VIETNAM NATIONAL UNIVERSITY OF AGRICULTURE FACULTY OF BIOTECHNOLOGY -------------***------------- GRADUATION THESIS TOPIC: RESEARCH ON IN VITRO PROPAGATION OF SCHISMATOGLOTTIS WALLICHII HANOI – 2022 VIETNAM NATION UNIVERSITY OF AGRICULTURE FACULTY OF BIOTECHNOLOGY – – – – – – – – – – – – GRADUATION THESIS TOPIC: “RESEARCH ON IN VITRO PROPAGATION OF SCHISMATOGLOTTIS WALLICHII” Supervisor : Dr. Nguyen Thi Lam Hai Student name : Le Thi Phuong Oanh Student code : 637061 Class : K63CNSHE HA NOI – 2022 DECLARATION OF AUTHORSHIP I hereby declare that this is my research work. The data, results and reports mentioned herein are honest and have not been used to be published in previous theses, dissertations, and scientific research works. I hereby certify that the citation information used in the thesis is clearly stated in the source, ensuring that the citation is correct as prescribed.
I take full responsibility for this promise! Hanoi, day month year 2022 Student Le Thi Phuong Oanh i ACKNOWLEDGEMENTS In order to complete this graduation thesis, I would like to thank the Faculty of Biotechnology of Vietnam National University of Agriculture for organizing and facilitating support and assistance in the process of studying and researching the topic. With the deepest gratitude, I would like to send to the teachers in the Department of Plant - Faculty of Biotechnology of Vietnam National University of Agriculture, especially Dr. Nguyen Thi Lam Hai, who followed me closely during the research and imparted valuable knowledge to us. throughout the time at school.
Thanks to the guidance and teaching of the teachers, my research topic can be completed well. I would like to sincerely thank Mr. Vu Cong Son and the staff at T.N Biotech Biological Joint Stock Company - who directly helped, cared and guided me to successfully complete this report in the past time. The final thesis was carried out over a period of nearly 6 months.
My initial step into practice is still limited and there are still many surprises, so it is inevitable that I will make mistakes, I look forward to receiving valuable comments from teachers so that my knowledge in this field can be improved. more complete and at the same time have conditions to supplement and raise their consciousness. Thank you sincerely! Hanoi, day month year 2022 Student Le Thi Phuong Oanh ii CONTENTS DECLARATION OF AUTHORSHIP. II LIST OF ABBREVIATIONS.
VI CONTENT OF FIGURE. VII CONTENT OF TABLE. General introduction about Schismatoglottis wallichii. Botanical characteristics of Schismatoglottis wallichii.
Some studies on tissue culture of species of the family Araceae. Introduction of Temporary Immersion Bioreactor system (TIB system) .Principle of Temporary Immersion Bioreactor system (TIB system). Some micropropagation studies of Temporary Immersion Bioreactor system (TIB system). RESEARCH MATERIALS AND CONTENTS AND METHODS .Object, material, location, time of culture.
Location, time of culture. Evaluation of the effect of growth hormones on the multiplication stage of Schismatoglottis wallichii. Comparison of growth of Schismatoglottis wallichii on the multiplication stage in different culture systems. Evaluation of the effect of growth hormones on the rooting stage of Schismatoglottis wallichii.
Comparison of growth of Schismatoglottis wallichii on the rooting stage in different culture systems. Method of proceeding. 24 PART 4: RESULTS AND DISCUSSION. Effect of growth hormones on the multiplication stage of Schismatoglottis wallichii.
Effect of auxins on shoot proliferation and growth of Schismatoglottis wallichii after 4 weeks of culture. Effect of IBA (3-Indolebutyric acid) in combination with BA on shoot proliferation and growth of Schismatoglottis wallichii. Effect of α-NAA (α-Naphthaleneacetic acid) in combination with BA on shoot proliferation and growth of Schismatoglottis wallichii. Comparison of efficiency on shoot proliferation and growth of Schismatoglottis wallichii between IBA and α-NAA in combination with BA supplemented medium after 4 weeks of culture.
Comparison of growth of Schismatoglottis wallichii on the multiplication stage in different culture systems. Effect of Auxin on the rooting stage of Schismatoglottis wallichii. Effect of IBA (3-Indolebutyric acid) on the rooting stage of Schismatoglottis wallichii. Effect of α-NAA (α-Naphthaleneacetic acid) on the rooting stage of Schismatoglottis wallichii.
