MINISTERY OF EDUCATION AND TRAINING VIETNAM ACADEMY OF SCIENCE AND TECHNOLOGY GRADUATE UNIVERSITY OF SCIENCE AND TECHNOLOGY NGUYEN NGOC HIEU STUDY ON THE ISOLATION AND BIOLOGICAL ACTIVITY OF NATURAL ACTIVE COMPOUNDS FROM PLANTS AND ENDOPHYTES Scientific Field: Organic Chemistry Classification Code: 62 44 01 14 DISSERTATION SUMMARY HA NOI - 2019 The dissertation was completed at: Institute of Chemistry Vietnam Academy of Science and Technology Scientific Supervisors: Dr Duong Ngoc Tu Institute of Chemistry - Vietnam Academy of Science and Technology Ass Prof Dr Duong Anh Tuan Institute of Chemistry - Vietnam Academy of Science and Technology st Reviewer: 2nd Reviewer: rd Reviewer: The dissertation will be defended at Institute of Chemistry, Vietnam Academy of Science and Technology, 18 Hoang Quoc Viet, Cau Giay District, Ha Noi City At … hour… date… month … 2019 The dissertation can be found in National Library of Vietnam and the library of Institute of Chemistry, Vietnam Academy of Science and Technology I INTRODUCTION Background Vietnam is still world famous for its biodiversity potentials, with over 12,000 species of higher plants, excluding fungi, algae and mosses Many species are endemic to Vietnam From the treasure of folk experience, we have had a lot of experience using and ingeniously combining these diverse plant materials into very precious, special and special folk remedies In the treatment of diseases, high health of human, protecting crops, eradicating pests, insects, harmful animals With the current level of scientific and technological development, it is necessary to continue continue to research, research, select from folk experiences in combination with the support of modern technology and equipment to create new products, bringing the value of using plant resources in Vietnam to reach High new, more valuable, more efficient, highly appreciated both in terms of science and technology as well as use value Plant endogenous fungi (endophytes) are currently being studied extensively and extensively in the world and are expected to be an unexplored resource for biotechnology and pharmaceuticals Recent statistical results, with an estimated 51% of active compounds isolated from endophytes are new compounds, have shown great potential for research and application of the endophyte Continuing the international cooperation program between the Institute of Chemistry (Vietnam Academy of Science and Technology) and the Institute of Biopharmaceuticals and Biotechnology (Heirich-Heine General University Duesseldorf, Germany) on the study of flora Vietnam to screen and detect natural bioactive compounds, potentially used to produce insecticides and fungal pathogens of plants, as well as expand to target new research subjects in the world as well as in Vietnam is NSTV, we propose the dissertation: "Study on the isolation and biological activity of natural active compounds from plants and endophytes" Objectives and aims of the dissertation The research object is species of plants including Aglaia duperreana Pierre, Aglaia oligophylla Miq., Piper betle L and Curcuma longa L and those endophytes The aims of the dissertation are: Extracting and determining the structure of organic compounds of four plant species with potential for insecticide and fungal diseases Isolating endogenous fungi from plant samples, extracting and determining the structure of component organic compounds Testing of insecticidal and fungal activity of extracts and component organic compounds New contributions of the dissertation 3.1 For the first time in Vietnam, the relationship between plants and plant endogenous fungi on Aglaia duperreana Pierre, Aglaia oligophylla Miq., Piper betle L and Curcuma longa L species in terms of chemical composition and biological activity has been studied in a systematic way There were differences between the chemical composition and biological activity of plant extracts and endogenous fungi This confirms the symbiotic and supportive relationship between host plants and endogenous fungi, as well as the potential of searching from endogenous plant fungi of alternative active ingredients to produce probiotics 3.2 A total of 19 compounds were isolated and structurally determined including compounds from A duperreana Pierre and A oligophylla Miq with known rocaglamide compounds (A, I, W, AB, J, rocaglaol) and new compound (rocaglamide AY), compounds known ar-tumeron, curcumin from C longa L., compounds known eugenol, chavicol, 4-Allylpyrocatechol from P betle L., known scopararane C compounds, diaporthein B from A duperreana Pierre endogenous fungi (M hawaiiensis), known compounds β-sitosterol, 4R, 4aS, 9aR) -1,9a-dihydronidulalin A, 4S, 4aR, 9aR) -4a-carbomethoxy-1,4,4a, 9atetrahydro-4,8-dihydroxy-6-methylxanthone and (24R) -methylcholesta-7.22 diene-3β, 5α, 6β-triol from endogenous fungi of Golden Turmeric (F oxysporum); and ergosterol from the P betle L endogenous (F solani) and identified 12 fatty acids from endogenous fungi of Golden Turmeric (F oxysporum) by the GC-MS data 3.3 A total of plant endogenous fungus have been isolated and identified These are the first announcements about the genome of endogenous fungal strains on the Aglaia duperreana Pierre, Piper betle L and Curcuma longa L plants in Vietnam 3.4 Extracts of leaves and bark of the Aglaia duperreana Pierre express 100% activity to inhibit to the growth of the Spodoptetra litura The extracted parts of Piper betle L and Curcuma longa L endogenous and curcumin essence inhibit 100% growth of the fungi causing the