Clonal fidelity testing_of_micropropagated_banana

1. BY SREEREMYA.S
M.phil
12/1/12
CLONAL FIDELITY ANALYSIS OF
MICROPROPAGATED BANANA ( Musa
Spp.cv.Grand Naine ) USING MOLECULAR
MARKER.

2. INTRODUCTION
 Banana belongs to the genus Musa.
 important crop plant.
 Musa acuminata & Musa balbisiana.
 Largest herbaceous flowering plant.
 Grand Nain bananas (also spelled Grande
Naine) are banana cultivars of Musa
acuminata. It is one of the most commonly
cultivated bananas
 Grand Naine is a monocot and belongs to
the genus Musa

3.  The Plant grows to a height of 6.5 to 7.5
Feet.
 RAPD- type of PCR reaction .
 Used to analyses the genome of an
individual using random primers.
 Clonal fidelity analysis is a technique to
analyse genetic stability of micro propagated
…. INTRODUCTION

4.  RAPD stands for Random Amplification
of Polymorphic DNA.
 RAPD has been tested to detect
somaclonal variation in cultivars of banana

5. PRACTICAL / SCIENTIFIC UTILITY
 Investigations on clonal fidelity will help to
distribute true-to-type plants of selected
variety to farmers in large scale, avoiding any
type of somaclonal variation

6. TECHNICAL PROGRAM
 Materials and Methods
 Sucker derived (1 plant) & micropropagated (15
plants) banana cv.Nendran (type Chengalikodan)
will be used for study.
 Isolation of plant genomic DNA will be standardized
as per standard protocols.
 Agarose Gel Electrophoresis will be carried out to
determine the purity and concentration of DNA.
 DNA samples isolated will be stored at 20ºC
temperature for RAPD and ISSR analyses.

7. ….TECHNICAL PROGRAM
RAPD Analysis
 RAPD primer screening will be carried out.
 Reaction mixture will be prepared with specific
template DNA, dNTPs, Taq DNA polymerase, MgCl
2 and primer in PCR buffer.
 PCR cycles .
 Agarose gel electrophoresis.

8. ….TECHNICAL PROGRAM
 Data analysis
 Markers visualized using Genotyper 2.5 software
 Scored manualy as present or absent for each
accession.
 Coefficient between each accession can be then
calculated using the SIMQUAL program in
NTSYS-pc version 1.70 (Rohlf 1992).
 Datas will be subjected to cluster analysis using
the UPGMA method in the SAHN program of
NTSYS to generate dendogram

9. Plant genomic DNA isolation
(CTAB Method)
Determination of DNA concentration and purity
(using AGE)
RAPD analysis
Data analysis
….Technical program

10. HYPOTHESIS
 If tissue cultured derived plant and sucker
derived plant show hundred percentage
similarity in RAPD profiles ,It can be
hypothesized that they maintain clonal
fidelity.

11. REFERENCE
 Gawal NJ, Jarret RL (1991). Cytoplasmic genetic
diversity in banana and plantain. Euphytica 52:19-
23.
 Bhat KV, Jarret RL, (1995) . Random amplified
polymorphic DNA and genetic diversity in Indian
Musa germplasm, Genet. Res. Crop Evol. 42: 107-
118.
 Qrtiz A,Renaud R,Calzada,Ritter R(1997). Analysis
of plum cultivars with RAPD markers.J.Hortic.Sci
72:1-9

12.  Porebski S, Bailey GL & Baum BR (1997)
Modifications of a CTAB DNA extraction protocol
for plant containing high polysaccharide and
polyphenol components. Plant Mol. Biol. Rep. 15:
8–15
 Atienzar F,Evender A,Savva D,Depledge
M(2000).Optmized RAPD analysis generates
high-quality genomic DNA. Profiles at high
annealing temperatures. Biotechniques 28:52-
54
REFERENCE