Summary of doctoral thesis in biology: Research on medicinal plant resources to propose solutions for conservation and sustainable use of some valuable species in Na Hang Nature Reserve,

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

MINISTRY OF EDUCATION VIETNAM ACADEMYOF SCIENCE AND AND TRAINING 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 TechnologyVietnam Academy ofScience and Technology Supervisors: PGS TS Trần Huy Thái Assoc Prof Dr Tran Huy Thai Assoc Prof Dr Nguyen Tien Dat 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 INTRODUCTION 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" 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 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 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 CHAPTER 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 $ trillion 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) 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 22014' - 22035' north latitude; East longitude 104017' - 105035 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 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 forest resources There is a district general hospital in Na Hang Town; commune health stations; there are 46 villages/52 villages having village health workers There are kindergartens with 26 school sites; primary schools with 27 school sites; junior high schools, 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 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… 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 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 times in 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 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 formula x replicates = 600 seeds Fibraurea tinctoria Lour.: Seeds are sown by methods of seed treatment (3 formulas) as follows: + Formula 1: Sowing seeds in moist sand + Formula 2: Soaking the seeds in warm water at 400C for 10 hours, then sowing in moist sand + Formula 3: Soaking the seeds in water for 10 hours and then sowing in moist sand Each formula has replicates, each repeats with 30 seeds Total seeds in experiment: 30 seeds/formula x formula x replicates = 270 seeds Care and maintenance: Daily watering, regular monitoring and careful protection 2.3.7.3 Method of data collection - Experiment on vegetative propagation: Supervising and recording: Number of live cuttings, number of cuttings with germ; number of cuttings producing roots - number of roots, average length of root, same effect of cuttings on survival rate, ability to germinate and produce root - Experiment on sexual propagation: Monitoring indicators (number of days seeds start to germinate, number of seeds germinated completely, number of seeds germinated…, number of days for seedlings to standard transplanted)… - Monitoring the emergence of young seedlings during the nursery stage: Indicators of measurement include: Statistics of live trees on the total number of trees arranged in each iteration Measuring the root diameter (D00) by panme ruler with a precision rate to 1/10mm, measuring the height by the meter carved to mm (topheight - Hvn) 2.3.8 Testing the in vitro biological activity (cytotoxic and α-glucosidase inhibitory activity, α-amylase) of some common and potential species 2.3.8.1 Creating a crude extract: Methanol extraction solvent The plant samples were soaked with solvent three times in ultrasonic tanks at 40°C for 30 minutes The extracts were then collected and stored in high yield crude extracts 2.3.8.2 Biological Activity Testing Method Evaluation of active cancer cell resistance: Cancer cell lines include: Human lung cancer (A-549, H1975), breast cancer (MCF-7, MDA-MB-231), pancreatic cancer PANC1), prostate cancer (DU145) is provided by GS Jeong-Hyung Lee, Kangwon National University, Korea Cancer cells are cultured in vitro by the method of Mosmann et al Evaluation of α-amylase inhibitory activity: α-amylase inhibitory activity was determined by the coloring reaction of starch Azure (blue starch) with water Evaluation of α-glucosidase inhibitory