The first report of myxomycetes amoebozoa in thai nguyen city northern vietnam rapid biodiversity assessment using tools in modern molecular era

THAI NGUYEN UNIVERSITY UNIVERSITY OF AGRICULTURE AND FORESTRY JOHN CARLO REDEÑA SANTOS THE FIRST REPORT OF MYXOMYCETES (AMOEBOZOA) IN THAI NGUYEN CITY, NORTHERN VIETNAM: RAPID BIODIVERSITY ASSESSMENT USING TOOLS IN MODERN MOLECULAR ERA BACHELOR THESIS Study Mode: Major: Faculty: Batch: Full-time Environmental Science and Management International Programs Office K45-AEP Thai Nguyen, 20/11/2017 Thai Nguyen University of Agriculture and Forestry Degree Program Bachelor of Science in Environmental Science and Management Student name Student ID Thesis Title Supervisor (s) Abstract: John Carlo Redeña Santos DTN1454290051 The first report of Myxomycetes (Amoebozoa) in Thai Nguyen City, Northern Vietnam: Rapid biodiversity assessment using tools in modern molecular era Dr Duong Van Thao & Dr Nikki Heherson A Dagamac Myxomycetes are phagotrophic, fungus-like protists abundantly occurring as microbial predators in soil ecosystems However, in spite of their cosmopolitan distributions, limited studies were conducted in Southeast Asia In particular, gaps are found in Northern Vietnam which has no single reported studies that annotates myxomycetes In order to fill the missing voids, a three-phase rapid biodiversity assessment employing purely moist chamber cultures was carried out in Thai Nguyen City, Northern Vietnam using (i) traditional taxonomic survey, (ii) classical diversity measurement and estimation utilizing sophisticated statistical programming pipelines and (iii) modern molecular technique using DNA barcoding (partial 18s rRNA gene) technology These efforts showed: (1) agricultural plantations with specified leaf textural morphology are good microhabitat for leaflitter inhabiting myxomycetes, (2) protected and unprotected forests shares myxomycetes assemblages, (3) anthropogenic activities plays a role in spore dispersal, (4) confirmation that both at community and genetic level, dispersal ecology of myxomycetes are not affected by management strategies or forest type, ii and (5) 54 species newly recorded for Northern Vietnam, of which are new records for the whole country, with possible species new to science As such, this study provided the very first baseline information on the biodiversity of myxomycetes for Northern Vietnam Keywords α and β diversity, barcoding, dispersal, forest management, slime molds, tropical plants Number of pages 143 pages Date of Submission September 25, 2017 Supervisor‘s signature iii ACKNOWLEDGEMENT This body of work was made possible by a lot of people who participated in this endeavor in very different but significant ways Firstly, I would like to acknowledge my whole family for the unconditional support, especially to my parents (Nanay Annie & Tatay Jun), to whom I am beyond grateful as they invested not just money but also time and sweat for my education To my brothers (PJ, Paulo, Third) and my sister (Darla) in blood, a humongous shout-outs to the four of you I would also like to include a special note of ―hindi matatawarang pasasalamat‖ to Dr Nikki Heherson A Dagamac for not just being an adviser and fostering me for three months, but also for being my mentor in every aspect of life Without his untiring efforts and never-ending patience, I would not have finished this beautiful piece of work I would also like to express my gratitude to Prof Dr Martin Schnittler (University of Greifswald), Prof Dr Wilhelm Steingrube (SusEnMan Project, University of Greifswald) and Frau Katharina Schmitt (International Office, University of Greifswald) for the research scholarship, internship and field collection fundings, and to Dr Duong Van Thao, Frau Anja Klahr (Laboratory coordinator, Allgemeine und Spezielle Botanik), Dr Manuela Bog, Dr Hoang Hai Thanh, Dr Nguyen Dang Cuong (TUAF), Oriana Sanchez-Mahecha and Nguyen Thuy Linh for the technical assistance Big thanks to my friends (Nicole, Jose, Hong, Thao, Keraia, Pons, Tina, Colleene, Kenneth, Alex, Mishel, Kathleen, Jamba, Bojo, Tintin, CJ, Edwin) for the personal support, and to my Greifswald friends (Rafa, Kasia, Kemani, Oleg, Tim, Lukas, iv Franzi, Jinny, Carlos, Brook, Christian, Paul, Sabine, Lorna, Dani, Julia) for keeping me stable during my research and internship