Nghiên cứu đa dạng loài và thành phần hóa học tinh dầu một số loài thuộc họ Sim (Myrtaceae Juss. 1789) ở tỉnh Hà Tĩnh.Nghiên cứu đa dạng loài và thành phần hóa học tinh dầu một số loài thuộc họ Sim (Myrtaceae Juss. 1789) ở tỉnh Hà Tĩnh.Nghiên cứu đa dạng loài và thành phần hóa học tinh dầu một số loài thuộc họ Sim (Myrtaceae Juss. 1789) ở tỉnh Hà Tĩnh.Nghiên cứu đa dạng loài và thành phần hóa học tinh dầu một số loài thuộc họ Sim (Myrtaceae Juss. 1789) ở tỉnh Hà Tĩnh.Nghiên cứu đa dạng loài và thành phần hóa học tinh dầu một số loài thuộc họ Sim (Myrtaceae Juss. 1789) ở tỉnh Hà Tĩnh.Nghiên cứu đa dạng loài và thành phần hóa học tinh dầu một số loài thuộc họ Sim (Myrtaceae Juss. 1789) ở tỉnh Hà Tĩnh.Nghiên cứu đa dạng loài và thành phần hóa học tinh dầu một số loài thuộc họ Sim (Myrtaceae Juss. 1789) ở tỉnh Hà Tĩnh.Nghiên cứu đa dạng loài và thành phần hóa học tinh dầu một số loài thuộc họ Sim (Myrtaceae Juss. 1789) ở tỉnh Hà Tĩnh.
MINISTRY OF EDUCATION AND TRAINING VINH UNIVERSITY TRAN HAU KHANH RESEACH DIVERSITY AND CHEMICAL COMPOSITION OF ESSENTIAL OILS OF SOME SPECIES OF MYRTACEAE JUSS 1789 IN HA TINH PROVINCE Major: Botany Code: 9420111 ABSTRACT OF Ph.D THESIS IN BIOLOGY NGHE AN – 2022 The thesis was fulfilled at Vinh University Scientific supervisors: Assoc Prof Dr Pham Hong Ban Assoc Prof Dr Tran Minh Hoi Reviewer 1: …………………………………………… Reviewer 2: …………………………………………… Reviewer 3: …………………………………………… The thesis will be defended before the Thesis Evaluation Board of Vinh University at … h… , … , … , 2022 The thesis can be found at: - Viet Nam National Library; - Information Centre, Nguyen Thuc Hao Library of Vinh University PREFACE Reason to choice the investigation subject Vietnam is one of the biodiversity centers as well as the crop centers of the world, lying in the cradle of angiosperms, There are many species were recognized locally sourced In the flora of Vietnam, The group of plants with essential oils is very rich and diverse Up to now, about 657 species have been recorded (accounting for 6.3% of the total known species in Vietnam) in 357 genere (15.8%) and 114 families (37.8%) Ha Tinh has a rich and diverse flora, Here, there are typical vegetation such as tropical evergreen closed forest vegetation, exist at elevations below 700m and lowland subtropical evergreen closed forests from 700-1,800m Myrtaceae is one of the large families of the Magnoliophyta, there are 5.950 species in 132 genera in the world, distributed mainly in the tropics, especially in Southeast Asia and Brazil In Vietnam, 15 genera with 107 species have been recorded Many species of Myrtaceae have meanings and give many different uses (wood, medicine, essential oil, ornamental, food, etc) and has been used by people in daily life for many potential applications in the pharmaceutical industry, has great significance in both ecological and economic terms Despite the great values and benefits mentioned above, Myrtaceae in Ha Tinh have not been studied for species diversity as well as for the chemical composition of essential oils and their activities Therefore, the author has chosen the subject “Research diversity and chemical composition of essential oils of some species of Myrtaceae Juss 1789 in Ha Tinh Province”, contribute to the assessment of species diversity, essential oil chemical composition and biological activity of some species in this family, contribute more to the practical application of people's lives, help local authorities to basis in conservation, rational exploitation, green and sustainable development Objectives 2.1 General objective Assess species diversity and biological characteristics, chemical composition and antibacterial activity of essential oils of some species of Myrtaceae in Ha Tinh Province 2.2 Detail objectives - Evaluation of the diversity of species composition and biological characteristics of species in Myrtaceaec in Ha Tinh Province - Chemical composition analysis of essential oils of leaves, flowers and fruits of some species of Myrtaceae in Ha Tinh Province - Test of antibacterial activity of essential oils of leaves of some species in Myrtaceae in Ha Tinh Province Scientific and practical significance of thesis This work has provided new data on the use value and diversity of Myrtaceae in Ha Tinh and provide scientific data for conservation, rational and efficient exploitation of plant resources containing essential oils of Myrtaceae, serving economic and social development in Ha Tinh Province New findings of the thesis - The first time, providing complete and systematic data on Myrtaceae in Ha Tinh Province, including 63 species in 13 genera Record more distribution in Ha Tinh for 38 species belonging to 09 genera of Sim Myrtaceae Record Feijoa genus with Feijoa sellowiana (O.Berg.) O.Berg species and Myrciaria genus with Myrciaria cauliflora (Mart.) O.Berg species have been added to the list of Vietnam - The first time providing data on essential oils of leaves of 09 species of Myrtaceae: Cleistocalyx nigrans (Gagnep.) Merr & Perry, Cleistocalyx retinervius Merr & Perry, Decaspermum parviflorum (Lamk.) J Scott), Rhodamnia dumetorum (Poir.) Merr & Perry, Syzygium bullockii (Hance) Merry & Perry, Syzygium hancei Merr.