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The Effects of Varying Concentrations of Growth Regulators

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The Effects of Varying Concentrations of Growth Regulators PH0400003 The Effects of Varying Concentrations of Growth Regulators Benzyladenine and Naphthalene Acetic Acid and Gamma Irradiation on the Gross Plantlet Morphology of Garcinia mangostana L. Ma. Fatima Nona M. Bonsol Jade Marie Edenvirg F. Lasiste Ma. Franchesca S. Quinio A Research Paper Submitted to the Faculty of the Philippine Science High School Diliman Campus in Partial Fulfillment Of the Requirements in Research 2 February 2004 APPROVAL SHEET This research work entitled "The Effects of Varying Concentrations of Growth Regulators Benzyladenine and Naphthalene Acetic Acid and Gamma Irradiation on the Gross Plantlet Morphology of Garcinia mangostana L.n by Ma. Fatima Nona M. Bonsol, Jade Marie Edenvirg F. Lasiste, and Ma. Franchesca S. Quinio, which was presented to the faculty of the Philippine Science High School Diliman Campus in partial fulfillment of the requirements in Science and Technology Research 2, is hereby accepted. Gerel Roa Research Adviser 11 ACKNOWLEDGEMENT The group would like to express their utmost gratitude: To the Almighty God; if not for the mental strength He supplied the group with they would have given up on this research long ago. To the parents, for their overwhelming support throughout the conduction of the project; for providing free transportation and lending a financial hand. To the Philippine Nuclear Research Institute's (PNRI) Nuclear Training Group (NTG), who helped in arranging all the papers needed to be able to work in the laboratory. To Mr. Apolinar B. Asencion, who tried to find a research that can be worked on. To Mrs. Avelina G. Lapade, who shared her written materials to increase the group's knowledge on the subject, and for allowing us to work on part of their research; and to Mrs. Marie Veluz and Kuya Manny, who were always around to give advice on the different laboratory techniques, and for guiding the members every step of the way. To Mr. Gerel Roa, who tried to make the members see their mistakes as best as he could; for the encouragement, support, and guidance. To Mr. Jun Samson, for allowing the group to maximize the use of the computer, and for lending them the glassware and chemicals needed. To. Mrs. Alma Calud and Mr. Ellery Jose, for being ready to give advice at all times. To Mr. Bert Barcelo, Mr. Rafael Santiago, Ms. Joanne Pe Benito, and Ms. Malou Capundag, for giving the group permission to juggle their class schedules in order to work at the PNRI whenever needed. To Mr. Alex Alix, Mr. Gerel Roa, Mr. Delfin Angeles, Mrs. Mercy Sanchez, Ms. Melissa Cardenas, Mrs. Alma Calud and Mrs. Sofia Docto, for buying the excess mangosteen from the group. To classmates and friends, whose little deeds aided us in getting through the year, amid the hustle. in ABSTRACT Mangosteen (Garcinia mangostana L.) is regarded as one of the best- flavored fruits in the world. Though this fruit yields high income and has great potential as an export, productivity in the Philippines is still limited due to its slow and difficult conventional propagation. To increase the plants' productivity, growth regulators can be added, the seeds can be exposed to irradiation to cause genetic mutations, and micropropagation techniques can be used. This study seeks to determine the effects of these three variables on the general morphological characteristics of the plantlets grown both in vitro and in vivo. The first set of mangosteen seeds was inoculated and grown in Murashige and Skoog (MS) basal culture medium with different treatment combinations of growth regulators, naphthalene acetic acid (NAA) and benzyladenine (BA). Treatment concentrations ranged from 0 parts per million (ppm) to 6 ppm. The treatment that best induced stem, leaf and root formation, and produced plantlets with long stems and roots, was MS with 6 ppm NAA combined with 6 ppm BA. A second set of seeds was then exposed to varying levels of gamma radiation and propagated in vivo and in vitro. For in vivo, it was observed that the length of stem of the plantlet decreases with increasing radiation dose; length of leaves of the mangosteen plantlets was best noted in seeds exposed to 10 Gy gamma rays. In in vitro, the two treatment concentrations that best helped seeds to develop callus, which were 0 ppm BA with 4 ppm NAA and 4 ppm BA with 6 ppm NAA, were added to MS basal medium. Seeds exposed to 5 Gy and grown in MS basal medium with 0 ppm BA and 4 ppm NAA were observed to have developed the most callus. IV TABLE OF CONTENTS TITLE PAGE Title i Approval Sheet ii Acknowledgement iii Abstract iv Table of Contents v List of Tables vi List of Figures vii Introduction 1 Background of the Study 1 Statement of the Problem 1 Significance of the