IN VITRO REGENERATION OF PINEAPPLE (Ananas comosus) L. MERR 'MD2' USING DIFFERENT CONCENTRATION OF INDOLE ACETIC ACID (IAA) AND KINETIN (KN) HORMONE NORMA ZIATI BINTI ISMAIL Final Year Project Report Submitted In Partial FulIJJ.ment ofthe Requirements for the Degree ofbachelor of Science (Bons.) Technology and Plantation Management in the Faculty of Plantation And Agrotechnology Universiti Teknologi Mara JULY 2015
DECLARATION This Final Year Project is a partial fulfilment of the requirements for a degree of Bachelor of Science (Hons.) Technology and Plantation Management, Faculty of Plantation and Agrotechnology, Universiti Teknologi MARA. It is entirely my own work and has not been submitted to any other University or higher education institution, or for any other academic award in this University. Where used has been made of the work of other people it has been fully acknowledged and fully referenced. I hereby assign all and every rights in the copyright to this Work to the Universiti Teknologi MARA ("UiTM"), which henceforth shall be owner of copyright in this Work and that, any reproduction or use in any form or by any means whatsoever is prohibited without a written consent ofuitm. '4 P1.t~U3"'" :2.015" Date : 1.. I hereby declare that I have checked this project and in my opinion, this project is adequate in term of scope and quality for the award of the degree of Bachelor of Science (Hons.) Technology and Plantation Management, Faculty of Plantation and Agrotechnology, Universiti Teknologi MARA. Name of Supervisor:...V.R: HS.H..A.H...R.W.R..~. Position: 8.EN.l.Q.B.LE.C.TU.a.EB. Date: l4:{.g:i."h.9.a5.. ii
TABLE OF CONTENTS ACKNOWLEDGEMENT TABLE OF CONTENTS LIST OF FIGURES LIST OF TABLES LIST OF ABBREVIATIONS ABSTRACT ABSTRAK iii iv v vi vii viii ix CHAPTER 1 : INTRODUCTION 1.1 Background ofstudy 1 1.2 Problem statement 3 1.3 Significance ofstudy 4 1.4 Objectives of study 4 1.5 Scope of study 4 1.6 Hypothesis testing 5 CHAPTER 2 : LITERATURE REVIEW 2.1 Pineapple (Ananas comosus) L. Merr 'MD2' 2.1.1 Origin and history ofmd2 pineapple 2.1.2 Taxonomy ofmd2 pineapple 2.1.3 Pineapple vernacular name 2.1.4 Characteristic of(ananas comosus) L. Merr 2.1.5 Pineapple (Ananas comosus) variety MD2 6 7 7 8 9 2.2 Propagation technique ofpineapple variety MD2 2.3 2.2.1 Conventional propagation ofmd2 pineapple 2.2.2 In vitro propagation ofmd2 pineapple 2.2.3 Source ofexplant used in in vitro regeneration Hormone used in micropropagation ofmd2 pineapple 2.3.1 Summarization ofhormone used from previous study 11 11 13 14 17 iv
CHAPTER 3 : METHODOLOGY 3.1 Location ofstudy 20 3.2 Material and apparatus used for in vitro regeneration ofmd2 pineapple 3.2.1 Plant material for in vitro regeneration ofmd2 pineapple 20 3.2.2 Medium for culture MD2 pineapple explant 3.2.3 Hormone used for culture MD2 pineapple explant 3.2.4 Equipment and apparatus used 20 21 21 3.3.1 Sterilization process for MD2 pineapple explant regeneration 3.3.1 Sterilization ofmedium for explant growth 3.3.2 Sterilization ofapparatus used 22 22 3.3.3 Sterilization ofmd2 pineapple explant and initiation of culture 22 3.4 Procedure for in vitro regeneration ofmd2 pineapple 3.4.1 Preparation of MD2 pineapple explant 23 3.4.2 Preparation ofmedia for explant tissue culture 23 3.5 MD2 pineapple explant culture on medium 25 3.6 Size ofplots and treatments 25 3.7 Parameter ofstudy 26 3.8 Experimental design 27 3.9 Statistical analysis 27 CHAPTER 4: RESULT 4.1 Overall observation 29 4.2 Effect number ofshoot 30 4.3 Effect length ofshoot 35 4.4 Effect height ofplantlet 41 4.5 Effect fresh weight ofplantlet 46 v
ABSTRACT In vitro regeneration of Ananas comosus L. Merr. 'MD2' (pineapple) was carried out using MD2 pineapple sucker as explant. In vitro regeneration was done by manipulating the combination of plant growth regulator between Indole Acetic Acid (IAA) and Kinetin (Kn). Eleven different combination of IAA and Kn hormone was applied in MS medium with vitamins supplemented with 8.0g/L agar and 30g/L sucrose at combination of different concentration ofiaa (0.00,0.05,0.10,0.15,0.20,0.25,0.30,0.35,0.40,0.45 and 0.5 mg/l) and Kn (0.0, 0.50, 1.00, 1.50,2.00,2.50,3.00, 3.50,4.00,4.50 and 5.00 mg/l) respectively and maintain at 25 C under 16 hours of photoperiod. The result shows that the sucker explant able to induce shoot proliferation on different combination of IAA and Kn. Optimum regeneration for the best result was obtained from MS medium supplemented with 0.35mg/L IAA and 3.50mg/L Kn. Shoot proliferation start undergo morphogenesis after 2 to 3 weeks culture on MS medium. In addition, this combination of plant growth regulator regenerated highest mean for the number of shoot regenerate per explant (2.98unit), the length of shoot (0.82cm), height of plantlet (1.01cm) and the weight of plantlet (1.42g) compared to other combination of IAA and Kn tested. However, all the explant showing different respond towards treatment given. x