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PROPAGATION of KAPUR BUKIT (Dryobalanops Beccarii DYER) THROUGH STE~ CUTTING and TISSUE CULTURE "

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PROPAGATION of KAPUR BUKIT (Dryobalanops Beccarii DYER) THROUGH STE~ CUTTING and TISSUE CULTURE • PROPAGATION OF KAPUR BUKIT (Dryobalanops beccarii DYER) THROUGH STE~ CUTTING AND TISSUE CULTURE ". ," ", •. - REBICCA EDWARD @ MAY ~ Bachelor of Science with Honours 647 (Plant Resource Science and Management) 1289 2005 2005 Pusat Khidmat Maklumat Akaden UNIVERSHI MALAYSIA SARAWA 94100 KOla Samarahao PROPAGATION OF KAPUR BUKIT (Dryoba/anops beccarii DYER) THROUGH STEM CUTTING AND TISSUE CULTURE P.KHIDMATMAKLUMA TAKADEMIK UIiIMAS 111111111111 1111111111111 1000128119 REBICCA EDWARD @ MAY This report is submitted in partial fulfillment of the requirements for the degree of Bachelor of Science with Honours (Plant Resource Science and Management) Faculty of Resource Science and Technology UNIVERSITI MALAYSIA SARAW AK 2005 I DECLARATION I hereby declare that no portion of the work referred to in this dissertation has been submitted in support of an application for another degree or qualification to this university or any other institution of higher learning. Rebicca Edward @ May Plant Resource and Science Management Faculty of Resource Science and Technology University Malaysia Sarawak 11 ACKNOWLEGEMENT First of all, I would like to thanks GOD for the continuous blessing that I have received during those challenging and wonderful times that I have gone through from the starting until the end of this project. Secondly, I wish to express my deepest gratitude and sincere appreciation to my supervisors, Madam Dayang Awa Abang Lingkeu and Assoc. Prof. Dr. Sim Soon Liang for their advices, guidance and supports from the very beginning until the completion of this project. Million of thanks are extended to Assoc. Prof. Dr. Isa Ipor, Dr. Ling Teck Yee and Plant Laboratory assistants for their help during the implementation of this project. I am also grateful to the Sarawak Forest Corporation (SFC) Semengoh for supplying and maintenance of the seedlings, invaluable suggestions, help and continuous interest in the study. I also would like to thank all my supportive friends; especially Malcolm Boxey Jilimin, Amelia Cynthia Bosi, Nor Hamirah, Muziah, Nadine William Norman, Khairulnaimah Khairudin, Wilhelmina Cluny, Kho Pei Ee, Freddy Yeo Kuok San, Mary Rose Laurence, Nooreffyanteh Jaflie, Norasiah, Rohana and my sister Lydia Kristen for their helping hand, companionship, understanding and willingness. Last but not least, my devoted appreciation and utmost thankfulness goes out to my loving and caring family, especially my parents: Mr. Edward Mongudal Koromol and Mrs. Bonah Garatam, brothers and sisters, relatives and Nelson Aloysius for their unconditional love, supports and prayers. III :'1) at UN! TABLE OF CONTENTS TITLE PAGE DECLARATION ii ACKNOWLEDGEMENT III TABLE OF CONTENTS iv LIST OF TABLES vi LIST OF PLATES viii LIST OF APPENDIXES viii ABSTRACT 1 CHAPTER 1 INTRODUCTION 2 2 LITERATURE REVIEW 5 2.1 Vegetative Propagation 5 2.1.1 Rooting of Dryobalanops lanceolata Burck (kapur paji) cuttings 6 2.2 Micropropagation (Tissue Culture) 8 2.2.1 Development of pre-somatic embryogenesis on Shore a pinanga 9 2.2.2 Somatic embryogenesis of Shorea macroptera 10 2.2.3 Browning exudates from explants 11 3 MATERIALS AND METHODS 12 3.1 Vegetative Propagation by Stem Cutting 13 3.1.1 Preparation of sand bed and plant growth regulator 13 3.1.2 Preparation and treatment of cuttings 13 3.1.3 Watering schedule 14 3.1.4 First Trial 14 3.1.5 Second Trial 15 3.1.6 Third Trial 15 3.1.7 Observation, Recording and Data Collection 16 3.2 Micropropagation (Tissue Culture) 17 3.2.1 Establishment of axenic culture 17 3.2.1.1 Surface sterilization 17 IV 3.2.1.2 Experiment 1 18 3.2.1.3 Experiments 2 to 8 18 3.2.2 Observation and Recording 19 3.2.3 Induction of multiple shoots formation 20 3.2.3.1 Induction of multiple shoots by BAP 20 3.2.3.2 Induction of multiple shoots by Kinetin and NAA 21 3.2.4 Induction of multiple shoot formation via callus 21 3.2.4.1 Induction of callus by 2,4-D 21 3.2.4.2 Induction of callus by 4 amino-3,5,6 trichloropicolinic acid (Picloram) 22 4 RESULTS 23 4.1 Vegetative Propagation by Stem Cutting 23 4.1.1 First Trial 23 4.1.2 Second Trial 25 4.1.3 Third Trial 26 4.2 Micropropagation (Tissue Culture) 28 4.2.1 Experiment 1. Establishment of axenic culture 28 4.2.2 Experiment 2 33 4.2.3 Experiment 3 34 4.2.4 Experiment 4 35 4.2.5 Experiment 5 36 4.2.6 Experiment 6 37 4.2.7 Experiment 7 38 4.2.8 Experiment 8 39 4.2.9 Induction of multiple shoot formation 40 4.2.10 Induction of multiple shoot formation via callus 40 5 DISCUSSION 43 5.1 Vegetative Propagation by Stem Cutting 43 5.2 Micropropagation (Tissue Culture) 47 6 CONCLUSION 49 REFERENCES 51 APPENDIXES 53 v LIST OF TABLES Table 4.1.1. Number of cuttings that have potential to root (first trial). 