Study 2 Scope and Limitations 2 Review of Related Literature 4 Methodology 13 Results and Discussion 22 Summary and Conclusion 33 Recommendations 35 Bibliography 36 Appendices 37 Appendix A: Complete Data Tables 37 Appendix B: T-Test Calculations 47 LIST OF TABLES TABLE TITLE PAGE 1 Chemicals Used in the Preparation of Stock Solutions 14 2 Other Chemicals Used in the Preparation of MS Basal Medium 15 3 Volume of BA and Distilled Water Added to Treatments 15 4 Volume of NAA and Distilled Water Added to Treatments 16 5 The Sixteen Treatments of Growth Regulators for Optimization 17 6 Treatment Combinations for Callus Formation 18 7 Averages Observed for the Optimization of Growth Regulators 22 8 Averages Observed for Callus Formation 23 9 Averages Observed for In Vivo Planting 23 10 Summary of Optimum Treatments 32 11 Complete Data for the Optimization of Growth Regulators 37 12 Complete Data for Callus Formation 40 13 Complete Data for Stem Length in In Vivo Planting 41 14 Complete Data for Leaf Length in In Vivo Planting 44 15 T-Test Calculations for In Vitro Root Length. 47 16 T-Test Calculations for In Vitro Stem Length. 47 17 T-Test Calculations for In Vitro Leaf Length. 48 18 T-Test Calculations for In Vivo Stem Length. 48 19 T-Test Calculations for In Vivo Leaf Length. 48 vi LIST OF FIGURES FIGURE TITLE PAGE 1 In Vivo Planting (Block 1) 19 2 In Vivo Planting (Block 2) 20 3 In Vivo Planting (Block 3) 20 4 Average Area of Seed Covered with Callus of In Vitro Plants in 24 Different Combinations of Growth Regulators 5 Average Root Length of In Vitro Plants in Different 24 Combinations of Growth Regulators 6 Average Stem Length of In Vitro Plants in Different 26 Combinations of Growth Regulators 7 Average Leaf Length of In Vitro Plants in Different 26 Combinations of Growth Regulators 8 Average Root Number of In Vitro Plants in Different 27 Combinations of Growth Regulators 9 Average Number of Stems of In Vitro Plants in 27 Different Combinations of Growth Regulators 10 Average Leaf Number of In Vitro Plants in 28 Different Combinations of Growth Regulators 11 Percentage Formation of the Parts of In Vitro Plants in Different 29 Combinations of Growth Regulators 12 Size of Callus for Two Growth Regulator Treatments at 29 Different Doses of Irradiation 13 Average Leaf and Stem Length of Mangosteen Exposed to 30 Different Amounts of Irradiation Vll INTRODUCTION Background of the Study One of the most praised of tropical fruits and certainly the most esteemed in the family GiUifferae, the mangosteen, Garcinia mangostana L, is almost universally known. It is usually eaten fresh or preserved into candies. It also has a variety of uses, since it can be used medicinally and at the same time as a raw material for building objects. Filipinos employ a decoction of the leaves and bark as a febrifuge to treat diarrhea, dysentery and urinary disorders. In Thailand, it has been used to make handles for spears, rice pounders and is employed in construction and cabinetwork. Though mangosteen yields high income, the Philippines still has limited productivity of its fruits due to the fact that this crop is seasonal and only bears fruits during its peak season. Furthermore, the Philippines is finding it hard to propagate mangosteen due to the fact that its conventional vegetative propagation is difficult while various methods of grafting have been unsuccessful. There is also a low yield of this fruits due to poor cultural management practices and lack of awareness of this crop. Statement of the Problem There are three ways of increasing the plants' productivity. One is through the use of growth regulators, the other involves exposing the seeds to irradiation to cause genetic mutations, and the last is by using micropropagation techniques. This study aims to determine the growth regulator concentrations that would best induce long and numerous roots, stems and leaves, and the most formation of callus, in seeds planted in vitro. It also seeks to establish the growth regulator treatment and irradiation dose combination that would produce seeds planted in vitro with the most callus formed, and to ascertain the 1 effects of irradiation on the stem and leaf lengths of the seeds grown in vivo. Suitability of tissue culture for mangosteen will also be examined. It is hypothesized that there is a significant difference in the morphological characteristics observed among the different treatments of growth regulators. A significant difference is also expected among the seeds exposed to varying levels of gamma irradiation, both in vivo and in vitro. Significance of the Study The morphological changes that are expected from the irradiation of the plants' seeds, and from the treatment with growth regulators, include development of plantlets with long and multiple roots, stems, and leaves, and the formation of the most callus.
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