in Germany Lastly, a very special mention to Ikkin, Ward and William for the Pizza party and Jenga session we shared during my stay in Germany v TABLE OF CONTENTS List of Figures List of Tables 10 List of Abbreviations 11 Part I Introduction 12 1.1 Research Rationale 12 1.2 Research Questions and Hypotheses 15 1.2.1 Occurrence of myxomycetes in homogenous vegetation 15 1.2.2 α and β diversity in a heterogeneous vegetation 16 1.2.3 Genetic diversity 17 1.3 Research Objectives 18 1.4 Scope and Limitations 20 1.5 Definition of terms 21 Part II Literature Review 25 2.1 Natural Classification history of myxomycetes 25 2.2 Economic and ecological importance 26 2.3 Cutting edge technologies applied in studying the diversity and ecology of myxomycetes 31 Part III Methodology 37 3.1 Occurrence of myxomycetes in homogenous vegetation 37 3.1.1 Collection of substrates and preparation of moist chambers 37 3.1.2 Characterization and identification of fruiting bodies 38 3.1.3 Calculation of productivity and myxomycete occurrence 39 3.2 α and β diversity in a heterogeneous vegetation 40 3.2.1 Collecting localities 40 3.2.2 Collection of substrates, laboratory isolation method and myxomycetes characterization 41 3.2.3 Data evaluation 43 3.3 Genetic diversity 46 3.3.1 Specimen acquisition 46 vi 3.3.2 DNA Extraction, amplification and sequencing 47 3.3.3 Sequence alignment, tree construction and genetic analysis 49 Part IV Results and Discussion 51 4.1 Occurrence of myxomycetes in homogenous vegetation 51 4.1.1 Results 51 4.1.2 Discussion 55 4.2 α and β diversity in a heterogeneous vegetation 57 4.2.1 Results 58 4.2.2 Discussion 73 4.3 Genetic diversity 78 4.3.1 Results 78 4.3.2 Discussion 80 PART V Conclusion 83 References 88 Appendices 104 vii LIST OF FIGURES Figure 1: The study area: Thai Nguyen City, Thai Nguyen Province showing the sampling location of the three agricultural plantations 37 Figure 2: Schematic diagram for the methodology of the first phase of this research study 40 Figure 3: Sampling locations for the two community forests in Thai Nguyen City, Thai Nguyen Province 41 Figure 4: Schematic diagram for the methodology of the second phase of this research study 46 Figure 5: Schematic diagram for the methodology of the third phase of this research study 50 Figure 6: The 2D bar graph shows (a) percentages of positive moist chambers between the aerial (AL) and ground leaf (GL) litter substrates in three different agricultural plantations and (b) percentages of positive moist chambers between Superorder Lucisporidia (bright-spored) and Superorder Fuscisporidia (dark-spored) in three different agricultural plantations 51 Figure 7: (a) Rarefaction curves for two different forest type and generated species accumulation curves from Chao estimator for the (b) both community forests; (c) for protected forest; (d) for unprotected forest .58 Figure 8: Box plot showing the comparison of six different diversity indices (Alpha = Fisher‘s alpha; Shannon = Shannon‘s H index; Simpson = Simpson‘s diversity index; N0 = species richness only, N1 = exponent of the Shannon diversity, N2= inverse of the Simpson diversity) 71 Figure 9: Non-metric multidimensional scaling (NMDS) of species occurrence between two community forests Black dots represent the position of myxomycetes species in the ordination space Colored circles represent the forest type; colored ellipses denote dispersion based on standard deviation of point scores 72 Figure 10: Phylogenetic tree of Diderma hemisphaericum: based on partial SSU sequences of the 13 successfully amplified specimens .79 LIST OF TABLES Table 1: Occurrence of myxomycetes: The table shows the list of myxomycetes and their average pH ±, - max values as measured on every positive moist chamber A species recorded as rare (1 record) uses only the pH value of the sole moist chamber where it was found The color on the left side of the species indicates their Superorder (yellow=bright-spored, Superorder Lucisporidia; brown=dark-spored, Superorder Fucisporidia) The abundance index (AI) in accordance to Stephenson et al (2013), and their total number of records is further reported on this table The distribution of records for substrates (AL or GL) and agricultural plants are also enumerated .52 Table 2: Computed values using Analysis of Molecular Variance 80 10 45 45 22 18 