Conservation of codonopsis javanica (blume) hook f et thomson and amomum longiligulare t l wu in son la province

MINISTRY OF AGRICULTURE AND RURAL DEVELOPMENT VIETNAM NATIONAL UNIVERSITY OF FORESTRY NGUYEN VU GIANG CONSERVATION OF CODONOPSIS JAVANICA (BLUME) HOOK F ET THOMSON AND AMOMUM LONGILIGULARE T.L.WU IN SON LA PROVINCE MAJOR: TROPICAL FORESTRY Code: 8620201 MASTER THESIS DISSERTATION TROPICAL FORESTRY ACADEMIC ADVISORS: ASSOC PROF DR LE XUAN TRUONG PROF DR HOANG VAN SAM Hanoi, 2022 i ABSTRACT This study was carried out within the framework of the scientific research and technological development project in Son La province with the ministerial level of Education and Training Ministry, code B2019-TTB-03 The research implement in districts such as Moc Chau, Van Ho, Song Ma, Thuan Chau, Sop Cop and Seed for propagation experiment were collected in study area as Van Ho district, Song Ma district, Thuan Chau For Codonopsis Javanica (Blume) Hook F et Thomson, the results show that the fruit diameter is from 1.2 to 1.3cm, the purity of the seeds is from 88.0 to 91.0%, the average weight of 1000 seeds is 0.194g, there are from 810 to 893 seeds/fruit 5,076,142 - 5,208,333 seeds/kg; germination rate from 7.4 to 7.5 days Germination rate did not differ significantly between the seed lots and seed samples Seeds germinate from 89.0 - 91.3%, the best seed soaking temperature is 40oC The best medium for direct sowing of C.Javanica seeds is 70% topsoil + 20% charred rice husks + 10% rotting manure Propagation by tubers gave survival rates from 87.8 to 91.1% and there was no difference between different tuber sizes After 45 days, the length of the tubers was from 8.9-11.5cm, the tubers were 1-1.5cm size for the best shoot growth Propagation by stem cuttings is not really suitable, the survival rate ranges from 23.33 to 63.33% The highest is NAA drug experiment with 400ppm concentrate with 20s drug immersion time IAA gave the lowest survival rate, followed by IBA and the highest was NAA For Amomum Longiligulare T L Wu, the species is being prioritized to focus on developing large-scale plantation according to Decision No 1976/QĐ-TTg This species is easy to plant, the roots spread quickly, fast grow and has high productivity After months, the results of the study indicated that shading has a significant effect on the survival rate, growth, diameter, and height of Sa mandarin purple seedlings The best intensity of shading at months of age is 50% Plants should not be sown in unshaded conditions because the plants are often yellowed, have poor growth and very low survival rates The composition of the real potting intestine had a ii significant effect on the survival rate, growth, diameter, and height of the seedlings The best potting composition is 70% topsoil +20% fine sand +10% farmyard manure (additional 2g phosphorus/pot) Survival rate reached 88.9%, root diameter reached 5.9mm, peak height reached 35.3cm Plants should not be sown with potting soil mainly on topsoil, the plants are often blocked with slow growth if there is additional manure iii CONTENTS ABSTRACT i CONTENTS iii LIST OF ABBREVIATIONS .v LIST OF TABLE .vi LIST OF FIGURES AND CHARTS vii Chapter INTRODUCTION Chapter REVIEW OF LITERATURE .4 2.1 Overview of research on medicinal plants .4 2.1.1 Review of medicinal plants in the world 2.1.2 History of using traditional medicinal plants in Vietnam 2.1.3 Research of Codonopsis javanica in the world and Vietnam 2.1.4 Research of Amomum longiligulare in the world and Vietnam .16 2.2 Study site, natural conditions and people 21 2.2.1 Study area 21 2.2.2 Natural conditions 21 2.2.3 Ethnic group .24 Chapter GOAL, OBJECTIVES AND METHODOLOGY 25 3.1 Goal .25 3.2 Objectives .25 3.3 Research questions 25 3.4 Methodology 25 3.4.1 Data inheritance method 25 3.4.2 Field investigation method .26 3.4.3 Data processing and testing propagation in the experimental nursery 28 Chapter RESULTS .34 4.1 The current status of distribution of C javanica and A longiligulare in Son La province 