPLOID

Gametic  cells  of species are called haploid cells which may be female gametic structure[ovum or egg] or may be male gametic structure[anther in plants and sperms in humans . production of haploid is called haploid production. Haploid cells i.e gametes having a single set of chromosome.

                                                                   When the haploid plant chromosome is doubled , normal chromosome no. of a particular species are obtained are called diploid plants these are homozygous in nature . for obtaining diploid plant from haploid plant using colchicine  . 

haploid plants only in nature generated from unfertilized egg or ovum, but  in laboratories we can produced haploid plant via using male and female gametes . anther and ovary both are the sexual part of any flower of angiosperm plants which are producing pollengrain[ male gamete] and when anthers are cultured in a suitable medium to producing a new plant called anther culture .

           

    GUHA AND MAHESHWARI  developed this technique in [1984] .used Dhatura innoxia  for anther culture .

Haploids are also can be formed by using ovaries and ovules. Firstly this techniques was used to produced haploid of DATURA.  This technique helpful for instantaneous expression of mutations and quick formation of pureline. 

meristem culture

MERISTEM CULTURE

MERISTEM CELLS ARE LOCALIZED AT THE TIP OF THE ROOT AND SHOOT PART OF PLANT BODY ,THESE CELLS ARE ACTIVELY DEVIDEDING CELLS WHICH ARE UNDIFFERENCIATING CELLS WHICH GIVES TO RISE TO THE PERMANENT TISSUE, MERISTEM CELLS GROWN INVITRO AFTER REMOVAL FOR OBTAINING VIRAL FREE PLANT .APICAL MERISTEM AND AXILLARY MERISTEM CELLS ARE GROWN IN VITRO CONDITIION IS KNOWN MERISTEM CULTURE . ITS INVOLVES THE IMPROVEMENT OF ALREADY EXITING SHOOT CELLS AND REGENERATION OF ADVENTITIOUS ROOT FROM THE DEVELOPING SHOOT.BUT NOT THE SHOOT NEW SHOOT MERISTEM . FOR MERISTEM CULTURE USUALLY APICAL SHOOT AND ROOT TIPS ARE USED . THESE ARE CULTURED IN THE PRESENCE OF CYTOKININ [ BAP].

MERISTEM CULTURRE IS USED IN :

1: PRODUCTION OF VIRUS FREE PLANT LIKE SUGARACANE , BANANA, APPLE, POTATO ETC.

2: GERMPLASM CONSERVATION

3: GENERATING TRANSGENIC PLANTS

4: FAST CLONAL GERMENATION.

endosperm culture

                                        ENDOSPERM CULTURE

                                                    Tissue culture techniques are also used to generate endosperms by culturing . it is a  unique because its supplies the nutrition to the developing embryonic cells . it is a tirloid cell is in its chromosomal constitution. These are used for the formation of seed less fruity plants like banana apples etc.

There are some techniques which is involve for the endosperm culturing are as follows :

a.     In aseptic conditions , immature seeds are taken from out side and dissected then remove  its endosperm along with embryo , mature seed can be taken.

b.     Then endosperm kept in a suitable medium and embryos are remove from its initial growth .

c.      Initially callus is developed . 

somatic embryo culture

                            Somatic embryo culture

Somatic embryo develops by the somatic cell [which may be takes from any part of body except sex cells . somatic embryo can generated at high concentration of auxin like 2,4 –D . somatic embryo develops into a matured embryo and it gives to rise to make a new plantlet or new plant organism which is similar to its parental in their genetic behavior as well as physiological or physical behavior.