& Perry, Syzygium lineatum (DC.) Merr & Perry, Syzygium tsoongii (Merr.) Merr & Perry and Syzygium zeylanicum (L.) DC The first time providing data on essential oils of flowers and fruits of 01 species of Myrtaceae: Decaspermum parviflorum (Lamk.) J Scott) - The bioactivity of essential oil of leaf of Syzygium tsoongii (Merr.) Merr & Perry, Syzygium bullockii (Hance) Merry & Perry and Syzygium zeylanicum (L.) DC has been determined The essential oils of the leaves of all three species above are resistant to Gram (+), including: Enterococus faecalis, Staphylococcus aureus and Bacillus cereus In addition, the essential oils of the leaves of Syzygium bullockii (Hance) Merry & Perry and Syzygium tsoongii (Merr.) Merr & Perry are also resistant to Candida albicans In particular, the essential oil of Syzygium bullockii (Hance) Merry & Perry species, in addition to the above abilities, is also resistant to Escherichia coli (Gram (-)) and reach MIC = 128 g/ml (IC50 = 46.67 g/ml) CHAPTER LITERATURE REVIEW 1.1 Myrtaceae researches 1.1.1 In the world In 1753, the Swedish botanist Linnaeus C has recorded genera with 13 species of Myrtaceae In 1789, Jussieu A named Myrtaceae with typus Myrta L., Later genera in this family had many changes, especially the Eugenia L genus was mostly converted into the Syzygium genus by Gaertner (1732-1791) In 1826, Blume splited from the Eugenia genus from Syzygium genus to build a new genus, Cleistocalyx Blume In addition to the overall research on Myrtaceae worldwide, There are also many individual research works such as: The flora of Indochina, episode (1908) divided Myrtaceae to 11 genera and 80 species In Australia, 75 genera and 1,500 species have been recorded In China, Merrill E D & Perry L M (1938a) and Chen J & Craven L A (2007) have built identification keys for 10 genera and 121 species of Myrtaceae In Thailand had 14 genera and 204 species Cambodia have recorded 27 species of the Syzygium genus Laos have recorded 14 genera and 118 species In Malaysia, 12 genera and 241 species have been recorded Philippine have recorded 12 genera and 250 species as a identification key of Myrtaceae According to Chen J & Craven L A (2007), the Myrtaceae had 130 genera and 4,500 to 5,000 species From 2008 to 2016, the Myrtaceae in the world were listed and published, including 132 genera and 5,950 species 1.1.2 In Vietnam Loureiro J is the first author to study Myrtaceae in Vietnam He described genera with 15 species of Myrtaceae in "FIora Cochinchinensis" (1788) Lecomte H (1908 - 1921) described 11 genera with 80 species distributed in Indochina, In which, 10 genera and 58 species have been recorded in Vietnam In 1965, Tamás Pócs recorded genera and 28 species of Myrtaceae in Vietnam In 1969, Le Kha Ke et al have built identification keys for Myrtaceae in Vietnam of genera and 15 species In 1993, Pham Hoang Ho published “Cây cỏ Viet Nam” have built identification keys for Myrtaceae in Vietnam of 14 genera and 101 species In 2003, Nguyen Tien Ban et al described 15 genera with 107 species In 1993, Tran Dinh Ly et al published “1,900 loài có ích Việt Nam”, 10 genera and 31 species have been recorded In 1997, Herbalist Le Tran Duc introduced and described genera with species used as medicine In 1999, Do Tat Loi described 11 species of genera used as medicine Do Huy Bich et al (2004) described genera and 30 species 11 genera and 32 species used as medicine described by Vo Van Chi (2012) 1.1.3 In Ha Tinh In the period from 1993 to 1998, FIPI, Forest Protection Department and WWF surveyed, investigated of plants in Vu Quang National Park, Ha Tinh Province and published genera with 17 species of Myrtaceae Le Trong Trai, Nguyen Huy Dung, Nguyen Cu, Le Van Cham and Eames J C (1996) published species of genera in Ke Go Nature Reserve, Ha Tinh Province Most recently (in 2020), Le Duy Linh in his doctoral thesis “Research on the composition of essential oil-bearing plant species in Vu Quang National Park, Ha Tinh Province” recorded genera and 14 species of Myrtaceae We find that myrtaceae in Ha Tinh Province have not been fully studied in terms of their species composition and biological characteristics Currently, only research works of general diversity are published sporadically 1.2 Studies on chemical composition of essential oil of Myrtaceae 1.2.1 In the world The work of Guenther E (1948) in "The Essential Oils", The author has described the biological characteristics and using of some essential oils of Myrtaceae Lawrence BM, in the works "Essential oils" (1992 - 1994), "Progress in essential oils" (1995 - 1997) and "Progress in essential oils" (2001) have enumerated over 1,000 plant species containing essential oils on world, including plants of the genus Eugenia genus, the Eucalyptus genus, the Melaleuca genus of Myrtaceae In 2006, Christophe Wiart’ “Medicinal plants of the Asia - Pacific: Drugs for the Future?” has been recognized that Myrtaceae's essential oil has therapeutic value, antioxidant, cytotoxic and antibacterial effects 1.2.2 In Vietnam In 1977, Vu Ngoc Lo published “precious essential oils trees”, The author introduced the biological, ecological and sowing characteristics as well as the technique of collecting, extracting and evaluating the quality of Melaleuca'essential oil Do Tat Loi (1985 - 1989) introduced some essential oil plants of the Eucalyptus genus Nguyễn Xuân Dũng (1996) in “Research on chemical composition contributing to chemical classification (chemotaxonomy) of some medicinal plants and essential oil plants in Vietnam” reported specie from the Cleitocalyx genus and 16 species from the Eucalyptus genus La Dinh Moi et al in “Essential oil plant resources in Vietnam” mentioned the general problems of essential oil plant resources in Vietnam, and introduced genera of Myrtaceae have essential oils in the flora of Vietnam, Besides, detailed description of the origin, use, production, characteristics of essential oils, etc of some species such as Melaleuca, Eucalyptus, Baeckea 1.2.3 In Ha Tinh Currently, we have not seen any published studies on the chemical composition of essential oils and biological activities of Myrtaceae in Ha Tinh Province 1.3 Biological characteristics of species in the Myrtaceae 1.3.1 Morphological