Nghiên cứu đa dạng sinh học của họ nấm túi Clavicipitaceae, Cordycipitaceae, Ophiocordycipitaceae tại khu bảo tồn thiên nhiên Copia và Vườn Quốc gia Xuân Sơn.Nghiên cứu đa dạng sinh học của họ nấm túi Clavicipitaceae, Cordycipitaceae, Ophiocordycipitaceae tại khu bảo tồn thiên nhiên Copia và Vườn Quốc gia Xuân Sơn.Nghiên cứu đa dạng sinh học của họ nấm túi Clavicipitaceae, Cordycipitaceae, Ophiocordycipitaceae tại khu bảo tồn thiên nhiên Copia và Vườn Quốc gia Xuân Sơn.Nghiên cứu đa dạng sinh học của họ nấm túi Clavicipitaceae, Cordycipitaceae, Ophiocordycipitaceae tại khu bảo tồn thiên nhiên Copia và Vườn Quốc gia Xuân Sơn.Nghiên cứu đa dạng sinh học của họ nấm túi Clavicipitaceae, Cordycipitaceae, Ophiocordycipitaceae tại khu bảo tồn thiên nhiên Copia và Vườn Quốc gia Xuân Sơn.Nghiên cứu đa dạng sinh học của họ nấm túi Clavicipitaceae, Cordycipitaceae, Ophiocordycipitaceae tại khu bảo tồn thiên nhiên Copia và Vườn Quốc gia Xuân Sơn.Nghiên cứu đa dạng sinh học của họ nấm túi Clavicipitaceae, Cordycipitaceae, Ophiocordycipitaceae tại khu bảo tồn thiên nhiên Copia và Vườn Quốc gia Xuân Sơn.Nghiên cứu đa dạng sinh học của họ nấm túi Clavicipitaceae, Cordycipitaceae, Ophiocordycipitaceae tại khu bảo tồn thiên nhiên Copia và Vườn Quốc gia Xuân Sơn.Nghiên cứu đa dạng sinh học của họ nấm túi Clavicipitaceae, Cordycipitaceae, Ophiocordycipitaceae tại khu bảo tồn thiên nhiên Copia và Vườn Quốc gia Xuân Sơn.Nghiên cứu đa dạng sinh học của họ nấm túi Clavicipitaceae, Cordycipitaceae, Ophiocordycipitaceae tại khu bảo tồn thiên nhiên Copia và Vườn Quốc gia Xuân Sơn.Nghiên cứu đa dạng sinh học của họ nấm túi Clavicipitaceae, Cordycipitaceae, Ophiocordycipitaceae tại khu bảo tồn thiên nhiên Copia và Vườn Quốc gia Xuân Sơn.BỘ GIÁO DỤC VÀ ĐÀO TẠO MINISTRY OF EDUCATION AND TRAINING HANOI NATIONAL UNIVERSITY OF EDUCATION NGUYEN DINH VIET STUDY ON DIVERSITY OF CLAVICIPITACEAE, CORDYCIPITACEAE, OPHIOCORDYCIPITACEAE IN CO.
MINISTRY OF EDUCATION AND TRAINING HANOI NATIONAL UNIVERSITY OF EDUCATION NGUYEN DINH VIET STUDY ON DIVERSITY OF CLAVICIPITACEAE, CORDYCIPITACEAE, OPHIOCORDYCIPITACEAE IN COPIA NATURE RESERVE AND XUAN SON NATIONAL PARK Major: Microbiology Code: 9.42.01.07 SUMMARY OF PhD THESIS IN BIOLOGY HaNoi - 2023 The thesis was completed at : HANOI NATIONAL UNIVERSITY OF EDUCATION Supervisors: Assoc Prof PhD Dương Minh Lam Prof PhD Trương Xuân Lam Reviewer 1: Assoc Prof PhD Dương Văn Hợp Vietnam National University, Hanoi Reviewer 2: Assoc Prof PhD Khuất Hữu Trung Agricultural Genetics Institute Reviewer 3: Assoc Prof PhD Bùi Thị Việt Hà Hanoi University of Science, Vietnam National University The thesis is defended at the doctoral thesis evaluation committee at At :00 on , 2023 INTRODUCTION Entomopathogenic fungi (EPF) or insect fungi refer to species of fungi that can infect, parasitize, harm, or destroy its insect host This group of fungi have high biodiversity But the most diverse are the families Clavicipitaceae, Cordycipitaceae, Ophiocordycipitaceae of the order Hypocreales, phylum Ascomycota Currently, families of fungi Clavicipitaceae, Cordycipitaceae, Ophiocordycipitaceae are recorded with 99 genera with 1468 species of parasites on plants, fungi, rotifers and animals; 40 genera and 1092 species of fungi parasitic on insects Despite of more than 120-year research history, the Clavicipitaceae, Cordycipitaceae, Ophiocordycipitaceae families are still attracting the attention of many scientists around the world because of their biological diversity and application potential Vietnam, where there is a tropical monsoon climate, there is a diversity of flora and fauna The natural conditions