activity: α-glucosidase inhibitory activity is based on the hydrolysis of 4-nitrophenyl-α-D-glucopyranoside (pNPG) to glucose and p-nitrophenol, the compound is yellow, under the catalytic enzyme α-glucosidase 2.3.8.3 Method of extraction and isolation of compounds - Isolation of purified substances by chromatographic methods: + Thin-layer chromatography was performed on DC-Alufolien 60 F254 silica gel and RP-18 F254 coated plates Detect the substance line by UV light 254 and 368 nm, and spray 10% H2SO4 on the plate and heat it slowly until the color appears + Common phase column chromatography (silica gel 230-400 mesh, Merck), reversed phase (RP-18, YMC ODS), ion exchange resin (Dianion HP20) or particle size adsorbate (Sephadex LH20) - Determine the structure of the molecules by modern spectroscopic methods including infrared absorption spectrophotometry (IR), ultraviolet (UV) absorption spectrometry, one-dimensional and two-dimensional magnetic resonance spectra (1D and 2D-NMR), mass spectrometry (MS) and high resolution (HRMS) 2.3.8.4 Method of determining the structure of compounds The general method for determining the chemical structure of compounds is the combination of the determination of physical parameters with modern spectroscopic methods 2.3.9 Data analysis All data processed on Excel, expressed as mean ± SE Student's t-test, F'test and one-way analysis of variance (one way ANOVA) were used to examine significant differences from the negative control, with P 50% at 100 μg/ml) including (TQ03, TQ04, TQ08, TQ09, TQ10, TQ13) For α-amylase, four samples exhibited an inhibitory effect of> 50% at 500 μg/ml including (TQ05, TQ15, TQ17, TQ18) Among these samples, sample TQ13 from (Miliusa sinensis Pax & K Hoffm), showed remarkable α-glucosidase inhibitory activity, and there’s not muchstudy for inhibitory activity of α-glucosidase So, this sample was selected for further study of the chemical composition 3.1.3.2 Study on chemical composition and biological activity of TQ02 from (Mallotus microcarpus Pax & K Hoffm) * Extract and isolation of compounds From the sample of (Mallotus microcarpus Pax & K Hoffm), the 11 compounds have been extracted and identified from MM1 to MM11, one new compound was first announced: 3-Methoxybenzensulfonic acid 4-O-β-D-glucopyranoside (MM4) Guaiacylglycerol1-(4-Hydroxy-3methoxyphenyl)-1,2,3-propanetriol (MM1) Zansimuloside A (MM3) Antiarol rutinoside (MM2) 3-Methoxybenzensulfonic acid 4-O-βD-glucopyranoside (MM4) Erigeside (MM5) 3,4’,-Dihydroxypropiophenone 3-Oglucoside (MM6) Methyl salicylate 2-rutinoside (MM7) Leonuriside A (MM8) 15 Coniferin (MM10) Syringin (MM9) Hovetrichoside A (MM11)  Evaluation of the cytotoxic effects of the clean substances Thus, clean substances isolated from TQ02 (Mallotus microcarpus Pax & K Hoffm) were evaluated for cytotoxic effects on H1975, A549 lung cancer lines and MCF-7 breast cancer Results from show that the new 3-Methoxybenzensulfonicacid 4-O-β-Dglucopyranoside (MM4) and 2-rutinoside methyl salicylate (MM7) compounds work on all three test cell lines Meanwhile, leonuriside A (MM8) has a very powerful effect on MCF-7 breast cancer cells 3.1.3.3 Study on the chemical composition and biological activity of TQ13 from (Miliusa sinensis Fin & Gagnep.) * Extract and isolation of compounds Specimens (Miliusa sinensis Fin & Gagnep.), were extracted and identified three compounds: MSW2.1, MSW2.2 and MSW5.1 MSW2.1 MSWW2.1 (Kaempferol 3-O-(2,6-diO-α-L-rhamnopyranosyl)-β-Dgalactopyranoside) MSW2.2 (Quercetin 3-O-(2,6-di-O-α-Lrhamnopyranosyl)-β-D-galactopyranoside) 16 MSW5.1 (Daucosteol) * Evaluation of the α-glucosidase inhibitor of the isolates Of the three compounds isolated from the TQ13 form (Miliusa sinensis Fin & Gagnep.), Daucosterol is a well-known compound that has been studied extensively and should not be selected for activity testing Both MSW2.1 and MSW2.2 have α-glucosidase inhibitory effects with an IC50 value of 557.3 and 692.5 μM This effect is compared with the activity of the antidiabetic drug acarbose (IC50 671.2 μM) Thus, these two flavonoid glycosides