25 22 12 12 43 17 43 17 41 26 46 46 22 18 26 22 12 12 44 16 43 16 42 27 10 10 10 47 47 22 18 26 22 12 12 43 15 43 15 41 27 48 48 22 19 26 22 12 12 43 14 43 14 41 27 49 49 23 19 26 23 12 12 43 14 43 14 42 27 98 10 50 50 23 19 27 23 12 12 43 13 43 13 42 28 51 51 23 19 27 23 12 12 44 13 44 13 43 28 10 10 52 52 23 20 27 23 12 12 44 13 44 13 42 28 53 53 24 20 27 24 12 12 44 13 44 12 42 54 54 24 20 28 24 13 13 45 12 45 12 55 55 24 20 28 24 13 13 45 12 45 56 56 24 21 28 25 13 13 46 11 46 16 41 26 100 16 33 40 27 41 40 22 17 17 16 42 27 104 16 34 41 27 41 41 22 17 17 16 41 27 100 16 34 41 27 41 41 22 17 17 15 41 27 98 15 34 41 27 41 41 22 17 17 16 42 27 102 16 34 41 28 41 41 23 17 17 16 42 28 103 16 35 41 28 41 41 23 16 17 16 43 28 103 16 35 42 28 41 41 23 17 17 99 15 42 28 99 15 35 42 29 41 41 23 17 17 28 98 15 42 28 98 15 36 43 29 42 42 24 17 18 43 29 15 43 29 99 15 37 44 29 42 42 24 17 18 12 43 29 99 10 15 43 29 100 15 37 44 30 42 42 24 17 18 11 43 29 99 15 43 29 99 15 37 44 30 42 42 24 17 18 57 57 25 21 28 25 13 13 46 12 46 12 43 30 96 14 43 30 96 14 38 45 30 42 42 25 17 18 58 58 25 21 29 25 13 13 46 13 46 13 43 30 94 14 43 30 94 14 38 45 31 42 42 25 17 18 59 59 25 21 29 25 13 13 46 13 46 13 44 30 93 14 44 30 93 14 38 45 31 42 42 25 17 18 60 60 25 22 29 25 13 13 46 13 46 13 43 30 92 14 43 30 92 14 38 45 31 42 43 25 17 18 61 61 26 22 29 26 13 13 46 13 46 13 44 31 94 14 44 31 94 14 38 45 31 42 43 25 17 19 62 62 26 22 29 26 13 13 46 13 46 13 43 31 91 13 43 31 91 13 38 45 31 43 43 26 17 3 19 63 63 26 22 30 26 13 13 47 13 47 13 44 31 94 14 44 31 94 14 39 46 32 43 43 26 17 3 19 64 64 26 22 30 26 13 13 47 14 47 14 44 31 14 44 31 94 14 39 46 32 43 43 26 17 3 19 31 94 10 65 65 26 23 30 26 13 13 47 14 47 14 45 15 45 31 100 15 39 46 32 43 43 26 17 3 19 66 66 27 23 30 27 13 13 47 13 47 13 45 31 98 14 45 31 98 14 39 46 32 43 43 26 17 3 19 67 67 27 23 31 27 13 13 47 12 47 12 44 31 93 13 44 31 93 13 40 46 32 43 43 27 17 3 19 68 68 27 23 31 27 13 13 47 12 47 12 44 31 93 13 44 31 93 13 40 46 33 43 43 27 17 3 19 69 69 27 23 31 27 13 13 47 11 47 11 44 32 93 13 44 32 93 13 40 46 33 43 44 27 17 3 19 70 70 27 24 31 27 13 13 47 11 47 11 45 32 94 13 45 32 94 13 40 47 33 43 44 27 17 3 19 71 71 27 24 31 27 13 13 47 11 47 11 45 32 96 14 45 32 96 14 40 47 33 43 44 27 17 3 19 72 72 28 24 31 28 13 13 47 11 47 10 45 32 93 13 45 32 93 13 40 47 33 43 44 28 16 3 19 73 73 28 24 32 28 13 13 47 10 47 10 44 32 87 12 44 32 87 12 41 47 34 43 44 28 16 3 19 74 74 28 24 32 28 13 13 47 10 47 10 44 32 87 12 44 32 87 12 41 47 34 44 44 28 16 3 19 75 75 28 24 32 28 13 13 47 10 47 10 44 32 87 12 44 32 87 12 41 47 34 44 44 28 16 3 19 76 76 28 25 32 28 13 13 47 10 47 10 44 32 87 12 44 32 87 12 41 47 34 44 44 28 16 3 19 77 77 29 25 32 28 13 13 47 11 47 11 44 33 87 12 44 33 87 12 41 48 34 44 44 29 16 3 20 78 78 29 25 33 29 13 13 47 11 47 11 44 33 87 12 44 33 87 12 41 48 34 44 44 29 16 3 20 79 79 29 25 33 29 13 13 47 11 47 11 45 33 88 12 45 33 88 12 42 48 35 44 45 29 16 3 20 80 80 29 25 33 29 13 13 47 11 47 11 44 33 86 12 44 33 86 12 42 48 35 44 45 29 16 3 20 81 81 29 25 33 29 13 13 47 10 47 10 45 33 87 12 45 33 87 12 42 48 35 44 45 29 16 3 20 82 82 29 26 33 29 13 13 47 10 47 10 45 34 85 11 45 34 85 11 42 49 35 44 45 29 16 3 20 83 83 30 26 33 30 13 13 47 10 47 10 45 34 86 11 45 34 86 11 43 49 35 44 45 29 16 3 20 84 84 30 26 33 30 13 13 47 47 45 34 85 11 45 34 85 11 43 49 36 44 45 30 16 3 20 85 85 30 26 34 30 13 13 47 10 47 10 45 34 86 11 45 34 86 11 43 49 36 44 45 30 16 3 20 86 86 30 26 34 30 13 13 48 10 48 46 34 87 12 46 34 87 12 43 49 36 45 45 30 16 3 20 87 87 30 26 34 30 13 13 47 47 45 34 86 11 45 34 86 11 43 49 36 45 45 30 16 3 20 88 88 30 27 34 30 13 13 47 47 45 34 84 11 45 34 84 11 43 49 36 45 45 30 16 3 20 89 89 30 27 34 30 13 13 47 47 45 34 83 11 45 34 83 11 43 49 36 45 45 30 16 3 20 90 90 31 27 34 30 13 13 47 47 45 34 82 10 45 34 82 10 43 49 36 45 46 31 16 3 20 91 91 31 27 35 31 13 13 47 47 45 34 82 10 45 34 82 10 43 49 36 45 46 31 16 3 20 92 92 31 27 35 31 13 13 48 47 45 35 83 11 45 35 83 11 44 50 37 45 46 31 16 3 20 93 93 31 27 35 31 13 13 47 47 45 35 81 10 45 35 81 10 44 49 37 45 46 31 16 3 20 94 94 31 27 35 31 13 13 47 47 45 35 81 10 45 35 81 10 44 50 37 45 46 31 16 3 21 95 95 31 28 35 31 13 13 47 47 45 35 79 10 45 35 79 10 44 50 37 45 46 31 16 3 21 96 96 31 28 35 31 13 13 47 47 44 35 77 44 35 77 44 49 37 45 46 31 16 3 21 97 