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Polypropylene .

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Oven 180
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.

- 12

10 20 .

- 13

) Balances ( 3 4

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- 14

Hot plate Magnetic stirrer

.

- 15

- 16

- 17

Refregerator .

- 18

Deep Freezer
.

- 19

Steamer .

- 20

pH meter pH
. pH 14
to 5.5 5.2 .to 5.8 5.6

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Autoclave
121 .to 1.5 1.2 /
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Forceps .

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Needels .

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Scalpels .13 - 12 - 11 - 10

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Binocular
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Air Condition

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Shaker ) (

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Centerfuge .

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Ultra violet lamp
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- 40

)( ELISA test .

- 41

Reader .Microplate

- 42

10

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Incubator 40 - 4
Timer 1000 10000
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Raks
12 24 36.

- 45

Water bath
.

- 46

Gloves

.

- 47

Over head .

- 48

Mask
.

- 49

Over shoes
.

- 50

Lux meter
.

- 51

.Chemicals .

- 52

- 53

Filter papers .

- Media preparation
-1 .
-2 .
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-4 .
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- General equipment
-1 .
-2 .
-3 .
-4 .
-5 .
-6 .
-7
.

- Growth room :
-1 27 17.
-2 .
-3 .
-4 .
-5 .
-6 .
-7 .
-8 .

.
12

 : ) (

Media

.

.


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- 1 Media Composition
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gelling agent
)(
.
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. .(White (1934
Murashige and Skoog
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N6 Chu
((1978 s Nitsch ( Nitsch and Nitsch ( 1969
.
.

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E1

N6

400
250
2100
600
450
-

185
400
2830
166
463

) /(
MS B5 Nitschs
185
68
950
720
-

250
150
2500
150
134

370
170
1900
1650
440
-

13

White
750
19
80
-

MgSO4. 7H2O
KH2PO4
NaH2PO4. H2O
KNO3
NH4NO3
CaCl2. 2H2O
(NH4)2 SO4

3
10
2
0.25
0.025
0.025
0.80
43

1.6
4.4
3.3
1.5
0.80
27.8
37.8
-

25
10
0.25
0.025
0.025
27.8
37.8
-

3
10
2
0.25
0.025
0.025
0.75
43

6.2
22.3
8.6
0.25
0.025
0.25
0.83
27.8
37.3
-

1.5
5.
3
0.01
0.75
-

25

25

20

20

30

20

10
1
1
250
5.5

1
0.5
0.5
5.8

0.5
0.5
5.0
100
2
0.5
0.05
5.8

10
1
1
100
5.5

0.5
0.5
0.5
100
2
5.8

0.01
0.01
0.05
3
5.8

H3BO3
MnSO4. 4H2O
MnSO4. H2O
ZnSO4. 7H20
NaMoO4. 2H2O
CuSO4. 5H2O
CoCl2. 6H2O
KI
FeSO4. 7H2O
Na EDTA. 2H2O
EDTA Na ferric salt
)Sucrose (g

Thiamine HCl
Pyridoxine HCl
Nicotinic acid
Myo-inositol
Glycine
Folic acid
Biotin
pH

:
) White: White (1953) Am. J. Bot. 40, 517 524). MS: Murashige and Skoog (1962
Physiol. Plant. 15 : 473). B5: Gamborg et al.( 1968) Exp. Cell Res. 50: 151). Nitschs:
(Nitsch and Nitsch 1969 ) Sci., N. Y., 163: 85).N6: Chu (1978); Pro. Symp. Plant
Tissue Culture, Sci. Press, Peking P. 43). E1: Gamborg et al. (1983). Plant Cell Rep. 2:
209).
-:Growth Regulators

.

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-1 Auxins

indole-3 acetic acid ) (IAA indole 3
butyric acid ) (IBA 4 2 )D) 2,4 4 ,2
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(NAA IAA
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to 10 mg /L 0.1
14


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100 1 = 1.