can be identified from TQ13 (Miliusa sinensis Fin & Gagnep.), an active substance that inhibits α-glucosidase associated with diabetes 3.2 Solutions for conservation, development and sustainable use of valuable medicinal plants in Na Hang Nature Reserve, Na Hang District, Tuyen Quang Province 3.2.1 Medicinal plants having potential for development 3.2.1.1 Criteria for selection of medicinal plants with potential for development Potential species are those that meet the following criteria: i/ Species with consuming markets; ii/ High economic value; iii/ There is a need to use in large and regular communities; iv/ Suitable for planting to collect materials 3.2.1.2 Proposing some potential medicinal plants to be researched in Na Hang Nature Reserve  Situation of exploiting and using medicinal herbs in Na Hang Nature Reserve Through research on medicinal plant resources combined with knowledge of harvesting and using medicinal plants of ethnic communities in Na Hang Nature Reserve, there were initially about 20 kinds of medicinal plants being exploited and having market Species are high and widely distributed in the area such as: Homalomena occulta, Asarum balansae, Tacca chantrieri, Curcuma longa, Curcuma zedoaria, Saururus chinensis, Gomphandra mollis and Tinospora sinensis At the same time, yearspecies are extirpated: Stephania rotunda Lour, Disporopsis longifolia Craib, Fibraurea tinctoria Lour and Ardisia silvestris Pitard, Fallopia multiflora (Thunb.) These species have been exploited in the past few years for sale to China At present, the Yellow Fibraurea tinctoria Lour, Ardisia silvestris Pitard; Disporopsis longifolia Craib.; Rauvolfia verticillata (Lour.) Baill.; Heterosmilax gaudichaudiana (Kunth) Maxim.), Stemona tuberola Lour are the most consumed medications Market and potential development of medicinal plants in Na Hang According to the results of the survey, the prices of some medicinal herbs such as Fallopia multiflora (Thunb.) Haraldson, Disporopsis longifolia Craib., Stephania rotunda Lour., Fibraurea tinctoria Lour., Ardisia silvestris Pitard are quite high So, these are 17 species that have been exploited indiscriminately while the conservation is not paid adequate attention With the above mentioned species, if exploited properly, along with the planting and protection, this will be an advantage to develop the economy and improve the lives of ethnic minorities in the Na Hang NR Species such as Curcuma longa L., Curcuma zedoaria (Berg.) Rosc., Fibraurea tinctoria Lour, Gynostemma pentaphyllum (Thunb.) Makino in the locality are in pretty big quantity, the people have planted them in the gardens, the trees grows well  Characteristics of medicinal plants with potential for development Based on the results above, 12 potential medicinal plants in the study area (Table 3.20) are to be tested for propagation, planting and development Table 3.20: Potential medicinal plants for development in Na Hang Nature Reserve No Vietnamese language Thiên niên kiện Tế tân nam Biến hóa Bình vơi Ethnic language Scientific name Flora family Vát vẹo (Tày); Sì nhàn chấn (Dao) Muầu đin (Tày, Dao) Mầu đin (Tày, Dao) Cà tom khao (Tày); Hùng mầu pẹ (Dao) Dây đau xương Khau bết phạ (Tày), Tiền mạy hoày (Dao) Homalomena occulta (Lour.) Schott Asarum balansae Franch Asarum caudigerum Hance Stephania rotunda Lour Araceae Tinospora sinensis (Lour.) Merr Menispermaceae Bách Stemona tuberosa Lour Stemonaceae Râu hùm Tacca chantrieri Andr Taccaceae Giảo cổ lam Cucurbitaceae Hà thủ ô đỏ 10 11 Hoàng đằng Nghệ đen 12 Nghệ vàng Gynostemma pentaphyllum (Thunb.) Makino Fallopia multiflora (Thunb.) Haraldson Fibraurea tinctoria Lour Curcuma zedoaria (Berg.) Rosc Curcuma longa L Rạng mạ (Tày), Tiệp phầy mua đòi (Dao) Bo thác lủa (Tày), Mào xạm đòi (Dao) Pyắc (Tày), Lạy im (Dao) Cốc gia vụ (Tày, Dao) Thau khem (Tày, Dao) Mịn đăm (Tày), Trằn đìa chịa (Dao) Mịn khao (Tày), Trằn đìa pẹ (Dao) Aristolochiaceae Aristolochiaceae Menispermaceae Polygonaceae Menispermaceae Zingiberaceae Zingiberaceae 3.2.2 Exploring the propagation potential of two medicinal plants that have