transgenic techniques ar describes in these slides which are mostily used in the feild of biotechnology

2. TISSUE CULTURE &
APPLICATIONS
PRESENT BY
NASEEM BHURGRI
PH.D. SCHOLAR (BIOTECHNOLGY DEP UNIVERSITY OF SINDH)

3. 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.

4. HISTORY OF TISSUE CULTURE
Tissue culture had its
origins at the beginning of
the 20th century with the
workof Gottleib Haberlandt
(plants) and Alexis Carrel
(animals

5. THE BACKGROUND,I
• the first commercial use of plant
clonal propagation on artificial
media was in the germination
and growth of orchid plants, in
the 1920’s
• in the 1950’s and 60’s there was
a great deal of research, but it
was only after the development
of a reliable artificial medium
(murashige & skoog, 1962) that
plant tissue culture really ‘took
off’ commercially

6. 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

8. “
”
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
room
Analytical
room
Aseptic
transfer
area
Culture
room
Media preparation room

9. 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

10. ASEPTIC TRANSFER AREA
Laminar air flow
Dissecting microscope
Dissection instruments
Gas outlets vacuum facility
Sterilization

11. CULTURE ROOM
•Environmentally
controlled
•Incubators with
controlled
temperature
•Rotary shakers
•Space for culturing
requiring complete
darkness
Culture
Room

12. ANALYTICAL ROOM
 COLORIMETER
 LOW SPEED
CENTRIFUGED
 INVERTED
CENTRIFUGED
 CHEMICAL REAGENT
RACKS
 VISCOSITY METER
 GAS OUTLETS

13. ACCLIMATIZATION ROOM
• HIGH ILLUMINATION
(4,000-10,000 LUX)
• HIGH HUMIDITY

14. OTHER ITEMS
• AIR CONDITIONS
• UNINTERRUPTED POWER SUPPLY
• BUNSEN BURNER
• ALUMINUM FOILS
• FLUORESCENTS LAMPS
• FIRE FIGHTING EQUIPMENT'S

18. MICRO ELEMENTS

19. ORGANIC COMPOUNDS

20. GROWTH REGULATORS

21. CONTI:

22. 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

24. 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.

25. 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

26. GIBBERELLINS
• it promotes stem elongation, bulb corn formation,
embryo maturation, but can inhibit callus and root
induction
• ga3 is most common gibberellins.

27. ABSCISIC ACID
it inhibit root growth and
germination of embryo
it is thermostable but light
sensitive.

28. TYPES OF PLANT
TISSUE CULTURE
Plant
tissue
culture
Embryo
culture
Meriste
ms
culture
Protopl
ast
culture
Organ
culture
Callus
culture
Cell
culture

29. CALLUS CULTURE
callus culture means an
organized proliferative mass
of cells produced rom
isolated plant cells, tissue of
organ when grown ascetically
on artificial nutrient media.
it produced when explant are
culture on the appropriate
solid media, with both of
auxin and cytokine in correct
condition. 2.4 d are
commonly used.

30. EMBRYO CULTURE
• DEFINATION :
• embryo culture can be defined as the in vitro culture
either of the polarized egg, zygote, proembryo or mature
embryo.

32. CELL CULTURE
• cell culture refers to the
removal of cells from an
animal or plant and their
subsequent growth in a
favorable artificial
environment.

33. 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

34. 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

35. Organ culture
Vegetative
organ
Root culture Leaf culture
Shoot tip
culture
Reproductive
organ
Complete
flower culture
Isolated ovary
culture
Isolated ovule
and embryo
culture

36. SUSPENSION CULTURE
 Can be initiated from any
part of the plant.
 Usually initiated from
friable callus already
growing in culture.
 Transferred into liquid
medium

37. 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

38. CONTI:

39. Isolation of protoplast
Separation of protoplast from
plant tissue
Enzymatic
method
Mechanic
al
method

40. MECHANICAL METHOD

41. ENZYMATIC METHOD

42. ANTHER CULUTRE

43. 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

44. 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

45. CONTI;

46. 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.

47. 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.

48. BENEFITS OF TISSUE
CULTURE

49. CONTI;

51. CONCLUSION

52. 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

53. CONTI:
• A REVISED MEDIUM FOR RAPID GROWTH AND BIOASSAY WITHTOBACCO TISSUE CULTURE. PHYSIOLOGIA
PLANTARUM 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