23 Table 4.1.2. Number of cuttings that have potential to root (second trial). 25 a Table 4.1.3. Number of cutting rooted , number of cutting develop at least one new shoot and the total length (cm)b of the new shoot, total c d number of new leaves and total length (cm) of roots . 26 Table 4.2.la. Mean number of axenic lamina explants after exposure for 15, 20,25 and 30 minutes in each of the 3 concentrations (35%, 40%, 45%) of 'Clorox'. 29 Table 4.2.1b. Mean number of contaminated lamina explants after exposure for 15,20,25 and 30 minutes in each of the 3 concentrations (35%, 40%,45%) of 'Clorox'. 29 Table 4.2.lc. Mean number of damaged lamina explants after exposure for 15, 20,25 and 30 minutes in each ofthe 3 concentrations (35%, 40%, 45%) of'Clorox'. 29 Table 4.2.1d. Mean number of axenic node explants after exposure for 15, 20, 25 and 30 minutes in each of the 3 concentrations (35%,40%, 45%) of 'Clorox'. 30 Table 4.2.1e. Mean number of contaminated node explants after exposure for 15, 20,25 and 30 minutes in each of the 3 concentrations (35%, 40%, 45%) of'Clorox' . 30 Table 4.2.1f. Mean number of damaged node explants after exposure for 15, 20, 25 and 30 minutes in each of the 3 concentrations (35%,40%,45%) of 'Clorox'. 31 Table 4.2.1 g. Mean number of axenic inter-nodal stem explants after exposure for 15,20,25 and 30 minutes in each of the 3 concentrations (35%, 40%,45%) of 'Clorox'. 32 Table 4.2.1h. Mean number of contaminated inter-nodal stem explants after exposure for 15, 20, 25 and 30 minutes in each of the 3 concentrations (35%, 40%, 45%) of'Clorox'. 32 Table 4.2.2. Mean number of axenic, contaminated and damaged lamina explants after exposure in 5% 'Clorox' for 15 minutes for 3 types of explants. 33 VI ,... ,.... Table 4.2.3. Mean number of axenic, contaminated and damaged lamina explants after exposure in 20% 'Clorox' for 15 minutes for 3 types of explants. 34 Table 4.2.4a. Mean number of axenic, contaminated and damaged lamina explants after exposure in 20% 'Clorox' for 15 minutes for 3 types of explants. 35 Table 4.2.4b. Mean number of axenic, contaminated and damaged shoot tips explant after exposure in 20% 'Clorox' for 15 minutes. 36 Table 4.2.5a. Mean number of axenic, contaminated and damaged lamina explants after exposure in 20% 'Clorox' for 15 minutes for 3 types of explants. 37 Table 4.2.5b. Mean number of axenic, contaminated and damaged shoot tips explants after exposure in 20% 'Clorox' for 15 minutes. 37 Table 4.2.7a. Mean number of axenic, contaminated and damaged lamina explants after exposure in 20% 'Clorox' for 15 minutes for 3 types of explants. 38 Table 4.2.7b. Mean number of axenic, contaminated and damaged shoot tips explants after exposure in 20% 'Clorox' for 15 minutes. 39 VII LIST OF PLATES I' Plate 1. Untreated cutting from the first trial and was used to monitor the rooting performance. 24 Plate 2. Three cuttings that survive for 15 weeks in the third trial. 27 Plate 3. Callus of D. beccarii shoot tip after 2 months of induction in Y2 MS + 1.5 mIll BAP + 1.0 mIll PPM. 41 Plate 4. New growth of axillary bud from D. beccarii stem node after 4 weeks of induction in Y2 MS + 1.0 mIll BAP + 1.0 mIll PPM. 41 Plate 5. Callus ofD. beccarii shoot tip after 2 months of induction in Yz MS + 1.0 mIll BAP + 1.0 mIll PPM. 42 LIST OF APPENDIXES Appendix I Univariate analysis ofvariance (ANOVA) Tables 53 Appendix n Post Hoc Tests Tables 56 Vlll Propagation of Kapur Bukit (Dryobalanops beccarii Dyer) through Stem Cutting and Tissue Culture Rebicca Edward @ May Plant Resource Science and Management Faculty Resource Science and Technology University Malaysia Sarawak ABSTRACT Kapur bukit (Dryobalanops beccarii Dyer) is a timber species of the Dipterocarpaceae. Kapur bukit is a good source of wood for construction purposes, particularly in plywood production. Vegetative propagation of this species through stem cutting and tissue culture was studied. In the study of vegetative propagation by stem cutting, rooting hormone were used and six treatments included were four different concentrations of indole­ 3-butyric acid (IBA), one with Seradix 2 and water as control. Three trials were conducted and the results showed that this species was difficult to be propagated by cuttings. An untreated cutting from the first trial developed root, shoot and new leaves, no cuttings rooted in second trial, while a cutting that has been treated with 2 gil IBA from the third trial developed root, shoot and new leaf.
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