are suitable for the growth and development of fungi However, studies on insect parasitic fungi of families Clavicipitaceae, Cordycipitaceae, Ophiocordycipitaceae are still few Most of the studies have been conducted in the direction of studying species diversity based on morphological classification and applied research on insect parasitic fungi in agriculture and medicine To date, there have been 16 genera, 82 species of insect parasitic ascomycetes belonging to families Clavicipitaceae, Cordycipitaceae, Ophiocordycipitaceae announced, with a number of studies at: Tam Dao National Park (National Park), Pu Mat National Park, Hoang Lien National Park, Cat Tien National Park, Central Highlands; Although in recent years, there have been more researches on insect parasitic fungi, but the data of Vietnam on this group of fungi is still limited Copia Nature Reserve, Son La Province and Xuan Son National Park, Phu Tho Province are two large forests with many high peaks In addition to Nguyen Thi Thuy Van's research on insect parasitic fungi capable of CODs biosynthesis, there are currently no studies on the biodiversity of insectparasitic species of the family Clavicipitaceae, Cordycipitaceae, Ophiocordycipitaceae to be announced Because of all reasons above, we decided to study the topic of “Study on diversity of Clavicipitaceae, Cordycipitaceae, Ophiocordycipitaceae in Copia Nature Reserve and Xuan Son National Park” Objectives Evaluation of diversity of Clavicipitaceae family, Cordycipitaceae, Ophiocordycipitaceae in Copia Nature Reserve (Son La) and Xuan Son National Park (Phu Tho) Scopes - Research subject: The fungi belong to the Clavicipitaceae family, Cordycipitaceae, Ophiocordycipitaceae - Region of interest: Copia Nature Reserve and Xuan Son National Park Research contents - Fungal sample collection in the study sites - Morphological descriptions of fungi - Fungal isolation using single spore isolation method - Morphological characteristics of the isolated strains - Fungal identification based on morphologies of samples and strains - DNA extraction and PCR using the three fragments of ITS, LSU, RPB1 - Sequencing and molecular analysis to investigate the phylogenetic relationship of the study sequences (Clavicipitaceae, Cordycipitaceae, Ophiocordycipitaceae) New contributions - The list of 33 taxon fungal of Clavicipitaceae, Cordycipitaceae, Ophiocordycipitaceae in Copia Nature Reserve (Son La) and Xuan Son National Park (Phu Tho) with detail description, pictures, samples and DNA sequences; - Identification of 25 species and new recorded species (Ophiocordyceps campes, Ophiocordyceps xuansonae, O multiperitheciata, Cordyceps bassiana, C cicada, Isaria fumosorosea, Lecanicillium kalimantanense, Leptobacillium leptobactrum, Metacordyceps yongmunensis) for the fungal flora in Vietnam, genus Simplicillium, Leptobacillium for the fungal flora in Vietnam - Ecological distribution of in Clavicipitaceae, Cordycipitaceae, Ophiocordycipitaceae species in Xuan Son and Copia 3 Structure of the thesis The thesis consists of 148 pages, including the following sections: Introduction: pages Chapter Overviews of research issues: 36 pages Chapter Materials and research methods: 13 pages Chapter Results and discussion: 93 pages Conclusions and Recommendations: pages References: The thesis cited 25 Vietnamese references, 189 foreign language references and 06 web pages Appendix: This thesis contains appendices including sample collection results and diversity indicators analysis in the study area 4 CHAPTER 1: OVERVIEWS OF RESEARCH ISSUES In this chapter, four issues related to the research in