Plant Transformation Methods_insiya Tankiwala_13bbt0201

8/16/2019 Plant Transformation Methods_insiya Tankiwala_13bbt0201

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PLANT BIOTECHNOLOGY

PLANT TRANSFORMATION

METHODS

PROF. V. PALANICHAMY

BY:

INSIYA TANKIWALA13BBT0201


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Direct Methods:

1. Particle Bombardment

A gene gn !" # $%!&%'(%) *#"(%)&e +e&%,e"- '-'(e/ !"%g%n#&&- +e'%gne+

!" *n( ("#n'!"#(%!n/ %' # +e,%)e !" %ne)(%ng )e&&' %( gene(%)

%n!"#(%!n4 (e %n'e"(e+ gene(%) #(e"%#& #"e (e"e+ ("#n'gene'. Te

*#-&!#+ %' #n e&een(#& *#"(%)&e ! # e#,- e(#& )!#(e+ %(

*'%+ DNA. T%' (e)n%5e %' !(en '%*&- "ee""e+ (!

#' $%!$#&&%'(%)' !" $%!&%'(%)'.

T%' +e,%)e %' #$&e (! ("#n'!" #&!'( #n- (-*e ! )e&&/ %n)&+%ng

*n('/ #n+ %' n!( &%%(e+ (! gene(%) #(e"%#& ! (e n)&e': %( )#n #&'!

("#n'!" !"g#ne&&e'/ %n)&+%ng *'(%+'.

https://en.wikipedia.org/wiki/Planthttps://en.wikipedia.org/wiki/Transformation_(genetics)https://en.wikipedia.org/wiki/Geneticshttps://en.wikipedia.org/wiki/Transgenehttps://en.wikipedia.org/wiki/Chemical_elementhttps://en.wikipedia.org/wiki/Heavy_metalshttps://en.wikipedia.org/wiki/Plasmidhttps://en.wikipedia.org/wiki/DNAhttps://en.wikipedia.org/wiki/Plastidshttps://en.wikipedia.org/wiki/Transformation_(genetics)https://en.wikipedia.org/wiki/Geneticshttps://en.wikipedia.org/wiki/Transgenehttps://en.wikipedia.org/wiki/Chemical_elementhttps://en.wikipedia.org/wiki/Heavy_metalshttps://en.wikipedia.org/wiki/Plasmidhttps://en.wikipedia.org/wiki/DNAhttps://en.wikipedia.org/wiki/Plastidshttps://en.wikipedia.org/wiki/Plant


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A gene gn %' 'e+ !" %ne)(%ng )e&&' %( gene(%) %n!"#(%!n/ %( %' #&'!6n!n #' $%!&%'(%) *#"(%)&e +e&%,e"- '-'(e. Gene gn' )#n $e 'e+

e7e)(%,e&- !n !'( )e&&' $( #"e #%n&- 'e+ !n *n( )e&&'.

Te !&&!%ng #"e (e '(e*' (! 'e # Gene Gn:

S(e* 1: Te gene gn #**#"#(' %' "e#+- (! 8"e.

S(e* 2: Wen (e gn %' ("ne+ !n #n+ (e e&% 9!' ("!g.

S(e* 3: Te e&% !,%ng (e +%'6 %( DNA )!#(e+ *#"(%)&e' (!#"+ (e

')"een.

S(e* : Te e&% #,%ng *'e+ (e *#"(%)&e' !,%ng ("!g (e ')"een

#n+ !,%ng (! (e (#"ge( )e&&' (! ("#n'!" (e )e&&'.

Te (#"ge( ! # gene gn %' !(en # )#&&' ! n+%7e"en(%#(e+ *n( )e&&'

g"!%ng !n ge& e+% %n # Pe("% +%'. A(e" (e g!&+ *#"(%)&e' #,e %*#)(e+

https://en.wikipedia.org/wiki/Callus_(cell_biology)https://en.wikipedia.org/wiki/Petri_dishhttps://en.wikipedia.org/wiki/Callus_(cell_biology)https://en.wikipedia.org/wiki/Petri_dish


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(e +%'/ (e ge& #n+ )#&&' #"e "ge&- +%'"*(e+. H!e,e"/ '!e )e&&' e"e

n!( !$&%(e"#(e+ %n (e %*#)(/ #n+ #,e '))e''&&- en,e&!*e+ # DNA )!#(e+

g!&+ *#"(%)&e/ !'e DNA e,en(#&&- %g"#(e' (! #n+ %n(eg"#(e' %n(! #

*n( )"!!'!e.

Ce&&' "! (e en(%"e Pe("% +%' )#n $e "e;)!&&e)(e+ #n+ 'e&e)(e+ !"

'))e''& %n(eg"#(%!n #n+ e


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!" DNA (! $e %n("!+)e+ %n(! (e )e&&. In %)"!$%!&!g-/ (e *"!)e'' !

e&e)("!*!"#(%!n %' !(en 'e+ (! ("#n'!" $#)(e"%#/ -e#'(/

!" *n( *"!(!*'(' $- %n("!+)%ng ne )!+%ng DNA. I $#)(e"%#

#n+ *'%+' #"e %


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"e*!"(e+. M!"e!,e" ("#n'%en( e


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# ''*en'%!n )!n(#%n%ng *'%+ DNA #n+ *n( (%''e >%#("e e$"-!'/

)#&&'/ )e&& )&'(e"?. I( %' (en %G!''-*% %"'(

L.? #n+ "egene"#(%!n ! '#&( (!&e"#n( *n(' e"e #&'! "e*!"(e+.