                                                                            After the germination of plantlet its transfer to out side the culture vessel and established in the field this transfer is done by a specific methods which is called hardening . at the time of hardening plantlets are kept into less lightened and high humid area . this procedure helps to tolerating this adverse condition relatively their environment conditions from the culture vessel.

shoot and root culturing

                 Shoot and root culturing 

For Shoot culturing cytokinin is used because its promotes the shoot elongation like BAP, and for root culturing Auxin is used because its promotes the growth of root like NAA [ naphthalic acetic acid ] when we want to develop both root and shoot we use the different ratio of the both growth regulators , excess amount of cytokinin promotes the shoot elongation and excess amount of auxin promotes the root elongation . and if the ratio of cytokinin and auxin are equal so root and shoot both are not form it form an unorganized structure which is called callus .

single cell culturing [ cell cloning]

                                     Single cell culturing   [ cell cloning]

Through mitosis single cell derived many cells which are similar to its parent cell which is called clone and this process of obtaining clones is called cloning . there are two popular techniques of single cell culturing are  as follows:

·       Bergsmann’s planting technique : mostly used technique ,cells are suspended into a liquid media on a particular cell density . sterilized agar medium kept in waterbath at 35c temp. liquid and agar media both are mixed in equal amount .

1mm thick layer of ca spread on petridish .

                                                                      then cells are embedded in the agar medium which is observable under microscope . large colonies are isolated and culture in a new media and separately.

·       Filter paper raft nurse tissue technique :

1.     Cells are placed over a piece of filter paper which is placed on the top of callus culture for many days . the single cell takes its nutrition from its callus and doing mitosis and increase its number . then colonies are formed these are isolated and further culture.

PLANT TISSUE CULTURE

                                                Plant tissue culture

Plant tissue culture is a technique which is used to maintain and growing plant parts and plant cells ,tissues and organs on an artificial medium in a suitable environmental condition.

That part is culture is called explants, that is , may be any part of plant which taken from out side the laboratory and grown under in a test-tube or in a glass flask in special sterile conditions in a special medium which is having nutritional substances for the plant.Ability to generate a new whole plant from any plant part [expalnt] and cells from the apex region of the shoot and root part is called cellular totipotency.  

Gottlieb haberlandt firstly started tissue culture technique in 1902.

Hormones are used in plant tissue culture are as follows :

·         Auxin [indole -3- acetic acid ]

·         Cytokinins [kinetin, zeatin]

·         Gibberellins [gebbrellic acid ]

·         Abscisic acid [abscisic acid ]

·         Polyamines 

For growing plant part environmental conditions are applying.

A.      Nutrient medium,

B.      Aseptic conditions ,

C.      Aeration of the tissue.

plant tissue culture methods

                                      Plant tissue culture methods

Two major methods which are includes in plant tissue culture are as follows:

1.     In vitro growth – callus and suspension culture .

2.     Types of explants – single cell culture, shoot and root culture ,  haploid culture , meristem culture, somatic embryo culture , cybrid production , protoplast culture , embryo culture , ovule culture, ovary culture etc.

                                                             

Callus and suspension culture : continuous  cell division in explants forms callus . callus is a unorganized structure and mass which formed by dividatio  of explants cells . for callus formation both auxin [2,4 dichlorophenoxy acetic acid ] and cytokinin [ BAP benzyl amino purine ] are used both are the growth regulators . they stimulates the plant growth , it is obtain 2-3 weeks.

Suspension culture  is a culture which is kept in a culture vessels . cells and tissues both are regularly transfer into new culture vessel which containing fresh media . this process is called subculturing . suspension culture is an liquid culture .

                                                These both culture are used for the protoplast isolation , production of transgenic plant , regeneration of plantlets and biomass production also.

environmental conditions for aseptic cultivation of plant part

ENVIRONMENTAL CONDITIONS FOR  ASPECTIC  CULTIVATION OF PLANT PART:

1.      NUTRIENT MEDIUM

2.      ASEPTIC CONDITIONS

3.      AREATION OF THE TISSUE

·         NUTRIENT MEDIUM: culture media is depending on the plant part which used for the culture . in the nutrient medium consisting inorganic salt sources[ both micro and macro elements ] , carbon and nitrogen sources [ usually sucrose and ammonium salts respectively] , vitamins [ nicotinic acid , pyridoxine , myoinositol , thiamine] , growth regulators [ Auxin , cytokinin  majorly , some other are  also used like gibberellins , adscisic acids , polyamines ], other compounds like casein , hydrolysate , coconut milk , malt extract, yeast extract, tomato juice etc. may be added.