97 32 28 35 32 13 13 47 47 44 35 78 44 35 78 44 50 38 45 46 32 16 3 21 3 3 3 3 3 3 3 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 4 3 3 2 2 2 2 2 3 3 3 3 3 3 3 3 3 3 3 2 2 2 2 2 2 2 129 98 98 32 28 35 32 13 13 47 47 44 35 78 44 35 78 44 50 38 45 46 32 16 3 21 99 10 10 10 10 99 10 10 10 10 32 28 36 32 13 13 47 46 44 35 77 44 35 77 44 50 38 45 46 32 16 3 21 32 28 36 32 13 13 47 47 45 35 79 10 45 35 79 10 45 50 38 46 46 32 16 3 21 32 28 36 32 13 13 47 47 45 36 79 10 45 36 79 10 45 50 38 46 46 32 16 3 21 32 29 36 32 13 13 46 46 45 36 79 10 45 36 79 10 45 50 38 46 46 32 16 3 21 32 29 36 32 13 13 46 46 45 36 77 45 36 77 45 50 38 46 46 32 16 3 21 10 10 10 10 10 10 10 10 10 11 10 10 10 11 11 11 11 11 11 11 11 11 11 11 11 11 11 11 11 11 12 11 11 12 12 12 12 12 12 12 12 12 12 12 12 12 12 12 12 12 13 13 12 12 13 13 13 13 13 13 13 13 13 13 13 13 13 14 14 13 13 13 13 13 14 14 14 14 14 14 14 14 14 14 14 14 14 14 14 14 14 15 15 14 15 15 32 29 36 32 13 13 47 46 45 36 77 45 36 77 45 50 38 46 46 32 16 3 21 33 29 36 32 13 13 47 46 45 36 77 45 36 77 45 50 38 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41 38 11 11 47 47 46 40 69 46 40 69 48 51 43 48 49 38 15 23 38 34 41 38 11 11 47 47 46 40 70 46 40 70 48 51 43 48 49 38 15 23 38 34 41 38 11 11 47 47 46 40 70 46 40 70 48 50 43 48 49 38 15 23 38 34 41 38 11 11 47 47 46 40 70 46 40 70 48 51 43 48 49 38 15 23 38 34 41 38 11 11 47 47 46 40 69 46 40 69 48 50 43 48 49 38 15 23 38 34 42 38 11 11 47 47 46 40 70 46 40 70 48 50 43 48 49 38 15 23 38 35 42 38 11 11 47 47 46 40 70 46 40 70 49 51 43 49 49 38 15 23 38 35 42 38 11 11 47 47 46 40 70 46 40 70 49 51 43 49 49 38 15 23 38 35 42 38 10 10 47 47 46 40 69 46 40 69 48 50 43 49 49 38 15 23 38 35 42 38 10 10 47 47 46 40 69 46 40 69 48 50 43 49 49 38 15 23 38 35 42 38 10 10 47 47 46 40 68 46 40 68 48 50 44 49 49 38 15 23 38 35 42 38 10 10 47 47 46 40 68 46 40 68 48 50 44 49 49 38 15 23 38 35 42 38 10 10 47 47 46 40 70 46 40 70 49 50 44 49 49 38 15 23 38 35 42 38 10 10 46 46 46 40 70 46 40 70 48 50 44 49 49 38 15 23 39 35 42 38 10 10 46 46 46 40 70 46 40 70 49 50 44 49 49 39 15 23 39 35 42 39 10 10 46 46 46 40 70 46 40 70 49 50 44 49 49 39 15 23 39 35 42 39 10 10 47 46 46 40 69 46 40 69 49 50 44 49 49 39 15 23 39 35 42 39 10 10 47 47 46 41 68 46 41 68 49 50 44 49 49 39 15 23 39 35 42 39 10 10 47 46 46 41 68 46 41 68 49 50 44 49 49 39 15 23 39 35 42 39 10 10 47 46 46 41 67 46 41 67 49 50 44 49 49 39 15 23 39 35 42 39 10 10 47 47 46 41 67 46 41 67 49 50 44 49 49 39 15 23 39 35 42 39 10 10 47 47 46 41 67 46 41 67 49 50 44 49 49 39 15 23 39 36 42 39 10 10 46 46 46 41 66 46 41 66 49 50 44 49 49 39 15 23 39 36 43 39 10 10 47 46 46 41 66 46 41 66 49 50 44 49 49 39 15 23 39 36 43 39 10 10 47 46 46 41 66 46 41 66 49 50 44 49 50 39 15 23 39 36 43 39 10 10 46 46 46 41 66 46 41 66 49 50 44 49 50 39 15 23 39 36 43 39 10 10 47 47 46 41 66 46 41 66 49 50 44 49 50 39 15 23 39 36 43 39 10 10 47 47 46 41 66 46 41 66 49 50 44 49 50 39 15 23 39 36 43 39 10 10 47 47 46 41 67 46 41 67 49 50 44 49 50 39 15 23 39 36 43 39 10 10 46 46 46 41 66 46 41 66 49 50 44 49 50 39 15 23 1 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 5 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 131 Appendix L.3: Values from the different estimators generated using EstimateS for both forest types Individuals (computed) S(est) S(est) 95% CI Lower Bound S(est) 95% CI Upper Bound S(est) SD S Mean (runs) Singletons Mean Singletons SD (runs) Doubletons Mean Doubletons SD (runs) Uniques Mean Uniques SD (runs) Duplicates Mean Duplicates SD (runs) ACE Mean ACE SD (runs) ICE Mean ICE SD (runs) Chao Mean Chao 95% CI Lower Bound Chao 95% CI Upper Bound Chao SD (analytical) Chao Mean Chao 95% CI Lower Bound Chao 95% CI Upper Bound Chao SD (analytical) Jack Mean Jack SD (analytical) Jack Mean Jack SD (runs) Bootstrap Mean Bootstrap SD (runs) MMRuns Mean MMMeans (1 run) Cole Rarefaction Cole SD (analytical) Alpha Mean Alpha SD (analytical) Shannon Mean Shannon SD (runs) Shannon Exponential Mean Shannon Exponential SD (runs) Simpson Inv Mean 4 1 2 0 1 3 0 1 0 0 1 3 0 1 0 0 1 0 1 0 1 17 26 4 17 26 4 0 0 0 2 4 0 1 10 35 35 36 1 2 5767 12456 14986 14599 309656 501639 492784 440894 1 0 0 3 0 1 3 0 1 8 5 4 1 1 5 5 1 1 1 1 1 1 5 1 1 1 1 1 1 12 17 21 26 2 12 17 21 26 2 10 15 18 21 42 61 78 95 11 12 12 14 10 13 17 10 15 18 21 10 13 17 11 12 1 10 12 14 16 2 