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TISSUE CULTURE & APPLICATIONS
PRESENT BY
NASEEM BHURGRI
PH.D. SCHOLAR (BIOTECHNOLGY DEP UNIVERSITY OF SINDH)
WHAT IS IT?
• THE TERM TISSUE CULTURE IS
COMMONLY USED IN A VERY WIDE
SENSE TO INCLUDE IN VITRO CULTURE
OF PLANT CELL, TISSUE AS WELL AS
ORGAN
• THE TISSUE CULTURE INVOLVES BOTH
ANIMAL AND PLANT CELL.
HISTORY OF TISSUE CULTURE
Tissue culture had itsorigins at the beginning ofthe 20th century with theworkof Gottleib Haberlandt(plants) and Alexis Carrel(animals
THE BACKGROUND,I
• the first commercial use of plantclonal propagation on artificialmedia was in the germinationand growth of orchid plants, inthe 1920’s
• in the 1950’s and 60’s there wasa great deal of research, but itwas only after the developmentof a reliable artificial medium(murashige & skoog, 1962) thatplant tissue culture really ‘tookoff’ commercially
THE BACKGROUND, II
•a more recent advance is the use of
plant and animal tissue culture
along with genetic modification
using viral and bacterial vectors and
gene guns to create genetically
engineered organisms
“
”
DESIGN AND LAB SETUP OF TISSUE CULTURE LABORATORY
SETTING OF OF A TISSUE CULTURE LAB REQUIRES PROPER PLANNING
IT IS DIVIDES INTO 5 AREAS
Acclimatization
roomAnalytical
room
Aseptic
transfer
area
Culture
room
Media preparation room
MEDIA PREPARATION ROOM
• REFRIGERATOR AND FREEZER
• WATER PURIFICATION AND STORAGE SYSTEM
• GLASS WARE WASHING FACILITY
• CONTINUOUS SUPPLY OF SINGLE AND DOUBLE DISTILLED
WATER
• CULTURE MEDIA, WASHING POWDER DISINFECTANTS
• CABINETS OR SHELVES
ASEPTIC TRANSFER AREA
Laminar air flow
Dissecting microscope
Dissection instruments
Gas outlets vacuum facility
Sterilization
CULTURE ROOM
•Environmentally
controlled
• Incubators with
controlled
temperature
•Rotary shakers
•Space for culturing
requiring complete
darkness
Culture
Room
ANALYTICAL ROOM
COLORIMETER
LOW SPEED
CENTRIFUGED
INVERTED
CENTRIFUGED
CHEMICAL REAGENT
RACKS
VISCOSITY METER
GAS OUTLETS
ACCLIMATIZATION ROOM
•HIGH ILLUMINATION
(4,000-10,000 LUX)
•HIGH HUMIDITY
OTHER ITEMS
• AIR CONDITIONS
• UNINTERRUPTED POWER SUPPLY
• BUNSEN BURNER
• ALUMINUM FOILS
• FLUORESCENTS LAMPS
• FIRE FIGHTING EQUIPMENT'S
MICRO ELEMENTS
ORGANIC COMPOUNDS
GROWTH REGULATORS
CONTI:
ROLE OF PLANT HORMONE IN GROTH OF PLANTS
A PLANT HORMONE CAN BE DEFINED AS A
SMALL ORGANIC MOLECULES THAT ELICITS
A PHYSIOLOGICAL RESPONSE AT VERY LOW
CONC.
PLANT GROWTH HORMONE PLAY AN
IMPORTANT ROLE IN THE PHENOTYPES
ACT AS MESSENGER BETWEEN
ENVIRONMENT AND THE GENOME
AUXIN
• essential for cell division, cell
elongation, cell differentiation,
organogenesis and embryo genesis,
callus formation.
• natural form auxin are iaa, iba,.
• synthetic form of auxin are naa,
2,4d.
CYTOKINIS
• cytokine promoting cell division, shoot
proliferation and influence the cell cycle
• embryogenesis and inhibit roof formation
• synthetic form is 2-ip which is most
active form of cytokine
• natural forms are bap and kinetin
GIBBERELLINS
• it promotes stem elongation, bulb corn formation,
embryo maturation, but can inhibit callus and root
induction
• ga3 is most common gibberellins.
ABSCISIC ACID
it inhibit root growth and
germination of embryo
it is thermostable but light
sensitive.
TYPES OF PLANT TISSUE CULTURE
Plant
tissue
culture
Embryo
culture
Meriste
ms
culture
Protopl
ast
cultureOrgan
culture
Callus
culture
Cell
culture
CALLUS CULTURE
callus culture means anorganized proliferative massof cells produced romisolated plant cells, tissue oforgan when grown asceticallyon artificial nutrient media.
it produced when explant areculture on the appropriatesolid media, with both ofauxin and cytokine in correctcondition. 2.4 d arecommonly used.
EMBRYO CULTURE
• DEFINATION :
• embryo culture can be defined as the in vitro culture
either of the polarized egg, zygote, proembryo or mature
embryo.
CELL CULTURE
• cell culture refers to the
removal of cells from an
animal or plant and their
subsequent growth in a
favorable artificial
environment.