characteristics of species in the Myrtaceae + Plant stem: Large tree, medium tree, small tree or shrub, often fragrant, wood of some species commonly used in construction and in household appliances + Leaves: Leaves simple, whole or toothed margin, most leaves opposite, except for species of the Melaleuca genus, Tristaniopsis, Eucalyptus, Callistermon and Baeckea have alternate leaves Leaves are usually glandular and without stipules, often fragrant and contain essential oils Leaves vary in shape and size Leaves have the following forms: oval, needle-shaped, ect, Leaf blade may be thick or thin, usually smooth, some species have glossy blade, may be white or white to light brown hairy The base of the leaves is balanced, petioles are short or 1-2 cm long, the tips of the leaves are often pointed, often with a main vein, very few species have main veins, the secondary veins usually come from the main veins, the veins are 1-3 mm from the cover + Flower: Inflorescences at the tips of branches or at the leaf axils, usually cymbal or clustered, rarely globose on the stem Bracts 1, easy to fall, bracts Flowers are even, hermaphroditic Tubular sepals completely attached to gourd or only a few, may have - sepals, arranged, very rarely tundra tubules Corolla - 5, arranged, often adherent to a spherical shape, when the flower completely opens and falls off like a cap Flowers often have numerous stamens, they are arranged in one or more rows, completely separate or slightly attached at the base of a short tube; anther attached to the back or to the base, cells, open in long slits, mesotropic with gland at the end The most not have bile discs, if present, they are rim-shaped, covering the base of the spout The gourd fully or partially lowered, may have to cells or more, attached to the central columnar ovule, rarely attached to the side; proboscis single, elongated, rounded tip, rarely - lobes; In the ovary, the ovules are arranged in two or more rows, attached in the corner of each cell, the ovule is inverted + Fruit: Capsule or fleshy, fibrous, with very short to long stalk, usually enclosed by calyx; Fruit globose, ovate, bell-shaped, open or unopened when ripe, often smooth or sometimes hairy Seeds single or multiple, usually without endosperm, embryo straight or curved, sometimes lean, with dicotyledons not far apart 1.3.2 Ecological characteristics, distribution of species in the Myrtaceae + Locality: The species often grows scattered or in clusters on the ground in forest, light forest, subforest, along streams, beside the road, limestone hills or coastal sandy areas Most mature trees are light-loving, young trees are shadetolerant There are many species of Myrtaceae in Vietnam that are imported and cultivated such as: Callistemon citrinus (Curt.) Skeels, Eucalyptus camaldulensis Dehnhart, Eucaluptus exserta F Muell, Eucalyptus maculata Hook and Melaleuca var minor (Smith) Duthie) are imported from Australia Psidium cujavillus Burm f.) and Psidium guajava L are imported from Central America, ect + Flowering season and fruit season: Species of Myrtaceae usually have a flowering season from January to October every year, depending on the specific species, there are different flowering and fruiting seasons Some species flower from January to March every year and bear fruit after 2-3 months, such as: Eucalyptus camaldulensis Dehnhart, Eucaluptus exserta F Muell, Eucalyptus maculata Hook Some other species have a flowering season from April to June such as: Melaleuca leucadendra L, Rhodamnia dumetorum (Poir.) Merr & Perry, Rhodomyrtus tomentosa (Ait.) Hassk, Syzygium jambos (L.) Alston, Syzygium odoratum (Lour.) DC), ect Many species flower from July to September such as: Cleistocalyx operculatus (Roxb.) Merr & Perry, Syzygium hancei Merr & Perry, ect There are also species that have a flowering season from September to October, such as: Syzygium sterrophyllum Merr & Perry In addition, some species have flowering and fruiting seasons almost all year round, such as: Callistemon citrinus (Curt.) Skeels, Myrciaria cauliflora (Mart.) O.Berg, Psidium guajava L CHAPTER MATERIALS, CONTENTS AND METHODOLOGY OF STUDY 2.1 Materials and scope of study - Objects of the study: Species in Myrtaceae in Ha Tinh Province (includes cultivated species and naturally distributed species) - Scope of the study: In the whole province of Ha Tinh, mainly focus is on Vu Quang National Park, Ke Go Nature Reserve, Huong Son district protection forest area, biodiversity conservation area upstream of Ngan Pho river, and forest area Nam Ha Tinh households, Hong Linh, Nghi Xuan protection forests and other areas where this family is distributed The study period was from May 2018 to May 2022 2.2 Content of study - Make a list and evaluate the diversity of species composition, diversity in stem form, diversity in use value, study the biological characteristics (morphology, ecology) and distribution of species in Myrtaceae in Ha Tinh Province - Determination of content, chemical composition in essential oils and testing of antibacterial activity of essential oils of some species in Myrtaceae in Ha Tinh Province 2.3 Study methodology 2.3.1 Botany methods 2.3.1.1 Data inheritance method - Researching documents, collecting and analyzing domestic and foreign documents on species in Myrtaceae Data on natural, socio-economic conditions in the study area Samples are kept in domestic and foreign museums The data on the area and environmental factors when implementing the research thesis Legal documents related to biodiversity conservation Scientific documents, relevant research results 2.3.1.2 Field survey method The field investigation process applied according to the method described by Nguyen