3 1 1 13 23 35 44 36 37 37 37 2 2 14601 18837 11836 7024 352722 464391 249949 129616 2 0 0 1 1 6 1 1 20 25 11 20 13 18 10 46 38 10530 211648 7 21 28 12 21 15 19 11 63 38 7906 142872 8 22 30 13 22 16 21 12 89 39 5280 86598 8 25 33 14 42 61 78 95 11 12 12 14 25 17 22 12 54 39 5279 79382 25 37 16 25 18 24 13 39 10 5277 73273 10 27 39 17 27 19 25 14 40 10 3967 62503 10 30 42 18 30 20 26 15 14 13 10 40 11 29 21 10 29 45 20 29 21 28 16 85 41 12 27 18 11 2 30 46 21 30 22 29 16 77 41 12 28 19 11 33 49 22 33 23 31 17 73 41 13 30 19 12 31 47 22 31 23 31 18 69 42 13 27 15 12 1 9 10 1 1 30 11 30 11 25 11 10 10 11 1 1 33 14 33 14 28 12 11 11 11 1 1 36 18 36 18 30 13 12 12 12 10 1 1 36 21 36 21 33 14 13 13 10 13 10 2 2 41 25 41 25 37 16 14 14 11 13 11 2 2 44 31 44 31 39 17 15 15 11 14 11 2 2 50 35 50 35 42 18 16 16 12 15 12 2 2 53 39 53 39 45 20 17 17 12 15 13 10 2 10 2 54 42 54 42 46 21 18 18 13 16 13 10 2 10 2 55 44 55 44 49 22 19 19 13 10 17 14 10 10 51 36 51 36 47 22 20 20 14 10 17 14 10 10 50 41 50 41 49 22 21 21 14 11 18 14 10 3 10 3 50 44 50 44 46 22 22 22 15 11 18 15 10 3 10 3 52 51 52 51 47 23 23 23 15 12 19 15 11 3 11 3 54 54 54 54 49 24 24 24 16 12 19 16 11 3 11 3 58 60 58 60 51 26 25 25 16 12 20 16 11 3 11 3 61 67 61 67 54 27 26 26 17 13 20 17 12 12 60 50 60 50 58 27 27 27 17 13 21 17 12 3 12 3 57 39 57 39 56 28 28 28 17 13 21 18 13 3 13 3 58 36 58 36 59 28 29 29 18 14 22 18 13 3 13 3 59 37 59 37 61 29 30 30 18 14 22 19 13 3 13 3 59 39 59 39 59 30 31 31 19 14 23 19 13 3 13 3 57 37 57 37 61 29 32 32 19 15 23 19 13 3 13 3 58 39 58 39 57 29 33 33 19 15 24 20 13 4 13 4 60 42 60 42 54 29 34 34 20 15 24 20 13 4 13 4 62 55 62 54 54 30 35 35 20 16 24 20 13 4 13 4 63 61 63 60 55 30 36 36 20 16 25 21 13 13 57 35 57 34 56 29 14 15 16 16 16 18 17 18 15 15 15 15 16 20 17 19 19 17 20 17 14 14 15 18 37 37 21 16 25 21 14 4 14 4 59 38 59 37 58 29 38 38 21 17 26 21 14 4 14 4 58 38 58 37 54 29 39 39 21 17 26 22 14 4 14 4 60 37 60 37 55 31 40 40 22 17 26 22 14 14 58 37 58 37 53 32 19 16 14 13 41 41 22 18 27 22 13 13 54 30 54 30 52 29 15 33 49 22 28 46 22 27 47 23 28 49 24 27 51 26 30 54 27 36 58 27 30 56 28 35 59 28 36 61 29 31 59 30 37 61 29 31 57 29 26 54 29 25 54 30 27 55 30 32 56 29 14 15 16 16 16 18 17 18 15 15 15 15 16 20 17 19 19 17 20 17 14 14 15 18 34 58 29 28 54 29 24 55 31 22 53 32 19 16 14 13 26 52 29 15 33 24 31 18 66 42 14 25 13 12 28 24 31 18 63 42 14 23 11 12 27 25 32 19 60 42 15 22 10 13 28 26 33 19 58 43 15 22 10 13 27 26 34 20 57 43 16 23 10 13 30 27 35 21 56 43 16 22 14 36 28 36 10 21 56 44 16 23 14 30 29 37 10 22 55 44 17 22 14 35 30 39 23 55 44 17 22 15 36 31 40 23 55 44 18 23 15 31 31 41 24 54 45 18 23 15 37 32 41 24 54 45 18 22 16 31 32 41 10 25 55 45 19 22 16 26 33 42 10 25 55 45 19 22 16 25 33 42 10 26 55 45 20 22 16 27 33 42 10 26 54 46 20 22 16 32 34 43 10 26 54 46 20 21 16 1 1 1 1 1 1 1 2 2 3 3 34 34 43 10 27 54 46 21 21 17 28 35 43 10 27 54 46 21 21 17 24 35 44 10 27 54 46 21 21 17 22 35 44 11 28 54 47 22 21 17 3 3 26 35 43 11 28 54 47 22 21 17 3 Simpson Inv SD (runs) Samples EstimateS (Version 9.1.0), Copyright R K Colwell: http://purl.oclc.org/estimates Diversity Output from Input File: All (30 July, 2017) 2 3 3 3 3 2 2 2 2 3 3 132 42 42 22 18 27 22 13 13 53 25 53 25 48 29 43 43 23 18 27 23 13 13 53 28 53 28 48 29 44 44 23 18 28 23 14 14 54 27 54 27 49 30 45 45 23 19 28 23 13 13 52 24 52 24 45 29 46 46 24 19 28 23 14 14 50 21 50 21 44 29 12 12 12 10 10 47 47 24 19 29 24 13 13 49 19 49 19 43 29 95 20 48 29 20 48 29 20 49 30 17 45 29 16 44 29 12 12 12 10 10 14 43 29 95 20 35 43 11 28 54 47 22 20 17 20 36 44 12 28 54 47 23 20 18 20 36 44 13 29 53 47 23 20 18 17 36 44 12 29 53 47 23 20 18 16 37 44 11 29 53 48 24 20 18 14 37 44 11 29 53 48 24 20 18 48 48 24 19 29 24 13 13 48 16 48 16 42 29 88 13 42 29 88 13 37 44 10 30 52 48 24 19 18 49 49 25 20 29 24 13 13 46 15 46 15 40 28 83 12 40 28 83 12 37 43 10 30 52 48 24 19 18 50 50 25 20 30 24 13 13 46 14 46 14 39 28 79 11 39 28 79 11 37 43 30 52 48 25 19 18 51 51 25 20 30 25 13 13 46 14 46 14 41 29 83 12 41 29 83 12 38 44 30 52 48 25 19 19 52 52 25 20 30 25 13 13 47 14 47 14 41 29 80 11 41 29 80 11 38 45 10 31 52 48 25 20 19 53 53 26 21 31 26 14 14 48 14 48 14 42 30 82 11 42 30 82 11 39 46 10 32 52 49 26 20 19 54 54 26 21 31 26 14 14 47 13 47 13 42 30 82 11 42 30 82 11 39 46 10 32 52 49 26 20 20 55 55 26 21 31 26 14 14 47 12 47 12 42 