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PLANT TISSUE CULTURE

MADE BYVISWANATHAN ROHIT

MSc BIOTECHNOLOGY


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CONTENTS

INTRODUCTION

PREPARATION OF MEDIA

SUB-CULTURED

SEED-CULTURED HARDENING

CONCLUSION


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INTRODUCTION

Broadly refers to technique of growing plant cells,tissues,organs, seeds or other plant parts in asterile environment on a nutrient medium

The first commercial use of plant propagation on

artificial media was in the germination and growthof orchid plants in the 1920s

It was only after the development of a reliableartificial medium by Murashige & Skoog in 1962that plant tissue culture really took offcommercially


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MURASHIGE & SKOOG MEDIA


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COMPONENT AMOUNT STOCK SOLUTION STOCK SOLUTION WORKING SOLUTION

mg/L mg/L mg/500ml ml/L 500ml 250ml

MACRO SALTS

NHNO 1650 33000 16500

KNO 1900 38000 19000

CaCl.2HO 440 8800 4400 50ml 25ml 12.5ml

MgSO.7HO 370 7400 3700KHPO 170 3400 1700

MICRO SALTS

KI 0.83 166 83

HBO 6.2 1240 620

MnSO.4HO 22.3 4460 2230

ZnSO.7HO 8.6 1720 860 5ml 2.5ml 1.25ml

NaMoO.2HO 0.25 50 25

CuSO.5HO 0.025 5 2.5

CoCl.5HO 0.025 5 2.5

Na SALTS (III)

FeSO.7HO 27.8 5560 2780 5ml 2.5ml 1.25ml

Na.EDTA 37.3 7460 3730

VITAMINS

MESO-INOSITOL 100 20000 10000

NICOTINIC ACID 0.5 100 50

PYRIDOXINE HCl 0.5 100 50 5ml 2.5ml 1.25ml

THIAMINE HCl 0.1 20 10

GLYCINE 2 400 200

SUCROSE 30g 15g 7.5g

AGAR 8g 4g 2gpH 5.8 5.8 5.8


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Preparation of media

i. MS basal + 0.1% A.C.

ii. MS basal

iii. MS + 0.25 mg /l NAA (alpha- naphthalene

acetic acid) + 0.2 mg/l BAP (benzyl amino purine).


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Sub-cultured

i. Two bottles ofCymbidiumSoul huntand one bottleofSleeping nymph iscultured in MS basal +0.1% A.C.

ii. One bottle each ofLunavian atlas andPineclash in MS basal + 0.1%

A.C.

iii. one bottle each of

Kalanchoe pinnata andAnoectochilus are sub-culture in MS basal.

iv. One bottle ofCitrusmedica is sub-cultured in

MS basal + 0.25 mg/lNAA + 0.2 mg/l BA.


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Seed-cultured

Cymbidium mastersiiis cultured in MS basal.

Citrus marcoptera is cultured in MS basal .


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Hardening

Taking out the plant from the bottleby forceps.

Cleaning the plants from media withtap water and then deep into Bavistinfor about 15 mins.

1:1 of charcoal and brick are collectedin a pot with small pores in it.

Plant is then taken out for Bavistinafter 15 mins and planted .

Then plastic is covered over a plant

and after that it is taken to hardeningroom.

Sleeping nymph.


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CONCLUSION

Plant tissue culture is a successful technique forgrowing endangered species & hybrids so that in thefuture there will be enough food for the generationsto come & saving the ornamental plants.


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