VIETNAM NATIONAL UNIVERSITY OF AGRICULTURE FACULTY OF BIOTECHNOLOGY -------------***------------- GRADUATION THESIS “COMPLETE THE MULTIPLICATION PROCESS OF PAULOWNIA FORTUNEI IN VITRO” HANOI, 3/2021 VIETNAM NATIONAL UNIVERSITY OF AGRICULTURE FACULTY OF BIOTECHNOLOGY -------------***------------- GRADUATION THESIS “COMPLETE THE MULTIPLICATION PROCESS OF PAULOWNIA FORTUNEI IN VITRO” Student’s name : Vu Duc Canh Class : K61CNSHE Faculty : Biotechnology Supervisor : Dr. Nguyen Thi Lam Hai HANOI, 03/2021 STATEMENT OF ORIGINAL AUTHORSHIP The work contained in this thesis has not been previously submitted to meet requirements for an award at this or any other education institution. To the best of my knowledge and belief, the thesis contains no material previously published or written by another person except where due reference is made. Hanoi, February 23rd, 2021 Student Vu Duc Canh i ACKNOWLEDGEMENTS To complete this graduation thesis, I would like to thank Nguyen Thi Lam Hai.Dr, and the teachers in the Department of Plant – Faculty of Biotechnology, who have always been helpful and guided me throughout the process of implementing the topic.
I would like to express my sincere thanks to all of the students working in the Department of Plant – Faculty of Biotechnology for creating the best conditions to help me during the time of the thesis. I would like to express my sincere thanks to the teachers of the Department of Biotechnology, Vietnam National University of Agriculture for imparting new knowledge to me, and having a passion for scientific research during my time studying at the university. Finally, I sincerely extend my gratitude to my family and friends who have always supported and encouraged me during my studies and scientific research. Hanoi, February 23rd, 2021 Student Vu Duc Canh ii CONTENTS STATEMENT OF ORIGINAL AUTHORSHIP.
ii Content of table. v Content of figure. vi LIST OF ABBREVIATIONS. The origin of the research object.
Botanical characteristics of Paulownia fortune. Advantages and application. Requires external scene. Reacher Paulownia fortunei trees in the world and in Vietnam.
Research overview in the world. Overview of domestic research. 12 PART 3: MATERIALS AND RESEARCH METHODS. Object, material, location, time of culture.
Location, time of culture. Content and reseach methods. 18 PART 4: CONCLUSION AND RECOMMENDATION. Effects of BA and Kinetin on shoot multiplication.
Effects of BA and IAA on shoot multiplication. Effects of BA and IBA on shoot multiplication. Effects of coconut milk on shoot multiplication. Effects of iron content in MS on shoot photosynthesis, thereby affecting the shoot multiplication.
Effects of Vitamin content in MS on shoot metabolism and shoot multiplication. Effects of Vitamin B2 on growth and development of cultured shoots. Effect of the MS concentration to the rooting (non-sterile - free sugar and liquid medium). Effect of α-NAA on rooting (non-sterile and liquid medium).
37 iv CONTENT OF TABLE Table 1: The statistic result of effects of BA and Kinetin on shoot multiplication. 21 Table 2: The statistic result of effects of BA and IAA on shoot multiplication. 23 Table 3: The statistic result of effects of BA and IBA on shoot multiplication. 25 Table 4: The statistic result of Effects of coconut milk on shoot multiplication.
27 Table 5: The statistic result of effects of iron content in MS on shoot photosynthesis, thereby affecting the shoot multiplication. 29 Table 6: The statistic result of effects of Vitamin content in MS on shoot metabolism and shoot multiplication. 31 Table 7: The statistic result of effects of Vitamin B2 on growth and development of cultured shoots. 33 Table 8: The statistic result of effect of the MS concentration to the rooting (non-sterile - free sugar and liquid medium).
35 v Content of figure Figure 1. Paulownia fortune shoot. Paulownia fortune leaves. The inflorescence of Paulownia fortune.