34 4.1.1 Results of the survey in Song Ma district 34 iv 4.1.2 Results of the survey in Sop Cop district 35 4.1.3 Results of the survey in Moc Chau district 37 4.1.4 Results of the survey in Van Ho district 38 4.1.5 Results of the survey in Thuan Chau district 39 4.2 Results of Codonopsis javanica in Son La experimental propogations 41 4.2.1 Result of the Propagation from C.javanica seeds 41 4.2.2 Result of the effect of root tuber size on the growth of C.janvanica seedlings in the nursery stage 46 4.2.3 Results of propagation technique of Codonopsis javanica from stem cuttings 48 4.3 Results of experimental propogations Amomum longiligulare 53 4.3.1 Result of the effect of shading level on the growth of seedlings of Amomum longiligulare from seeds in the nursery stage 54 4.3.2 Result of the effect of potting soil composition on seedling growth from seed in the nursery stage 58 DISCUSSIONS 62 CONCLUSION 64 ACKNOWLEDGEMENT 66 REFERENCES 67 APPENDIX PHOTOS v LIST OF ABBREVIATIONS VNUF Vietnam National University of Forestry NWFP Non-Wood Forest Products NTFP Non-Timber Forest Product vi LIST OF TABLE Table 2.1 Statistical data of Amomum species in Vietnam .17 Table 3.2 Investigating canopy cover form .27 Table 4.1 Indigenous knowledge of people about the growth of species 41 Table 4.2 Results of the experiment quality testing seeds .42 Table 4.3 Result of the germination rate, germination rate by seeds in districts 43 Table 4.4 Result of the germination rate, speed germination rate based on seed treatment temperature 44 Table 4.5 Result of the ratio of seedlings follow the formula of potting soil 45 Table 4.6 The results of budding in the experiments Effect of root tuber size on the growth of C.javanica seedlings 46 Table 4.7 Result of the survival rate and root length of root cuttings with immersion during 10s 48 Table 4.8 Result of the survival rate and root length of the cuttings with immersion time 20s .50 Table 4.9 Result of the survival rate and root length of the cuttings with immersion time 30s .51 Table 4.10 Result of the Effect of shading intensity on survival rate 54 Table 4.11 Results of the effect of shading on the diameter and hieght of A longiligulare seedlings 56 Table 4.12 Results of One-way analysis of variance Test 57 Table 4.13 Result of the effect of shading intensity on survival rate 58 Table 4.14 Results of the effect of potting components on the diameter and height of 9-month-old seedlings 59 Table 4.15 Results of testing the One-way analysis of variance on growth in diameter and height in the experimental pot .61 vii LIST OF FIGURES AND CHARTS Figure 4.1 The map of investigate points in Song Ma, Son La province 34 Figure 4.2 The map of investigate points in Sop Cop, Son La province 36 Figure 4.3 The map of investigate points in Moc Chau, Son La province 37 Figure 4.4 The map of investigate points in Van Ho, Son La province 38 Figure 4.5 The map of investigate points in Thuan Chau, Son La province 40 Chart 4.1 The chart of average survival rate of shaded test after months .55 Chart 4.2 The chart of the growth of the root diameter after 9-month-old seedling in the shading experiments 56 Chart 4.3 The chart of the growth and height after 9-month-old seedlings in the shading experiments 57 Chart 4.4 The chart of the growth of the root diameter after the 9-month-old seedling in the potting component experiments 60 Chart 4.5 The chart of the growth of the height after the 9-month-old seedling in the potting component experiments 61 Chapter INTRODUCTION Vietnam is a Southeast-Asia country which located at the tropical regions, due to its high and globally significant biodiversity values is recognized as one of the nations where the conservation of biodiversity should be prioritized In 1994, Vietnam became a signatory to the United Nations Convention on Biological Diversity (CBD) and ranked 16th in 25th countries in the world that have the most biodiversity (MONRE, 2005) Biodiversity contributes a crucial role to develop and maintain the socioeconomic