gray rot disease (Botrytis cinera) For the first time, the Golden turmeric and curcumin are systematically studied to be used as raw materials for processing biological fungicides The layout of the dissertation The dissertation consists of 141 pages and contains 159 references The layout of the dissertation includes the following sections: Preface (4 pages), Chapter 1: Overview (32 pages), Chapter 2: Objects and methods (13 pages), Chapter 3: Experimentals (19 pages), Chapter 4: Results and discussion (44 pages), Conclusions (1 pages), Recommendations (1 page), Publications (1 page), References (16 pages), and Appendix (43 pages) II DISSERTATION CONTENTS Preface This part discusses the background, the scientific and practical significance, and the objectives of the research project Chapter Literature review 1.1 The fungi, insects harmful and the role of plant protection drugs 1.2 The trend of replacing chemical pesticides with bio-pesticides 1.3 Biological pesticides extracted from plant materials 1.4 Plant endogenous fungi and the prospect of searching for new generation biologically active substances 1.5 Introduction of the species of Aglaia duperreana Pierre, Aglaia oligophylla Miq., Piper betle L and Curcuma longa L Chapter Methods 2.1 Isolation and purification methods Compounds were isolated and purified by using a combination of various chromatographic methods including thin-layer chromatography (TLC), column chromatography (CC) on different stationary phases such as Silicagel and Sephadex 2.2 Methods for the determination of the chemical structures The chemical structures of isolated compounds were elucidated by a combination of physical parameters (melting point), modern spectroscopic methods (IR, UV, CD, MS, 1D-NMR, and 2D-NMR) with chemical conversion, and by comparing with literature data 2.3 Methods for isolation and biomass of the endogenous fungi 2.4 Method for screening insecticidal and fungal activity Chapter EXPERIMENTALS 3.1 Result of isolation of endogenous fungi from plant samples + Four (04) endogenous fungal strains were isolated from Curcuma longa L.: Fusarium solani, Fusarium sp., Trichoderma atroviride and Fusarium oxysporum + Three (03) endogenous fungal strains were isolated from Aglaia duperreana Pierre: Colletotrichum gloeosporioides, Colletotrichum crassipes and Microdiplodia hawaiiensis + Two (02) endogenous fungal strains isolated from the Piper betle L are Colletotrichum sp and Fusarium solani 3.2 Result of isolation of plant compounds 3.2.1 Isolation of compounds from the Aglaia duperreana’s bark 3.2.1.1 Processing plant samples Bark dried samples (3kg) was extracted three times with methanol in an ultrasonic device at room temperature Translate the total amount of distilled solvent in the pressure drop, the temperature of 45 oC obtained 115g of methanol residue The residue of methanol is added with water and extracted with an increasing solvent of n-hexane and ethyl acetate After removal of the solvent, obtain the residue of n-hexane (25g), ethyl acetate (20g) and methanol (65g), respectively 3.2.1.2 Isolation of compounds from ethyl acetate residue Ethyl acetate residue (AD.E, 20g) is separated by column chromatography VLC with the solvent elution of n-hexane gradient: EtOAc: MeOH (4: 2: to 0: 1: solvent) segments denoted from ADE1 to ADE8 Diagram 3.2.1 Diagram to isolate compounds from Aglaia duperreana’s bark Run the chromatographic column of ADE3 segment (5.4 g) on silica gel (4063µm) with the gradient CH2Cl2-MeOH solvent system (from 100: to 0: 100) to obtain segments, symbols is ADE3.1-ADE3.9 Segment from ADE3.1 (1.29 g) run column CC with solvent CH2Cl2: isopropanol obtained segments (ADE3.1.1 to ADE3.1.9) Collect segments ADE3.1.4-ADE.1.1.7 (412mg) and run sephadex column with methanol solvent, collecting 36 small segments Use TLC and HPLC to collect tubes 1-36 to obtain clean substances obtained in the form of amorphous white powder The process of separating compounds from the bark of Aglaia dupperreana Pierre is described in the diagram 3.2.1  Compound 1: Compound (3.9 mg) was isolated from the bark of the Aglaia dupperreana in white amorphous form, [α] 20D-90.5 (c, 0.25, CHCl3) UV (MeOH) λmax 219.7 and 273.0 nm ESI-MS spectrometer (positive mode): m/z 561,1 (M+H)+, 528,4 (M+Na)+ 1H-NMR (MeOD): δ ppm 4,95 (d, J = 6.9 Hz, H-1), 4,11 (dd, J = 6.9 Hz, 13,8 Hz , H-2), 4,36 (d, J =13,8 Hz, H-3), 6,30 (d, J =1,9Hz, H-5), 6,17 (d, J =1,9 Hz, H-7), 7,12 (d, J=8,8 Hz, H-2’), 6,64 (d, J=8,8 Hz, H-3’), 6,64 (d, J =8,8 Hz, H-5’), 7,12 (d, J = 8,8 Hz, H-6’), 6,86 (m, H-2”), 6,98 (m, H-3”), 6,98 (m, H-4”), 6,98 (m, H-5”), 6,86 (m, H-6”), 3,81 (s, OMe-6), 3,84 (s, OMe-8), 3,66 (s, OMe-4’), 3,34 (s) & 2,86 (s) NMe  Compound 2: Compound (3,8 mg) was isolated from the bark of the Aglaia dupperreana in white amorphous form, [α]20D-80 (c, 0.45, CHCl3) UV (MeOH) λmax 209 and 279 nm ESI-MS spectrometer (positive mode): m/z 564,1 (M+H)+, 586,4 (M+Na)+ 1H-NMR (MeOD): δ ppm 6,03 (d, J = 5,0 Hz, H1), 4,29 (dd, J = 5,0 Hz, 14,5 Hz , H2), 4,29 (d, J =14,5 Hz, H3), 6,26 (d, J =1,9 Hz, H5), 6,11(d, J =1,9 Hz, H7), 6,78 (d, J=1,9 Hz, H-2’), 6,62 (d, J =8,2 Hz, H-5’), 6,70 (d, J = 6,9 Hz, H-6’), 7,02 (m, H-2”), 6,98 (m, H-3”), 6,98 (m, H-4”), 6,98 (m, H-5”), 7,02 (m, H-6”), 3,81 (s, OMe-6), 3,73 (s, OMe-8), 3,71 (s, OMe-4’), 3,37 (s) & 2,79 (s) NMe, 1,81 (s, OCOCH3) • Compound 3: Compound (2,1 mg) was isolated from the bark of the Aglaia dupperreana in white amorphous form, [α]20D-55,0 (c, 0.45, CHCl3) UV (MeOH) λmax 210 and 272,5 nm ESI-MS spectrometer (positive