Plant hormone are also plays an important role in growth  and differentiation of cultured cells and tissues . the pH of culture medium is should be 5.7 . mostly MS medium is used for the plant tissue culturing which is developed by murashige and skoog in 1962. Usually a gelly substance AGAR  is used to solidify the culture which gives a substrate or basement to grow the plant part and plant cells.

·         

ASEPTIC CONDITIONS : it contains sugar which helpful for the growth of microorganisms such as bacteria and fungi . these microbes are competing with the plant cells and tissues and kill them . so its important to maintain the aseptic conditions of tissue culture. Complete removal of microorganisms from any surface is called sterilization.

·       

  AREATION OF TISSUE : it is also an important aspect of culture technique. It is taken by occasionally stirring the medium by sterring or by automatic shaker .Plant part which taken out is called explant 

PLANT TISSUE CULTURE

                                                Plant tissue culture

Plant tissue culture is a technique which is used to maintain and growing plant parts and plant cells ,tissues and organs on an artificial medium in a suitable environmental condition.

That part is culture is called explants, that is , may be any part of plant which taken from out side the laboratory and grown under in a test-tube or in a glass flask in special sterile conditions in a special medium which is having nutritional substances for the plant.Ability to generate a new whole plant from any plant part [expalnt] and cells from the apex region of the shoot and root part is called cellular totipotency.  

Gottlieb haberlandt firstly started tissue culture technique in 1902.

Hormones are used in plant tissue culture are as follows :

·         Auxin [indole -3- acetic acid ]

·         Cytokinins [kinetin, zeatin]

·         Gibberellins [gebbrellic acid ]

·         Abscisic acid [abscisic acid ]

·         Polyamines 

For growing plant part environmental conditions are applying.

A.      Nutrient medium,

B.      Aseptic conditions ,

C.      Aeration of the tissue.                                               

role of biotechnolgoy in improvement of vegetable crops

        Role of biotechnology in improvement of                           vegetable crop

 Biotechnology having an important role as a modern tool for improving crops since last 30 years. By this a single commercially important crops are develop by using biotechnological tools eg: somatic hybrids, cybrids, GE variety in india .

Vegetable production : modern biotechnology which develop to use high yielding product and more nutritive crop varieties also , improve their resistance power to  diseases , and also decrease the use of fertilizers and other chemical substances. Crop varieties are improve by their clonal propagation and their improve their genetic material as well .To affects the plant and animal traits by traditional methods like cross pollination, grafting , and cross breeding but now biotechnology helps to made quickly , on molecular level through additional and deletion of any gene or introduction of any new foreign gene .

1.      For eg: Bacillus thuringiensis microbe [ pest resistant crop ] which produce toxic protein which is harmful for the insects . that gene produced protein by the microbes is isolated and transfer into the crop by genetic engineering and its transfer into by potato and cotton.

                       Bt protein is not harmful for human so its easier to transgenic crop for farmers .

2.      To increase their ripening power of tomatos . these are also genetically modified .

3.      Many rice varieties also. Like golden rice.

4.      Nicotina tobaccum it is also genetically modified

Brief account on human genome project

                                                      HUMAN GENOME PROJEECT

HGP is an global methodical research project with a main goal of formative the sequences of chemical base pair which modify the structure of DNA . francis Collins directeddd the national institutes of health national human genome research institute labors.

A working draft of the genome was announced in 2000 and complete in 2003 research centre from the united states the united kingdom ,japan, frances ,germany, china , and Pakistan. Gene mapping is an important step in the growth of medicine and other aspects of physical condition care.

HGP appreciate the genetic structure of the human species, the project has also listening carefully on several non-human organisms such as E.coli , fruit ,fly and laboratory mouse . it is largest single investigative projects in modern science.

Its first aimed to map contained in a human haploid references genome [more than three billion ] and several expand tjis to diploid human genome including the global HapMap project. 

Appliaction and benefits of biotechnology in envrionment

Application and benefits of biotechnology in environment : there are four different types of applications of environmental biotechnology . they are as follows:

1.       Biomarker: It gives the response to a chemical that help to measure the level of damaging caused or the exposure of the toxic or the pollution effect caused. These are also called biological marker.majorly it helps to relate the connection between oils and its sources.