the thesis have been reviewed, namely: (1.1) Diversity and taxonomy of fungi and insect parasitic fungi; (1.2) Research on insect parasitic fungi of Clavicipitaceae family, Cordycipitaceae, Ophiocordycipitaceae in the world; (1.3) Research on insect parasitic fungi of Clavicipitaceae family, Cordycipitaceae, Ophiocordycipitaceae in Vietnam; (1.4) Characteristics of the study area The section (1.1) Diversity and taxonomy of fungi and insect parasitic fungi introduces the kingdom of fungi and entomopathogenic (section 1.1.1), The role of morphology, molecular biology and chemistry in the identification of ascomycetes and insect fungi (section 1.1.2) has been presented from page to page 11 In the section (1.2) Research situation on insect parasitic fungi of Clavicipitaceae family, Cordycipitaceae, Ophiocordycipitaceae in the world will be mentioned, the contents of taxonomic position and number of genera and species of the families Clavicipitaceae, Cordycipitaceae, Ophiocordycipitaceae (section 1.2.1) Morphological characteristics of the parasitic species of the Clavicipitaceae family, Cordycipitaceae, Ophiocordycipitaceae (section 1.2.2) Ecological characteristics of the Clavicipitaceae, Cordycipitaceae, Ophiocordycipitaceae (section 1.2.3) Chemical compounds were extracted from species of the Clavicipitaceae family, Cordycipitaceae, Ophiocordycipitaceae (section 1.2.4) Research on diversity of insect fungi in the Clavicipitaceae family, Cordycipitaceae, Ophiocordycipitaceae in the world (section 1.2.5) is presented in detail from page 11 to page 34 In section (1.3) there will be the research on the insect fungi belong to Clavicipitaceae, Cordycipitaceae, Ophiocordycipitaceae in Vietnam, research directions in Vietnam will be discussed from page 34 to 38 Section (1.4) will discuss about the characteristics of study areas in Copia natural reserve and Xuan Son National Park from page 38 to 40 5 CHAPTER II: STUDY AREAS, PERIOD, MATERIALS AND METHODS In this chapter, the Study areas, period, materials and methods subjects were presented in section (2.1) including: Sample collection routes, study time, research materials, tools and equipment, types of media used in the study were presented Research Methods (Section 2.2) are divided into the main groups as follows: 2.2.1 Sample collection 2.2.1.1 Forest habitats 2.2.1.2 Study altitudes 2.2.1.3 Sample collection routes 2.2.1.4 Sample collection and processing 2.2.2 The microbiological method 2.2.2.1 Isolation method 2.2.2.2 Breeding method 2.2.2.3 Freeze-drying method 2.2.2.4 Activation method of selected fungal strains 2.2.2.5 Morphological descriptions and sample identification 2.2.3 Molecular biology methods 2.2.3.1 Extraction of total DNA 2.2.3.2 DNA multiplication 2.2.3.3 Sequencing methods 2.2.4 Mathematical methods CHAPTER III: RESULTS AND DISCUSSIONS 3.1 Diversity of entomopathogenic fungi belong to the Cordycipitaceae family, Clavicipitaceae, Ophiocordycipitaceae in study area 3.1.1 Isolation and identification During the period from December 2015 to December 2019, we conducted 57 sampling times, of which 35 times were collected at Copia NR and 22 times at Xuan Son NP A total of 1881 research samples were classified into 13 genera of fungi, 120 fungal strains were isolated to be used for morphological analysis, 41 fungal strains were used in molecular biology 3.1.2 Morphological characteristics A total of 1881 research samples were classified into 13 genera of fungi with specific descriptive data on: original description, synonym synonyms, sample symbols, location of sampling coordinates, descriptive characteristics of substrate, fruit