Condi

nubient
tions. hureproliferaton
dUxin cela lusedtool
ing tIssuecl ofPormahion
nd
prupugation4D
2
and
Cun mulkiply
to
inhocducing
the be
viFau tAgar
Co.2mg)|+ pH-5.7
leqves leqves mesponseinto
plant imducion: Aga
may ml. |00d
a 200
(TAA),thecallus
it chi¡n:t 1
ccum tabuccumcnd in making mgli) aclded-
taba
pper
them wndiferentiated
by
Vicling in callus encoaging
accomplished Kineinindu
lceicin The plant callus
acid
addeal
-
odled
0T esulFirg C0. soluion
partsi callus cwlaions goluion
C2mglml)e-stock
Stock
solu
ion Co.2m
lL)
Nicoiang N. cuOund
of
tissue. C2mg/)+BAP
as Ficn
solution :.Final
c¡mc. sdukion
Co.2.mglL)
um inducion such
leaves
(mq|)+
89
ingion,plant coyl Solu
cultue a indole-3-
enHy explan genetic
ransformatiem
shudies. C2mg)1) lm)e-soc
Experiment around ial cal - stock stocle
from ov +NAAhupo
dpplications
tabaccum L Stock
of essen
ciCferenti
numerus l - .:.
Finel
conc.
carota masS ThePreue atiom For lakicnS
Curota
Cultwe conlus the
suurose
24
aota Ime ILMedi
growS as c¡nc. Media
media
is medic
SHmuli. Auxins
are from N. prepar Co2mg (g
Duuus guch +
o.
establishingthqt studies reventingof D- Sucrose 4.4glL Final IL
Estabishing
callus A
callus
is reqenerate for 5ucrDse-
g°lo CalFor
medid
I
of massS vuriey MS+ fe L
Hypocols hormones,
hormona) I medid HoYmone I :. Kinetin-
Fox r
of cells
Media 2 + For 240-
coyls Cultre Media Agqr
- NAA
-
Callus MS :.
plant a FoY MS
CAnd
HypoTo to in
"
Requirement
Preparalions:
Aivn Principle "
and
 FoY
media 2:

2,49 CI mgm) stocle solukion

i. Fr dde- Iml
IL med id Stock 5oluticn a
Findl conc. C1mg /L)
( l mglml) stock 5olutiom
aclded - |00
. For IL mediq stock soluticn
: . Fincl con C. Co- mglL)
Media Preparation
and Sucrose according to given
JWeiqh Ms
culcicns cund pu them in clecund ¢ dy schôt
Ccl
amownt of cwam.
durn botHle then add requrel
hormones Por
ii Mix it well cund then cdd
media 1 - NAA cnd 24D wert add
before qutoclaue f
before cutoclave.
medica 2 - 21uDcUnd BAP were add
hormones
Ii Set the pH ot medig alter addinq
a 5..
iJ Autoclave the media. After Quho claving in Mediq l
after cool ing it don add Kinelin. nd
weu plaks. htet oi lee
VJPour media in deep
ler it soliliy. This is medig recely Por Callus induction.
Stock solution Preparation:
- NAA, Kinetin, 240 cand BAP ll cwere 13F dissolved
in g5 lo ethanol then Sinal volume madle up cwi th
distilleel war.
- er Kinetin need to heat ct 250C.
- Then syringe Cilleree al.
Stred ct cool c¡nditi¡ns.

Requiremenks Por Steri lizatHon of leqves of Ntubuccum

and D. Caota seeds
Stexile Pore ceps
$o °lo ethanol SBerle iler pCuper
- 4 °lo 5odium hypochloride
o-1,tween 20
ocedure: Sterilialion of,, leuves
leu ent
Ntubucewn
stete blude
i|N taba ccum ut in to 1%1 Cn by
leaves
Ond he forecers.
were
ranstered in
ii Fo Steilization leaf bits were
surface
5o mi cenriuqe tube. uith disilled oaker 2-3 rmes
"J Leaf bils u)#e u30shed
ethnol s
nen discad Diskilled usae and gave o fe
wash foy | min. sEe
was given tuice.
nrtr removing to % etarcl Dw uash
ere hreaeel win
V3 AFte Temoving disiled uge leat bts
o-I 'lo fen
4"> So dium hypochlorde soluion containing
fr 10 rnin.
tuosen 20 & eaf birs
ViJ To Temove Godiun yochorid and
Frther dried onto Sherile fil er papen tor s-10 min.
Vii After Sufuce serilizaion, l2af bits wee ppied cn MS

