Research on genetic diversity of ngoc linh ginseng (panax vietnamensis ha et grushv) based on morphological traits and ssr markers

VIETNAM NATIONAL UNIVERSITY OF AGRICULTURE FACULTY OF BIOTECHNOLOGY - oOo - GRADUATION THESIS TITLE: RESEARCH ON GENETIC DIVERSITY OF NGOC LINH GINSENG (Panax vietnamensis Ha et Grushv.) BASED ON MORPHOLOGICAL TRAITS AND SSR MARKERS HANOI, 2022 VIETNAM NATIONAL UNIVERSITY OF AGRICULTURE FACULTY OF BIOTECHNOLOGY - oOo - GRADUATION THESIS TITLE: RESEARCH ON GENETIC DIVERSITY OF NGOC LINH GINSENG (Panax vietnamensis Ha et Grushv.) BASED ON MORPHOLOGICAL TRAITS AND SSR MARKERS Student name : Tran Huyen Trang Student code : 637435 Class : K63CNSHE Department : Biotechnology Supervisor : Associate Professor Dr Le Hung Linh Dr Nguyen Thi Thuy Hanh HANOI, 2022 COMMITMENT I hereby declare that this is my research project under the guidance of Associate Professor Dr Le Hung Linh - Department of Molecular Biology, Institute of Agricultural Genetics and Dr Nguyen Thi Thuy Hanh - Department of Biology, Vietnam National University of Agriculture The results and data in this thesis are truthful have not been published by anyone in any way I confirm that all information and data from articles and sources of other authors contain full citations and references from official sources I take full responsibility for this guarantee Hanoi, 05th December, 2022 Student Tran Huyen Trang i ACKNOWLEGEMENTS During the process of studying, researching and completing the thesis, I have received the help of many individuals First, I would like to express my thanks to the lecturer in the Faculty of Biotechnology of the Vietnam National University of Agriculture for their knowledge and dedication to conveying valuable knowledge to me during my time studying at the university Secondly, I am using this opportunity to express my deep and sincere gratitude to Associate Professor Dr Le Hung Linh - Department of Molecular Biology, Institute of Agricultural Genetics to and Dr Nguyen Thi Thuy Hanh Department of Biology, Vietnam National University of Agriculture, who took time out of their hectic schedules to listen, guide, and keep me on the right track Their passionate guidance aided me throughout the research and writing of this thesis I am also grateful to the Department of Molecular Biology, Institute of Agricultural Genetics, for technical support during implementation I would not forget to express special thanks to my Dr Khuat Thi Mai Luong and Ms Nguyen Thi Thuy Ngoan whose guidance, superior knowledge, and experience supported me during my study time at the thesis Last but not least, I am grateful to my family and my friends for supporting me spiritually throughout writing this thesis and my life in general I sincerely thank you! Hanoi, 05th December, 2022 Student Tran Huyen Trang ii CONTENTS COMMITMENT i ACKNOWLEGEMENTS .ii LIST OF TABLES v LIST OF FIGURES v LIST OF ABBREVIATIONS .vii ABSTRACT viii INTRODUCTION PART I LITERATURE REVIEW 1.1 Introduction about Panax genus 1.2 Introduction about Ngoc Linh ginseng 1.2.1 The original of Ngoc Linh ginseng 1.2.2 Natural features of Ngoc Linh ginseng 1.2.3 Vegetable traits, growth, and development of ginseng Ngoc Linh 1.2.4 Evaluation of Genetic Diversity of Plant Species – Ginseng PART MATERIAL AND METHODS 14 2.1 Material 14 2.2 Methods 14 2.2.1 Morphological Evaluation 14 2.2.2 Molecular Method 14 2.2.3 Phylogenetic