mode): m/z 534,1 (M+H)+, 556,4 (M+Na)+ 1H-NMR (MeOD): δ ppm 5,99 (d, J = 6,3 Hz, H1), 3,94 (dd, J = 5,9 Hz, 14,5 Hz , H2), 4,19 (d, J =14,5 Hz, H3), 6,26 (d, J =1,9 Hz, H5), 6,12 (d, J =1,9 Hz, H7), 7,17 (d, J=8,8 Hz, H-2’), 6,61 (d, J =8,8 Hz, H-3’), 6,61 (d, J = 8,8 Hz, H-5’), 7,17 (d, J=8,8 Hz, H-6’), 6,91 (m, H-2’), 7,00 (m, H-3”), 7,00 (m, H-4”), 7,00 (m, H5”), 6,91 (m, H-6”), 3,74 (s, OMe-6), 3,81 (s, OMe-8), 3,65 (s, OMe-4’), 2,57 (s, NMe), 1,84 (s, OCOCH3) • Compound 4: Compound (7,2 mg) was isolated from the bark of the Aglaia dupperreana in white amorphous form, [α]20D-110,0 (c, 0.45, CHCl3) UV (MeOH) λmax 210,4 and 272,6 nm ESI-MS spectrometer (positive mode): m/z 548,2 (M+H)+, 570,4 (M+Na)+ 1H-NMR (MeOD): δ ppm 5,95 (m, H1), 4,21 (m, H2), 4,21 (m, H3), 6,18 (d, J =1,9 Hz, H5), 6,03 (d, J =1,9 Hz, H7), 7,08 (d, J=8,8 Hz, H-2’), 6,54 (d, J =8,8 Hz, H-3’), 6,54 (d, J = 8,8 Hz, H-5’), 7,08 (d, J=8,8 Hz, H-6’), 6,80 (m, H-2”), 6,92 (m, H-3”), 6,92 (m, H-4”), 6,92 (m, H-5”), 6,80 (m, H-6”), 3,64 (s, OMe-6), 3,72 (s, OMe-8), 3,56 (s, OMe-4’), 3,27 (s) & 2,69 (s) NMe, 1,71 (s, OCOCH3) • Compound 5: Compound (1,9 mg) was isolated from the bark of the Aglaia dupperreana in white amorphous form, [α]20D-41,1 (c, 0.22, CHCl3) UV (MeOH) λmax 211,3 and 278,7 nm ESI-MS spectrometer (positive mode): m/z 509,0 (M+H)+, 531,2 (M+Na)+ 1H-NMR (MeOD): δ ppm 5,00 (d, J =5,7 Hz, H1), 3,96 (dd, J =5,7 Hz & 13,9 Hz, H2), 4,21 (d, J = 13,9, H3), 6,27 (d, J =1,9 Hz, H5), 6,15 (d, J =1,9 Hz, H7), 6,70 (d, J=1,9 Hz, H-2’), 6,64 (d, J = 8,8 Hz, H-5’), 6,64 (d, J=8,8 Hz, H-6’), 6,91 (m, H-2”), 7,00 (m, H-3”), 7,00 (m, H-4”), 7,00 (m, H-5”), 6,91 (m, H-6”), 3,81 (s, OMe6), 3,82 (s, OMe-8), 3,67 (s, OMe-4’), 3,61 (s, OCOCH3) • Compound 6: Compound (10 mg) was isolated from the bark of the Aglaia dupperreana in white amorphous form, [α]20D-125 (c, 0.48, CHCl3) UV (MeOH) λmax 212,8 and 272,3 nm ESI-MS spectrometer (positive mode): m/z 457,10 (M+H)+, 890,9 (2M+Na)+ 1H-NMR (MeOD): δ ppm 4,69 (d, J =5,5 Hz, H1), 2,80 (ddd, J =6,3 Hz & 13,5 Hz, 14, Hz, H-2α), 2,06 (ddd, J =1,1 Hz & 6,2 Hz, 11,8 Hz, H-2β) 3,89 (dd, J = 13,5 & 14,0 Hz, H3), 6,28 (d, J =1,9 Hz, H5), 6,17 (d, J =1,9 Hz, H7), 7,10 (d, J=8,8 Hz, H-2’), 6,61 (d, J = 8,8 Hz, H-3’), 6,61 (d, J=8,8 Hz, H-5’), 7,10 (d, J = 8,8 Hz, H-6’), 7,00 (m, H-2”), 7,00 (m, H-3”), 7,00 (m, H-4”), 7,00 (m, H-5”), 7,00 (m, H-6”), 3,87 (s, OMe-6), 3,85 (s, OMe-8), 3,81 (s, OMe-4’) 3.2.2 Isolation of compounds from leaves of Aglaia oligophylla 3.2.2.1 Processing plant samples The leaf sample of Aglaia oligophylla (3kg) was extracted times with methanol in the ultrasonic device at room temperature Translate the total amount of distillate solvent collected under reduced pressure, temperature 45 ° C, obtained 100g residue of methanol The residue of methanol is added with water and extracted with an increasing solvent of n-hexane, dichloromethane and ethyl acetate After removal of the solvent, obtain the residue of n-hexane (20g), dichloromethane (3.6g), ethyl acetate (18g) and methanol (55g), respectively 3.2.2.2 Isolation of compounds from diclometane residue Diclomethan extract (AO.D, 3.6 g) conducted with VLC silicagel 60 column obtained segments (AOD1 to AOD7) The OAD3 segment continues to run CC using a solvent system CH2Cl2: MeOH (10: 1) to obtain segments (OAD3.1 to OAD3.3) Compound is obtained by running preparative HPLC to OAD3.2 segment, detector λ = 210 nm with solvent system MeOH: H2O (3: 7) Diagram 3.2.2 Diagram to isolate compounds from leaves of Aglaia oligophylla  Compound (New Compound) Compound (3,3 mg) was isolated from the leaf of the Aglaia oligophylla Muq in white amorphous form, [α]20D-50,5 (c, 0.45, CHCl3) UV (MeOH) λmax 210,4 and 271,1 nm ESI-MS spectrometer (positive mode): m/z 528,1650 (M+Na)+ similar with C28H27NO8Na Spectrometer data of Compound showed at Table 4.3.1.1 3.2.3 Isolation of compounds from golden turmeric (Curcuma longa) 3.2.3.1 Processing plant samples The dried golden turmeric is finely ground, extracted with ethyl acetate solvent, then the solvent is then attracted to attract the essential oil 3.2.3.2 Isolation of compounds Turmeric essential oil (TDN, 30.8g) is separated on silica gel column chromatography with gradient n-hexan-ethyl acetate solvent system with 12 segments Segment is re-purified by sephadex LH20 with elution solvent MeOH obtained compound (2.8mg) Diagram 3.2.3 Diagram to isolate golden turmeric compounds The sludge residue after distillation entails extracting the vapors to extract the essential oil times with ethyl acetate or alcohol 960 The extract is vacuumed until only a concentrated solution is left in the heat room temperature to Compound (1.9 mg) was isolated from the bark of Aglaia dupperreana in white amorphous form, [α] 20D-41.1 (c, 0.22, CHCl3) UV spectra (MeOH) indicate λmax 211,3 and 278,7 nm Electronic atomic mass spectrometer ESI-MS (positive mode) for m / z 509,0 (M + H) +, 531,2 (M + Na) + corresponds to the molecular formula C28H28NO9 On the 1H-NMR spectrum of 5, two protons of H-5 and H-7 of ring A resonate at δH 6.27 ppm (d, J = 1.9Hz) and 6,15 ppm (d, J = 1.9 Hz) ) The hydroxyl group at C-3 'changed the spin system of AA'BB' characteristic of the B ring in the ABC spin rocglamide compounds as in compound This group has caused a change in the degree of transformation Study between methoxyl groups: OCH3-6 at δH 3.81 ppm, OCH3-8 at 2H 3.82 ppm and OCH3-4 'at δH 3.67 ppm Three protons of H-1, H-2 and H-3 resonate at δH 5.0 ppm (d, J = 5.7 Hz), 3.96 ppm (dd, J = 5.7 & 13.9 Hz) and δH 4.21 