“ASSESSMENT OF GENETIC

STABILITY OF INVITRO CONSERVED GERMPLAST OF TARO

(COLOCASSIA ESCULENTA)”

Submitted by- Monika sharmaM.Sc 4th sem

Govt. kamala raja girls post graduate (Autonomous College) Under the guidance of Dr. Sangeeta Yadav Senior scientist (NBPGR)

TISSUE CULTURE AND CRYOPRESERVATION UNIT, NATIONAL BUREAU OF PLANT GENETIC RESOURCES, ICAR PUSA CAMPUS, NEW DELHI

Goals/Objectives:

This project aims to: A. Develop an efficient micro propagation system

for taro varieties.

B. Evaluate field performance of tissue cultured and conventionally propagated taro.

C. Establish and maintain an invitro germplasm collection of taro.

CONTENTS

Introduction Materials and methods Results Conclusion

CLASSIFICATION: Kingdom: Plantae

Phylum: AngiospermClass: MonocotyledonOrder: AlismatalesFamily: AraceaeGenus: ColocasiaSpecies: C. esculenta Variety: esculenta

INTRODUCTION

Taro is a common name for the corms and tubers of several plants in the Araceae family. Colocasia esculenta is the most widely cultivated. Taro is native to Southern India and Southeast Asia. It is a perennial, tropical plant primarily grown as a root vegetable for its edible starchy corm, and as a leaf vegetable. It is a food staple in African, Oceanic and South Indian cultures and is believed to have been one of the earliest cultivated plants.

MATERIALS AND METHODS

Preparation of culture media as per requirements.

Preparation of stock solution. Invitro establishment of explants of taro. Maintenance of culture. Subculture.

Invitro establishment of explants of taro

Explants preparation

Shoot tips with a little piece of corm tissue were excised

Surface sterilization

Washing of explants with 0.1% solution of tween-20 detergent.

Shaking and mixing for 5 min.

Washed thoroughly with distilled water

Cleaned explants were subjected to 0.1% mercuric chloride solution treatment for 10 min.

Rinsed with sterilized distilled water 4-5 times.

Treatment with bavistin streptomycin and washed with sterilized distilled water.

Inoculation of surface-sterilized explants on Z10

Medium under aseptic condition.

Incubate, maintain the culture and grown for 30 days.

Subculturing on Z10 medium and then transfer to

field for plant growth.

DNA EXTRACTION CTAB METHOD

DNA PURIFICATION

There are several methods for purifying DNA. The one you choose depends on the nature of your DNA sample.

For eg:- By centrifugation at 15000 rpm for 15 min etc.

DNA QUANTIFICATIONBecause DNA and RNA absorb ultraviolet light, with a absorption peak at 260nm wavelength, spectrophotometers are commonly used to determine the concentration of DNA in a solution. Inside a spectrophotometer, a sample is exposed to ultraviolet light at 260 nm, and a photo-detector measures the light that passes through the sample. The more light absorbed by the sample, the higher the nucleic acid concentration in the sample.

PCR STEPS

PCR PROCESS

AGAROSE GEL ELECTROPHORESIS

2% agarose gel is prepared

Sample is loaded in the wells of the gel

DNA bands under trans illuminator

After the electrophoresis of the DNA sample the result is visualizes under UV transilluminator.

The bands are then observed under UV light.

The ladder is loaded along with the samples to verify that the resultant band is the desired

DNA sample.

OBSERVATION

Bands under UV

RESULTS:-

This project will result in the production of numerous disease-free plant materials of elite.

High yielding varieties of taro which will be distributed to local farmers and for international exchange.

Establishment of an invitro germplasm collection will ensure the protection of different taro varieties from environmental stresses.

It will also ensure conservation of these valuable genetic resources for future breeding activities and for the younger generation.

THANK

YOU