trees 19 PART RESULTS 19 3.1 Evaluation of morphological traits of Ngoc Linh Ginseng 20 3.2 Building a morphological table of Ngoc Linh Ginseng 23 3.3 Build a phylogenetic tree based on the data encryption table and UPGMA subgroup type 29 3.4 Evaluation of genetic relationships of 24 Ngoc Linh ginseng samples by molecular markers 31 PART DISCUSSION AND SUGGESTION 39 iii DISCUSSION 44 SUGGESTION 39 REFERENCES 40 APPENDIX 45 iv LIST OF TABLES Table 1.1: Panax species in the world Table 2.1: List SSR markers 16 Table 2.2: PCR reaction components 17 Table 2.3: 8% Polyacrylamide gel components 18 Table 3.1: Classification of morphology describing and Encoding table of Ngoc Linh ginseng leaves over years-old 23 Table 3.2: The morphological evaluation traits of subgroups in 120 individuals of Ngoc Linh ginseng 30 Table 3.3: Total band and polymorphisms of SSR markers 34 Table 3.4: Summary of the results of evaluating the homology between 24 samples of Ngoc Linh ginseng 38 v LIST OF FIGURES Figure 3.1 Leaves and petioles color of Ngoc Linh ginseng 21 Figure 3.2 Inflorescence morphology and inflorescence bloom 22 Figure 3.3: Ngoc Linh ginseng seed 23 Figure 3.4 Shape of seeds of Ngoc Linh ginseng 23 Figure 3.5: Phylogenetic tree showing the relationship of 120 individuals of Ngoc Linh ginseng based on Euclidean distance A, B, C are subgroups 29 Figure 3.6 : The results of a survey of 28 molecular markers in two representatives of Ngoc Linh ginseng 32 Figure 3.7 : Results PCR of 24 samples of Ngoc Linh ginseng using primer, 1kb ladder 33 Figure 3.8 Aphylogenetic tree describes the genetic diversity relationship between24samples of Ngoc Linh ginseng 35 vi LIST OF ABBREVIATIONS Abbreviation Abbreviation PCR Polymerase chain reaction SSR Simple-sequence repeat µl Microliter ml Milliliter bp Base pairs DNA Deoxyribonucleic Acid vii ABSTRACT The most recently identified Panax species is Ngoc Linh Ginseng (Panax vietnamensis Ha et Grushv) Plants are renowned in Vietnam for their medical properties and are primarily employed in medicine (Le et al 2015) Ngoc Linh ginseng has been overexploited and is practically extinct in the wild due to its beneficial effects on human health and high commercial worth The genetic diversity of Ngoc Linh ginseng was assessed using morphological traits and SSR markers in this study This is to maintain, select, and produce new kinds as well as proper propagation methods to satisfy the needs of the pharmaceutical processing industry's development The first step was to evaluate 15 morphological traits of 120 Ngoc Linh ginseng individuals, create a trait coding table in Excel, and create a taxonomy tree in NTSYSpc 2.1 The second step, 24 individuals were chosen based on the stem length, petiole length, and flower stalk length parameters The genetic diversity of the 24 Ngoc Linh ginseng samples was assessed using eight SSR molecular markers in the second stage This suggests that some samples had morphological and genetic similarities Keywords: Ngoc Linh ginseng, morphological traits, genetic diversity, molecular marker viii homology of 0.63, 24 samples of Ngoc Linh ginseng were separated into three branches from a single original branch The first branch has two Ngoc Linh ginseng samples (1 and 119), whereas the second branch contains 17 Ngoc Linh ginseng samples (2, 6, 114, 152, 151, 397, 180, 187, 337, 497, 276, 272, 108, 205, 262, 194 