. Poleth!lene "l!col# Protoplast F$sion

T%' (e)n!&!g- %' #**&%)#$&e !" *"!(!*'( !n&-. Te )e%)#& 'e+ %'

*!&-e(-&ene g&-)!&. I( '(%(e' en+!)-(!'%' #n+ (e"e$- )#'%ng (e

*(#6e ! DNA. In (%' e(!+ *"!(!*'( #"e 6e*( %n *!&-e(-&ene g&-)!&

>PEG? '!&(%!n. Te )!n)en("#(%!n ! PEG 'e+ %' 1@ #,%ng 000 +#&(!n

!&e)" e%g(. A(e" e

*"e'en)e ! PEG #n+ !(e" )e%)#&'/ PEG %' "e!,e+ #n+ %n(#)( *"!(!*'(

#"e (en )&("e+ (! !" )e&&' %( #&&' #n+ )!&!n%e' %n("n. Se&e)(%!n*"e''"e %' (en #**&%e+ (! ge( (e ("#n'!"#n('. Te ("#n'e" ! gene

#)"!'' (e *"!(!*'( e$"#ne )#n $e %n%(%#(e+ $- # n$e" ! )e%)#&'

! %) *!&-e(-&ene g&-)!& %' (e !'( %*!"(#n(. I( #' $e)!e (e !'(

%+e&- 'e+ +e (! (e #,#%$%&%(- ! '%*&e ("#n'!"#(%!n *"!(!)!&. Me(!+

#' +e,e&!*e+ '%ng )#&)% #&g%n#(e %)"! $e#+' (! %!$%&%=e DNA

!&e)&e' %n )!$%n#(%!n %( *!&-e(-&ene g&-)!& ("e#(en( #&'!.


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%. &iposome Mediated "ene 'rans(er

L%*!'!e' #"e )%")" &%*%+ !&e)&e' %( #n #5e!' %n(e"%!" (#( )#n )#""-

n)&e%) #)%+'. I( en)#*'(e' (e DNA "#gen(' #n+ (en #+e"e' (! (e

)e&& e$"#ne' #n+ 'e %( (e (! ("#n'e" DNA "#gen('. T'/ (e

DNA en(e"' (e )e&& #n+ (en (! (e n)&e'. I( %' # ,e"- e)%en( (e)n%5e

'e+ (! ("#n'e" gene' %n $#)(e"%#&/ #n%#& #n+ *n( )e&&'. V#"%!' "e*!"(' !n(e %n(eg"#(%!n ! gene' %n("!+)e+ $- e#n' ! &%*!'!e' !&&!e+ $-

("#n'gen%) *n( "egene"#(%!n !" (!$#))! #n+ e#( #,e $een *$&%'e+ '!

#".


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)ndirect Methods:

1. *+robacteri$m mediated

A!ng (e ,#"%!' ,e)(!"' 'e+ %n *n( ("#n'!"#(%!n/ (e T% *'%+

! Ag"!$#)(e"% (e#)%en' #' $een %+e&- 'e+. T%' $#)(e"%# %'

6n!n #' n#("#& gene(%) eng%nee" ! *n(' $e)#'e (e'e $#)(e"%#

#,e n#("#& #$%&%(- (! ("#n'e" T;DNA ! (e%" *'%+' %n(! *n(

gen!e *!n %ne)(%!n ! )e&&' #( (e !n+ '%(e #n+ )#'e #n

n!"g#n%=e+ g"!( ! # )e&& #'' 6n!n #' )"!n g#&&. T% *'%+' #"e

'e+ #' gene ,e)(!"' !" +e&%,e"%ng 'e& !"e%gn gene' %n(! (#"ge(

*n( )e&&' #n+ (%''e'. Te !"e%gn gene %' )&!ne+ %n (e T;DNA "eg%!n

! T%;*'%+ %n *)e ! n#n(e+ 'e5en)e'.


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T! ("#n'!" *n('/ &e# +%')' >%n )#'e ! +%)!('? !" e$"-!gen%) )#&&'

>%n )#'e ! !n!)!('? #"e )!&&e)(e+ #n+ %ne)(e+%( Ag"!$#)(e"% )#""-%ng "e)!$%n#n( +%'#"e+ T%;*'%+ ,e)(!".

Te %ne)(e+ (%''e %' (en )&("e+ >)!;)&(%,#(%!n? !n '!!(

"egene"#(%!n e+% !" 2;3 +#-' +"%ng %) (%e (e ("#n'e" ! T;

DNA #&!ng %( !"e%gn gene' (#6e' *)e. A(e" (%'/ (e ("#n'!"e+

(%''e' >&e# +%')')#&&%? #"e ("#n'e""e+ !n(! 'e&e)(%!n ) *n(

"egene"#(%!n e+% '**&een(e+ %( '#&&- &e(#& )!n)en("#(%!n

! #n #n(%$%!(%) (! 'e&e)(%,e&- e&%%n#(e n!n;("#n'!"e+ (%''e'. A(e"

3;@ ee6'/ (e "egene"#(e+ '!!(' >"! &e# +%')'? #"e ("#n'e""e+ (!

"!!(;%n+)%ng e+%/ #n+ #(e" #n!(e" 3; ee6'/ )!*&e(e *n('#"e ("#n'e""e+ (! '!%& !&&!%ng (e #"+en%ng >#))&%#(%=#(%!n? !

"egene"#(e+ *n('. Te !&e)" (e)n%5e' &%6e PCR #n+ '!(e"n

-$"%+%=#(%!n #"e 'e+ (! +e(e)( (e *"e'en)e ! !"e%gn gene' %n (e

("#n'gen%) *n('.


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Plant Transformation Methods_insiya Tankiwala_13bbt0201