Fruit of Paulownia fortune. Root system of Paulownia fortune. 7 Figure 6: Effects of BA and Kinetin on shoot multiplication (after 4 weeks). 23 Figure 7: Effects of BA and IAA on shoot multiplication (after 4 weeks).
24 Figure 8: Effects of BA and IBA on shoot multiplication (after 4 weeks). 26 Figure 9: Effects of coconut milk on shoot multiplication (after 4 weeks). 28 Figure 10: Effects of iron content in MS on shoot photosynthesis, thereby affecting the shoot multiplication. 30 Figure 11: Effects of Vitamin content in MS on shoot metabolism and shoot multiplication.
32 Figure 12: Effects of Vitamin B2 on growth and development of cultured shoots. 34 Figure 13: Effect of the MS concentration to the rooting (non-sterile - free sugar and liquid medium). 36 vi LIST OF ABBREVIATIONS Abbreviations Annotate MS Murashige and Skoog medium CT Formula CV Coefficient of variation LSD5% Least significant differences BA Benzylidenes IAA Indole-3-acetic acid IBA Indole-3-butyric acid NAA 1-naphthaleneacetic acid vii ABSTRACT Paulownia fortunei is a tree that has a lot of economic value and has high growth rate, good wood quality, and less pest and disease. The project was conducted to find the best formula for multiplication in vitro paulownia fortunei and the best rooting recipe with low economic cost.
With the use of disease-free samples and in vitro culture, transfer to different culture media obtained the following results: the highest shoot multiplier was obtained on MS medium supplemented with 0.31 cm / plant and 2.14 shoots /plant) tall, fat, strong shoots. With the MS formula supplemented with 0.5 mg / L BAP and coconut milk, the multiplier was low, however green plants, and healthier and fresher roots were suitable for complete tree formation. With the formula of iron supplementation by changing the amount of iron in the MS ingredient by 1.5 times, the multiplier ratio of 2.53 shoots / plant and the average height of 1.98 cm / plant is obtained, the plants are greener and healthier. With the formula of changing the amount of Vitamins by increasing their content in MS and Vitamin B2 supplementing formula, the shoot multiplication efficiency is not increased, the multiplier factor is not high.
culture plants from culture different medium to non-sterile liquid root medium, medium containing 1/3 of the non-sterile liquid MS is best. The best substrate for planting plants in the garden is peat moss. Preface Paulownia fortunei is one of 9 species of tree species in the genus Paulownia Paulownia fortunei commonly called the dragontree, dragon tree or Fortune's empress tree, is a deciduous tree in the family Paulowniaceae, native to southeastern China (including Taiwan), Laos and Vietnam. It is an extremely fast-growing tree, due to its use of C4 carbon fixation, and is planted for timber harvesting.
Paulownia fortunei grow rapidly, after 6 years, they can be harvested. The wood is very light and hard, resistant to termites, does not deform when the weather changes, so it is often used as interior decoration, interior parts of aircraft, yachts, handicrafts, and musical instrument. The easy-to-dissolve Paulownia fortunei leaves have good soil improvement effects. In addition, the Paulownia fortunei leaves have a high protein content to make animal feed effectively.
The flowers of this species are beautiful, fragrant, and are used to grow honeybees, or to prepare some medicines such as asthma (Pham Hoang Ho). In addition, Paulownia fortunei root is distributed 30-40cm deep below the ground, so it is less competitive with short-term plants, and it also holds good soil and high economic value, so it is very suitable for afforestation (Nguyen Nhan Hoai et al, 1999). Due to many preeminent properties, Paulownia fortunei has been paid attention to research and development in many countries.8 million hectares of species of the genus Paulownia have been planted in China. Some other countries such as the US, Canada, and Australia also pay attention to research and develop this plant.