and 213) The third branch contained the final five Ngoc Linh ginseng samples, 11, 233, 19, 53, and 94 When evaluating the genetic diversity of 24 samples of Ngoc Linh ginseng based on morphological and molecular traits, it was found that sample belonged to group A and branched separately from the rest of the ginseng samples Group contained 11/17 ginseng samples belonging to subgroup B, including samples 2, 108, 152, 187, 194, 205, 213, 272, 276, 337, and 297 Group had only 19 samples belonging to subgroup C It shows that there are some genetic similarities in both morphology and molecule Evaluation of the diversity and genetic distance of 24 ginseng samples from Ngoc Linh provided the most general view of the kinship relationship between the varieties The results of the molecular genetic evaluation of 24 samples of Ngoc Linh ginseng varieties are shown in Table 3.5 According to row 24 in table 3.5, the homology of Ngoc Linh ginseng samples Sample 497 with the remaining 23 ginseng samples oscillating from 0,41 – 0,74; the highest with ginseng sample number 337 (reaching 0,74); the lowest with ginseng sample 1(0,41) Sample 397 with the remaining 22 ginseng samples oscillating from 0,49 – 0,71; the highest with ginseng sample numbers 337 and 187 (reaching 0,71); the lowest with ginseng sample 119 (0,49) Sample 337 with the remaining 21 ginseng samples oscillating from 0,56 – 0,77; the highest with ginseng sample numbers 276 and 272 (reaching 0,77); the lowest with ginseng sample 119 (0,56) Sample 276 with the remaining 20 ginseng samples oscillating from 0,49 – 0,71; the highest with ginseng sample number 205 (reaching 0.71), the lowest with ginseng sample 108 (0,49) Sample 272 with the remaining 19 36 ginseng samples oscillating from 0,49 – 0,74, the highest with ginseng sample number (reaching 0,74); the lowest with ginseng sample 119 (0,49) Sample 262 with the remaining 18 ginseng samples oscillating from 0,49 – 0,77; the highest with ginseng sample number 205 (reaching 0,77); the lowest with ginseng sample 119 (0,49) Sample 233 with the remaining 17 ginseng samples oscillating from 0,55 – 0,77; the highest with ginseng sample number (reaching 0,77); the lowest with ginseng sample 119 (0,55) Sample 213 with the remaining 16 ginseng samples oscillating from 0,43 – 0,77; the highest with ginseng sample numbers 194 and (reaching 0,77); the lowest with ginseng sample 119 (0,43) Sample 205 with the remaining 15 ginseng samples oscillating from 0,47 – 0,77; the highest with ginseng sample numbers 180 and 94 (reaching 0,77); the lowest with ginseng sample 119 (0,47) Sample 194 with the remaining 14 ginseng samples oscillating from 0,47 – 0,76; the highest with ginseng sample number (reaching 0,76); the lowest with ginseng sample 119 (0,47) Sample 187 with the remaining 13 ginseng samples oscillating from 0,58 – 0,8; the highest with ginseng sample number 152 (reaching 0,8); the lowest with ginseng sample 53 (0,58) Sample 152 with the remaining 12 ginseng samples oscillating from 0,46 – 0,77; the highest with ginseng sample number (reaching 0,77); the lowest with ginseng sample 119 (0,46) Sample 151 with the remaining 11 ginseng samples oscillating from 0,55 – 0,83; the highest with ginseng sample number 180 (reaching 0,83), the lowest with ginseng sample (0,55) Sample 180 with the remaining 10 ginseng samples oscillating from 0,5 – 0,79; the highest with ginseng