Nghia Thin (2007) Based on the topographic map and the map of ecosystems in Ha Tinh Province, delineate the route and study site Research points and routes pass through different habitats that characterize the study area Using compass, GPS satellite positioning machine to determine the location of the sampling point in the field In each 0.9: The constant applies to essential oils with a density lighter than water (0.9) 2.3.2.3 Methods of analyzing chemical composition of essential oils Preparation of analytical sample for gas chromatography: Dissolve 1.5 mg of dried essential oil with anhydrous Na2SO4 in ml of pure hexane of the type used for chromatographic analysis + Gas chromatography (GC) with FID probe: Made on Agilent Technologies HP 7890A Plus with FID detector, HP-5MS capillary column at length of 30 m, inner diameter (ID) = 0.25 mm, a layer of 0.25 m with hydrogen as carrier gas The injection chamber temperature is 250°C Detector temperature is 260oC Program the temperature of 60°C (2 minute), increase by 4°C/minute until 220°C, stop at this temperature for 10 minutes + Gas chromatography (GC/MS): This is performed on Agilent Technologies HP 7890A / HP 5973 MSD combined gas chromatography system with separation column and chromatographic operating conditions as mentioned above and with Helium as carrier gas 2.4 Test method for antibacterial activity Testing the antimicrobial activity of essential oils on some strains of Gram (+), Gram (-) bacteria and Candida albicans Step Test the antibacterial effect by diffusion method on agar plate Test samples are considered positive when a sterile ring is greater than or equal to 20 mm Step Samples with positive activity in step will be tested in step to calculate the minimum inhibitory concentration (MIC) according to Vanden Berghe A D & Vlietinck A J (1991); Vlietinck A J (1998) and McKane L & Kandel J (1996) carried out on a 96-well microplate Tested antibiotics include: Streptomycin for Gram (+) bacteria, Tetracyclin for Gram (-) bacteria, Nystatin or Amphotericin B for Candida albicans Samples with MIC ≤ 200µg/ml are considered as antimicrobial 2.5 Statistical analysis method Data were analyzed statistically using Microsoft Office Excel 2016 11 CHAPTER RESEARCH FINDINGS AND DISCUSSION 3.1 The diversity of the Myrtaceae in Ha Tinh procince Research results of Myrtaceae in Ha Tinh Province have identified 63 species and 13 genera Of the 63 species, there are 33 species distributed naturally, 23 imported species and cultivated species In the 13 genera, the distribution of species in the genus is uneven, The most diverse is the genus Syzygium with 31 species (accounting for 49.21%), of which 24 species are naturally distributed, 06 species are cultivated and 01 species is imported; next is the genus Eucalyptus with 14 species (accounting for 22.22%), of which all 14 species are imported; genus Cleistocalyx 04 species (accounting for 6.35%), of which 03 species are naturally distributed and 01 species are cultivated; genus Psidium 03 species (accounting for 4.76%), of which all 03 species are imported; genus Decaspermum 02 species (accounting for 3.17%) and both species are distributed naturally; genus Melaleuca 02 species (accounting for 3.17%), of which 01 imported species and 01 natural species; genera Baeckea, Rhodamnia, Rhodomyrtus 01 species each (shared 1.59%), all are naturally distributed; genera Callistemon, Eugenia, Feijoa, Myrciaria each genus 01 species (accounting for 1.59%), all imported From Table 3.1, we see that the number of imported species is quite high (23 species), of which the genus Eucalyptus is the genus with the second largest number of species (14 species) and 100% of the species in the genus are imported Besides, there are 07 species in the family that are cultivated So, we see that the origin of the species in the Myrtaceae family is quite diverse, the species in the family have many meanings and have many practical applications in life, so they are imported and planted with a large number of species (table 3.1) Table 3.1 Number of genera and plant species of Myrtaceae in Ha Tinh Province Percentage Sources No Genus Species (%) Baeckea Callistemon 1 1.59 1.59 12 Natural tree Import tree Cleistocalyx 6.35 Decaspermum Eucalyptus Eugenia Feijoa 14 1 3.17 22.22 1.59 1.59 Melaleuca 3.17 10 11 12 Myrciaria Psidium Rhodamnia Rhodomyrtus 1 1.59 4.76 1.59 1.59 13 Syzygium 31 49.21 13 63 100 Total - 03 Natural tree - 01 Crops Natural tree Import tree Import tree Import tree - 01 Natural tree - 01 Import tree Import tree Import tree Natural tree Natural tree - 24 Natural tree - 06 Crops - 01 Import tree 63 species To further clarify the diversity of Myrtaceae in Ha Tinh Province, the results were compared with Myrtaceae in Vietnam (Nguyen Kim Dao (2003)), the results are presented in table 3.2 Table 3.2 Comparison of the number of species in genera studied in Ha Tinh with the number of species currently known in Vietnam No Genus Ha Tinh (1) Vietnam (2) 10 11 12 13 Baeckea Callistemon Cleistocalyx Decaspermum Eucalyptus Eugenia Feijoa Melaleuca Myrciaria Psidium Rhodamnia Rhodomyrtus Syzygium 1 14 1 1 31 1 25 61 Note: (2): According to Nguyen kim Dao (2003) 13 Percentage (%) between (1) and (2) 100.00 100.00 80.00 66.67 56.00 100.00 Unable to determine 100.00 Unable to determine 100.00 50.00 100.00 50.82 Table 3.2 shows, Species composition of Myrtaceae in Ha Tinh Province is quite diverse with 63 species compared to 107 species and varieties, accounting for 58.87% of the total number of species and varieties known in Vietnam and 13 genera compared to 15 genera, accounting for 86.67% of total genera in Vietnam Among the 13 genera, there are new genera with new species that have