potential for exploitation and use in Na Hang Nature Reserve 3.2.2.1 Propagation of (Tacca chantrieri Andre) in Na Hang Nature Reserve  Experiment on vegetative propagation * Study the effects of season and type of cuttings on budding growth rate and rooting rate of Tacca chantrieri Andre The results showed that in the winter and spring, the rate of bud emergence and rooting of all three types are higher than the two seasons of Summer and Fall 18 Among the three types of cuttings, 60 days after cutting, the rate of budding and rooting of the cuttings from the stem is the highest; in the spring (86,7% budding, 77,0% rooting), followed by cuttings from the top of tuber (67.8% budding and rooting), the lowest rate is cuttings from root (60,4% budding, 58,1% rooting) * Study on the effect of growth promoters and their concentration on bud emergence and rooting rates of cuttings of Tacca chantrieri Andre After treatment with growth regulators IAA, α-NAA and IBA at different concentrations (1.000ppm, 1.500ppm and 2.000ppm) for cuttings from the Tacca chantrieri Andre, cuttings were made in the fall (01/9/2016) The results showed that the rate of budding and rooting of cuttings was very high after 60 days (100%) at 1.500ppm with both IBA and IAA  Seed propagation experiment After 90 days, the seed germination rate in outdoor sand was 14.3% and the room condition was 11.0% Lobster (Tacca chantrieri Andre) after germination of leaf growth, tree height; The development of roots and the average length of roots are very slow Tacca chantrieri Andre in the nursery period from to months have relatively high survival rate after months reached 83.33% Seedlings from the time when transplanted into the pot until months old leaves have reached 5.44 leaves per tree, soaring to very low height reached 8.15 cm After months of nursery care in the Tacca chantrieri Andre nursery did not meet the criteria for further nursing and continued monitoring This indicated that seedling with Tacca chantrieri Andre Difficult and inefficient compared to propagation by cuttings from tubers 3.2.2.2 Propagation of (Fibraurea tinctoria Lour.) in Na Hang Nature Reserve  Experiment on vegetative propagation * Study on the effects of season and type of cuttings on budding growth rate and rooting rate of (Fibraurea tinctoria Lour.) The experiment used three types of cuttings: top cuttings, trunk cuttings and old cuttings from the stem of the Fibraurea tinctoria Lour were conducted in the Spring, Autumn, Fall and Winter Results showed that after 60 days in all seasons, trunk cuttings have the highest rate of budding and rooting and the lowest rate belongs to top cuttings Trunk cuttings have 78.9% survival rate (in the Summer) and 76.7% budding rate in the spring Whereas, top cuttings only reached the survival rate of 41.1% (spring) and 14.1% budding rate (spring) This can be explained by the fact that the top cuttings are still young and nutrients stored in the cuttings are not sufficiently enough to supply, resulting in poor survival and growth Cuttings in the summer had the highest survival rate of 78.9%; followed by spring cuttings with a survival rate of 71.1% Cuttings in winter and fall showed 28.9% and 37.8% respectively Thus, the rate of budding of cuttings in spring is highest, followed by the summer and the lowest is cuttings in winter 19 * Study on the effects of growth promoters and their concentrations on the (Fibraurea tinctoria Lour.) The time for cuttings in the Fall (1/09/2016) The results showed that the use of IBA concentration 1.500ppm gave high rooting rate and early rooting time (96.7% after 60 days) IAA concentrations of 1.500ppm and IBA of 2.000ppm with highest results (94.4% after 60 days) For root numbers and root length, IAA concentrations of 1.500ppm gave very high results  Seed propagation experiment Results of experiments showed that different treatment methods of seeds, germination start and finish time as well as seed germination ratio were also different The germination time and germination rate of seed are also different In CT1, if sowing seeds in the