sample number 94 (reaching 0,79); the lowest with ginseng sample (0,5) Sample 119 with the remaining ginseng samples oscillating from 0,53 – 0,68; the highest with ginseng sample numbers and 11 (reaching 0,68); the lowest with ginseng sample (0,53) Sample 114 with the remaining ginseng samples oscillating from 0,52 – 0,8; the highest with ginseng sample number 108 (reaching 0,8); the lowest with ginseng sample 11 (0,52) Sample 108 with the remaining ginseng samples 37 oscillating from 0,53 – 0,73; the highest with ginseng sample number 19 (reaching 0,73); the lowest with ginseng sample (0,53) Sample 94 with the remaining ginseng samples oscillating from 0,53 – 0,8; the highest with ginseng sample number 53 (reaching 0,8); the lowest with ginseng sample (0,53) Sample 53 with the remaining ginseng samples oscillating from 0,52 – 0,83; the highest with ginseng sample number 19 (reaching 0,83); the lowest with ginseng sample (0,52) Sample 19 with the remaining ginseng samples oscillating from 0,5 – 0,77; the highest with ginseng sample number (reaching 0,77); the lowest with ginseng sample (0,5) Sample 11 with the remaining ginseng samples oscillating from 0,61- 0,73; the highest with ginseng sample number (reaching 0,73); the lowest with ginseng sample (0,61) Sample with the remaining ginseng samples oscillating from 0,61 – 0,73; the highest with ginseng sample number (reaching 0,73); the lowest with ginseng sample (0,61) Table 3.4: Summary of the results of evaluating the homology between 24 samples of Ngoc Linh ginseng 38 PART CONCLUSION AND SUGGESTION CONCLUSION 15 morphological traits of stems, leaves, flowers, and fruits were evaluated When constructing phylogenetic trees, samples of the Ngoc Linh ginseng variety were divided into three subgroups: Subgroup A had 19 members, while Subgroup B had 85 members and Subgroup C had the remaining 16 members The main stem length, leaflet length, and peduncle length of the Ngoc Linh ginseng breeds tested were quite diverse, and can be utilized as various morphological criteria to evaluate and split plant kinds into subgroups 24 samples of Ngoc Linh ginseng were prepared using SSR molecular markers on an 8% polyacrylamide gel The results showed homology from 0.55 to 0.86 At 65% homology, 24 ginseng samples were divided into groups Group included two individual samples and 119, in which these individuals belong to subgroup A in morphology, which is different from the rest Group included 17 individuals, of which 11/17 belonged to subgroup B in morphology, including samples 2, 108, 152, 187, 194, 205, 213, 272, 276, 337, and 297 Group included samples, but only sample 19 belonged to Subgroup C in terms of morphology This showed that some samples were genetically similar in both morphology and molecules SUGGESTION Further research with multiple SSR primers and other DNA techniques is needed for a good data set, and sources - selecting new varieties and developing high-quality plants for production 39 REFERENCES References in English Bang K.H., et al (2012) Variations of agronomic traits of cultivars and breeding lines in Korean ginseng (Panax ginseng C.A Mey.) Korean J Med Crop Sci., 20 (4), pp 231237 Bijlsma, P., Van Arendonk, J.A.M And Woolliam, J.A., 2000 A general procedure for predict rates of inbreeding in populations undergoing mass selection Genetics, vol 154, no 4, pp 1865-1877 https://doi.org/10.1093/genetics/154.4.1865 