body, sporangium, sporangium There are species of genera (Ophiocordyceps campes, Ophiocordyceps sphecocephala, Ophiocordyceps nutans, Ophiocordyceps pseudolloydii, Ophiocordyceps unilateralis, Ophiocordyceps myrmecophila, Purpureocillium lilacinum), 10 species of genera (Cordyceps bassiana, Cordyceps cicada, Cordyceps militaris, Cordyceps takaomontana, Beauveria bassiana, Isaria fumosorosea, Isaria amoene-rosea, Isaria tenuipes, Isaria cicadae, Gibellula pulchra), the Clavicipitaceae family has species of genera (Metacordyceps yongmunensis, Metarhizium anisopliae, Conoideocrella luteorostrata, Hypocrella calendulina) 3.1.3 Morphological characteristics of unidentified taxons There are 13 strains of fungi identified to the genus, including: Ophiocordyceps sp.1, Ophiocordyceps sp.13, Ophiocordyceps sp.2, Ophiocordyceps sp.3, Cordyceps sp.4, Cordyceps sp.5, Cordyceps sp.6, Cordyceps sp.7, Isaria sp.8, Simplicillium sp.9, Lecanicillium sp.10, Leptobacillium sp.11, Hypocrella sp.12 3.1.4 General discussion on morphological characteristics and microscopic structure of species of Cordycipitaceae, Clavicipitaceae, Ophiocordycipitaceae in study area In the process of collecting and analyzing samples in research areas, the subject has drawn some comments on the morphological characteristics and microscopic structure of the Cordycipitaceae, Clavicipitaceae, Ophiocordycipitaceae families as follows: Where the host occurs; Location of substrate appearance; The size of the substrate; The color of the substrate; The substrate shape; The location of the fruit body; Fruit body shape; Spores (Asci), sporangia (Ascospores) 3.2 Genetic relationships of the species of entomopathogenic fungi of Clavicipitaceae, Cordycipitaceae, Ophiocordycipitaceae in study area 3.2.1 Genetic relationships taxon of Cordycipitaceae According to phylogenetic analysis based on gene segments ITS, LSU, RPB1, in Copia NR and Xuan Son NP, there are the species of Beauveria bassiana XS39, Beauveria bassiana XS35, Beauveria bassiana CPA66 of Beauveria; Isaria cicadae XS56, Isaria cicadae CPA60, Isaria amoene-rosea CPA44, Isaria fumosorosea XS69, Isaria tenuipes XS26, Isaria amoene-rosea CPA44, Isaria sp.6 Isaria sp.8 of Isaria; Gibellula pulchra XS100 of Gibellula; Cordyceps bassiana CPA13, Cordyceps cicadae CPA80, Cordyceps militaris CPA70, Cordyceps takaomontana CPA61 Cordyceps takaomontana XS76, Cordyceps sp.5, Cordyceps sp.7, Cordyceps sp.4 of Cordyceps, Leptobacillium leptobactrum of Leptobacillium, Lecanicillium kalimantanense of Lecanicillium, Simplicillium sp.9 of Simplicillium the Cordycipitaceae 3.2.2 Genetic relationships taxon of Ophiocordycipitaceae From morphological characterization and phylogenetic analysis, we concluded that in the study areas, there are: Ophiocordyceps sphecocephala XS78, Ophiocordyceps nutans CPA50, Ophiocordyceps nutans XS58, Ophiocordyceps campes CPA51, Ophiocordyceps pseudolloydii XS65, Ophiocordyceps unilateralis XS70, Ophiocordyceps xuansonae XS154, Purpureocillium lilacinum XS77, Purpureocillium lilacinum CPA7, The sample CPAV2, CPAV3 are the teleomorph and anamorph state of O multiperithecia, Ophiocordyceps sp.2 3.2.3 Genetic relationships taxon of Clavicipitaceae Phylogenetic tree of the fungal strain Hypocrella calendulina XS147, Hypocrella sp.12, Conoideocrella luteorostrata XS18, Metacordyceps yongmunensis XS155, Metarhizium anisopliae XS68 belong to Clavicipitaceae 8 3.2.4 List taxon of Cordycipitaceae, Clavicipitaceae, Ophiocordycipitaceae Table 3.2 List taxon of Cordycipitaceae, Clavicipitaceae, Ophiocordycipitaceae samples Order Species Xuan Copia Son Ophiocordycipitaceae