2.       Bioenergy: biogas, biomass,fuel ,and hydrogen are collectively called bioenergy.  It is used in the sector of industrial , domestics , and space environmentally. One of the pioneer examples of green energy are the wastes collected from organic and biomass waste ,these waste helps use to over the pollution issue caused .

3.       Bioremediation : conversion of hazardous substances into non- toxic compounds are called bioremediation process. It majorly use to clearifying the environment using microorganism.

4.       Biotransfosrmation: changes in the biology of environment which are changes the complex compound to simple , non-toxic to toxic compounds ,  or other way round is called biotransformation. It is mostly used in the manufacturing sector where the toxic substances are convert into bioproducts.

Application of biotechnology in medicine

Application of biotechnology in medicine :

  Biopharmaceutical :

 biotechnology is a techniques which helps to synthesize drugs .there are no any inclusion of chemicals but only microorganisms have completely it likely to expand them. Heavy protein molecules are the typically source of biopharmaceutical . they internally attack in the body by a hidden mechanism of the diseases and wipe out them . now a days scientists are expand such  biopharmaceuticals drugs are creating which are helps to treated against the diseases like hepatitis , cancer, and heart diseases also.          

 Gene therapy:

Gene therapy is the technique of  biotechnology which helps to diagnose diseases like cancer and Parkinson’s diseases. The apparatus which used to introduced the fit genes into the body which obliterate the injured cell and replace them. These fit genes correct the genetic information and how the genes are start performance in the favor of body.

Pharmacogenomics:

It is an additional genetic engineering modified method which is used to learn the genetics information of a person . it is the mixture of pharmaceutical and genomics. This field is to expand such drugs which are inserted into the person according to the genetic information there in the individual .

Genetic testing:

 genetic testing is a technique which conclude the heredity the genetics diseases in parents , sex, and carrier screenings . in this technique DNA probes which are having a genetic information alikely mutant sequences . this techniques is also used to identify the criminals and child parenthood.

Biotechnology in vaccine and antibiotics

Use of biotechnology in vaccine and antibiotics :

Vaccine : oral vaccines are likely a solution which works more extensively to increase the diseases in immature countries , where the cost is very excessive . when vaccines are injected in the hereditarily engineered crop , fruits vegetables , planned to carry antigenic proteins from transferable pathogens these are replying to activate their immune system .

                           For eg: vaccine treating for cancer is anti-lymphoma vaccine has been made using tobacco plant which carries RNA from cloned malignant B-cells.so the vaccine increase the strength of immune system beside the cancer.

Antibiotics : plants are used to isolation of antibiotics for both humans and animals also. The stock of synthetic antibiotics which are more expensive rather then natural antibiotics products .but it has many bioethics issues which encourage the expansion of Antibiotics-resistant bacterial strain . the large quantity of antibiotic product plant versus fermented unit is easy to purify and condensed risk of contamination as compared to that of using mammalian cells and culture media.

                                                                                  

 

Application of biotechnology in other field

Four major applications that are includes in biotechnology are as follows:

1.       In Industrial areas ,  including health care [medical field] .

2.       In agricultural field.

3.       Uses in crop Improvement.

4.       In environmental field and other products like biodegradable plastics , vegetables and oil [vanaspati ghee]

:  Advantage of biotechnology in medicine

                                                                               It is very broad field and its applications are used in many varieties of science like agriculture field ,and drug designing and other fields also biotechnology introduce many techniques for genetic engineering like gene therapy  ,[ RDT] recombinant DNA technology, and PCR [polymerase chain reaction or retort which are helps to increase the  number of DNA segments and manipulation in DNA  segments which helps to diagnose many diseases and helps to improve the body cells by introducing strong genes .

There are some applications of biotech which having very important place in turf of medicine and gives good result.

·         Biopharmaceutical

·         Gene therapy

·         Pharmogenomics

·         Genetics testing

Application of biotechnology in agriculture:

                                                                               Biotechnology  is vast deliberately similar with the biomedical investigate , but some other groups of industries take advantages of biotech metheology for studying, cloning and varying gene therapies .