Bijlsma, R., Bundgaard, J., Boerema, A.C And Van Putten, W.F., 1997 Genetics and environmental stress, and the persistence of populations In: R Bijlsma and V Loeschcke, eds Environmental stress, adaptation and evolution Basel: Birkhauser Verlag, pp 193207 http://dx.doi.org/10.1007/978-3-0348-8882-0_11 Bruford, M.W., Davies, N., Dulloo, M.E., Faith, D.P and Walters, M., 2017 Monitoring changes in genetic diversity In: M Walters and R.J Scholes, eds The GEO handbook on biodiversity observation networks New York: Springer, pp 107-128 http://dx.doi.org/10.1007/978-3-319-27288-7_5 Chien, H.X., Tai, N.T., Truc, N.B., Tinh, T.X., Thao, L.B., Luan, T.C And Nhut, D.T., 2011 Factors effecting in vitro microrhizome formation of Panax vietnamensis Ha et Grushv and quantitation of saponin content of there from generated plantlets grown in Ngoc Linh Mountain Journal of Biotechnology, vol 9, pp 317-331 Choi K.T (2008) Botanical traits, pharmacological effects and medicinal components of Korean Panax ginseng C A Meyer Acta Pharmacologica Sinica, 29(9), 1109–1118 Christensen, R K., & Gazley, B (2008) Capacity for public administration: analysis of meaning and measurement Public Administration and Development, 28(4), 265– 279 doi:10.1002/pad.500 Elena A V., Iury Yu A., Reunova G D., T P T Nguyen, Yuri N Z., 2018 A comparative Analysis of genetic Variability and differentiation in Panax vietnamensis Ha et Grushv and P gienseng C.A Meyer using ISSR markers Russian journal of genetics, 54(2): 262−265 Flower, C.E., Fant, J.B., Hoban, S., Knight, K.S., Steger, L., Aubihl, E., Gonzalez-Meler, M.A., Forry, S., Hille, A And Royo, A.A., 2018 Optimizing conservation strategies for a threatened tree species: in situ conservation of white ash (Fraximus americana L.) genetic 40 diversity through insecticide treatment Forests, vol 9, no 4, p 202 http://dx.doi.org/10.3390/f9040202 Frankham, R., Bailou, J.D And Briscoe, D.A., 2002 Introduction to conservation genetics Cambridge: Cambridge Univeristy Press Galina D Reunova, Irina L Kats, Tamara I Muzarok, Trang N T P, The Dang Tat, Yuri N Zhuravlev, 2010 Genetic diversity of Panax ginseng C A Meyer, inferred from amplified fragment length polymorphism markers” Gijbels, P., Hert, K.D., Jacquemyn, H And Honnay, O., 2015 Reduced fecundity and genetic diversity in small populations of rewarding versus deceptive orchid species: a metaanalysis Ecology and Evolution, vol 148, pp 153-159 Grazina, L., Amaral, J S., Costa, J., & Mafra, I (2021) Towards authentication of Korean ginseng-containing foods: Differentiation of five Panax species by a novel diagnostic tool LWT, 151, 112211 Ha, T.D And Grushvitzky, I.V., 1985 New species in Panax (Araliaceae) in Vietnam Le Journal de Botanique, vol 70, pp 518-522 Hammer, K and KHOSHBAKHT, K., 2005 Towards a ‘red list’ for crop plant species Genetic Resources and Crop Evolution, vol 52, no 3, pp 249-265 http://dx.doi.org/10.1007/s10722-004-7550-6 Hamrick, J.L and GODT, M.J.W., 1996a Conservation genetics of endemic plant species In: J.S AVISE and J.L HAMRICK, eds Conservation genetics: case histories from nature New York: Chapman and Hall, pp 281-304 http://dx.doi.org/10.1007/978-1-47572504-9_9 Hamrick, J.L., Godt, M.J.W And Sherman-Broyles, S.L., 1992 Factors influencing levels of genetic diversity in woody plant species New Forests, vol 6, no 1-4, pp 95-124 http://dx.doi.org/10.1007/BF00120641 