G.H.Sung, J.M.Sung, Hywel-Jones & Spatafora (2007) Ophiocordyceps Petch (1931) Ophiocordyceps sphecocephala (Klotzsch ex Berk.) 478 G.H Sung, J.M Sung, Hywel-Jones & Spatafora 2007 Ophiocordyceps nutans (Pat.) G.H Sung, J.M Sung, 52 Hywel-Jones & Spatafora Ophiocordyceps pseudolloydii (H.C Evans & Samson) G.H Sung, J.M Sung, Hywel-Jones & Spatafora 2007 Ophiocordyceps unilateralis (Tul & C Tul.) Petch Ophiocordyceps campes Tasan, Thanakitp & Luangsa-ard (2020)* Ophiocordyceps myrmecophila (Ces.) G.H Sung, 10 J.M Sung, Hywel-Jones & Spatafora 2007 O multiperitheciata* 10 Ophiocordyceps sp 10 Ophiocordyceps xuansonae** Purpureocillium Luangsa-ard, Hywel-Jones, Houbraken & Samson (2011) 10 Purpureocillium lilacinum (Thom) Luangsa-ard, 51 135 Houbraken, Hywel-Jones & Samson Cordycipitaceae Kreisel ex G.H.Sung, J.M.Sung, Hywel-Jones & Spatafora (2007) Beauveria Vuill (1912) 194 38 11 Beauveria bassiana (Bals.-Criv.) Vuill 1912 Cordyceps Fr., 12 Cordyceps bassiana Z.Z Li, C.R Li, B Huang & M.Z Fan 2001* 13 Cordyceps cicada (Miq.) Massee* 14 15 16 Cordyceps militaris (L.) Fr Cordyceps takaomontana Yakush & Kumaz Cordyceps sp.4 42 27 50 15 Order 17 18 19 20 21 22 23 24 25 26 Species Cordyceps sp.5 Cordyceps sp.7 samples Xuan Copia Son Gibellula Gibellula pulchra (Sacc.) Cavara Isaria Persoon in Roemer, 1794 30 Isaria amoene-rosea Henn 1902 30 45 Isaria cicadae Miq 1838 18 Isaria fumosorosea Wize 1904 * 395 111 Isaria tenuipes Peck 1878 Isaria sp.6 Isaria sp.8 Simplicillium W Gams & Zare, Nova Hedwigia 73(1-2): 38 (2001) Simplicillium sp.9 Lecanicillium W.Gams & Zare (2001) 27 Lecanicillium kalimantanense* Leptobacillium 28 Leptobacillium leptobactrum* Clavicipitaceae (Lindau) Earle (1901) Metarhizium Sorokīn, 1879 29 Metarhizium anisopliae (Metschn.) Sorokīn Metacordyceps G.H.Sung, J.M.Sung, Hywel-Jones & Spatafora (2007) 30 Metacordyceps yongmunensis G.H Sung, J.M Sung & Spatafora, in Sung, Hywel-Jones, Sung, Luangsa2 ard, Shrestha & Spatafora, (2007)* Hypocrella Sacc., 1878 31 Hypocrella calendulina Hywel-Jones & Mongkols 85 Mycol (2009) 32 Hypocrella sp.12 10 Conoideocrella 33 Conoideocrella luteorostrata (Zimm.) D Johnson, G.H Sung, Hywel-Jones & Spatafora (2009) Total samples 15 25 Total taxon Note: * New species recorded for Vietnam ** New species 3.2.5 New records of fungal in Vietnam In this study, we recorded genera found for the first time in Vietnam 10 including Leptobacillium Zare & W Gams; Simplicillium W Gams & Zare As a consequence, the total number of genera of Cordycipitaceae, Clavicipitaceae, Ophiocordycipitaceae found in Vietnam increased to 19 (Table 3.2) New species recorded for Vietnam: Ophiocordyceps campes, Ophiocordyceps xuansonae, O multiperitheciata, Cordyceps bassiana, C cicada, Isaria fumosorosea, Lecanicillium kalimantanense, Leptobacillium leptobactrum, Metacordyceps yongmunensis 3.3 Ecological diversity of Cordycipitaceae, Clavicipitaceae, Ophiocordycipitaceae in study areas 3.3.1 Diversity in taxon The results on the number and percentage of species in the genera of the fungus family Cordycipitaceae, Clavicipitaceae, Ophiocordycipitaceae in the study area are shown in Table 3.4 From the research results (Tables 3.2, 3.4), we have identified genera: Purpureocillium, Beauveria, Leptobacillium, Lecanicillium, Gibellula, Conoideocrella, Metacordyceps, Metarhizium (1 taxon, 3.03%); Hypocrella (2 taxon, 6.06%), Cordyceps (7 taxon, 21.21%), Isaria (6 taxon, 18.18%), Ophiocordyceps (9 taxon, 27.27%) Comparison with research on insect parasitic fungi in Vietnam Comparison of the number of species in 13 genera of families Cordycipitaceae, Clavicipitaceae, Ophiocordycipitaceae