not been listed in Vietnam, the genus Feijoa with Feijoa sellowiana (O.Berg.) O.Berg and the genus Myrciaria with Myrciaria cauliflora (Mart.) O Berg are now commonly imported and grown in Vietnam There are genera with the maximum number of species and equal to the number of species known in Vietnam, including: Baeckea, Callistemon, Eugenia, Rhodomyrtus (each genus has species), the genus Melaleuca has species and the genus Psidium has species With the high diversity of Myrtaceae in Ha Tinh Province, we proceed to identify species in this family with additional distribution areas in Ha Tinh Province 3.2 Species in the family Myrtaceae were recorded with an additional distribution area in Ha Tinh Province Research results on plant diversity Myrtaceae in Ha Tinh Province have added 38 species and 09 genera with distribution in Ha Tinh Province With 38 species above, there are species distributed in the Northern region: Bồ đề liễu (Eucalyptus exserta F.Muell.), Trâm dầu (Syzygium balsamineum Wall.), Trâm Ba (Syzygium baviense (Gagnep.) Merr & Perry), Trâm bois (Syzygium boisianum (Gagnep.) Merr & Perry) Trâm (Syzygium imitans Merr & Perry) There are species distributed in the Central region, including: Trâm gân mạng(Cleistocalyx retinervius Merr & Perry) Trâm finet (Syzygium finetii (Gagnep.) Merr & Perry) There are species distributed in the Central Highlands, including: Khuynh diệp ướt (Eucalyptus botryoides Smith), Bạch đàn tro (Eucalyptus cinerea F Muell.), Khuynh diệp to (Eucalyptus grandis Hill ex Maiden), Bạch đàn maiden (Eucalyptus Maidenii F Muell.) Khuynh diệp dẻo (Eucalyptus viminalis Labill.) There are 14 species distributed in the Southern region, including: Trâm vối ô (Cleistocalyx circumcissa (Gagnep.) Phamh.), Bạch đàn trắng (Eucalyptus alba Reinw ex Blume), Bạch đàn dài (Eucalyptus longifolius Link), Bạch đàn xoan (Eucalyptus populifolia Hook.), Bạch đàn liễu (Eucalyptus tereticornis Smith), Ổi cảnh (Psidium cujavillus Burm f.), Trâm hoa mảnh (Syzygium leptanthum (Wight) Niedenz.) Trâm pierre (Syzygium pierrei (Gagnep.) Merr & Perry) There is species distributed from the North to the Central region: Trâm bon (Syzygium bonii (Gagnep.) Merr & Perry) There are species distributed from the North to the Central Highlands, including: Thập tử mảnh (Decaspermum gracilentum (Hance) Merr & Perry) Tiểu diệp an (Eucalyptus resinifera Smith) There is species distributed from the Central region to the Central Highlands region: Trâm hoa xanh (Syzygium chloranthum (Duthie) Merr & Perry) There are species distributed from the Central region to the Southern region, including: Bạch đàn úc (Eucalyptus camaldulensis Dehnhart), Trâm bôi (Syzygium corticosum (Lour.) Merr & Perry) Trâm kiền kiền (Syzygium syzygioides (Miq.) Merr & Perry) There are species distributed from the Central Highlands to the South, including: Tràm đỏ (Callistemon citrinus (Curt.) Skeels), Trâm trắng (Syzygium chanlos (Gagnep.) Merr & Perry), Trâm hoa dài (Syzygium lineatum (DC.) Merr & Perry) Trâm nam (Syzygium cochinchinensis (Gagnep.) Merr & Perry) There are species with a wide distribution, almost stretching across the country from the North to the South, including the following species: Ổi dứa (Feijoa sellowiana (O.Berg.) O.Berg), Thập tử hoa nhỏ (Decaspermum parviflorum (Lamk.) J Scott), Trâm sơ ri (Eugenia uniflora L.), Bạch đàn chanh (Eucalyptus citriodora Hook f.), Nho thân gỗ (Myrciaria cauliflora (Mart.) O.Berg), Trâm chụm ba (Syzygium formosum (Wall.) Masam) Roi (Syzygium malaccense (L.) Merr & Perry) 3.3 Diversity in stems form The results of investigation and analysis of diverse stem forms of Myrtaceae in 15 Ha Tinh Province with main stem forms, the results are presented in table 3.3 Table 3.3 Stem form of species of Myrtaceae in Ha Tinh Province No Stems form The large groups of trees (GOL) The average groups of trees (GOT) The small groups of trees (GON) The shrubs groups of trees (BUI) Total Species 11 16 26 10 62 Percentage (%) 17.46 25.40 41.27 15.87 100.00 Table 3.3 shows, in the four stems form, the large groups of trees with 11 species, (accounting for 17.46%) of the genera Eucalyptus and Syzygium; The average groups of trees with 16 species (accounting for 25.40%) mainly of the genera Cleistocalyx, Eucalyptus, Myrciaria Syzygium; the small groups of trees with 26 species (accounting for 41.27%) mainly of the genera Cleistocalyx, Melaleuca, Syzygium and the shrubs groups of trees with 10 specie (accounting for 15.87%) of the genera Feijoa, Baeckea, Decaspermum, Eugenia, Psidium, Rhodamnia Rhodomyrtus In four stems form is the small stems form and average stems form which are the predominant stems forms 3.4 Diverse in uses The use value of the tree groups of Myrtaceae is shown in table 3.4 Table 3.4 The uses of plants in Myrtaceae in Ha Tinh No Uses Plants for essential oils Plants for timber Plants for medicinal Plants for edible Plants for ornamental Plants for different uses Plants for tannin, dye Symbol CTD LGO THU ANQ CAN # TAN Species 63 48 32 25 11 Percentage (%) 100 76.19 50.79 39.68 17.46 11.11 6.35 Plants for essential oils: With 63 species for essential oils, accounting for 100% of the total species The topic currently has studied about essential oils of some species such as: Trâm vối đen (Cleistocalyx nigrans (Gagnep.) Merr & Perry), Trâm gân 16 mạng(Cleistocalyx retinervius Merr & Perry), Thập tử hoa nhỏ (Decaspermum parviflorum (Lamk.) J Scott.), Tràm gió (Melaleuca quinquenervia (Cav.) S.T.Blake), Sim rừng lớn (Rhodamnia dumetorum (Poir.) Merr & Perry), Đinh hương (Syzygium aromaticum (L.) Merr & Perry), Trâm bullock (Syzygium bullockii (Hance) Merry & Perry), Trâm hance (Syzygium hancei Merr & Perry), Trâm hoa dài (Syzygium lineatum (DC.) Merr & Perry), Trâm trắng (Syzygium tsoongii (Merr.) Merr & Perry) and Trâm tích lan (Syzygium zeylanicum (L.) DC.) Plants for timber: With 48 species are used to build housewares, boats, construction and tools, mainly belongs to the genus Syzygium with typical species such as: Trâm (Syzygium imitans Merr & Perry), Trâm núi (Syzygium levinei (Merr.)Merr & Perry), Trâm hoa dài (Syzygium lineatum (DC.) Merr & Perry), Trâm nam (Syzygium cochinchinensis (Gagnep.) Merr & Perry), Trâm hance (Syzygium hancei Merr & Perry),… Plants for medicinal: With 32 species of 11 genera that are used as health tonics, treating digestive diseases, burns, asthma, liver diseases, colds, typically: Ổi (Psidium guajava L.), Sim (Rhodomyrtus tomentosa (Ait.) Hassk.), Vối (Cleistocalyx operculatus (Roxb.) Merr & Perry), Chổi xể (Baeckea frutescens L.), Bạch đàn úc (Eucalyptus camaldulensis Dehnhart), Plants for edible: With 25 species, such as: Vối (Cleistocalyx operculatus (Roxb.) Merr & Perry), Ổi (Psidium guajava L.), Sim (Rhodomyrtus tomentosa (Ait.) Hassk.), Nho thân gỗ (Myrciaria cauliflora (Mart.) O.Berg), These are species that are being widely grown in different regions of the country Especially species such as: Ổi (Psidium guajava L.), Ổi dứa (Feijoa sellowiana (O.Berg.) O.Berg) and Nho thân gỗ (Myrciaria cauliflora (Mart.) O.Berg) Currently, these are widely popular fruit trees, fruits with very high vitamin C content, delicious taste, can be planted on an industrial scale for export, bringing high profits Plants for ornamental: With 11 species which are widely grown and traded in the market such as: Sắn thuyền (Syzygium polyanthum (Wight) Walp.), Roi (Syzygium 17 jambos (L.) Alston), Roi (Syzygium semarangense (Blume) Merr & Perry), Tràm đỏ (Callistemon citrinus (Curt.) Skeels), Nho thân gỗ (Myrciaria cauliflora (Mart.) O.Berg), Ổi cảnh (Psidium cujavillus Burm f.), Trâm vối ô (Cleistocalyx circumcissa (Gagnep.) Phamh.) and Trâm mùi (Syzygium oleinum Wall ex Walp) Plants for different uses: With species, in addition to the main uses mentioned above, it also has some of other uses such as: making firewood, making paper, making brooms, Plants for tannin, dye: With species including: Vối (Cleistocalyx operculatus (Roxb.) Merr & Perry), Trâm hance (Syzygium hancei Merr & Perry), Roi (Syzygium jambos (L.) Alston) and Trâm tích lan (Syzygium zeylanicum (L.) DC.) So, with 63 species with use value (accounting for 100%) of the total number of species distributed in Ha Tinh Province belonging to different groups; 63 species for essential oils (accounting for 100%), 48 species for timber plants (accounting for 76.19%), 32 species for medicinal plants (accounting for 50.79%), 25 species for edible (accounting for 39.68%), 11 species for ornamental plants (accounting for 17.46%), species for different uses (accounting for 11.11%) and species for tannin plants (accounting for 6.35%) To get more information and scientific bases to contribute to the orientation of conservation, planning, planting, exploitation and use of plant resources in Ha Tinh Province and in Vietnam We conducted a study on the biological characteristics of species in Myrtaceae in Ha Tinh Province 3.5 Characteristics of species in Myrtaceae in Ha Tinh Province This section describes the identification, ecology, distribution, sample and uses Research results have determined the coordinates of 63 species of Myrtaceae distributed in Ha Tinh, Simultaneously, samples were collected and described some biological characteristics (morphological, biological and ecological characteristics, distribution and use value) of 63 studied species The results of the study are the basic document on the taxonomy of plant species in 13 genera and 63 species of Myrtaceae, 18 contribute to add some data for the major of plant taxonomy 3.6 Chemical composition of essential oils of some species of Myrtaceae in Ha Tinh Province Results of analysis of 11 samples of leaf essential oil, 01 flower essential oil and 01 fruit essential oil sample of 11 species in Myrtaceae were synthesized (table 3.5) The yield of essential oils varied from 0.10% to 0.96% fresh weight Essential oils are usually pale yellow or pale green and are lighter than water The chemical components were determined to account for 74.47% - 100% of the total amount of essential oils The number of compounds identified ranged from 16 to 56 compounds, of which β-caryophyllene compounds were present in most essential oils of the species analyzed and accounted for a high proportion (ranging from 9.11% to 64.53%) Besides, it was also found that the essential oil of the leaves of the Rhodamnia dumetorum (Poir.) Merr & Perry) has a very high proportion of αpinene compounds (accounting for 90.47%) In the essential oils of the species analyzed are mainly monoterpenes and sesquiterpenes Table 3.5 Chemical composition of essential oils of some species of Myrtaceae in Ha Tinh Province Sample The main chemical Number of Content components account for compounds (%) more than 5% of the identified essential oil Trâm vối đen (Cleistocalyx Leaves nigrans (Gagnep.) Merr & Perry) n-octanal (18.05%), decanal (17.29%), 4Zdecanal (15.14%) Trâm gân mạng(Cleistocalyx Leaves retinervius Merr & Perry) No Species Thập tử hoa nhỏ (Decaspermum parviflorum Leaves 0.10 39 0.21 38 0.96 21 19 caryophyllene oxide (15.48%), myrcene (15.29%), βcaryophyllene) (8.90%), (E)-β-ocimene (8.39%) caryophyllene (43.98%), humulene (10.74%), copaene (8.27%), α- (Lamk.) J Scott) Tràm gió (Melaleuca quinquenervia (Cav.) S.T.Blake) Sim rừng lớn (Rhodamnia dumetorum (Poir.) Merr & Perry) Đinh hương (Syzygium aromaticum (L.) Merr & Perry) Trâm bullock (Syzygium bullockii (Hance) Merr & Perry) Trâm hance (Syzygium hancei Merr.