wet sand, seeds have germinated to 98.9% only after 51 days In CT3, the seeds of Fibraurea tinctoria Lour were sown after soaking in cold water for 10 hours with a longer germination period and a lower germination rate of 58 days at 82.2% Thus, the seeds of Fibraurea tinctoria Lour., after harvesting and processing, it can be sown directly in the wet sand without any treatment method and still achieve a very high germination rate  Experiment on growing young seedlings of (Fibraurea tinctoria Lour.) in the nursery garden Each follow-up of 30 trees / iterations Seedlings from seeds after reaching the average height of 7cm, the average diameter of 0.2cm, about - leaves will actually be transplanted into the pot to nourish in the nursery Survival rate: Survival rate of Fibraurea tinctoria Lour seedlings werevery high After months, the survival rate has decreased but still high at 98% On diameter growth: It is shown that during the period of the nursery, the Fibraurea tinctoria Lour plant grows in relatively short root diameter From the tree measurement data show that after months new trees meet the standards of the garden The coefficient of variation in diameter tends to decrease with age On growth of height: So after months of nursing, the trees reach the height needed for afforestation The coefficient of height variation of the seedlings in nursery period was also low at 1.23 - 2.22% Thus, it can be concluded that the growth of seedlings of Fibraurea tinctoria Lour in the nursery period from to months is very uniform 3.2.3 Proposed solutions for conservation and sustainable development of some medicinal plants in Na Hang Nature Reserve, Tuyen Quang province 3.2.3.1 Preservation of medicinal plant resources of people in Na Hang Nature Reserve + The stock of medicinal plants in Na Hang Nature Reserve is decreasing (showing the frequency of encounter and quantity of products being exploited, traded ).Some medicinal plants in Na Hang Nature Reserve bring high economic value, creating a significant source of income for local people such as: Stephania rotunda Lour., Fibraurea 20 tinctoria Lour., Ardisia silvestris Pitard, Rauvolfia verticillata (Lour.) Baill., Heterosmilax gaudichaudiana (Kunth) Maxim + The species are welldeveloped in the area, which are suitable for local conditions, topography, fast growing, good development, easy-to-find seedlings and simple planting techniques can be planted in gardens, forest gardens and also bring economic value to people such as: Curcuma longa, Curcuma zedoaria, Tacca chantrieri, Gynostemma pentaphyllum… + To preserve and develop medicinal plants in Na Hang Nature Reserve in order to meet current and future needs, it is neccesarry to exploit rationally to ensure (in - situ) regeneration combined with the research on plantation (ex - situ) + Many species of medicinal plants are overexploited by the local people, leading to exhaustion in nature, some species are currently very difficult to see such as: Paris polyphilla Sm., Stephania rotunda, types of mistletoe At present, many species have been purchased by Chinese traders in large quantities through the small quota such as: Fibraurea tinctoria, Stephania rotunda, Rauvolfia verticillata, Heterosmilax gaudichaudiana…This is also the cause of decrease of medicinal plant resources + Currently, the knowledge of using medicinal plants, herbal medicines of local physicians are only oral, passing from the past generation to the next generation without being recorded for long-term preservation The young generation in the locality has not really paid much attention to the inheritance of using medicinal plants from the previous generation, so causing the fact that knowledge of medicinal plants become gradually lost 3.2.3.2 Solutionsfor conservation and sustainable development of some medicinal plants in Na Hang Nature Reserve With the aim of managing, protecting and developing medicinal plant resources, we propose some adjustments and additions to improve existing solutions and plans such as:  Technical solutions: This is the most important scientific-technical