31 83 11 42 31 83 11 40 47 32 52 49 26 21 20 56 56 26 21 31 27 14 14 48 11 48 11 43 32 84 12 43 32 84 12 41 48 33 52 49 26 21 20 57 57 27 22 32 27 14 14 49 12 49 12 44 32 87 12 44 32 87 12 41 49 33 52 49 27 21 21 58 58 27 22 32 27 14 14 48 11 48 11 43 32 83 11 43 32 83 11 41 49 34 52 49 27 21 21 59 59 27 22 32 28 14 14 48 12 48 12 44 32 84 11 44 32 84 11 41 48 34 52 49 27 21 21 60 60 27 22 32 28 14 14 48 12 48 12 44 32 84 11 44 32 84 11 42 48 34 52 50 27 21 21 61 61 28 23 33 28 14 14 48 12 48 12 44 33 83 11 44 33 83 11 42 49 34 53 50 28 21 21 62 62 28 23 33 28 14 14 47 11 47 11 44 33 82 11 44 33 82 11 42 49 35 53 50 28 21 21 63 63 28 23 33 29 14 14 49 12 49 12 45 33 85 11 45 33 85 11 43 50 35 53 50 28 21 21 64 64 28 23 33 29 14 14 49 12 49 12 45 34 85 11 45 34 85 11 43 50 35 53 50 28 21 22 65 65 29 23 34 29 14 14 49 12 49 12 45 34 86 11 45 34 86 11 43 50 35 53 50 28 21 22 66 66 29 24 34 29 14 14 48 12 48 12 46 34 86 12 46 34 86 12 43 50 10 35 53 50 29 20 22 67 67 29 24 34 29 14 14 48 12 48 12 46 34 86 12 46 34 86 12 43 50 36 53 50 29 20 22 68 68 29 24 34 30 14 14 47 12 47 12 46 34 86 11 46 34 86 11 44 50 36 53 50 29 20 22 69 69 29 24 35 30 14 14 47 12 47 12 45 34 84 11 45 34 84 11 43 50 36 53 50 29 20 22 70 70 30 24 35 30 14 14 47 12 47 12 45 34 83 11 45 34 83 11 44 51 36 53 51 30 20 22 71 71 30 25 35 30 14 14 47 10 47 10 45 34 81 10 45 34 81 10 44 51 36 53 51 30 20 22 72 72 30 25 35 30 14 14 48 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32 27 38 32 14 14 48 48 48 37 84 11 48 37 84 11 46 53 39 53 52 32 20 3 23 85 85 32 27 38 33 14 14 48 48 47 37 82 10 47 37 82 10 46 53 39 53 52 32 20 3 23 6 6 6 6 86 86 33 27 38 33 14 14 48 48 47 37 83 10 47 37 83 10 47 53 39 53 52 33 19 3 23 3 3 3 3 3 3 4 4 4 4 4 4 4 4 4 3 3 3 3 3 3 133 96 96 34 29 40 34 14 14 50 10 50 10 49 39 86 10 49 39 86 10 49 55 10 41 54 53 34 19 3 24 97 97 34 29 40 34 14 14 50 50 49 39 85 10 49 39 85 10 49 55 10 41 54 53 34 19 3 24 98 98 35 29 40 35 14 14 50 50 50 39 86 10 50 39 86 10 49 55 10 41 54 53 34 19 3 24 99 99 35 29 40 35 14 14 50 50 49 39 85 10 49 39 85 10 49 55 10 41 54 53 35 19 3 24 100 100 35 29 40 35 14 14 50 10 50 10 49 39 84 10 49 39 84 10 49 55 10 42 54 53 35 19 3 24 101 101 35 29 40 35 14 14 50 10 50 10 49 39 84 10 49 39 84 10 49 55 10 42 54 53 35 19 3 24 102 102 35 30 41 35 14 14 51 10 51 10 49 39 82 10 49 39 82 10 49 55 11 42 54 53 35 19 3 24 103 103 35 30 41 36 14 14 51 11 51 11 49 40 81 49 40 81 50 56 11 42 54 53 35 19 3 24 104 104 35 30 41 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116 116 37 31 43 37 14 14 52 10 52 10 51 41 83 51 41 83 51 57 44 55 54 37 19 3 25 117 117 37 32 43 37 14 14 52 10 52 10 51 41 84 51 41 84 52 57 44 55 54 37 19 3 25 118 118 37 32 43 37 14 14 52 10 52 10 50 41 82 50 41 82 51 57 44 55 54 37 19 3 25 119 119 37 32 43 38 14 14 53 53 51 42 83 51 42 83 52 57 44 55 54 37 19 3 25 120 120 37 32 43 38 14 14 53 53 51 42 83 51 42 83 52 58 45 55 54 37 19 3 25 121 121 38 32 43 38 14 14 53 53 51 42 82 51 42 82 52 57 45 55 54 38 19 3 25 122 122 38 32 43 38 14 14 53 52 51 42 81 51 42 81 52 57 45 55 54 38 19 3 25 123 123 38 32 43 38 14 14 52 52 50 42 79 50 42 79 52 57 45 55 54 38 19 3 25 124 124 38 32 44 38 14 14 52 52 50 42 80 50 42 80 52 57 45 55 54 38 19 3 25 125 125 38 32 44 38 14 14 52 52 51 42 81 51 42 81 52 58 45 55 54 38 19 3 25 126 126 38 33 44 38 14 14 52 52 51 42 80 51 42 80 52 57 45 55 54 38 19 3 25 127 127 38 33 44 39 14 14 52 52 51 42 81 51 42 81 52 58 45 55 54 38 19 3 25 128 128 38 33 44 39 14 14 52 52 51 42 80 51 42 80 52 57 45 55 54 38 19 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10 63 69 57 59 59 50 17 29 54 99 10 63 68 57 59 59 50 17 29 64 54 99 10 63 69 57 59 59 51 17 29 64 54 99 10 63 69 57 59 59 51 17 29 318 318 51 45 56 51 12 12 59 59 64 54 98 10 64 54 98 10 63 69 57 59 59 51 17 29 319 319 51 45 56 51 12 12 59 59 63 54 98 10 63 54 98 10 63 69 57 59 59 51 17 29 320 320 51 45 56 51 12 12 59 59 64 54 99 10 64 54 99 10 63 69 57 59 59 51 17 29 321 321 51 45 56 51 13 13 60 60 64 54 10 64 54 10 63 69 57 59 59 51 17 29 322 322 51 45 56 51 13 13 60 60 64 55 10 64 55 10 63 69 57 59 59 51 17 29 323 323 51 45 56 51 13 13 60 60 65 55 11 65 55 11 63 69 57 59 59 51 17 29 324 324 51 45 56 51 12 12 60 60 65 55 11 65 55 11 63 69 57 59 59 51 17 29 325 325 51 45 56 51 12 12 60 60 65 55 326 326 51 45 56 51 12 12 59 59 65 55 327 327 51 45 57 51 12 12 60 59 65 55 