& Perry) Flowers 0.66 32 Fruits 0.38 32 selinene (6.41%), eudesm-4(14), (11)diene (6.18%) γ-elemene (37.02%), caryophyllene (14.53%), ocimene (11.87%) caryophyllene (23.44%), eudesma-4(14), (11)-diene (17.46%), α-selinene (13.60%), humulene (7.15%) Leaves 0.69 38 1,8 cineole (22.69%), ocymene (14.81%), γterpinene (5.66%) Leaves 0.82 16 α-pinene (90.47%) Leaves 0.18 39 β-caryophyllene (42.53%), (E)-β-ocimene (19.38%), αhumulene (5.37%) Leaves 0.36 27 β-caryophyllene (49.65%) 0.15 50 γ-guaiene (11.07%), βcaryophyllene (9.11%), cis-calamenene (7.46%), αcopaene (6.97%), transcadina-1,4-diene (5.09%) 0.15 32 β-caryophyllene (64.53%), αpinene (6.14%) 46 β-caryophyllene (23.40%), bicyclogermacrene (21.23%), (Z)-β-ocimene (10.61%), αhumulene (6,33%) Leaves Trâm hoa dài (Syzygium Leaves lineatum (DC.) Merr & Perry) Trâm trắng (Syzygium tsoongii 10 Leaves (Merr.) Merr & Perry) 0.27 20 11 Trâm tích lan (Syzygium zeylanicum (L.) DC.) Leaves 0,12 56 bicyclogermacrene (25%), β-caryophyllene (20.14%), α-pinene (7.66%) 3.7 Antimicrobial activity of essential oils of some species in Myrtaceae Sample of leaves essential oil of Trâm trắng species (sample symbol: TQT-TD), leaves essential oil of Trâm bullock (sample symbol: TBL-TD) and leaves essential oil of Trâm tích lan (sample symbol: TTL-TD) which were tested for antibacterial activity against some strains of Gram (+) bacteria.: Enterococus faecalis (ATCC 299212), Staphylococcus aureus (ATCC 25923) and Bacillus cereus (ATCC 14579); Strains of Gram (-) bacteria: Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853) and Salmonella enteric (ATCC 13076); Nấm men train: Candida albicans (ATCC 10231) (table 3.6) Table 3.6 Results of determination of MIC and IC50 of leaf essential oil samples Minimum inhibitory concentration MIC (g/ml) No Samples Gram (+) bacteria Gram (-) bacteria E.faecalis S.aureus B.cereus E coli C.albicans P.aeruginosa S.enteric C.albicans TQT-TD 64 64 64 (-) (-) (-) 128 TBL-TD 32 32 64 128 (-) 256 64 TTL-TD 32 128 128 (-) (-) (-) 256 Giá trị IC50 (g/ml) TQT-TD 19.10 17.89 19.87 (-) (-) (-) 45.67 TBL-TD 10.23 6.78 29.78 46.67 (-) 120.78 28.78 TTL-TD 10.34 45.78 48.87 (-) (-) (-) 78.78 Notes: TQT-TD: Samples of essential oil from leaves of Trâm Trái Trăng species; TBL-TD: Samples of essential oil from leaves of Trâm bullock species; TTL-TD: Samples of essential oil from leaves of Trâm Tích Lan species; (-): Unknown The results in Table 3.6 show that the essential oils of the leaves of the studied species are also resistant to strains of Gram (+) bacteria, including: (Enterococus faecalis (ATCC 299212), Staphylococcus aureus (ATCC 25923) and Bacillus cereus (ATCC 14579)), in which sample TBL-TD has stronger resistance than the other two samples, reaching MIC = 21 32 g/ml (IC50 = 10.23 g/ml) for the strain Enterococus faecalis and MIC = 32 g/ml (IC50 = 6.78 g/ml) for the strain Staphylococcus aureus and reaching MIC = 64 g/ml (IC50 = 29.78 g/ml) for the strain Bacillus cereus In addition, this sample is also resistant to the strain Escherichia coli (MIC = 128 g/ml, IC50 = 46.67 g/ml) ) at the same time resistant to additional strain Candida albicans reaching MIC = 64 g/ml (IC50 = 28.78 g/ml) Besides, we see that the TQT-TD sample is not only resistant to strains of Gram (+) bacteria, but also resistant to Candida albicans strains and reaching MIC = 128 g/ml (IC50 = 45.67 g/ml) Sample TTL-TD in addition to being resistant to Enterococus faecalis strain at MIC = 32 g/ml (IC50 = 10.34 g/ml) they were also resistant to Candida albicans strain and reached MIC = 256 g/ml (IC50 = 78.78 g/ml) The results of this study show the potential for bioactivity of the essential oils of the three studied species to be used to treat a number of human diseases This result tells us why local people often use the leaves of these plants to treat some common diseases such as: Tuberculosis, toothache, hepatitis, diarrhea, dysentery, rheumatism, syphilis, worm treatment,….From the essential oils of the leaves of these three species can be applied to prepare some common medicines related to the pathogens tested above The research results have provided more information on the biological activities of the essential oils of the leaves of Trâm trắng, Trâm bullock and Trâm tích lan distributed in Ha Tinh With this result, the results of the study have evaluated the diversity of species composition and biological characteristics of species in Myrtaceae in Ha Tinh Province (morphological, biological and ecological characteristics, distribution, form) body and use value); at the same time analyze the chemical composition in the essential oils of leaves, flowers and fruits of some species in Myrtaceae; besides, the antibacterial activity of leaf essential oil of some species of this family was tested in Ha Tinh Province The results of the project are the basic documents on taxonomy of plant species in Myrtaceae in Ha Tinh, contributing to some additional data for the major of plant taxonomy; At the same time, it provides more information on natural compounds and antibacterial activities of some species in the family, as a scientific basis for orientation for further studies on chemistry, in order to 22 exploit and develop effectively develop, conserve and use biologically active ingredients from species in Myrtaceae in Ha Tinh Province In addition to the results achieved, due to limited funds and time constraints, we still have many problems that we cannot research, specifically: Research on the chemical composition of essential oils in different parts of species in Myrtaceae, from which there are data and scientific bases for a more complete assessment of their medicinal value and medicinal properties; Research on the