ppm (d, J = 13.9 Hz) In addition, the methoxyl group in the singlet form of the acetate fraction at C-2 appears the resonant signal at δH 3,61 (s) aromatic protons of C-ring observed at δH 6.91-7.00 ppm, are the same as in compounds 2, and Analysis of 1H-NMR, MS, UV spectra data of combined with comparison with spectral data and physical constants in reference [65] compound was identified as rocaglamide J with chemical structure as follows: 4.3.1.6 Compound (Rocaglaol) Compound (10mg) was isolated from the bark of Aglaia dupperreana in white amorphous form, [α] 20D-125 (c, 0.48, CHCl3) The UV spectrum (MeOH) of indicates 212max 212,8 and 272,3 nm Electron dust mass spectrometry ESI-MS (positive mode): m / z 457.10 (M + H) +, 890.9 (2M + Na) + corresponds to the molecular formula C26H26NO6 The 1H-NMR spectrum of compound shows protons of H-5 and H-7 of ring A resonating at δH 6.28 ppm (d, J = 1.9 Hz) and at δH 6.17 (d, J = 1.9Hz) Very special AA'BB spin system 'of round B was observed at δH 7.10 (2H, d, J = 8.8Hz) and at δH 6.61 (2H, d, J = 8.8 Hz) Three groups of methoxyl in the form of singlet have the resonant signal at δH 3.87 (OMe-6), at δH 3.85 (Ome-8) and at δH 3.81 (Ome-4 ’) Comparing with compound 1, compound has a change in the aliphatic region The resonance of the methylene proton appears as a pair of multilayered geminal interactions (m) at δH 2.06 ppm (ddd, J = 1,1, 6,2 & 11,8 Hz) and 2,80 ppm (ddd , J = 6.2, 11.8 & 14.0 Hz), both protons also exhibit vicinal interaction with methyl groups surrounded by phenyl group and hydroxyl group at δH 3.89 and 4.69 ppm Thus the methylene signals have a link between the methyl protons The signal of methylene group appeared at δH 2.06 and 2.80 ppm of H-2α and H-2β was explained with relatively small spin interaction constant (J = 1.5 Hz) in equatorial-equatorial form and a large interaction constant (14.0 Hz) axial-axial form for H-1β and H-3α respectively 19 Analysis of 1H-NMR, MS, UV spectra data of combined with comparison with spectral data and physical constants in reference [65] compound was identified as rocaglaol with the chemical structure as follows : 4.3.1.7 Compound (Rocaglamide AY) - new substance Compound (3.3 mg) was isolated from leaves of Aglaia oligophylla Muq In the amorphous white form, [α] 20D-50.5 (c, 0.45, CHCl3) UV spectra (MeOH) of indicate λmax 210.4 and 271.1 nm High resolution mass spectrometry HRESIMS for molecular dummy ion peaks at m / z = 528,1650 [M + Na] +, corresponding to the molecular formula C28H27NO8 1H and 13C-NMR spectra of compound are similar to the spectrum of rocaglamide J (compound 5) with the signal of methoxy groups at δH 3.90 (8OMe); 3.84 (6-OMe) and 3.71 (4'-OMe), together with two fragrant proton signals with meta-interaction at δH 6.15 and 6.26 (each signal d, J = 1.9Hz) , protons of aromatic rings were replaced once at δH 6.99 - 7.12 ppm and acetate groups (δC 57.5 and 170.0ppm) The difference of compound compared to compound is shown in two points The first is that the hydroxy group at C-1 in compound has been converted to the C-1-oxime substituent in the molecule, which is demonstrated by the low-field shift of C-1 (δC 153.0 ppm) compared to compound Secondly, the characteristic signal appears for the spin AA'BB 'system of ring B at δH 7.13 (2H, d, J = 8.8Hz) and 6.71 (2H, d, J = 8.8Hz), this proves that the hydroxy substituents did not exist in the compound The 2DNMR spectrum analysis of substance combined with the literature leads to the conclusion that its structure is C-3'-demethoxy derivative of rocaglamide T [48,66] This is a new substance and is named rocaglamide AY Table 4.3.1.1 NMR data of compound (CDCl3) Vị trí *δC (ppm) 153,0 57,0 57,1 δC (ppm) δH (ppm)J(Hz) 153,0 57,1 3,80 (d,13,5) 57,2 3,67 (d,13,5) 3ª 4ª 8ª 105,1 160,0 88,9 164,0 93,0 158,3 107,7 105,7 161,3 89,9 165,3 93,8 160,3 110,0 HMBC COSY 3,8b, 9, 1” 2,3a, 9, 1’, 1”,2”/6” 6,26 (d,1,9) 4a,6,7,8a 6,15 (d,1,9) 5,6,8,8a 20 Vị trí 8b 1’ 2’b 3’c 4’ 5’c 6’b 1” 2”/6” 3”/5” 4” (CO) 8-OCH3 6-OCH3 4’-OCH3 COOCH3 *δC (ppm) 115,0 125,6 113,1 127,8 158,8 126,8 125,6 134,8 127,7 127,8 δC (ppm) δH (ppm)J(Hz) 117,0 128,7 113,2 7,13 (d,8,8) 149,3 6,71 (d,8,8) 149,5 11,5 6,71 (d,8,8) 121,4 7,13 (d, 8,8) 136,7 129,4 6,99 (m) 128,7 7,12 (m) 127,8 170,0 128,0 171,7 56,1 56,3 56,3 57,5 HMBC COSY 3a 2’,6’,4’,1” 3’/5’ 2’/6’ 2’/6’,4’,1” 3a 2’/6’ 3’/5’ 3”/5” 1”,2”/6” 3”/5” 2”/6”, 4” 3”/5” 7,12 (m) 3,90 (s) 3,84 (s) 3,71 (s) 4’ (C=O) * δC: 13C-NMR spectral data of rocaglamide T measured in CDCl3 [66] 4.3.2 Chemical composition of yellow turmeric (Curcuma longa L.) From the dichloromethane extract residue of yellow turmeric, two compounds were isolated and determined chemical structure including ar-turmerone (8) and curcumin (9) The chemical structure of these compounds is determined based on 1H-, 13C-NMR spectral data, and compared with previously published documents for known compounds 4.3.2.1 Compound (Ar-turmerone) 1H and 13C-NMR spectra of substance showed that the molecule contained an aromatic ring in position and 4, expressed by the signal of aromatic protons at δH = 7.1 (m, 4H, H-2, 3,5,6) and aromatic carbon, including CH and carbon 4; methyl groups include three tertiary groups [δH = 1.84; 2.1 and 2,3 (each signal 3H, s)], a quadratic group [δH = 1.23 (3H, d, J = 7Hz), H-15CH3] Besides, NMR spectrum also shows the presence of a ketone group at δC = 199.8 (C-9); olefinic proton in δH 6.02 (H-10) and aliphatic proton resonates in the region from 2.61 to 3.28ppm The spectral data analyzed above are completely consistent