328 328 51 45 57 51 12 12 60 60 65 55 329 329 51 45 57 51 12 12 60 60 65 55 330 330 51 45 57 51 12 12 60 60 65 55 331 331 51 46 57 51 12 12 60 60 65 55 332 332 51 46 57 51 12 12 60 60 65 55 333 333 51 46 57 51 12 12 60 60 65 55 334 334 51 46 57 51 12 12 60 59 64 55 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 11 65 55 10 65 55 10 65 55 10 65 55 11 65 55 11 65 55 11 65 55 10 65 55 10 65 55 10 64 55 335 335 51 46 57 51 12 12 60 60 64 55 336 336 51 46 57 51 12 12 60 60 64 55 337 337 51 46 57 51 12 12 60 60 65 55 338 338 51 46 57 51 12 12 60 60 64 55 339 339 51 46 57 51 12 12 60 60 64 55 340 340 51 46 57 51 12 12 60 60 65 55 341 341 51 46 57 51 12 12 60 60 64 55 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 10 11 63 69 57 59 59 51 17 29 10 63 69 57 59 59 51 17 29 10 63 69 57 59 59 51 17 29 10 64 69 57 59 60 51 17 29 11 64 69 57 59 60 51 17 29 11 64 69 57 59 60 51 17 29 11 64 69 57 59 60 51 17 29 10 63 69 57 59 60 51 17 29 10 63 69 57 60 60 51 17 29 10 63 69 57 60 60 51 17 29 10 64 55 10 63 69 57 60 60 51 17 29 10 64 55 10 63 69 57 60 60 51 17 29 10 65 55 10 64 69 57 60 60 51 17 29 10 64 55 10 64 69 57 60 60 51 17 29 10 64 55 10 65 55 10 64 55 10 10 10 10 64 69 57 60 60 51 17 29 10 64 69 57 60 60 51 17 29 64 55 10 10 64 69 57 60 60 51 17 29 10 64 69 57 60 60 52 17 29 342 342 52 46 57 51 12 12 60 60 64 55 10 343 343 52 46 57 52 12 12 60 60 64 55 98 64 55 98 64 69 57 60 60 52 17 29 344 344 52 46 57 52 12 12 60 60 63 55 96 63 55 96 64 69 57 60 60 52 17 29 345 345 52 46 57 52 12 12 60 60 64 55 97 64 55 97 64 69 57 60 60 52 17 29 346 346 52 46 57 52 12 12 60 60 64 55 97 64 55 97 64 69 57 60 60 52 17 29 347 347 52 46 57 52 12 12 60 60 64 55 97 64 55 97 64 69 57 60 60 52 17 29 348 348 52 46 57 52 12 12 60 60 64 55 97 64 55 97 64 69 57 60 60 52 17 29 349 349 52 46 57 52 12 12 60 60 64 55 97 64 55 97 64 69 58 60 60 52 17 29 350 350 52 46 57 52 12 12 60 60 64 55 96 64 55 96 64 69 58 60 60 52 17 29 351 351 52 46 57 52 12 12 60 60 64 55 98 64 55 98 64 69 58 60 60 52 17 29 352 352 52 46 57 52 12 12 60 60 64 55 98 64 55 98 64 69 58 60 60 52 17 29 353 353 52 46 58 52 12 12 60 60 64 55 99 64 55 99 64 69 58 60 60 52 17 29 354 354 52 46 58 52 12 12 60 60 64 55 99 10 64 55 99 10 64 69 58 60 60 52 17 29 355 355 52 46 58 52 12 12 60 60 64 55 99 10 64 55 99 10 64 69 58 60 60 52 17 29 356 356 52 46 58 52 12 12 60 60 64 55 97 64 55 97 64 69 58 60 60 52 17 29 357 357 52 46 58 52 12 12 60 60 64 55 97 64 55 97 64 69 58 60 60 52 17 29 358 358 52 46 58 52 12 12 60 60 64 55 97 64 55 97 64 69 58 60 60 52 17 29 359 359 52 46 58 52 12 12 60 60 64 55 98 64 55 98 64 69 58 60 60 52 17 29 360 360 52 47 58 52 12 12 60 60 64 55 98 64 55 98 64 69 58 60 60 52 17 29 361 361 52 47 58 52 12 12 60 60 65 55 99 65 55 99 64 70 58 60 60 52 17 29 362 362 52 47 58 52 12 12 60 60 64 55 98 64 55 98 64 69 58 60 60 52 17 29 363 363 52 47 58 52 12 12 60 60 64 55 98 64 55 98 64 69 58 60 60 52 17 29 364 364 52 47 58 52 12 12 60 60 64 55 97 64 55 97 64 69 58 60 60 52 17 29 365 365 52 47 58 52 12 12 60 60 64 55 97 64 55 97 64 69 58 60 60 52 17 29 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 0 1 0 0 0 138 366 366 52 47 58 52 12 12 60 60 64 55 97 64 55 97 64 69 58 60 60 52 17 29 367 367 52 47 58 52 12 12 60 60 64 55 97 64 55 97 64 69 58 60 60 52 17 29 368 368 52 47 58 52 12 12 60 60 64 55 97 64 55 97 64 69 58 60 60 52 17 29 369 369 52 47 58 52 12 12 60 60 64 55 96 64 55 96 64 69 58 60 60 52 17 29 370 370 52 47 58 52 12 12 60 60 64 55 95 64 55 95 64 69 58 60 60 52 17 29 371 371 53 47 58 52 12 12 60 60 64 55 95 64 55 95 64 69 58 60 60 53 17 29 372 372 53 47 58 52 12 12 60 60 64 55 96 64 55 96 64 69 58 60 60 53 17 29 373 373 53 47 58 52 12 12 60 60 64 55 96 64 55 96 64 69 58 60 60 53 17 29 374 374 53 47 58 52 12 12 60 60 64 56 97 64 56 97 64 70 58 60 60 53 17 29 375 375 53 47 58 53 12 12 60 60 64 56 97 64 56 97 65 70 58 60 60 53 17 29 376 376 53 47 58 53 12 12 61 61 64 56 97 64 56 97 65 70 58 60 60 53 17 29 377 377 53 47 58 53 12 12 60 60 64 56 97 64 56 97 65 70 58 60 60 53 17 29 378 378 53 47 58 53 12 12 61 60 64 56 97 64 56 97 65 70 58 60 60 53 17 29 379 379 53 47 58 53 12 12 60 60 63 56 93 63 56 93 65 69 58 60 60 53 17 29 380 380 53 47 58 53 12 12 60 60 64 56 94 64 56 94 65 69 58 60 60 53 17 29 381 381 53 47 58 53 12 12 60 60 63 55 92 63 55 92 65 69 58 60 60 53 17 29 382 382 53 47 59 53 12 12 60 60 63 56 92 63 56 92 65 69 