                                                                                                   Our everyday lives we have turn out to be familiar tio the throught of enzymes and many peoples are recognizable GMOs [genetic modified organisms ] are used for their food .since the days of “GEORGE WASHINGTON CARVE”agricultural has been producing special products which are altering to improving us life .

Are as follows :  

·         Vaccine

·         Antibiotics

·         Flowers

·         Biofuels

·         Plant and animal reproduction

·         Pesticide resistant crop

·         Nutrient supplementation

·         Abiotic strain contamination

·         Manufacturing power of fibers

·          

Application of biotechnology in food processing : 

                                                                                              In developing countries application of food processing is an issues of debates and discussion for a long time . study of biotechnology is as practical to be bioprocessing to increase the size of countries and growth of products substances and development of highly productive products through fermentation processes.

                                                                        There are some issues while usingb technology for various applications to increasing country development.

: infrastructural and logistical factors

: socio economics in additional to cultural factors

: nourishment and food stuff security

: intellectual property rights [IPRs]

Application of biotechnology in environment factors :

                                                                                                   Environment biotechnology is that which is solve the problems of environment and in the ecosystem is called environmental biotechnology . it is required for the study of environmental factors and only applying on it .

                                                           According to international society – environment biotechnology is that which helps to develop , regulate the biological system and keep safe the environment from pollution and contamination of air , water , and land also.

 Four major applications that are includes in biotechnology are as follows:

1.       In Industrial areas ,  including health care [medical field] .

2.       In agricultural field.

3.       Uses in crop Improvement.

4.       In environmental field and other products like biodegradable plastics , vegetables and oil [vanaspati ghee]

:  Advantage of biotechnology in medicine

                                                                               It is very broad field and its applications are used in many varieties of science like agriculture field ,and drug designing and other fields also biotechnology introduce many techniques for genetic engineering like gene therapy  ,[ RDT] recombinant DNA technology, and PCR [polymerase chain reaction or retort which are helps to increase the  number of DNA segments and manipulation in DNA  segments which helps to diagnose many diseases and helps to improve the body cells by introducing strong genes .

There are some applications of biotech which having very important place in turf of medicine and gives good result.

·         Biopharmaceutical

·         Gene therapy

·         Pharmogenomics

·         Genetics testing

Application of biotechnology in agriculture:

                                                                               Biotechnology  is vast deliberately similar with the biomedical investigate , but some other groups of industries take advantages of biotech metheology for studying, cloning and varying gene therapies .

                                                                                                   Our everyday lives we have turn out to be familiar tio the throught of enzymes and many peoples are recognizable GMOs [genetic modified organisms ] are used for their food .since the days of “GEORGE WASHINGTON CARVE””    agricultural has been producing special products which are altering to improving us life .                        

Are as follows :  

·         Vaccine

·         Antibiotics

·         Flowers

·         Biofuels

·         Plant and animal reproduction

·         Pesticide resistant crop

·         Nutrient supplementation

·         Abiotic strain contamination

·         Manufacturing power of fibers

·          

Application of biotechnology in food processing : 

                                                                                              In developing countries application of food processing is an issues of debates and discussion for a long time . study of biotechnology is as practical to be bioprocessing to increase the size of countries and growth of products substances and development of highly productive products through fermentation processes.

                                                                        There are some issues while usingb technology for various applications to increasing country development.

: infrastructural and logistical factors

: socio economics in additional to cultural factors

: nourishment and food stuff security

: intellectual property rights [IPRs]

Application of biotechnology in environment factors :

                                                                                                   Environment biotechnology is that which is solve the problems of environment and in the ecosystem is called environmental biotechnology . it is required for the study of environmental factors and only applying on it .

                                                           According to international society – environment biotechnology is that which helps to develop , regulate the biological system and keep safe the environment from pollution and contamination of air , water , and land also.

 v

BRANCHES OF BIOTECHNOLOGY

FEW BRANCHES Of BIOTECHNOLOGY :