Research on medicinal plant resources toproposesolutions forconservation and sustainable useof some valuable species in na hang nature reserve, Tuyen Quang province

MINISTRY OF EDUCATION AND TRAINING VIETNAM ACADEMYOF SCIENCE AND TECHNOLOGY GRADUATE UNIVERSITYOF SCIENCE AND TECHNOLOGY NGUYEN THI HAI NGUYỄNTHỊ HẢI RESEARCH ON MEDICINAL PLANT RESOURCES TOPROPOSESOLUTIONS FORCONSERVATION AND SUSTAINABLE USEOF SOME VALUABLE SPECIES IN NA HANG NATURE RESERVE, TUYEN QUANG PROVINCE Major: Botany Code: 9.42.01.11 SUMMARY OF DOCTORAL THESIS IN BIOLOGY HaNoi – 2018 The dissertationwas completed at the Graduate University of Science and Technology- Vietnam Academy ofScience and Technology Supervisors: 1. PGS. TS. Trần Huy Thái 1. Assoc. Prof. Dr. Tran Huy Thai 2. Assoc. Prof. Dr. Nguyen Tien Dat 2. PGS. TS. Nguyễn Tiến Đạt Reviewer1: Reviewer2: Reviewer3: The dissertation will bedefended atthe Council ofPhD. Thesis Assessment held at the Graduate University of Science and Technologyat ... , on.. 2018 Thesis can be further referredat National Library of Vietnam, Library of Graduate University of Science and Technology 1 INTRODUCTION 1. The necessity of the dissertation Na Hang Nature Reserve was established under Decision No. 274/UB-QD dated 9th May 1994 by the People's Committee of Tuyen Quang province. About 68% of the area is tropical moist forests. Of these, about 70% are limestone forest, and the rest are lowland evergreen forest. In 2006, Nguyen Nghia Thin and his colleagues identified and listed 1,162 species of plants in the Na Hang Nature Reserve, many of them were listed in the Vietnam Red Data Book (2007) such as Garcinia fragraeoides, Excentrodendron tonkinense, Chukrasia tabularis, Markhamia stipulata etc. Beside the research resultsof some authors at the Forest Inventory and Planning Institute, Birdlife International program; Nguyen Nghia Thin and Dang Quyet Chien (2006); Nguyen Anh Tuan et al. (2012); so far, there is no comprehensive research on medicinal plant resources, especially the promising species in Na Hang Nature Reserve. On the other hand, medicinal plant resources in Na Hang Nature Reserve are becoming increasingly scarce, some of which are endangered due toindiscriminate and planless exploitation and. The potential and prospects of medicinal plant resources and indigenous knowledge of indigenous peoples in Na Hang Nature Reserve are plentiful and varied; But these studies are few and far from complete. Research on the scientific basis for conserving, restoring and sustainably using the diversity of medicinal plants in Na Hang Nature Reserve, Tuyen Quang province, is an urgent, critical issue. scientific significance and high economic and social value. Based on this situation, the author conducted the study "Research on medicinal plant resources to propose solutions for conservation and sustainable use of some valuable species in Na Hang Nature Reserve, Tuyen Quang province". 2. The goal of the dissertation To study and evaluate the current status of medicinal plant resources in Na Hang Nature Reserve, Tuyen Quang provinceto propose solutions for conservation and sustainable development of some valuable species. 3. The scientific and practical significance of the dissertation * Scientific significance: The results of the research will contribute to supplement the data on the current distribution of medicinal plants and propose measures to preserve biodiversity of plant resources. * Practical significance: The result of the research is the scientific basis for the agencies in management, production and business to make policies for development, investment in production, creating a stable source of pharmaceutical raw materials and sustainable use; at the same time, effective conservation of species of high potential and value; Research results on screening biologically active species will contribute to the development of new dmedicineproduction. 4. The composition of the dissertation The dissertation consists of 146 pages: Introduction - 02 pages; Chapter 1: Overview of research issues - 32 pages; Chapter 2: Subjects, contents and methods of research - 11 pages; Chapter 3: Research Results and Discussion - 86 pages; Conclusions and recommendations - 02 pages; References - 11 pages 2 CHAPTER 1 OVERVIEW OF RESEARCH ISSUES 1.1. Resource of medicinal plants in the world 1.1.1. History of research on medicinal plant resources in the world Medicinal herbs were developed as a Chinese cultural tradition from about 5000 years ago (2,737 BC - 2,697 BC). The earliest known Sumarian genealogical record-writing knowledge was written in hieroglyphics in 2000 BC "Materia Medica" recorded details of the effects of 250 medicinal plants. India is also a country with a long tradition of using medicinal herbs. The earliest recorded record of the use of medicinal plants was found in the book of Rig - Vedas around 4500 - 1600 BC, which is considered to be the oldest book on the use of medicinal plants in human history. Today, more than 8.000 plant species are known to be used in India. Not only in Asia, using medicinal herbs is appeared in European countries long time ago. In Africa, the earliest written records on the use of medicinal plants were recorded by ancient Egyptians over 3.600 years ago, with about 800 remedies and over 700 medicinal plants, including Aloe vera, Cannabisetc. 