Having hormcnes- NAA C2mg1) + Kine tin Co2ralLD+ 2 . 4 (

i Tncubain condiions Por in viho Culhsre ere 25c ardeiK

fr seed steslrahion.
íi After Sreriliratin seeds were put on rmedia MS4rse
+Agar (o-3) ie. bass tnedia r

intergry 2500t 3000 lu.

23 nd dt,.
 Frem leqves a
UServations .Aucticn

Conclusion'
 tcbau
Experimen. explcs of N
Establishing hoot
Aim T6
induction from leaves and

eshablish shoot induction Frm leaves and

Sem

Sem
explauks of

N tubacum
sem CUnd leut
Inhoduen: In plunt tissue culke Shoof in duchion em
growth
egulaks.
precise balcunce of
explaus is dependut on a and mopenglhy
merislematic acHviry Cypiccy
merisem
Because of ther shung pticularty in
to Creae shootsI Stem expleunts,
leqveg a t pre fened
Yonger icl. The
r Shoot tip Cnt She chosen. ion polent
hiqher cell divis dids in the
leat explants cue to their componeut5
particulcmy ils vasculqr oot induction.The
leaF lamin gbusequent sh
chien of calluses and
within the shem or leaf explan
Pudu lo cations
Selectiom of pertlculcr cion ecticjency. The
cing shoot indu Such as NAA
for enhan Quxins
IS Critical beginh ing callus
wltth
qrowth with
PrÜcedure entails stimulating shoot netueen boosing
and then |ins potentially
such as 6AP with qibbereconcenhuticnS are nmehtu
Cytokinins
elongaion. These homones species, necessikahing
Shoot plcunt
adjusted based on the experimental hrials.
lously
iterative optimization through
Renuiemenk Selectiom of explcunts
and
sem -
Pepanatins " For Shoot induction hem Chosen.
meristem or shoot tip cne
Particulanty apical
For sh oot in chion bem leqve5 -
Yownger leaes.
of explcs
Requiemns fo sterilizaticn

- 4 lo Sodium Hypochloridle
40% Ehunol
Slerle Distilled waer
0. °% tween 0
- Autoclueel Porcceps
Serile Pillers papers
 bemleaves :
Media kr shoot incuchicn
MS - 4i4|
SUcose - 3° -

Agor o.o -
PH - g.1

PGRS Slock 6oln ((mglml)

B4P ¬Img|| ) mecdia.
Sojn ddoed in | L
Stock
: Tml um
stock so)n (2 mg/mi) meclic1.
NAA Co4 mqll) qdded in I L
.". 50 Fam stock Soluticn
induction em stem:
Media fr shoot

o shen and leaves

Serilzation cppo.
Pro cedure: cund leaf bils of Ix| cm
Cut sho Stem of lcm Size seperely,
i cenhifuge tubes
Take both exp lauus in 5oml
iJ Disilled cuaer.
washes o slerile
ilij Give 2-3
wash with o "o
ehanol for Imin.
iV Give q washes eaile disilled waler.
give 2
VJ To Yemove ethanol waer heat e*plcuts with 4°/
distilled
ViJ After removal ot for lo min.
hypochleride with o- °lb tween 20.
Soclium 3-4
Sodiwm Hypochleride and tueeh 20 give
vii] To emove wakr.
washes of sleie dishilled on sierile Ciller paper. (Smin)
pu explants
Vi Aften Slerilization Forceps curing hcnd lng
malke sue to property het Slerilie
of explauts.
ix] Pate explcs on especkive shoo} nduchion medlia
 und
Conelusion -