International union for the protection of new varieties of plants (UPOV) Ginseng.; 2017 Jo I.H., Lee S.H., Kim Y.C., Kim D.H., Kim H.S., Kim K.H., Chung J.W., Bang K.H (2015) De novo transcriptome assembly and the identification of gene-associated singlenucleotide polymorphism markers in Asian and American ginseng roots Mol Genet Genomics, 290(3):1055–1065 Khan, S., Tosun, A., & Kim, Y S (2015) Ginsenosides as Food Supplements and Their Potential Role in Immunological and Neurodegenerative Disorders Bioactive Nutraceuticals and Dietary Supplements in Neurological and Brain Disease, 303–309 41 Kim J., Jo B H., Lee K L., Yoon E S., Ryu G H., Chung K W., 2007 Identification of new microsatellite markers in Panax ginseng Molecules and Cell, 24: 60−68 Kim S., Park K (2003) Effects of Panax ginseng extract on lipid metabolism in humans Pharmacol Res., 48(5):511–513 Laurance, W.F., 2004 Forest-climate interactions in fragmented tropical landscapes Philosophical Transactions of the Royal Society of London Series B, Biological Sciences, vol 359, no 1443, pp 345-352 http://dx.doi.org/10.1098/rstb.2003.1430 PMid:15212089 Le T.H., Lee G.J., Vu H.K., Kwon S.W., Nguyen N.K., Park J.H., Nguyen M.D (2015) Ginseng saponins in different parts of Panax vietnamensis Chem Pharm Bull, 63 (11): 950-954 Liu, H., Xia, T., Zuo, Y.J., Chen, Z.J And Zhou, S.L., 2011 Development and characterization of microsatellite markers for Panax notoginseng (Araliaceae), a Chinese traditional herb American Journal of Botany, vol 98, no 8, pp e218-e220 http://dx.doi.org/10.3732/ajb.1100043 PMid:21821584 Ministry of Science and Technology And Vietnam Academy Of Science And Technology, 2007 Red book of Vietnam Part II - plants Hanoi, Vietnam: House for Science and Technology Nguyen Thi Hong Mai, Le Thanh Son, Nguyen Thi Phuong Trang, Genetic traits of Panax vietnamensis Ha & Grushv Populations based on SSR, Tap chi sinh hoc 2020, 42(1): 11– 19 Reed, D.H And Frankham, R., 2003 Correlation between fitness and genetic diversity Conservation Biology, vol 17, no 1, pp 230-237 http://dx.doi.org/10.1046/j.15231739.2003.01236.x Reunova, G.D., Koren, O.G., Muzarok, T.I And Zhuravlev, Y.N., 2014 Microsatellite analysis of Panax ginseng natural population in Russia Chinese Medicine, vol 5, no 4, pp 231-243 http://dx.doi.org/10.4236/cm.2014.54028 Ryu K.R., Yeom M H., Kwon S.S., Rho H.S., Kim D.H., Kim H.K., Yun K.W (2012) Influence of air temperature on the histological traits of ginseng (Panax ginseng C A Meyer) in six regions of Korea AJCS 6(12): 1637 – 1641 Sebbenn, A.M., Licona, J.C., Mostacedo, B And Degen, B., 2012 Gene flow in an overexploited population of Swietenia macrophylla King (Meliaceae) in the Bolivian 42 Amazon Silvae Genetica, vol 61, no 1-6, pp 212-220 http://dx.doi.org/10.1515/sg-20120027 Sharma, S.K & Pandit, M.K (2011) A morphometric analysis and taxonomic study of Panax bipinnatifidus Seem (Araliaceae) species complex form Sikkim Himalaya, India Plant System Evolution 297: 87–98 Templeton, A.R., Shaw, K., Routman, E And Davis, S.K., 1990 The genetic consequences of habitat fragmentation Annals of the Missouri Botanical Garden, vol 77, no 1, pp 13-27 http://dx.doi.org/10.2307/2399621 Tran, Q.L., Adnyana, I.K., Tezuka, Y., Harimaya, Y., Saiki, I., Kurashige, Y., Tran, Q.K And Kadota, S., 2002 Hepatoprotective effect of majonoside R2, the major saponin from Vietnamese ginseng (Panax vietnamensis) Planta Medica, vol 68, no 5, pp 402-406 http://dx.doi.org/10.1055/s-2002-32069 PMid:12058314 Van Dan, N., Ramchiary, N., Choi, S.R., Uhm, T.S., Yang, T.