Comparison of efficiency on quality of Schismatoglottis wallichii between IBA and α-NAA supplemented medium on the rooting stage after 4 weeks of culture. Comparison of growth of Schismatoglottis wallichii on the rooting stage in different culture systems. CONCLUSIONS AND RECOMMENDATIONS .46 v LIST OF ABBREVIATIONS Abbreviations Annotate MS Murashige and Skoog medium CV Coefficient of variation LSD5% Least significant differences BA Benzyl Aminopurine TIB Temporary Immersion Bioreactors IBA Indole-3-butyric acid α-NAA α-Naphthaleneacetic acid vi CONTENTS OF FIGURE Figure 2. 2: The Herb of Schismatoglottis wallichii.
3: The blade (leaves) of Schismatoglottis wallichii. 4: The Inflorescences of Schismatoglottis wallichii. 5: Technological design and operational principle of Temporary Immersion Bioreactor system. 1: Schismatoglottis wallichii shoots in BA supplemented mediums after 4 weeks of culture …………………………………………………………….
2: Schismatoglottis wallichii shoots in BA supplemented mediums combined with IBA after 4 weeks of culture. 3: Schismatoglottis wallichii shoots in BA supplemented mediums combined with α-NAA after 4 weeks of culture. 4 Schismatoglottis wallichii shoots between BA supplemented medium combined with IBA and α-NAA after 4 week of culture. 5: Schismatoglottis wallichii shoots in vitro culture systems after 4 weeks of culture.
6: Schismatoglottis wallichii in IBA (3-Indolebutyric acid) supplemented mediums on the rooting stage after 4 weeks of culture. 7: Schismatoglottis wallichii in α-NAA (α-Naphthaleneacetic acid) supplemented mediums on the rooting stage after 4 weeks of culture. 8: Schismatoglottis wallichii in α-NAA (α-Naphthaleneacetic acid) and IBA (3-Indolebutyric acid) supplemented mediums on the rooting stage after 4 weeks of culture. 9: Schismatoglottis wallichii in vitro culture systems on the rooting stage after 4 weeks of culture.
42 vii CONTENTS OF TABLE Table 4. 1: Effect of BA (Benzyl Aminopurine) on shoot proliferation and growth of Schismatoglottis wallichii after 4 weeks of culture. 2: Effect of IBA (3-Indolebutyric acid) in combination with BA on shoot proliferation and growth of Schismatoglottis wallichii after 4 weeks of culture 28 Table 4. 3: Effect of α-NAA (α-Naphthaleneacetic acid) in combination with BA on shoot proliferation and growth of Schismatoglottis wallichii after 4 weeks of culture.
4: Efficiency of in vitro culture systems of Schismatoglottis wallichii after 4 week of culture. 5: Effect of IBA (3-Indolebutyric acid) on the rooting stage of Schismatoglottis wallichii after 4 weeks of culture. 6: Effect of α-NAA (α-Naphthaleneacetic acid) on the rooting stage of Schismatoglottis wallichii after 4 weeks of culture. 7: Efficiency of in vitro culture systems of Schismatoglottis wallichii on the rooting stage after 4 week of culture.
40 viii ABSTRACT To aid with fresher air and offer beauty to living spaces, the usage of decorative plants has grown in popularity in recent years. As a result, decorative plants like Schismatoglottis wallichii are quite popular. Because it has a significant economic impact, Schismatoglottis wallichii is one of the most significant ornamental plants. The objective of this study was to compare the efficacy of the temporary imersion bioreactor (TIB), a semi-solid culture system, and liquid culture systems for Schismatoglottis wallichii in order to determine the most ideal culture system for in vitro multiplication of Schismatoglottis wallichii.
The best outcomes for plants were likewise achieved when cytokines and auxins were added during the rapid multiplication and roots stages. Using in vitro cultures and disease-free samples, the following outcomes were attained by rotating between several cultures: On MS + 1 mg/l BA + 0.75 mg/l IBA + 6 g/l agar, the maximum shoot multiplication yield (3.9 leaves/plant, and 100% rooted) was attained. In the rooting stage, MS + 0.75 mg/l α-NAA + 6 g/l agar was the best medium for chimatos plants (average root length: 3.35 roots/plant, mass plant weight: 10. the temporary imersion bioreactor (TIB)is the most effective in vitro culture technology for Schismatoglottis wallichii during both rapid multiplication and rooting.