solution Based on the findings of the study conducted in Na Hang Nature Reserve, we propose the following recommendations: + Conservation of resources (in - situ): It was conducted in Thac Mo natural forest (400 to 600 m above sea level) in Son Phu commune, Na Hang district, Tuyen Quang province; choosing the area near the stream, under the forest canopy to cultivate the plants such as: Asarum balansae, Asarum caudigerum, Tacca chantrieri, Homalomena occulta, Gynostemma pentaphyllum, Stephania rotunda (area 1) Fibraurea tinctoria, Stemona tuberosa, Tinospora sinensis, Fallopia multiflora, Curcuma zedoaria, Curcuma longa L are planted in the valley where there is light (area 2) The total area of 02 planted garden areas is 500m2, with the number of 1200 trees (average 100 plants/species) Supervising the growth and development of the plants in the area, the initial results showed that the species: Curcuma zedoaria, Curcuma longa, Asarum balansae, Tacca chantrieri, Homalomena occulta, Fibraurea tinctoria have the ability to grow and develop, 21 showing in high survival rate, some plantswere flowering and/or fruiting (Curcuma zedoaria, Curcuma longa, Tacca chantrieri); And for the species: Gynostemma pentaphyllum, Stephania rotunda, Tinospora sinensis, Fallopia multiflora: they have low survival rate, slow growth, only Fallopia multiflora is flowering; Particularly, species such as: Asarum caudigerum, Stemona tuberosa, Tinospora sinensis have relatively low survival rate, many plants after being cultivatedare gradually dying + Transposition conservation (ex - situ): Implemented in the forest gardens of 02 households in Ban Bung, Thanh Tuong commune, Na Hang district with an area of 1000 m2/household with the number of 100 seedlings per species In general, thanks to the selection of suitable natural conditions, favorable conditions and good care, the growth and development of those species are relatively equal, high rate of survival Some other solutions requiring community participation:  Regarding awareness + It is necessary to promote propaganda and advocacy for all levels, sectors, communes and ethnic communities in Na Hang Nature Reserve to raise awareness about the value of medicinal plant resources and medicinal remedy + To organize training courses for local people on methods of exploiting, using, expanding cultivation and sustainably collecting valuable medicinal plants  Regarding policy + It is necessary to improve the quality of human resources To adopt policies and preferential regimes to attract investment from enterprises, scientists and people to study and produce experimentally the varieties of medicinal plants + District People's Committee should pay attention to investment in material facilities, equipment and funds to preserve and develop gene sources of medicinal plants to meet the requirements and tasks such as: ▪ Planning of land areas with favorable natural conditions for the development of medicinal plants for propagation and planting of medicinal plants The study found that there are areas such as Khau Tinh, Ban Bung, Nam Pen, Nam Trang, Thac Mo, Phieng Bung having natural conditions very favorable for growing and developing medicinal plants ▪ Developing and implementing specific policies to assist poor people in forest villages in finding new and more sustainable live lihoods  Regarding organization: + Units and social organizations should actively develop programs and projects on the development of medicinal plants from indigenous medicinal plants while ensuring safety and quality while preserving and developing their sources All medicinal plants are attached to the target of socio-economic development of the locality + To build medicinal plant gardens in each commune in the Nature Reserve in order to preserve precious and rare medicinal plants, and to develop them into seed sources in service of local medicinal plant development programs To care and invest in research for 22 conservation and development of medicinal plant resources + Establishing Oriental medicinecooperatives in the communes will be effective in the