1.1.2. Assessment of the value and economic value of medicinal plant resources in the world 1.1.2.1. Assessment of the use value of medicinal plant resources Worldwide, it is estimated that up to 70.000 species of plants are used in folklore. WHO reports that more than 21.000 plant species have been used for health care. India used about 7.500 species. By 1997, China used more than 6.000 species. In Africa, more than 5.000 plant species are used for medical purposes. In Europe, with a long tradition in the use of plants, about 2.000 medicinal herbs and aromatic spices are used in commerce ... 1.1.2.2. Assessment of the economic value of medicinal plant resources On a global scale, sales of medicinal plants are estimated to be around 16 billion euros a year. In the 1990s, the annual import turnover of medicinal plants averaged over 4.000 tons, valued at $ 1.224 million. Of this figure, 80% of import and export value is from 12 countries in Asia and Europe. Japan and South Korea are the two countries that consume the most medicinal plants. China and India are the leading providers of medicinal plants; Hong Kong and the US are important trade centers. Many medicinal plants are not only exploited for local use but also for exporting throughout the world. A large number of medicinal plants in Asia and Africa, in addition to being harvested for domestic use, are also used for export. Up to 80% of medicinal plants are exported from Asian countries. Demand for medicinal plants increases by 15 - 25% annually and according to the World Health Organization (WHO), it is estimated that by 2050 the demand for medicinal plants will increase by more than $ 5 trillion. 3 1.1.3. Research on the experience of using medicinal plants of the peoples of the world Survey and study of medicinal plants together with traditional knowledge on medicinal plants, indigenous experience in the use of medicinal plants associated with biodiversity conservation, preservation of cultural valueswhich are attracting attention in most countries around the world. 1.2. Overview of medicinal plant resources in Vietnam 1.2.1. History of research on medicinal plant resources in Vietnam Vietnam has many researches such as: Hai Thuong Lan Ong - Le Huu Trac (1720 - 1791), Crévost and Pétel (1928 - 1935), Do Tat Loi (1957), Vu Van Chuyen (1966), Nguyen Nghia Thin, La Dinh Moi et al (2005) 1.2.2. Overview of use value and economic value of medicinal plant resources in Vietnam According to the statistics of the health sector, each yearwe consume from 30.000 - 50.000 tons of different medicinal herbs. In Vietnam in recent years, each year has exported from 5.000 to nearly 10.000 tons of pharmaceuticals, valued at about $15 million ... In addition, export some semi-active ingredients as active ingredients such as Berberin, palmatin, rotundin, rutin, ... Some businesses have exported drug substance such as Artemisinin, artesunate, ... and many other Oriental drugs. 1.2.3. Study on the experience of using medicinal plants of different ethnic groups in Vietnam In Vietnam, there are researches on medicinal plants that specialize the area such as: Peoples of Tay, Dao, H'Mong in Ba Be National Park (Bac Kan); San Diu National Park in Tam Dao National Park (Vinh Phuc); Dao in Ba Vi (Ha Tay); Muong in Cuc Phuong; Tay and Dao in Yen Ninh and Yen Do communes, Phu Luong district (Thai Nguyen); Cao Lan in Doi Can, Yen Son district (Tuyen Quang province); Muong (Thanh Hoa); Thai (Nghe An), ... 1.2.4. Studies on the flora and medicinal plants in Na Hang The studies on botanyin Na Hang are not abundant. In addition to the publication of Mike Hill & Nevill Kemp (1996), Nguyen Nghia Thin, Dang Quyet Chien (2006); Nguyen Anh Tuan and Tran Huy Thai (2012); Phan Ke Loc, Pham Van The, L.V. Averyanov et al. (2013) almost no analysis on the systematic research on medicinal plants. 