In Vietnam, Paulownia fortunei tree has been planted in recent years. Trees grow quite quickly in the first few years and show a wide development potential for the purpose of planting raw material forests, creating environmental landscape. Paulownia fortunei is also a perennial tree, so it is also used for planting protective forests. In the nature, it is often intercropped with some pioneer species, it is easy to intercrop with some other plants such as bamboo, intercropped to shade agricultural crops or coffee, tea etc.
In vitro propagation of Paulownia fortunei have been and is being researched by 1 scientists in many places. In Vietnam, there are also in vitro propagation projects of this plant. However, these works are heavy on basic research and the multiplication coefficient is not high. Stemming from the fact, with a density of planting about 800 trees / ha, with an area of up to hundreds of thousands of hectares, it is very important to take initiative in the source of Paulownia fortunei seedlings with the need of millions of trees at the same time.
Therefore, in order to meet the requirements of the production of high-quality Paulownia fortunei in vitro with good multiplier, I conducted a research project: "Complete the multiplication process of Paulownia fortunei in vitro". Objectives The project aims to research and build a process to improve the Paulownia fortunei variety using in vitro culture technology as the basis and easy to apply in the production of the Paulownia fortunei variety in large quantities, uniformity, good growth, suitable for the afforestation in Vietnam. Requirements - Study on the effects of growth regulators on shoot multiplier. - Study the effect of organic compounds on shoot multiplier.
- Effects of Vitamins, iron on growth and development in Paulownia fortunei in vitro culture. - Optimized in vitro root formation of cultured (non-sterile) Paulownia fortunei shoots. - Determine the suitable substrate to crown Paulownia fortunei in vitro. The origin of the research object The Scrophulariaceae Paulownia is native to Asia.
In China, it has been known for over 2,000 years, while Japan and north Korea have appeared about 1000 years ago. In the 19th century, the Paulownia was found in some countries in Europe, South America and Australia. In New Zealand, China Paulownia has been very successful in using for reforestation. China's Paulownia acreage so far has grown to more than 5 million hectares.
In Vietnam, Paulownia fortunei is one of 9 tree species of the genus Paulownia which was from the 2000s. Distribution Due to many good properties, Paulownia fortunei has been noticed in research and development in many countries. Paulownia fortunei is distributed naturally in many Asian countries such as China, Laos, Cambodia,. In China, there are about 1.8 hectares of species belonging to the genus Paulownia, some other countries such as the US, Australia also pay attention to research.
development of this plant. In Vietnam, Paulownia fortunei is grown in some provinces in the North, Central Highlands, some southwestern provinces with great potential for planting this plant such as An Giang, Hau Giang, Tra Vinh, Soc Trang. Scientific classification Kingdom: Plantae Tracheophytes Angiosperms Eudicots Asterids Order: Lamiales 3 Family: Paulowniaceae Genus: Paulownia Species: P. fortunei Binomial name: Paulownia fortune (Seem.
Synonyms: Basionym Campsis fortunei Seem. Heterotypic Paulownia duclouxii Dode, Bull. Paulownia longifolia Hand. Paulownia meridionalis Dode, Bull.
Some of the species recognized by Plants of the World Online and The Plant List: • Paulownia catalpifolia • Paulownia elongata • Paulownia fargesii • Paulownia fortunei (synonyms: P. mikado) • Paulownia kawakamii (synonyms: P. viscosa) • Paulownia tomentosa (synonyms: P. Botanical characteristics of Paulownia fortune Shoot Figure 1.
Paulownia fortune shoot Bark: Thin-plate, light gray, smooth, with slight cracks in adult trees.The diameter of the trunk: the circumference of a 1.5-2-year-old tree is 8-14 cm, the circumference of a 3-4 year-old tree is 20-24 cm, and the circumference of an adult 18-year-old tree is up to 80 cm. Wood of Paulownia accumulates the substance of tannin, which makes it resistant to eating by termites and wood fretters. Paulownia feels good in urban conditions. Paulownia fortune leaves Features of the plant: in the first year of life, the tree has large fibrous leaves, which can reach 50-85 cm in diameter.