1. “ASSESSMENT OF GENETIC
STABILITY
OF INVITRO CONSERVED
Govt. GERMPLAST kamala raja girls OF post TARO
graduate
(COLOCASSIA ESCULENTA)”
Submitted by-
Monika sharma
M.Sc 4th sem
(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
2. 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.
5. 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.
6. Preparation of culture media as per
requirements.
Preparation of stock solution.
Invitro establishment of explants of taro.
Maintenance of culture.
Subculture.
7. 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
8. 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.
9.
10. 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 QUANTIFICATION
Because 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.
11.
12.
13. 2% agarose gel
is prepared
Sample is loaded in
the wells of the gel
DNA bands under trans illuminator
14. 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.
16. 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.