with the Ar-turmerone data in reference [146] Recent research shows that this compound exhibits the activity of repelling the two insects Sitophilus zeamais and Spodoptera frugiperda [146] 4.3.2.2 Compound (Curcumin) 21 Essence of curcumin (compound 9) is purified by preparative thin-plate chromatography with the solvent dichloromethane: methanol (98: 2) Nuclear magnetic resonance data show that this is a 50:50 mixture of two enol and ketone profiles of curcumin 4.3.3 Chemical composition of Piper betle L From the dichloromethane extract residue, there are compounds isolated and determined chemical structure including eugenol (10), chavicol (11), 4allylpyrocatechol (12) The chemical structure of these compounds is determined based on 1H-, 13C-NMR spectral data, and compared with previously published documents for known compounds 4.3.3.1 Compound 10 (Eugenol) On 1H-NMR spectrum of substance 10 showed that the signal of aromatic protons of one aromatic ring was replaced times in δH 6.65 (dd, J = 2; 8.5 Hz, 1H, H-3), 6, 75 (brs, 1H, H-5), 6.77 (d, J = Hz, 1H, H-6) and signals of the allyl group: olefinic protons in δH 5.03 (H-9); 5.93 (H-8) and a methylene group at 3.28 (H-7) There are also methyl groups attached to aromatic rings at 3.83 (s, 3H, H-2-OCH3) Analyzing 1H-NMR spectral data in combination with comparing spectral data reference [147] allows to confirm that compound 10 is named eugenol 4.3.3.2 Compound 11 (Chavicol) Signals of 11 protons were observed on 1H-NMR spectrum of compound 11 In which, there are aromatic protons of an aromatic ring replaced twice in ortho δH position 6.77 (m, 2H, H-2, 6); 7.04 (m, 2H, H-3,5) and signals of allyl group: olefinic protons in δH 5.05 (m, 2H, H-9); 5.93 (m, 1H, H-8) and a methylene group at δH 3.3 (d, J = 6.5, 2H, H-7) 13C-NMR spectrum of 11 indicates that there is a total of carbon molecules in the molecule, including methylene groups at δC 39.5 (C-7) and 115,2 (C-9), methin groups at nhómC 115.4 (C2.6); 129,6 (C-3,5) and 137,8 (C-8), carbon at level at δC 132,2 (C-4) and 153,9 (C-1), there is carbon at the fourth level with oxygen at δC 153.9 (C-1) Analysis of 1H- and 13C-NMR spectral data in combination with reference spectral data reference [148] allowed to confirm that compound 11 is named chavicol with the chemical structure below 4.3.3.3 Compound 12 (4-allylpyrocatechol) Signals of protons were observed on the 1H-NMR spectrum of 12 There were aromatic protons of an aromatic ring at δH 6.62 (dd, J = 2.0 and 8.1Hz, 1H, H- 5); 6.70 (d, J = 2.0Hz, 1H, H-6); 6.78 (d, J = 8.1Hz, 1H, H-3) and signals of allyl group: olefinic protons in ởH 5.03 (m, 1H, H-9cis); 5.06 (m, 1H, H-9trans); and 5.90 (m, 1H, H-8) and one methylene group at δH 2.05 (2H, d, J = 6.7 Hz, H-7) In addition, the spectrum also showed signals of two hydroxy groups at δH 5,48 (1-OH); 5.53 (2-OH) (br s, 1H signal) 22 The spectral data of 12 completely coincide with 4-allylpyrocatechol in reference [148] 4.3.4 Chemical composition of M hawaiiensis from Aglaia From the residue of diclometan fragment of M hawaiiensis, two compounds were isolated and identified chemical structure including Scopararane C (13) and Diaporthein B (14) The chemical structure of these compounds is determined based on 1H-, 13C-NMR spectral data, and compared with previously published documents for known compounds 4.3.4.1 Compound 13 (C Scopararane) Compound 13 is obtained in the form of a white needle-shaped crystal, Rf = 0.87 (solvent n-hexane-ethyl acetate 3: 1), melting point 171,6 0C 1H NMR spectrum of substance 13 shows the signal of olefinic protons resonating at δH 7.04 (1H, d, J = 1.5Hz, H-14); 5.87 (1H, dd, J = 17.5; 10.5Hz, H15a) and 5.05 (1H, dd, J = 11.0; 0.5Hz, H-15b); 5.11 (1H, dd, J = 17.5; 0.5Hz, H16) of two double bonds, one double bond is replaced once and one double bond is replaced times; methyl groups in the form of singlet at δH 1.20 (s, 6H, H17,20); 1.29; 1.40 (each 3H, s, H-18,19) and 10 aliphatic proton signals resonate in the region from 1.4 to 2.0 ppm 13C NMR spectrum of compound 13 gives the signal of 20 carbon, including groups CH3, groups CH2, CH groups and carbon ranks Besides signals suitable for 1H spectrum, 13C NMR spectrum also gives see the presence of carbonyl group (δC 181,6, C-7), double bond alternate times at δC 142.7 (C-5), in which the signal is shifted toward the low field at δC 144 , (C-6) showed a double bond hydroxy substituent and a carbon aliphatic linked to oxygen shown in signal ởC 74.1 (C-9) NMR spectral data of 13 see table 4.3.4.1 The 1H and 13C NMR spectroscopic data analyzed here are in comparison with the data in the reference [149] to confirm that compound 13 is called Scopararane C The chemical structure is as follows 4.3.4.2 Compound 14 (Diaporthein B) Substance 14 is obtained in the form of a white square-shaped crystal, Rf = 0.33 (dm: n-hexane-ethyl acetate 3: 1), mm 189.6 0C Mass spectrometry ESI-MS for molecular ion peaks at m / z 363 [M - H] - Combined with preliminary analysis of MS spectrum data, 1H and 13C NMR spectra, it can be confirmed that 14 is a diterpen with molecular formula C20H28O6 The 1H and 13C NMR spectra of 14 are almost similar to those of substance 13 only in the following three points: The first point is the signal of the C-20 methyl group at δH 1.20 in substance 14 replaced by the signal of the oxygen group methylene at δH 3.71 and 4.14 (each signal d, J = 10.0Hz, H-20) The second difference is the signal loss of carbon olefinic and the emergence of quadratic aliphatic carbon attached to oxygen at 81 81.9 (C-5) and 104.1 (C-6) in substance 23 14 Significantly low field-shift signals at 104.1 ppm (C-6) are due to the association with oxygen atoms These two differences can be explained by the formation of ether bonds between C-20 and C-6 Thirdly, the spectrum of substance 14 also showed the signal of oxygen-methine group at δH 4.03 (1H, dd, H-11) and δC 67.7 (C-11) Analysis of 1D-NMR, MS spectroscopic data, of 14 combined with comparison with spectral data in reference [150], compound 14 was identified as diaporthein B with the chemical structure as follows: Table 4.3.4.1 NMR spectral data of compounds 13 and 14 C-NMR (CDCl3) (δ ppm) 13 14 30,06 25,2 17,7 17,7 41,4 37,5 35,4 37,3 13 142,7 81,9 142.8 81.9 144,4 104,1 144.4 104.1 181,6 196,3 181.6 196.2 133,7 134,7 133.7 34.7 74,1 76,2 74.1 76.2 45,1 51,1 45.1 51.1 25,2 67,7 25.2 67.7 29,6 81,92 29.5 39.9 38,7 6,81 (d; 2) 147,8 5,82 (dd, 17,5; 11) 112,5 40,01 150,5 144,0 38.8 147.9 145.4 40.1 150.4 144.1 5,1 (m) 145,4 113,1 112.6 113.1 Vị trí C H-NMR (CDCl3) (δ ppm) 13 14 1,53 (m); 2,0 (m) 1,97 (m); 2,04 (m) 1,66 (m) 1,63 (m); 1,68 (m) 1,45 (m); 1,56 (m) 1,27 (m); 1,60 (m) 10 11 1,59 (m); 1,8 (m) 12 1,94 (m); 1,97 (m) 13 14 15 4,03 (dd) 1,75 (m) 2,07 (m) 13 C-NMR (CDCl3) (δ ppm) 13* 14* 30.6 25.2 17.7 17.6 41.1 37.5 35.4 37.3 17 7,04, (d; 1,5) 5,87 (dd, 17,5; 10,5) 5,05 (dd; 11,0; 0,5); 5,11 (dd; 17,5; 0,5) 1,20 (s) 1,22 (s) 23,3 25,97 23.3 25.9 18 1,40 (s) 1,19 (s) 30,6 26,97 30.1 26.9 19 1,29 (s) 26,9 23,73 26.9 23.7 20 1,20 (s) 29,4 68,66 29.4 68.6 -OH 6,72 (s) 1,44 (s) 3,71 (d, J=10,0); 4,14 (d, J=10,0) 4,97 (s) 16 13 *: Scopararane C measured in CDCl3; 1H-NMR (500 MHz); 13C-NMR (125 MHz) [149] 14 *: Diaporthein B measured in CDCl3; 1H-NMR (500 MHz); 13C-NMR (125 MHz) [150] 4.3.5 Chemical composition of F oxysporum from Golden turmeric tree From the residue of methanol extract of F oxysporum, compounds 15-26 were identified by the method of mass spectrometry chromatography (GC / MS) and compounds were isolated and identified chemical structure includes β-sitosterol 24 (27), 4R, 4aS, 9aR) -1,9a-dihydronidulalin A (28), (4S, 4aR, 9aR) -4acarbomethoxy-1,4,4a, 9a-tetrahydro -4,8-dihydroxy-6 methylxanthone) (29) and (24R) -methylcholesta-7,22-diene-3β, 5α, 6β-triol (30) The chemical structure of these compounds is determined based on 1H-, 13C-NMR spectral data, and compared with previously published documents for known compounds 4.3.5.1 Compounds from 15 to 26 Compounds from 15 to 26 are identified by the technique of Gas Chromatography Mass Spectometry GC / MS results of substances 15-26 are described in Table 4.3.5.1 Table 4.3.5.1 GC-MS results of substances 15-26 TT 15 16 17 18 19 20 21 22 23 24 25 26 Time 10.39 13.57 16.46 17.22 19.69 24.07 24.28 24.49 25.34 33.75 41.92 49.68 Fatty acids 14: 15:0 16:1n-7 16: 17: 18: 2(n-6) 18:1(n-9) 18:1(n-7) 18: 20:0 22:0 24:0 Hàm lượng % 0.93 0.20 2.55 29.04 2.82 27.98 28.63 0.27 6.9 0.34 0.17 0.17 Tên khoa học Tetradecanoic acid Pentadecanoic acid 9-hexadecenoic acid Hexadecanoic acid Heptadecanoci acid Octadecadienoic acid Octadecenoic acid Octadecenoic acid Octadecanoic acid Eicosanoic acid Docosanoic acid Tetracosanoic acid 4.3.5.2 Compound 27 (β-sitosterol) Compound 27 is obtained in the form of needle-shaped crystals, white, melting temperature is 132-133 OC, Rf = 0.26 (solvent system deploys n-hexane-axeton 9: 1) Then use TLC chromatography with standard β-sitosterol and determined compound 27 is β-sitosterol, with the chemical structure as follows: 4.3.5.3 Compound 28 (4R, 4aS, 9aR) -1,9a-dihydronidulalin A) 1H NMR spectrum of substance 28 gives signals of olefinic protons in (H-3), 6.06 (H-2), 6.29 (H-5) and 6.38 (H5Hz, H-4), three proton aliphatic at ởH 2.67 (H-1a), 2.76 (H-1b) and 3.64 (H-9a) 3H, 611.52 s, 1H, 8-9a) was also observed on 1H NMR spectrum On the 13C-NMR spectrum, compound 28 also -9 - 4a), 105.1 (C-8a), 149.9 (C-6), 157.6 (C-10a), 161.4 (C-8) The above analytical spectra data of compound 28 completely coincide with the data of the compound (4R, 4aS, 9aR) 1,9a-dihydronidulalin A in reference [151] Therefore, compound 28 is defined as 25 4R, 4aS, 9aR) -1,9a-dihydronidulalin A The spectral data of compound 28 are shown in Table 4.3.5.2 with the chemical structure as follows: 4.3.5.4 Compound 29 (4S, 4aR, 9aR) -4a-carbomethoxy-1,4,4a, 9a-tetrahydro4,8-dihydroxy-6-methylxanthone) NMR spectrum of compound 29 is very similar to that of compound 28, which is also a 4a-carbomethoxy-4,8-dihydroxy-6-methylxanthone derivative The difference is expressed in a low -4 and a high field -4a compared to compound 28 This allows the prediction of compound 29 as optical isomer of substance 28 The structure of 29 is determined to be (4S, 4aR, 9aR) -4a-carbomethoxy-1,4,4a, 9a-tetrahydro-4.8dihydroxy-6-methylxanthone when comparing its spectral data with reference [151] Spectrum data of compound 29 are shown in Table 4.3.5.2 Table 4.3.5.2 NMR spectral data of compounds 28 and 29 STT Hợp chất 28 13 H C 2.67 m, 1H 24.5, CH2 2.76 m, 1H 6.06 m, 1H 132.6, CH 5.88 m, 1H 123.7, CH 4.67 d, 5, 1H 66.1, CH 4a 85.2, C 6.29 s, 1H 108.1, CH 149.9, C 6.38 s, 1H 111.2, CH 161.4, C 8a 105.1, C 197.7, C 9a 3.64 m, 1H 40.3, CH 10a 157.6, C 6-CH3 2.28 s, 3H 22.4, CH3 OH-8 11.52 s, 1H COOCH3 168.4, C COOCH3 3.64 s, 3H 52.9 CH3 Hợp chất 29 13 C 25.6, CH2 13 C-28* 24.5 132.6 123.7 66.0 85.2 108.0 149.8 111.2 161.3 105.1 197.7 40.3 157.6 22.4 40.3 168.3 52.9 5.96 m, 1H 5.79 m, 1H 4.62 m, 1H 6.43 s, 1H 6.49 s, 1H 3.34 dd (7&11), 1H 2.36 s, 3H 11.34 s, 1H 3.77 s, 3H H 127.5, CH 127.3, CH 69.3, CH 82.7, C 108.4, CH 151.1, C 