preservation and development of medicinal plants, and will improve the market access for people, as the premise for guiding agencies in the process of implementation and maintenance, development of sustainable models in the locality + Combining solutions for pharmaceutical companies and the traditional medicine Associations in the districts, communes to develop medicine from medicinal herbs in the area CONCLUSIONS AND RECOMMENDATIONS Conclusions 1/ Medicinal plant resources in Na Hang Nature Reserve, Tuyen Quang province are abundant and diverse; Through surveys and study 647 species belonging (accounted for 55.7% of total known species) to 433 genera of 137 families in the four vascular plant divisions were identified Comparing with the previous reports, the dissertation has added 180 species 2/ In Na Hang Nature Reserve, Tay and Dao ethnic minorities have traditional customs of using medicinal plants for health care and treatment The medicinal plants that Tay people often exploit and use are quite abundant (223 species/176 genera/84 families) that aremore than Dao people (164 species/136 genera/72 families) Of these, 151 species/130 genera/71 families were used by both ethnic groups 3/ Results of screening biological activity of 18 medicinal plantsamples collected from Na Hang Nature Reserve include: 07 samples (TQ02, TQ05, TQ07, TQ10, TQ11, TQ15, TQ16) have effects of anti-lung cancer cell H1975, 03 samples (TQ05, TQ14, TQ15) can inhibit breast cancer cells MCF-7, 01 sample (TQ05) can inhibit breast cancer cells MDA-MB-231, 01sample (TQ13) can be capable of resisting pancreatic cancer cells PANC1; 06 samples (TQ03, TQ04, TQ08, TQ09, TQ10, TQ13) have strong α-glucosidase effects (inhibition>50% at 100μg/ml) For α-amylase, 04 samples (TQ05, TQ15, TQ17, TQ18) have an inhibitory effect of> 50% at 500μg/ml 4/ From sample of (Mallotus microcarpus Pax & K Hoffm) of TQ02 the chemical structure of 11 compounds were isolated and determined, including new compound 3-Methoxybenzensulfonic acid 4-O-β-D-glucopyranoside The new compound 3-Methoxybenzensulfonic acid 4-O-β-D-glucopyranosideand compound Methyl salicylate 2-rutinoside have effects on all three lines of lung cancer cells H1975, A549 and breast cancer cell MCF-7; Leonuriside A substance has a very strong effect on breast cancer cell MCF-7 with value IC50 = 0,48 µM experimental conditions 5/ From sample of (Miliusa sinensis Fin & Gagnep.) of TQ13, the chemical structure of 02 flavonoid compounds were isolated and determined, including: Kaempferol 3-O-(2,6-diO-α-L-rhamnopyranosyl)-β-D-galactopyranoside; Quercetin 3-O-(2,6-di-O-α-L-rhamnopyranosyl)-βD-galactopyranoside and 01 sterol compound Daucosterol Both flavonoid compounds inhibited α-glucosidase with IC50 values of 557.3 and 692.5 μM 23 6/ From the results of study, evaluation of the potential, the situation of exploitation, trading and use of 20 valuable medicinal plants in Na Hang Nature Reserve, the dissertation preliminary selects and proposes to select 12 potential species of medicinal plants that need to be researched on planting and developing to create a source of pharmaceutical materials for local economic and social development as well ascontribute to sustainable preservation 7/ Results ofpropagation for two species (Tacca chantrieri Andre) and (Fibraurea tinctoria Lour.) showed that: - The propagation by cuttings from tubers of (Tacca chantrieri Andre) and from the stem of (Fibraurea tinctoria Lour.) with the treatment of stimulants gave remarkable results: number of seedlings germinated, rooted is quite high - Seedling of (Tacca chantrieri Andre) hada very low germination rate (only 11.0 - 14.3%) Seedlings in the nursery period from to months have leaf numbers of 5.44 leaves per tree, height up to 8.15 cm Seedlings grow slowly, after months of care in lobster nurseries are not eligible for export Meanwhile, in the case of (Fibraurea tinctoria Lour.), the seed germination rate was very high (98.9% when sown in moist sand) The seedlings in the planting bag grow well, with a high survival rate of up to 98% after months, less pestilent