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PLANT TISSUE CULTURE :
MICROPROPAGATION
TECHNIQUES
Department of biotechnology
N.C.College of Engineering,Israna(Panipat)
Work carried ot nder
Dr. !bash "a#la
!enior !cientific $fficer
Pro#ect %ide,&icropropagation
Pro#ect 't
Centre for Plant iotech.,CC!'*,isar
y
'shish "atyal
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$tline$tline
IntroductionIntroduction
NeedNeed
Stages of MicropropagationStages of MicropropagationExperimental WorkExperimental Work
Applications of Plant Tissue cultureApplications of Plant Tissue culture
Results & DiscussionsResults & Discussions
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IntrodctionIntrodction
What is Plant TissueWhat is Plant Tissue
CultureCulture
It is the technique of growing plant cells,tissues, organs, seedsor other plant parts in a sterile environmenton a nutrient medium
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Plant Tissue Culture = Plant Cloning
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W! We Need Tissue "ulture#
Quick multiplication of plants that are rare, elite orface extinction
Progen! is disease free " true to t!pe
#igh multiplication rates
$xport of high qualit! planting material
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% Totipotent Cell
&organ in '()' gave the term *Totipotent
Cell+ impl!ing that a cell is capale ofdeveloping into a whole organism$
Plant Cells are Totipotent
In '()- #aerlandt .
/ave Practical shape to
Totipotenc! Concept
%ttempted to culture cells
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0tages ) to 1
Stage 1: $xplants in initiation medium
Stage 2: 2ewl! formed ud, shootlet or callusare cultured on multiplication media
Stage 3: 0hootlets that are formed aretransferred to rooting media
Stage 4: 3ooted plantlets are hardened
0tage ). /row the parent plants in sterileconditions or glasshouse
Stages of MicropropagationStages of Micropropagation
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Tissue Culture 0tages
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/$2$3%4 P35C$673$
$xplants Collected
Inoculated in *Initial ud proliferation medium+
Transferred to &ultiplication medium
0u8cultured, rooted, hardened and planted
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&icropropagation
4arge scale multiplication of high value plantmaterial
2ormall! use meristem as explant
meristem is the growing point of a plant
consists of cells which are undifferentiated
%ll regenerated material are clones
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&icropropagation
%llows for continuous production
Permits seasonal production
%llows for long term preservation of plants
0mall space requirements
9ecause the meristem is starting material,products are virus and disease free
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Experimental WorkExperimental Work
TI007$ C74T73$ I2 07/%3C%2$
STEPS IN%'%ED(
Wasing
Explant Preparation
Media Preparation
Sterili)ation
Inoculation*ardening
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Washing
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$xplant Preparation
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&edia Preparation
C i i f &0 di
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Composition of &0 media:&urashige and 0koog, '(;-
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Macronutrients mg/1
(NH4)NO3 1,650K NO
31,900
CaCl2.2H
2O 440
MgSO4.6H
2O 370
KH2PO4 170
Iron mg/l
Na2!"# 33.6
$%SO4.7H2O 27.8
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Vitamins mg/l
n&'o&n&' a'& 0.05*+&o-&n%.HCl 0.05
&a/&n%.HCl 0.01
Cytokinin mg/l
K&n%&n 0.1
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mg/l l+'&n% 0.2
myo-&no&ol 100
## 10Su'o% 30,000
#ga (0.8 )
*H 5.7
Other components
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0terili>ation
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0terili>ed &edia 0torage
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Inoculation
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Primar! 0torage
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0econdar! 0torage
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#ardening
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#ardening 5f Tissue Culture Plants
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%pplications of Plant Tissue culture%pplications of Plant Tissue culture
#as een referred to as#as een referred to as iotechnolog! of developingiotechnolog! of developing
worldworld
4ow capital4ow capital costs relative to other technologiescosts relative to other technologies
#igh laor input#igh laor input :laor is cheap in most developing:laor is cheap in most developing
countries
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3esults
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5f Course@Challenges and
5pportunitiesare alwa!s
two sides
of the same coin
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