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100 2 1 100
1 = 1 . pH
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=
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IAA DNA

-:
-1
(chlorophenoxy acetic acid (4- CPA 4

(P chlorophenoxy acetic acid (PCPA
-2

5 4 2
(Trichlorophenoxy acetic acid (2, 4,5 T 2,4,5

-3

2 4
2 methy 4 chlorophenoxy acetic acid (MCPA).

-4

4 6 5 3
)4 amino - 3, 5, 6- trichloropicolinic acid (picloram.

-5

6 3 2
3,6 dichloro 2 methoxy benzoic acid. (dicamba).

15


.
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-2

Cytokinins

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:
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6(4hydroxy3methyltrans-2 butanylamino) purine

)Zeatin

- 2 Zeatin Riboside .
- 3 ip-2 y y dimethylamino) purine) 6 .
- 100
2 1 ) 10- 5( 100 3 .
to 10 mg/L 0.01

:
1 Kinetin (6 ( N (2 furfurylamino)1 H purine 6 amine
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- 3 BA 6 benzyladinine .



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diphenylurea


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BA D 2,4 mg/ l-1 15
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RNA
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-3

Gibberellins and Abscisic Acid

16

 .
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.
.
.
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.
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.
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pH

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5.8 pH Buffer solution
pH pH pH meter :
pH 4.5 7 .
pH 4.5 :
- 1 IAA .
- 2 .
- 3 .
- 4 .
pH ) ( 8 :
- 1 .
- 2 .
- 3 .
- 4 .
- 5 .
- 6 .
pH pH 5 3
.

) (

Media types

17

 - 1

Solid media

7 2 / 2 /
pH 5.7 45
.
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Liquid media

pH 5.5


Shaker )(
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Semi-solid media

Solidifying Agents

.

.
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24 60 24
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25
Gelrite,
Phytagel, alginate, methacel
.
.
.
2/ ) El-Shobaky
.(1991

Media Preparation

.

. .

1
.

18



.
) (Stocks
.
Stocks
100 )
( :
Micro nutrients

100 X

1X

mg/100L

mg/L

83

0.83

KI

620

6.2

H3BO3

860

8.6

ZnSO4.7H2O

2230

22.3

MnSO4.4H2O

25

0.25

NaMoO4.2H2O

2.5

0.025

CuSO4.5H2O

2.5

0.025

CoCl2.6H2O

3730

37.3

Na2.EDTA

2780

27.8

FeSO4.7H2O

FeSO4. 7H2O
Na EDTA. 2H2O 45
100.
85 100.
70 100..
1
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.
.
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20
)MS) Stocks and Media Preparation

Stocks ) ( - 4
5 . ) (:
30 - 20 .
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100 .
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4
5
6
7

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-

Explant

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Age of Explant
-


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.
-

Size of Explant

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Season of Explant

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20

 Quality of the Explant Source

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: :

Sterilization of the Explant

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(1

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%70 .
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Culture of the explant

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Incubation of the cultures

Growth Chamber
Stands

. :
- Light requirement :
- 1

Light intensity

Initiation Morphogenetic processes


1000 3000
10000 .
).(El-Shobaky 1991
- 2

Photoperiod

16 8 12

.
-

Heat requirement

27 - 22

35 - 27
18 . 25-20
.(El-Shobaky (1991
21

 : :

Acclimatization

10000
Under mist

4 - 3
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1 : 1

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Greenhouse and Field


:
- 1 .
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60
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23

 :

Production of Banana (Mousa sp.) through tissue culture

.
58 18


) Banana Bunch Top Virus Banana
(Mosaic
.


.

.

- 1
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:

stage Starting

-:
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- 11 2 27
1500 16 .
0.1 / .

.
1 2 .
: Multiplication stage
:
MS salt+5mg/LBAP + 30 g/L Sucrose + 100 mg/L Inositol + 2g/L Gelrite or 8 g/L Agar

.

MS salt+1mg/LBAP + 30 g/L Sucrose + 100 mg/L Inositol + 2g/L Gelrite or or 8 g/L Agar

: Rooting stage

) ( ) (
) ( . :
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:
2 27 16 8+ -3000
4000 .


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.

:
. . & . ) (2002 21
. _
. 601 .

26

27