1.3. The issues of conservation and research on propagation of medicinal plant resources Biodiversity conservation, especially of plant species, is a global issue, not just for nations but for international organizations (IUCN, WWF, FAO, WHO, etc.). According to Akerele (1991), the problem of preserving medicinal plants in countries is the recognition and preservation of the value of using them in the national medicine. Referring to the preservation of threatened medicinal plants, Hamann (1991) also argues that there is no other way but to understand the distribution of the medicinal plants to establish internal conservation areas (in - situ) conservation and external conservation or (ex - situ) 4 conservation. Biodiversity conservation in general and conservation of medicinal plants in particular, associated with the conservation of indigenous knowledge in Vietnam have been receiving special attention. * Propagation by cuttings Cutting is a methodof asexualpropagation which has a large multiplication coefficient, which maintains good character of the mother plant and is relatively inexpensive. It is widely used in breeding of plants, ornamental trees and fruit trees. * Seed propagation Seed propagation is a traditional and natural breeding method, which gives high propagation and can be stored and transported easily. The decisive factor for seed propagation efficiency is the quality of seed. 1.4. Medicinal plants and anti-cancer activity from medicinal plants The therapeutic effects of medicinal plants are fromnatural compounds that were synthesized and accumulated. Currently, around 3.000 plant species in the world are reported to contain compounds that have anti-cancer activity. The American Cancer Research Institute collected 35.000 plant samples from 20 different countries and screened about 114.000 extracts with anti-cancer activity. In Vietnam, in recent years, research on medicinal plants which are capable of treating cancer has also been considered. Numerous studies on antitumor activity have been published; however, studies on two species of Mallotus microcarpus and Miliusa sinensis are very few. 1.5. Natural conditions, socio - economic conditions of Na Hang Nature Reserve Geographic coordinates: From 22 0 14' - 22 0 35' north latitude; East longitude 104 0 17' - 105 0 35. The total area of this special use forest is 22,401.5 hectares and is located in four communes of Thanh Tuong, Son Phu, Khau Tinh, Con Lon and Na Hang town. The climate is in the tropical monsoon region of Northeast Vietnam and is characterized by the high mountain climate. The year is divided into two distinct seasons, with the following characteristics: Summer usually starts from April to September, hot and humid climate, heavy rain; winter is cold and dry, lasts from October to March next year; average annual temperature is 23.5°C; Average rainfall: 1.400 – 1.600 mm. There are two large river systems: Nang River and Gam River. There are 52 villages in the 4 communes and Na Hang town, 3,916 households/10,081 households (accounting for 38.84% of the total households in the district). There are four main ethnic groups: Tay, Kinh, Dao and H'mong. The remaining ethnic groups such as Cao Lan, Han ... Agricultural production is mainly one-crop rice production, average grain is low. The area also develops grass-feeding animals and make use of local sources to feed breeding pigs, chicken, ducks. Forestry production is mainly managing the protection of existing 5 forest resources. There is a district general hospital in Na Hang Town; 5 commune health stations; there are 46 villages/52 villages having village health workers. There are 5 kindergartens with 26 school sites; 5 primary schools with 27 school sites; 5 junior high schools, 1 high school. Na Hang town is a political, economic and cultural center of Na Hang district, which has been invested with many projects for economic, cultural and social development of the locality. CHAPTER 2 SUBJECTS, CONTENTS AND METHODOLOGY OF RESEARCH 2.1. Subjects, location and time of research 2.1.1. Research subjects Medicinal plants in Na Hang Nature Reserve and its vicinity. 2.1.2. Location and time of research Investigation, study and survey in Na Hang Nature Reserve, mainly in Thanh Tuong, Son Phu and Khau Tinh communes, Na Hang district, Tuyen Quang province. Specimens were researched, analyzed and stored in the Plant Propagation Chamber of the Institute of Ecology and Biological Resources - Vietnam Academy of Science and Technology.Empirical studies on chemistry and biological activity were conducted in the laboratory of bioactive compounds, Institute of Marine Chemistry - Vietnam Academy of Science and Technology. Time of research: From 2014 to 2016. 2.2. Contents of research - Research on medicinal plant resources in Na Hang Nature Reserve, Na Hang district, Tuyen Quang province: Diversity of medicinal plant resources; Diversity of medicinal plants used by Tay and Dao ethnic communities in Na Hang Nature Reserve. - Biological Activity Test and Chemical Structure Analysis: Screening biological activity; Research on chemical composition and biological activity of sample TQ02 (Mallotus microcarpus Pax & K. Hoffm) and biological activity of sample TQ13 (Miliusa sinensis Fin. & Gagnep.). - Solutions for conservation, development and sustainable use of valuable species at Na Hang Nature Reserve, Na Hang District, Tuyen Quang Province: Medicinal plants with potential for development; Propagation potentials of two medicinal plants with potential of exploitation and use; Proposing solutions for preservation and development of some medicinal plants in Na Hang Nature Reserve. 