111.1, CH 162.5, C 103.6, C 197.1, C 45.2, CH 159.0, C 22.6, CH3 170.7, C 53.4, CH3 13 C-29* 25.6 127.4 127.3 69.3 82.7 108.4 151.1 111.1 162.5 103.6 197.1 45.2 159.0 22.6 170.7 53.4 28 *: 4R, 4aS, 9aR) -1,9a-dihydronidulalin A measured in CDCl3; 13C-NMR (125 MHz) [151] 29 * :( 4S, 4aR, 9aR) -4a-carbomethoxy-1,4,4a, 9a-tetrahydro-4,8-dihydroxy-6methylxanthone measured in CDCl3; 13C-NMR (125 MHz) [151] 4.3.5.5 Compound 30 (24R-methylcholesta-7,22-diene-3β, 5α, 6β-triol) Compound 30 is obtained in the form of soluble white powder in hot methanol, melting heat is 221-224oC, Rf = 0.61 (solvent dichloromethane-methanol 9: 1) 1H-NMR spectrum shows signals of methyl groups, including two single signals (singlet) at δH 0.91 (H-18); 0.54 (H-19) and four doublet at δH = 0.99 (J = 6.5Hz, H-30); 0.88 (J = 7.0 Hz, H-28); δH = 0.81 (J = 6.5Hz, H-27) and δH = 0.80 (d, J = 6.5Hz, H-26) In addition, the presence of two double bonds is expressed in three signals, including two protons at the trans position of a double bond substituted 26 twice at δH 5.23 (dd, 1H, J = 7.5 & 15.5Hz, H- 23); 5.17 (dd, 1H, J = & 15.5Hz, H-22) and a proton of double coupling is replaced three times at δH = 5.08 (1H, m, H-7); signals of three hydroxyl groups at δH 4.48 (1H, d, J = 5.5Hz, H-29), 4.22 (1H, d, J = 5.5Hz, H-30) and 3.58 (1H, s, H-31) ; signals of two methine groups attached to the hydroxyl group at δH 3.76 (1H, m, H-3), 3.37 (1H, s, H-6) and 20 aliphatic protons in the area from 1.2 to 2.0ppm were also observed in 1HNMR spectrum 13C-NMR spectrum shows that molecule 30 has 28 carbon atoms, perfectly suited to the 1H-NMR spectrum data, including two double bonds at δC 139.6 (C-8), 135.3 (C-22), 131.3 (C-23) and 119.4 (C-7); three carbon is attached to oxygen at δC 74.4 (C-5), 72.1 (C-6), 65.9 (C-3); methyl groups at δC 12.0 (C-19), 17.2 (C-28), 17.6 (C18), 19.4 (C-26), 19.7 (C-27), 20.9 (C-20) and 15 carbon resonate in areas from 21.30 to 55.30 ppm Analysis of 1D-NMR spectroscopic data allows the estimation of compound 30 as a sterol in the form of trihydroxyergostane The structure of compound 30 is concluded as (24R) methylcholesta-7,22-diene-3β, 5α, 6β-triol when compared with literature [18] This compound was previously isolated from a soft coral Sinularia sp The chemical structure of compound 30 is as follows: Table 4.3.5.3 Spectrum data 1H- and 13C-NMR of substance 30 27 Hợp chất 30 (1H-NMR: 500Hz 13C-NMR: 125 MHz) STT 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 H (DMSO) 1.6 m & 1.2 m 1.4 m & 1.3m, 1H 3.76 m, 1H 1.9 m & 1.4 m 3.37 m, 1H 5.08 m, 1H 1.92m, 1H 1.42 m 1.95 m & 1.23 m 1.80 m 1.48 m & 1.3 m 1.7 m & 1.2 m 1.25 m 0.91 s, 3H 0.54 s, 3H 0.99 d (6.5 , 3H) 2.0 m 5.17 dd, 1H, & 15.5 5.23 dd, 1H 7.5 & 15.5 1.82 m, 1H 1.4 m, 1H 0.80 d (6.5, 3H) 0.81 d (6.5, 3H) 0.88 d (7.0, 3H) 4.48 d (5.5, 1H) OH 4.22 d (5.5, 1H) OH 3.58 s, 1H OH 13 C* (DMSO) 32.6 33.9 67.6 41.9 76.1 74.2 120.4 141.6 43.8 38.1 22.4 39.9 43.8 55.3 23.5 28.5 56.2 18.8 12.5 21.5 40.8 136.2 132.1 43.1 33.4 19.8 20.2 17.8 13 C (DMSO) 31.1 CH2 32.4 CH2 65.9 CH 40.0 CH2 74.4 C 72.1 CH 119.4 CH 139.6 C 42.2 CH 36.6 C 21.3 CH2 39.0 CH2 42.9 C 54.1 CH 22.5 CH2 27.6 CH2 55.3 CH 17.6 CH3 12.0 CH3 20.9 CH3 39.9 CH 135.3 CH 131.3 CH 41.9 CH 32.4 CH 19.4 CH3 19.7 CH3 17.2 CH3 30 *: 4R, 4aS, 9aR) -1,9a-dihydronidulalin A measured in CDCl3; 13C-NMR (125 MHz) [18] 4.3.6 The chemical composition of F sonani from the Piper betle From the residue of ethyl acetate extract, we isolated and determined the chemical structure of ergosterol compound (31) The chemical structure of this compound is determined based on 1H-, 13C-NMR spectral data, and compared with previously published documents Compound 31 (Ergosterol): Compound 31 (7.1mg), is a needle-shaped crystal, melting temperature 168-169 ° C The 13C-NMR spectrum showed that the compound 31 has a total of 28 carbon, including methyl groups, methylene groups, 11 metine groups and quadratic carbons NMR spectroscopic data of compound 31 allow this prediction to be a sterol framing agent, shown by the characteristic signal of a hydroxy metine group at - 3), methyl groups, including singlet -18), 0.92 (H-27), 0.97 (H-28) and 1.09 (H-26) with many resonant aliphatic protons in the region 28 from about 1.2 to 2.5 ppm In addition, the 1H-NMR spectrum of compound 31 also showed the presence of three double bonds, expressed by the signal of four -H-22,23), 5.43 (1H , H-7) and 5.62 (1H, H-6) -7), 119.6 (C-6), 132.0 (C-23), 135.5 (C-22), 139.8 (C-5) and 141.3 (C-8) Comparing the spectral data of compound 31 with reference [152], it can be concluded that compound 31 is called ergosterol Synthetic biosynthesis ergosterol can inhibit pathogenic fungi, pathogenic microorganisms, make chemicals produce agricultural medicines and potentially cytotoxic NMR spectral data of compound 31 are presented as table 4.3.6.1 Chemical structure of compound 31: Table 4.3.6.1 1H- and 13C-NMR spectral data of substance 31 Vị trí C 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 H (ppm) 3.69 (m) 5.62 (dd, 2.4, 5.4) 5.43 (dd, 2.4, 5.4) 0.61 (s) 0.92 (s) 1.01 (d, 6.6) 5.20 (dd, 7.8, 15.0) 5.20 (dd, 7.8, 15.0) 1.09 (d, 6.6) 0.88 (d, 6.6) 0.97 (d, 6.6) C (ppm) 38.4 32.0 70.4 40.8 139.8 119.6 116.3 141.3 46.2 37.0 21.1 39.1 42.8 54.6 23.0 28.3 55.7 12.0 17.6 40.4 21.1 135,6 132.0 42.8 33.1 19.6 19.9 16.3 C (ppm) 31* 38.4 32.0 70.4 40.8 139.8 119.6 116.3 141.3 46.2 37.0 21.1 39.1 42.8 54.6 23.0 28.3 55.7 12.0 17.6 40.4 21.1 135,6 132.0 42.8 33.1 19.6 19.9 16.2 DEPT CH2 CH2 CH CH2 C CH CH C CH C CH2 CH2 C CH CH2 CH2 CH CH3 