insects, and at months of age can be grown on a commercial scale 8/ The study initially proposed some solutions (identification of potential medicinal plants, techniques, organization, management, education ) to develop, cultivate, exploit, use and conserve the diversityof medicinal plant resources in Na Hang Nature Reserve, Tuyen Quang province Recommendations It is proposed further research, especially the investigation, evaluation and selection of species with development potential From then testing thepropagation of precious and rare medicinal plants with potential for conservation and development in Na Hang Nature Reserve Tacca chantrieri Andre has just been researching two treatments of Tacca chantrieri Andre so the seed germination rate is very low Further research into technical measures to improve seed germination should be continued; Tacca chantrieri Andre, during the nursery stage, until the seedlings meet the requirements of the nursery Continue to investigate and look for species containing biological active compounds from medicinal plant resources in Na Hang Nature Reserve, especially potential active compounds for treatment of cancer, cardiovascular disease, etc 24 NEW POINTS OF THE DISSERTATIONS  For the first time, the list of medicinal plant species in Na Hang Nature Reserve was relatively sufficiently reported, including 647 species of 433 genera and 137 families of four vascular plant divisions, of which adding 180 new species; The number of species of medicinal plants commonly used in Na Hang Nature Reserve has been collected: Tay people use 223 species and Dao people use164 species Among them, there are 151 species, 130 genera and 71 families used by both ethnic groups for medicine  Initially, the research on asexual and sexual propagation for two species of (Tacca chantrieri Andre) and (Fibraurea tinctoria Lour.) in Na Hang Nature Reserve  From samples of (Mallotus microcarpus Pax & K Hoffm) the chemical structure of 11 compounds were isolated and determined, including new compound 3-Methoxybenzensulfonic acid 4-O-β-D-glucopyranoside From samples of (Miliusa sinensis Fin & Gagnep.) the chemical structure of 02 flavonoid compounds were isolated and determined, including: Kaempferol 3-O-(2,6-di-O-α-L-rhamnopyranosyl)-β-Dgalactopyranoside; Quercetin 3-O-(2,6-di-O-α-L-rhamnopyranosyl)-β-D-galactopyranoside and 01 sterol compound Daucosterol Both flavonoid compounds inhibited α-glucosidase with IC50 values of 557.3 and 692.5 μM in active test LIST OF PUBLIC RELATED WORKSHOP RELATED TO THE DISSERTATION Nguyen Thi Hai, Nguyen The Cuong, Tran Huy Thai, Chu Thi Thu Ha, Nguyen Anh Tuan (2015) Some research results on medicinal plant resources in Na Hang Special Use Forest, Tuyen Quang province The 6th National Conference on Ecology and Environment, 2015, p 1093-1099 Nguyen Thi Hai, Chu Thi Thu Ha, Nguyen The Cuong, Nguyen Anh Tuan, Tran Huy Thai (2016), Research on the use of medicinal plants by Tay people in Na Hang Nature Reserve, Tuyen Quang province - J Viet Env., 2016, Vol 8, No 5, pp 227-283 Tran Huy Thai, Nguyen Thi Hai, Nguyen Thi Hien, Chu Thi Thu Ha, Nguyen The Cuong, Pham Thanh Binh, Nguyen Hai Dang, Nguyen Tien Dat (2017), Cytotoxic constituents of Mallotus microcarpus Pax & Hoffm J Natural Product Communications, 2017, Vol 12 No 3, p 407-408 (SCIE) Nguyen Thi Hai, Tran Huy Thai, Pham Huu Hanh, Nguyen Thi Hong Hanh, Doan Thi Phuong Ly (2017), Fibraurea tinctoria Lour., in Na Hang Nature Reserve, Tuyen Quang province MARD, No 19, October 1, 2017, pp 151-157 Nguyen Thi Hai, Tran Huy Thai, Nguyen The Cuong, Tran Thi Thanh Van (2017), Diversity of medicinal plants in Na Hang Nature Reserve, Tuyen Quang province (2017) Journal of Tan Tao University, No 6, September 2017, pp 45-50 ... 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" The goal of the... establish internal conservation areas (in - situ) conservation and external conservation or (ex - situ) conservation Biodiversity conservation in general and conservation of medicinal plants in particular,... 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