in the research area and studies in Tam Dao National Park, Cat Tien National Park, Pu Mat National Park, Hoang Lien National Park, Chu Yang Sin National Park, Langbian Mountain - Da Lat, Muong Phang Nature Reserve, Cat Tien National Park (Table 3.5) Comparison on the number of species with the studies in some areas having similar climatic and topographical conditions to the study areas about the family Cordycipitaceae, Clavicipitaceae, Ophiocordycipitaceae (Table 3.6) Although the time and scope of the study is less than that of publications in Southeast Asia and different regions of the world, our research results are completely consistent with previous studies The results obtained at the study areas once again identified a high diversity of insect parasitic cysts in the Asia 11 region In addition, the research results here have identified the genus Cordyceps, Ophiocordyceps with the highest number of species, followed by the fungal genera Hypocrella, Metarhizium This is completely consistent with research results in tropical and subtropical regions of the world 3.3.2 Species biodiversity In this study, we use the species biodiversity index (H’) and dominance index (Cd) to evaluate the species biodiversity The species biodiversity index (H’- Shannon index) was used to evaluate the degree of species diversity of each genus based on the number of species and the density of individuals in that genus Index (H’) is proportional to the biodiversity The dominance index (Cd) was used to assess the predominance of each genus in the family The higher index (Cd) score, the predominance genus is The results of biodiversity indexes are shown in the table 3.7 Table 3.7: Biodiversity indices of each genus in the study areas Orde r 10 11 12 13 Number Index (Cd) Number of Index Scientific Name of species (H’) samples Ophiocordycipitaceae G.H Sung et al Ophiocordyceps 574 0.56 0.0657 Purpureocillium 186 0.23 0.0098 Cordycipitaceae Kreisel ex G.M Sung et al Beauveria 232 0.26 0.0152 Cordyceps 145 0.29 0.0021 Gibellula 1 0.004 0.0000003 Isaria 631 0.53 0.074 Lecanicillium 1 0.004 0.0000003 Simplicillium 1 0.004 0.0000003 Leptobacillium 1 0.004 0.0000003 Clavicipitaceae (Lindau) Earle ex Rogerson Hypocrella 95 0.17 0.0021 Conoideocrella 0.02 0.00002 Metarhizium 0.013 0.000005 Metacordyceps 0.0073 0.000001 Tổng 1881 33 Note: H’ is the species biodiversity index Shannon Cd is the dominance index 12 So, biodiversity results in table 3.7 are completely consistent with the level of diversity in publications around the world (Table 3.6) In addition, this result once again confirms the wide distribution of the species Ophiocordyceps, Cordyceps, Beauveria, Purpureocillium, Hypocrella and confirms the biodiversity in study areas when the new genera Leptobacillium, Simplicillium, Lecanicillium occured 3.3.3 Biodiversity according to study area During the research period from December 2015 to June 2019, the study collected 1881 samples of insect parasitic fungi of families Cordycipitaceae, Clavicipitaceae, Ophiocordycipitaceae, of which 834 samples were collected in Copia NR, 1047 samples were collected in NP Xuan Son With the results on the topic of assessing biodiversity by indices: fi - Percentage of sample (species) occurrence, Shannon species diversity index (H’), dominance index (Cd) and the index assessing the similarity of the species between sample collection place (SI) The results of the diversity indices are shown at (Table 3.8, Table 4.7 - 4.12PL4) Copia: Isaria (fi=54.56), Beauveria (fi=23.26), Cordyceps (fi=13.43), chi Purpureocillium (fi = 6.12), Ophiocordyceps (fi = 2.64) H’= 