MERISTEM CULTURE
• a cell culture
developed from a
small portion of the
meristem (growing
tip) tissue of a plant.
either a stem shoot or
root meristem can be
used
ORGAN CULTURE
• organ culture is a development
from tissue culture methods of
research, the organ culture is able
to accurately model functions of
an organ in various states and
conditions by the use of the
actual in vitro organ itself
Organ culture
Vegetative
organ
Root culture Leaf cultureShoot tip
culture
Reproductive
organ
Complete
flower culture
Isolated ovary
culture
Isolated ovule
and embryo
culture
SUSPENSION CULTURE
Can be initiated from any part of the plant.
Usually initiated from friable callus already growing in culture.
Transferred into liquid medium
PROTOPLAST CULTURE
• ISOLATED PROTOPLAST HAVE BEEN DESCRIBED AS A NAKED CELLS BECAUSE THE
CELL WALL HAS BEEN REMOVED BY EITHER A MECHANICAL AL AND ENZYMATIC
METHOD
CONTI:
Isolation of protoplast Separation of protoplast from
plant tissue
Enzymatic method
Mechanical
method
MECHANICAL METHOD
ENZYMATIC METHOD
ANTHER CULUTRE
APPLICATION OF TISSUE CULTURE
Medicine
and
research
There are various application of tissue
culture in a wide array of scientific
industries
Cancer cells,, are analyzed in vitro to
produced appropriate medicines that will
eliminate the abnormal cells
Skin grafting,, where severely burned
tissues are replaced with new ones using
culture cells
APPLICATION
Plant fiber culture
One of the most common types of tissue
culturing is conducted in botanical and
agricultural research.
Plant tissue is much easier to grow in a
nutrient rich suspension than human or
animal tissue, and with the right nutreints
different tissue samples can often be
encouraged to develop as they would on a
normal plant suspension
CONTI;
APPLICATION OF PLANT TISSUE CULTURE IN INDUSTIRES
PLANT CELL CULTURE IS USED FOR BIOTRANSFORMATION (MODIFICATION OF
FUNCTIONAL GROUPS OF ORGANIC COMPOUNDS BY LIVING CELLS.
FOOD AND AGRICULTURAL BIOTECHNOLOGISTS ARE INVOLVED IN USING TOOLS
OF MOLECULAR BIOLOGY TO ENHANCE THE QUALITY AND QUANTITY OF FOODS
AND ECONOMIC CROPS.
CONTI:
• FOR EXAMPLE, GOLDEN RICE WAS GENETICALLY ENHANCED WITH ADDED BETA
CAROTENE, WHICH IS A PRECURSOR TO VITAMIN A IN THE HUMAN BODY. PLANT
CELLS CAN BE CULTURED IN FERMENTERS FOR THE INDUSTRIAL PRODUCTION OF
SECONDARY METABOLITES USING CELL CULTURE.
BENEFITS OF TISSUE CULTURE
CONTI;
CONCLUSION
REFERENCES
REFERENCES D.DUMET,A.ADEYEMI,O.B.OJUEDERIE (2008). YAM INVITRO
GENEBANKING. GENEBANK MANUAL.
HTTP://WWW.IITA.ORG/GENEBANK/MANUAL
D.DUMET,A.ADEYEMI,O.B.OJUEDERIE (2008).
CASSAVA IN VITRO PROCESSING AND THE GENEBANKING.GENEBANK
MANUAL. HTTP://WWW.IITA.ORG/GENEBANK/MANUAL HORT689/AGRO689
BIOTECHNIQUES IN PLANT BREEDING H.S CHAWLA .2002 INTRODUCTION TO
PLANT BIOTECHNOLOGY 2ND EDITION. OXFORD & IBH PUBLISHING C./ PVT. LTD
NEW DELHI INDIA MURASHIGE T. AND SKOOG F. (1962). A
CONTI:
• A REVISED MEDIUM FOR RAPID GROWTH AND BIOASSAY WITHTOBACCO TISSUE CULTURE. PHYSIOLOGIAPLANTARUM 15: 473-497.
• HTTP://AGGIE-HORTICULTURE.TAMU.EDU /TISSCULT/PLTISSUE/PLTISSUE.HTML
• HTTP://WWW.LIV.AC.UK/~SD21/TISSCULT/WHAT.HTM
• WWW-PLB.UCDAVIS.EDU/COURSES/S99/PLB111I/TCMEDIUM.HTML
• HTTP://MEMBERS.AOL.COM/MRDJREED/PRIVATE/PTC.HTML
• HTTP://WWW.ACCESSEXCELLENCE.ORG/LC/ST/ST2BGPLANTPREP.HTML
• WWW.OSMS.OTAGO.AC.NZ/MAIN/BURSARY
• HTTP://WWW.KITCHENCULTUREKIT.COM/HISTORYTC.HTM
• HTTP://WWW.SIGMAALDRICH.COM/AREA_OF_INTEREST/LIFE_SCIENCE/CELL_CULTURE/HELPFUL_RESOURCES/CE
LL_CULTURE_HANDBOOK,HTM