2.3. Research Methodology 2.3.1. Inheritance method Inheriting documents related to the field of study: List of plant species in Vietnam; Dictionary of Vietnamese medicinal plants; Medicinal plants and medicinal animals in Vietnam; Plant diversity of Na Hang Nature Reserve, Tuyen Quang province 6 2.3.2. Method of collection, processing and identification of samples - Method of collection and processing of samples: According to Nguyen Nghia Thin (1997, 2007). - Identification of scientific names: Using the morphologiccomparison method, based on: Handbook for searching and recognizing angiosperm families in Vietnam; Flora of Vietnam; Flora in Vietnam; List of plants of Vietnam, Volume I, II, III; Dictionary of medicinal plants in Vietnam... 2.3.3. Method of assessing the diversity of medicinal plant resources To assess the diversity of medicinal plant resources of Na Hang Nature Reserve, based on Nguyen Nghia Thin (1997) assessment method, such as: Diversity of taxon levels (divisions, classes, families, species), at the same time identifying the diverse indicators, as well as their life forms. Disease groups are classified according to Le Tran Duc (1995) "Vietnamese medicinal plants for the processing and initial treatment" and Decision No. 3465/QD-BYT dated July 08, 2016... 2.3.4. Evaluation method for preserving gene source of medicinal plants The status of species is evaluated according to the criteria of Decree 32/2006/ND-CP, Red List of medicinal plants (2006), Red Book of Vietnam (2007), IUCN Red List (2014)... 2.3.5. Method of mapping the distribution of rare and precious plant species The geographic map of Na Hang Nature Reserve is based on a 1:50.000 topographical map; National coordinates VN2000. The data is updated to 2015 from satellite imagery including information layers such as topography (road level), traffic, residential areas, hydrology, administrative boundaries... 2.3.6. Method of researching plant medicine of the minority groups In the process of community research, an ethnobotanical survey was used, with tools such as the Rapid Rural Appraisal (RRA) and the Participatory Rural Appraisal (PRA). PRA methodology is conducted in two ways: using some questions for selected group of people or researchers and interviewers. For the survey of medicinal plants, medicinal products used by two Tay and Dao ethnic groups in Na Hang Nature Reserve: Survey sites of three typical communes: Thanh Tuong, Son Phu and Khau Tinh were selected. In each commune, 15 people of Tay and Dao ethnic groups were selected to conduct interviews, including: the elderly, middle-aged and young, both men and women; Tay ethnic group: interviewed over 30 people, Dao ethnic group: Interviewed over 15 people. Each responded to 20 votes. Investigating the exploitation, yield, market price, harvesting time ... ofmedicinal plants used for medicine:It was conducted with 30 people: those who go to forest for exploiting and gathering medicinal plants; local traders or local marketers (local traders); local physicians who use medicinal plants to make medicines (members of the Traditional Medicine Association of Na Hang district)... in the study area. 7 2.3.7. Propagation method of some main medicinal plants 2.3.7.1. Asexual propagation method Vegetative propagation is done by cutting different types from tubers. Vegetative propagation (Tacca chantrieri Andre) and branches (Fibraurea tinctoria Lour.) collected from naturally grown trees. Experiments include: a. Effects of season and cuttings on survival capacity and rooting from cuttings: seasons are defined as Spring, Summer, Fall, Winter; Cuttings include: top cuttings, trunk cuttings and cuttings from tubers and branches. Each test formula was 30 cuttings; Repeat 3 times in 3 different gardens. Data were recorded once in 60 days with both species. b. Effects of growth regulators and their concentrations on the survival and rooting rate of cuttings: Growth regulators used in the experiments were: α-NAA (α-napthilene acetic acid), IBA (Indol butyric acid), IAA (Indole Acetic Acid); with 3 types of concentration: 1.000ppm; 1.500ppm and 2.000ppm, the control plot not using growth regulator. The experiment was arranged in 10 formulas. 2.3.7.2. Method of sexual propagation Tacca chantrieri Andre: Seeds are sown on moist sand, avoid direct sunlight; Simultaneously with the experiments on petri dishes under laboratory conditions. Each experiment sowed 100 seeds. The experiments were repeated three times, monitoring seed germination time. Total number of seeds in the experiment: 100 seeds/formula x 2 formul