activities of the main compounds in essential oils, at the same time, needs to study the unique combination of the main compounds and the combination of the main compounds with the minor compounds These studies offer hope that they may become the basis for new bioactives of commercial significance; Research on antiviral activity, antioxidant activity, inhibitory activity on the growth and development of cancer cells; studying and applying essential oils of species in Myrtaceae to the production of medicines to treat human diseases; The studies and overall assessment of essential oil reserves, economic value of some essential oils have potential for practical application, which can help managers in formulating strategies and making policies conservation, exploitation, green and sustainable development CONCLUSIONS AND SUGGESTIONS Conclusions Identified 63 species of 13 genera in Myrtaceae in Ha Tinh Province Of which the most diverse is the genus Syzygium with 31 species Additional 38 species and 09 genera in Myrtaceae distributed in Ha Tinh Province Recorded Feijoa genus with Feijoa sellowiana (O.Berg.) O.Berg species and Myrciaria genus with Myrciaria cauliflora (Mart.) O.Berg species have been added to the list of Vietnam Described some biological characteristics (morphological, biological, ecological, distribution and use) of 63 species All of the above species are useful: Plants for essential oils with 63 species, Plants for timber with 48 species, Plants for medicinal with 32 species, Plants for edible with 25 species, Plants for ornamental with 11 species, Plants for different uses with species and Plants for 23 tannin, dye with species The content and chemical composition have been determined in the essential oils of leaves of 11 species of 05 genera, essential oils of flowers and essential oils of fruits of species For the first time providing data on essential oils of leaves of 09 species of Myrtaceae For the first time providing data on essential oils of flowers and fruits of 01 species of Myrtaceae Tried the antibacterial activity of the essential oils of the leaves of the Trâm trắng, the Trâm bullock species and the Trâm tích lan species The results of the study showed that the essential oils of the leaves of all three species were resistant to Gram (+) strains of bacteria, including: Enterococus faecalis (ATCC 299212), Staphylococcus aureus (ATCC 25923) and Bacillus cereus (ATCC 14579 In addition, the essential oils of the leaves of Trâm bullock and Trâm trắng species also showed resistance to Candida albicans In particular, the essential oil of Trâm bullock species, in addition to the above abilities, is also resistant to the Escherichia coli strain (Gram (-)) and reached MIC = 128 g/ml (IC50 = 46.67 g/ml) Suggestions It is necessary to continue to study the chemical composition of essential oils in different parts of the species in Myrtaceae, on that basis, there are scientific bases to help more fully evaluate their medicinal value and medicinal properties Continue to further study the bioactivity of essential oils (antibacterial activity, antiviral activity, antioxidant activity, inhibitory activity on the growth and development of cancer cells, etc.) of the following species: Thập tử hoa nhỏ (Decaspermum parviflorum (Lamk.) J Scott), Đinh hương (Syzygium aromaticum (L.) Merr & Perry), Trâm bullock (Syzygium bullockii (Hance) Merry & Perry), Trâm hoa dài (Syzygium lineatum (DC.) Merr & Perry) and Sim rừng lớn (Rhodamnia dumetorum (Poir.) Merr & Perry) On that basis, studies on the application of essential oils of these species to the production of medicines for treating human diseases are carried out 24 PUBLISHED WORKS Khanh T H, Ban P H & Hoi T M (2020), Constituents of essential oils from the leaf, fruit, and flower of Decaspermum parviflorum (Lam) J Scott Arch Pharma Pract, 11 (1), 88-91 (ESCI) Tran Hau Khanh & Pham Hong Ban (2020), Analysis of Essential Oils from Leaf of Syzygium hancei Merr & Perry, Syzygium caryophyllatum (L.) Alston and Syzygium lineatum (DC.) Merr & Perry from Vietnam, Journal of Essential Oil Bearing Plants, 23 (3), 548-558 (SCIE) Tran Hau Khanh, Pham Hong Ban & Tran Minh Hoi (2020), Diversity of family Myrtaceae Juss 1789 in Ke Go Nature Resever, Ha Tinh Province, The Journal of Agriculture and Rural Development, Vol 12, 120-124 Tran Hau Khanh, Pham Hong Ban & Tran Minh Hoi (2020), Diversity on the family Myrtaceae of the Vu Quang National Park, Ha Tinh Province, Science Journal, Vinh university, Vol 2A (49), 30-37 Tran Hau Khanh, Pham Hong Ban, Tran Minh Hoi (2020), Chemical composition of essential oils of Rhodamnia dumetorum (Poir.) Merr & Perry and Syzygium zeylanicum (L.) DC., Vietnam Journal of Science and Technology, Vol 62 (11), 12-16 Tran Hau Khanh, Pham Hong Ban & Tran Minh Hoi (2020), Chemical composition of essential oils of Cleistocalyx nigrans and Cleistocalyx retinervius species collected in Vu Quang National Park, Ha Tinh Province, The Journal of Agriculture and Rural Development, Vol 21, 63-67 Tran Hau Khanh, Pham Hong Ban, Tran Minh Hoi (2021), Chemical Composition of Essential Oils from the Leaves of Syzygium Bullockii and Syzygium Tsoongii in Ke Go Nature Reserve, Ha Tinh Province, VNU Journal of Science: Natural Sciences and Technology, Vol 37 (2), 18-23 Tran Hau Khanh, Pham Hong Ban, Tran Minh Hoi, Nguyen Thị Ha (2021), Diversity of Family Myrtaceae Juss 1789 in Ha Tinh Province, VNU Journal of Science: Natural Sciences and Technology, Vol 37(4) 01-10 Tran Hau Khanh, Pham Hong Ban & Tran Minh Hoi (2021), Diversity of Myrtaceae Juss 1789 in the Ngan Pho river Watershed Biodiversity Conservation area, Ha Tinh Province, The Journal of Agriculture and Rural Development, Vol 21, 104-110