-J., Ahn, I.-O And Lim, Y.P., 2010 Development and characterization of new microsatellite markers in Panax ginseng (C.A Meyer) from BAC end sequences Conservation Genetics, vol 11, no 3, pp 1223-1225 http://dx.doi.org/10.1007/s10592-009-9924-y Wen, J (2001) Species diversity, nomenclature, phylogeny, biogeography, and classification of the ginseng genus (Panax L., Araliaceae), In: Punja, Z.K (Ed.) Utilization of biotechnological, genetic and cultural approaches for north American and Asian Ginseng improvement: Proceedings of the International Ginseng Workshop Simon Fraser University Press, Vancouver, pp 67–88 Xiang, Q.B & Lowry, P.P Araliaceae, In: Wu, C.Y., Rawen, P.H & Hong (2007) 43 References in Vietnamese Khuất Thi Mai Lương, Nguyễn Thi Minh Nguyệt, Chu Đức Hà, Trần Thị Hoa Mỹ, Đinh Văn Phê, Lê Hùng Lĩnh, Tạp chí Khoa học Cơng nghệ Nơng nghiệp Việt Nam - Số 11(96)/2018 Lê Hùng Lĩnh, Báo cáo tổng hợp kết quả dề tài, Nghiên cứu xây dựng thị phân tử phục vụ giám định, khai thác phát triển Sâm Ngọc Linh (Panax vietnamensis), MÃ SỐ: ĐTĐL.CN-29/16 Lê Thanh Sơn và Nguyễn Tập (2006), Những đặc điểm sinh thái bản Sâm Ngọc Linh Tạp chí dược liệu, tập 11, số 4, trang 145 – 147 Nguyễn Thượng Dong, Trần Công Luận Nguyễn Thị Thu Hương (2007) Sâm Việt Nam số thuốc họ nhân sâm Nhà xuất bản Khoa học Kỹ thuật Nguyễn Văn Đạt, Trần Thị Phương Anh, Vũ Tiến Chính, Phan Kế Long, Hồng Lê Tuấn Anh Chi Sâm – Panax L (Họ Ngũ Gia Bì - Araliaceae) Ở Việt Nam) Pham Thi Ngoc, Pham Thanh Huyen, Nguyen Quynh Nga, Phan Van Truong, Nguyen Minh Khoi, Dinh Doan Long, A Molecular Phylogeny of Panax L Genus (Araliaceae) Based on ITS-rDNA and matK Support for Identification of Panax Species in Vietnam, VNU Journal of Science: Medical and Pharmaceutical Sciences, 2020; 91-99 Trương Thị Hồng Hải, Dương Thanh Thủy, Đặng Thanh Long, Hồ Thị Huyền Trân, Nguyễn Mạnh Tuấn, Đa Dạng Di Truyền Dựa Trên Đặc Điểm Hình Thái Của Quần Thể Sâm Ngọc Linh (Panax Vietnamensis Ha et Grushv.) Ở Nam Trà My, Quảng Nam, Tạp trí khoa học Đại học Huế, Tập 127, số 1C, 2018 tr.203-210 Trương Thị Hồng Hải, Trần Viết Thắng, Nguyễn Đức Phước, Võ Văn Tư, Ngiên cứu xây dựng bảng mơ tả tính trạng sâm Ngọc Linh huyện Nam Trà My, Quảng Nam, Tạp trí khoa học Đại học Huế, Tập 127, số 3A, 2018, Tr.19-36 44 APPENDIX APPENDIX 1: Summary table of the traits of 120 individuals of Ngoc Linh ginseng by Excel 2010 Morphology Ngoc Linh ginseng seed garden in Kon Tum supplied a raw data sheet in Excel on how to measure 120 samples (UPOV - International Union for the Protection of New Varieties of Plants) + Length of main stem: from the root to the petiole + Count the number of leaflet and stipule + Stem color, level of purple on the stem, petiole color, distribution of purple color on the stem + Leaf shape, serrated edge 45 + Measure size of leaflet: Length: measure from petiole to leaf top; Width: measure the widest part (between leaf) of the leaf + Measure the length of the petiole: measured from the transition between the stem and the leaf blade + Measure the length of peduncle: measured from the transition between the petiole and the peduncle + Inflorescence shape, flowering time, fruit shape 46 47 Subgroup A 48 Subgroup C Subgroup B 49 APPENDIX 2: Experiment 50