Clonal fidelity testing_of_micropropagated_banana__(musa

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Clonal fidelity testing_of_micropropagated_banana

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Clonal fidelity testing_of_micropropagated_banana__(musa

  1. 1. BY SREEREMYA.S M.phil 12/1/12 CLONAL FIDELITY ANALYSIS OF MICROPROPAGATED BANANA ( Musa Spp.cv.Grand Naine ) USING MOLECULAR MARKER.
  2. 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. 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. 4.  RAPD stands for Random Amplification of Polymorphic DNA.  RAPD has been tested to detect somaclonal variation in cultivars of banana
  5. 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. 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. 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. 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. 9. Plant genomic DNA isolation (CTAB Method) Determination of DNA concentration and purity (using AGE) RAPD analysis Data analysis ….Technical program
  10. 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. 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. 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

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