58 60 60 53 17 29 383 383 53 47 59 53 12 12 60 60 63 56 92 63 56 92 65 69 58 60 60 53 17 29 384 384 53 47 59 53 12 12 60 60 63 56 92 63 56 92 65 69 58 60 60 53 17 29 385 385 53 47 59 53 12 12 60 60 63 55 91 63 55 91 65 69 58 60 60 53 17 29 386 386 53 47 59 53 12 12 60 60 63 56 91 63 56 91 65 69 58 60 60 53 17 29 387 387 53 47 59 53 12 12 60 60 63 56 91 63 56 91 65 69 58 60 60 53 17 29 388 388 53 47 59 53 12 12 60 60 63 56 91 63 56 91 65 69 58 60 60 53 17 29 389 389 53 47 59 53 12 12 60 60 63 56 90 63 56 90 65 69 59 60 60 53 17 29 390 390 53 47 59 53 12 12 60 60 63 56 91 63 56 91 65 69 59 60 60 53 17 29 391 391 53 47 59 53 12 12 61 60 63 56 91 63 56 91 65 69 59 60 60 53 17 29 392 392 53 47 59 53 12 12 61 60 63 56 91 63 56 91 65 69 59 60 60 53 17 29 393 393 53 48 59 53 12 12 61 61 63 56 91 63 56 91 65 70 59 60 60 53 17 29 394 394 53 48 59 53 12 12 61 61 63 56 91 63 56 91 65 70 59 60 60 53 17 29 395 395 53 48 59 53 12 12 61 61 63 56 90 63 56 90 65 69 59 60 60 53 17 29 396 396 53 48 59 53 12 12 61 60 63 56 90 63 56 90 65 69 59 60 60 53 16 29 397 397 53 48 59 53 12 12 61 61 63 56 90 63 56 90 65 69 59 60 60 53 16 29 398 398 53 48 59 53 12 12 61 61 63 56 90 63 56 90 65 69 59 60 61 53 16 29 399 399 53 48 59 53 12 12 61 61 63 56 90 63 56 90 65 69 59 60 61 53 16 29 400 400 53 48 59 53 12 12 61 61 63 56 91 63 56 91 65 70 59 60 61 53 17 29 401 401 53 48 59 53 12 12 61 61 63 56 91 63 56 91 65 70 59 60 61 53 16 29 402 402 53 48 59 53 12 12 61 61 63 56 90 63 56 90 65 70 59 60 61 53 17 29 403 403 54 48 59 53 12 12 61 61 63 56 90 63 56 90 65 70 59 60 61 54 17 29 404 404 54 48 59 54 12 12 61 61 63 56 90 63 56 90 65 70 59 60 61 54 17 29 405 405 54 48 59 54 12 12 61 61 63 56 89 63 56 89 65 70 59 60 61 54 17 29 406 406 54 48 59 54 12 12 61 61 63 56 90 63 56 90 65 70 59 60 61 54 17 29 407 407 54 48 59 54 12 12 61 61 63 56 90 63 56 90 65 70 59 60 61 54 17 29 408 408 54 48 59 54 12 12 61 61 63 56 91 63 56 91 65 70 59 60 61 54 17 29 409 409 54 48 59 54 12 12 61 61 64 56 91 64 56 91 65 70 59 60 61 54 16 29 410 410 54 48 59 54 12 12 61 61 63 56 91 63 56 91 65 70 59 60 61 54 16 29 411 411 54 48 59 54 12 12 61 61 63 56 90 63 56 90 66 70 59 60 61 54 17 29 412 412 54 48 60 54 12 12 61 61 64 56 91 64 56 91 66 70 59 60 61 54 17 29 413 413 54 48 60 54 12 12 61 61 64 56 91 64 56 91 66 70 59 60 61 54 17 29 414 414 54 48 60 54 12 12 61 61 64 56 91 64 56 91 66 70 59 61 61 54 17 29 415 415 54 48 60 54 12 12 61 61 64 57 92 64 57 92 66 71 60 61 61 54 17 29 416 416 54 48 60 54 12 12 61 61 64 57 91 64 57 91 66 71 60 61 61 54 17 29 417 417 54 48 60 54 12 12 61 61 64 57 91 64 57 91 66 71 60 61 61 54 16 29 418 418 54 48 60 54 12 12 61 61 64 57 91 64 57 91 66 70 60 61 61 54 16 29 419 419 54 48 60 54 12 12 61 61 64 57 92 64 57 92 66 71 60 61 61 54 16 29 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 7 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 139 420 420 54 48 60 54 12 12 62 62 64 57 92 64 57 92 66 71 60 61 61 54 16 29 140 Appendix M Computed values of fisher‘s alpha, Shannon index, and Simpson‘s index for both protected and unprotected forests Fisher’s alpha Protected Unprotected 13.96386 14.33703 Minimum 9.45 12.26 1st Quarter 12.80 12.46 Median 14.32 13.61 Mean 13.96 14.34 3rd Quarter 15.48 15.48 Max 17.77 17.87 Protected Unprotected Minimum 2.539 2.630 1st Quarter 2.628 2.660 Median 2.677 2.774 Mean 2.680 2.771 3rd Quarter 2.729 2.886 Max 2.828 2.907 alpha Shannon’s index 141 Simpson’s index Protected Unprotected Minimum 0.8767 0.8792 1st Quarter 0.8883 0.8926 Median 0.9080 0.9153 Mean 0.9067 0.9178 3rd Quarter 0.9264 0.9345 Max 0.9343 0.9375 142 Appendix N Certificate of completion given by the Institute of Botany and Landscape Ecology, University of Greifswald for research internship and final thesis 143 ... Redeña Santos DTN1454290051 The first report of Myxomycetes (Amoebozoa) in Thai Nguyen City, Northern Vietnam: Rapid biodiversity assessment using tools in modern molecular era Dr Duong Van Thao &... Although, the number of myxomycetes records for Vietnam is rapidly increasing, much of these myxomycetes species are reported only in Southern Vietnam However, no profiling of myxomycete species was reported... September 2017 The collection of samples was carried out in the month of March – April 2017 in Thai Nguyen City, Vietnam while the process of setting up the myxomycetes culture, molecular methods,