CH3 CH CH3 CH CH CH CH CH3 CH3 CH3 31 *: Ergosterol measured in CDCl3; 1H-NMR (500 MHz); 13C-NMR (125 MHz) [152] The results of chemical composition analysis of compounds in plants and plant endogenous fungus show many interesting things: 29 + The chemical composition of natural compounds in plants and in plant endogenous fungi is not the similar + The endogenous fungal extracts of A duperreana Pierre , C longa L and P betle L have the different biological activity than the host plant extract These survey results provide interesting guesses about the extremely diverse role of endogenous fungi on host plants and should be sought to better understand these coordination mechanisms, Chapter CONCLUSIONS AND PROPOSAL 5.1 Conclusion For the first time in Vietnam, the relationship between plants and plant endogenous fungi on the species of A duperreana Pierre , C longa L and P betle L on the chemical composition and biological activity has been studied in a systematic way There were differences between the chemical composition and biological activity of plant and endogenous fungi extracts This confirms the symbiotic and supportive relationship between host plants and endogenous fungi, as well as the potential of searching from endogenous plant fungi of alternative active ingredients to produce probiotics A total of 19 compounds were isolated and structurally determined (including: compounds from A duperreana Pierre and A oligophylla Miq with compounds know rocaglamide A, I, W, AB, J, rocaglaol and new compound rocaglamide AY, compounds known ar-tumeron, curcumin from C longa L., compounds known eugenol, chavicol, 4-Allylpyrocatechol from P betle L., known scopararane C compounds, diaporthein B from endogenous fungi M hawaiiensis, known βsitosterol, 4R, 4aS, 9aR) -1,9a-dihydronidulalin A, 4S, 4aR, 9aR) -4a-carbomethoxy1,4,4a, 9a-tetrahydro-4,8-dihydroxy- 6-methylxanthone, (24R) -methylcholesta-7 , 22diene-3β, 5α, 6β-triol from F oxysporum; and ergosterol from the F solani and identified 12 fatty acids from endogenous fungus of Golden Turmeric (F oxysporum) by GC-MS data A total of plant endogenous fungus strains have been isolated and identified These are the first announcements about the genome of endogenous fungal strains on the Vietnamese C longa L and P betle L plants in Vietnam 30 Extracts of leaves and bark of the A duperreana Pierre express 100% activity to inhibit cavity growth of Spodoptetra litura The extract of the C longa L plant, endogenous fungus F oxysporum and F solani and pure Curcumin compound essence inhibit 100% growth of fungi causing gray rot disease (Botrytis cinera) For the first time, yellow turmeric C longa L and curcumin are systematically studied to be used as raw materials for processing biological fungicides 5.2 Proposal Further studies should be implemented for the biological fungicides production 31 PUBLICATIONS Articles Duong Ngoc Tu, Duong Anh Tuan, Le Dinh Minh, Trinh Duc Cong, Dang Xuan Quy, Nguyen Ngoc Hieu, Dang Duc Quyet, Vu Duy Hien, Dang Vu Thi Thanh (2013), Research on manufacturing Funbv preparations except Botrytis cinerea fungi damage plants from golden turmeric extract (Curcuma longa) Chemistry and Applications, (19), 29-32 ISSN 1859-4069 Duong Ngoc Tu, Nong Thien Sang, Le Dinh Minh, Nghiem Ngoc Duc, Duong Anh Tuan, Nguyen Thanh Thuy, Nguyen Thi Hien, Nguyen Ngoc Hieu, Dang Duc Quyet, Vu Duy Hien, Dang Vu Thi Thanh (2013) Study on the ability to inhibit Botrytis cinerea fungi causing gray rot disease in vegetables from golden turmeric (Curcuma longa) of Vietnam Chemistry and Applications, (21), 10-13 ISSN 1859-4069 Ngoc Tu Duong, RuAngelie Edrada-Ebel, Rainer Ebel, Wenhan Lin, Tuan Tuan, Xuan Quy Dang, Ngoc Hieu Nguyen and Peter Proksch (2014) New rocaglamide derivatives from Vietnamese Aglaia species Natural Product Communications, (6) pp 833-834 ISSN 1934-578X Nguyen Ngoc Hieu, Nghiem Van Duc, Nguyen Thi Thanh Thuy, Nguyen Thi Hien, Duong Anh Tuan, Bui Thi Hai Yen, Nguyen Viet Ha, Duong Ngoc Tu (2014) Study on the effectiveness of fungicides and harmful effects of FUNBV in Me Linh, Hanoi Journal of Chemistry T.52 (6A) 93-97 ISSN 0866-7144 Nguyen Ngoc Hieu, Nghiem Van Duc, Pham Thi Dung, Duong Ngoc Tu (2016) Study on chemical composition and activity of endogenous fungi isolated from golden turmeric Chemical journal, 54 (3) 382-386 ISSN 0866-7144 Poster 32 Nguyen Ngoc Hieu, Nghiem Van Duc, Nguyen Thi Thanh Thuy, Nguyen Thi Hien, Duong Anh Tuan, Bui Thi Hai Yen, Nguyen Viet Ha, Duong Ngoc Tu (2015) Study on ability of Funbv botanical product to control fungal diseases on vegetables in Me Linh, Hanoi.The fourth youth scientific Conference, Institute of Chemistry, VAST, Feb 4th 2.Nguyen Ngoc Hieu and Duong Ngoc Tu (2016) Stuty on structure of endophytic fungi substances isolated from medical Vietnamese plants Molecular biology and its applications in health and food/feed production in South-East Asia, IPMB Alumni Meeting, Sep 18–21, Can Tho University 33 ... turmeric is finely ground, extracted with ethyl acetate solvent, then the solvent is then attracted to attract the essential oil 3.2.3.2 Isolation of compounds Turmeric essential oil (TDN, 30.8g)... 4.3.5.1 GC-MS results of substances 15-26 TT 15 16 17 18 19 20 21 22 23 24 25 26 Time 10.39 13.57 16.46 17.22 19.69 24.07 24.28 24.49 25.34 33.75 41.92 49.68 Fatty acids 14: 15:0 16:1n-7 16: 17: 18:... compound 29 are shown in Table 4.3.5.2 Table 4.3.5.2 NMR spectral data of compounds 28 and 29 STT Hợp chất 28 13 H C 2.67 m, 1H 24.5, CH2 2.76 m, 1H 6.06 m, 1H 132.6, CH 5.88 m, 1H 123.7, CH 4.67