Prefaceace Schismatoglottis wallichii is a species of Angiosperms (Flowering Seed Plants) (Monocotyledon) in the Araceae family. This species was acquired by Hook. first scientific description in 1893. The leaf surface is wrinkled and swollen, parallel to the leaf veins.
The primary leaf color is green, often variegated with dark-green and grey strips. When the need to love bonsai is more concerned, Schismatoglottis wallichii is also one of the favorite plants in the bonsai collection of people who are passionate about bonsai. In recent years in Vietnam, the hobby of ornamental plants is also very developed. Schismatoglottis wallichii has also become a sought-after favorite.
Exploitation in the wild is limited, so the propagation of Schismatoglottis wallichii by plant tissue culture is of more interest. The advantage of systems is that they increase multiplication rates, allow greater availability of culture medium components, and reduce production costs. Plant tissue culture has been applied to production since the 20th century when economics and science developed, many plant tissue culture systems were applied and achieved high efficiency. Temporary immersion bioreactor system, in recent years, this system has begun to be applied to large-scale production of micropropagation in some countries such as Brazil, and the Netherlands.
is considered to help plants grow quickly and save a lot of labor costs in culture. Therefore, this system has great potential for the production of micropropagation. In Vietnam, there have been several studies on the application of bioreactor systems for tissue culture, but they are few and the applicability is not high. Therefore, to meet the requirements for the production of high-quality Schismatoglottis wallichii in vitro, I conducted a research project: “Research on in vitro propagation of Schismatoglottis wallichii”.
Objectives The aim of the present study is: - To find-out the optimal medium for the morphogenesis of Schismatoglottis wallichii - Comparison of micropropagation performance of Schismatoglottis wallichii on several different culture systems during rapid propagation and rooting stage 1. Requirements - Study the effect of growth regulators on shoot proliferation and rooting stage. - Compare the efficiency of in vitro culture systems (the temporary immersion bioreactor (TIB), the semi-solid medium culture system and the liquid medium culture system). General introduction about Schismatoglottis wallichii Schismatoglottis wallichii is a plant of the family Araceae.
It is found in islands around Southeast Asia, In Malesia, from Peninsular Malaysia, Singapore, Sumatra, Java, Borneo, Philippines, and east New Guinea. Habitat of terrestrial native Schismatoglottis wallichii (Primary Rainforest, Secondary Rainforest, Freshwater Swamp Forest). The preferred climate zone is Tropical. Schismatoglottis wallichii is a light-loving plant, so it is suitable for planting in the garden, terrace, balcony, or in the window that receives light every day.
with a highlight with a white stripe on the leaves as a decorative accent to create attraction in the bonsai industry. Scientific Classification Science name: Schismatoglottis wallichii Hook.f Common name in Viet Nam: ‘Mường Thái’ Regnum Plantae Unclassified Angiospermae Unclassified Monocots Ordo Alismatales Familia Araceae Genus Schismatoglottis Species S. wallichii Now, Schismatoglottis has more 100 recognized species from tropical and subtropical Asian peninsulas, especially it is found in primary rainforests. Around the world, many growers and researchers on Schismatoglottis species as well as choose to create new lines and varieties to serve human needs.
Botanical characteristics of Schismatoglottis wallichii Herb: Up to 50 cm, but typically 20 cm or so. Stem epigeal, upright to decumbent, 1., pleonastic, hapaxanthic, 1-2 cm thick, and bright green modules. Petiole 12-48 cm long, glabrous, sheathing in the lower 1/3 to 3/5; wings of sheath fully attached, tapering, persistent to slowly degrading in the marginal distal part; blade (broadly oblanceolate to) oblong-lanceolate to narrowly ovate, usually rather distinctly asymmetric, bright to mid-green, occasionally suffused with purple and/or purple-backed, adaxially sometimes (to very shallowly cordate), midrib slightly prominent abaxially (dry) or not, secondary venation arising mainly from the midrib; tertiary venation obscure. 2: The Herb of Schismatoglottis wallichii Blade: Blade is moderately tough, elliptic to ovato-sagittate, semi-glossy mid-green, occasionally variegated with a single grey-green central stripe, or scattered grey green throughout, or (rarely) entirely grey, measuring 10-29 cm long by 3.