1.66, Cd= 0.291 Xuan Son: Ophiocordyceps (fi = 52.72), Isaria (fi = 16.81), Purpureocillium (fi = 12.89), Beauveria (fi = 3.63), Cordyceps (fi = 3.15), Hypocrella (fi = 9.07), Conoideocrella (fi = 0.76), Metarhizium (fi = 0.38), Metacordyceps (fi = 0.19), Gibellula, Simplicillium, Leptobacillium, Lecanicillium có fi = 0.10 H’= 1.91, Cd= 0.25 3.3.4 Seasonal diversity At Copia NR, the results of sampling during the period from December 2015 to June 2019, are shown in the data table (Table 4.7PL4, 4.8PL4) and fig 3.34, fig 3.35 13 Fig 3.34 The number of samples collected from 2015 to 2019 in Copia Fig 3.35 Number of samples collected by months of the year in Copia At Xuan Son National Park, the results are shown in tables 4.9, 4.10 – PL4 and fig 3.36, fig 3.37 with the time of sampling in 2017, 2018, months of 2019 14 Fig 3.36 The number of samples collected from 2017 to 2019 in Xuan Son Fig 3.37 Number of samples collected in the following months in Xuan Son 3.3.5 The influence of habitat on species diversity in the study areas The results of the study on the number of samples collected of the genera in the habitats are shown in Table 3.9, Table 4.12PL4 15 Table 3.9 Effects of habitats on species diversity in study areas Habitat Genus Ophiocordyceps No samples fi No samples fi No samples fi No samples fi No samples 22 1.17 0 0 411 21.84 141 7.49 574 51 2.71 120 6.38 15 0.8 186 Purpureocillium fi Beauveria 115 6.11 12 0.64 67 3.56 28 1.49 10 0.53 232 Cordyceps 55 2.93 0 57 3.03 29 1.55 0.21 145 0.05 152 8.08 Simplicillium 0.05 Lecanicillium 0.05 Leptobacillium 0.05 Hypocrella 40 2.13 Conoideocrella 0.43 Metarhizium 0.21 Metacordyceps 0.11 Gibellula Isaria 11 0.58 0 444 23.59 24 1.28 631 55 2.92 95 Total number of species of each habitat 10 22 10 Total number of samples 203 12 619 798 249 188 Ratio % 10.79 0.64 32.91 42.42 13.24 100 H' index 0.38 0.03 0.8 1.16 0.49 Cd index 0.005 0.00004 0.045 0.046 0.005 Note: (1) - Natural forest in Copia; (2) - Upland farming in Copia; (3) - Forest recovery after fire in Copia; (4) - Natural forest on the mountain in Xuan Son; (5) - Natural forest on Xuan Son limestone mountain; (6) - Total number of samples collected at Copia and Xuan Son 16 3.3.6 Diversity of entomopathogenic fungi according to the altitude in study areas Table 3.10: Diversity of according to the altitude in study areas Order 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 Species Ophiocordyceps sphecocephala Ophiocordyceps nutans Ophiocordyceps pseudolloydii Ophiocordyceps unilateralis Ophiocordyceps campes Ophiocordyceps myrmecophila O.multiperitheciata Ophiocordyceps sp.2 Altitude (1) (2) (3) (4) Number of collected samples (6) (1) 149 41 1 478 61 1 10 10 10 186 232 69 50 15 1 30 75 18 506 1 1 215 82 10 Ophiocordyceps xuansonae Purpureocillium lilacinum Beauveria bassiana Cordyceps bassiana Cordyceps cicada Cordyceps militaris Cordyceps takaomontana Cordyceps sp.4 Cordyceps sp.5 Cordyceps sp.7 Gibellula pulchra Isaria amoene-rosea Isaria cicadae Isaria fumosorosea Isaria tenuipes Isaria sp.6 Isaria sp.8 Simplicillium sp.9 Lecanicillium kalimantanense Leptobacillium leptobactrum Metacordyceps yongmunensis 75 50 14 50 55 27 42 32 10 10 10 11 163 50 15 1 45 10 41 30 30 18 112 1 1 343 17 Species Order 30 31 32 33 Altitude (6) (1) (2) (3) (4) (1) Number of collected samples 4 10 10 80 85 11 14 13 14 286 504 450 641 1881 15.2 26.79 23.92 34.08 0.48 0.85 0.85 0.84 0.01 0.02 0.01 0.04 Metarhizium anisopliae Hypocrella sp.12 Hypocrella calendulina Conoideocrella luteorostrata Total number of species Total number of samples Tỷ lệ % H’ index Cd index Note: (1) - 100 < altitude