and environment It is very important in the supply of pharmaceutical raw materials also In the past, our ancestors knew how to take advantage of the wild plant for healing and making medicine However, Vietnam has great potential for medicinal plants but has not been promoted Currently, those species are being exploited in the wild by local people to maintain their livelihood, this leads to degrading in a number of species and narrow the distribution area Medicinal plant resources are being depleted, many valuable medicinal herbs are in danger of extinction, natural forests are degraded in terms of both area and forest volume, conditions The forest's ecology is no longer suitable for a number of tree species to grow and develop Overexploitation leads to many types of trees that are no longer able to regenerate Management has not had a macro policy to conserve and develop medicinal resources in natural forests as well as genetic resources of these plants If this situation continues, these precious medicinal species are in danger of extinction in nature The non-wood forest products (NWFP) sector (collection, cultivation, trade and processing) in Viet Nam affords employmentto hundreds of thousands of people, including the inhabitants of urban areas (Luong Van Tien 1991) NWFP exports reached $40 million USD annually between 1986 and 1990 (NTFP Project Document 2000) After 1995, People moving from the rural areas tend to maintain their consumption patterns in the cities and the urbanization of the population has increased the demand for NWFP Currently, deregulation and more free market access for NWFP is taking place It is about 4000 different types of medicines ranging from traditional herbal cures to modern remedies introduced from the west are produced in Viet Nam The low productivity and obsolete technology of the Vietnamese pharmaceutical industries can currently meet only 30 percent of the total demand, while 70 percent of the domestic demand has to be supplied by imports (VFFSCP 1997) It is about 300 species are harvested for domestic and commercial purposes (including domesticated medicinal trees) Valuable plants include Acathopanax spp (Ngũ gia bì gai), Schefflera spp (Ngũ gia bì chân chim), Polygonum multiflorum (Hà thủ ô đỏ), Lonicera spp (Kim ngân), Polygonatum and Disporopsis (Hoàng tinh), Strychnos wallichiana (Hoàng nan), Draceana spp (Huyết giác), Codonopsis spp (Đẳng sâm), Artemisia annua (Thanh hao hoa vàng), Stephanis rotunda (Bình vôi) and Cibotium barometz (Cẩu tích) (Nguyen Tap 2001) It has been estimated that 80–90 percent of the medicinal plants produced in northern Viet Nam are exported in the form of dried plants or extracts and that only 10–20 percent of the medicinal plants are consumed or processed domestically (Lecup 1996) Son La is a mountainous province located in the Northwest of Vietnam which has the total land area of this province is 14,125 km², in which, the total natural area of the province with 357,000 of forest It has a richness in biodiversity and ethnic diversity with twelve ethnics There are three the most prevalent ethnics such as Muong, Thai, Mong Through thousands of year, the progenies has been indoctrinated by word of mouth their traditional customs and uniquely indigenous knowledge of folk cure by using medicinal plants Son La province is rich and potential in herb plants because of its large natural forest area, such as Morinda officinalis (Ba kích), Codonopsis javanica (Đẳng Sâm), Polygonum multiflorum (Hà thủ ơ), Drynaria fortunei (Cốt tối bổ), Amomum villosum (Sa nhân đỏ), and Amomum longiligulare Based on biological characteristics, distribution, efficiency, consumptional and commercial value Appendix 11: The results of the survival rate and root length of C javanica stem cuttings under the drugs /drug concentrates under the drug immersion time 20s ET Experimental stem cutting Number of rooting (stem cutting) 16.IAA100ppm(20S) 90 23 67 25.56 90 7.2 17.IAA200ppm(20S) 90 24 66 26.67 90 8.1 18.IAA300ppm(20S) 90 35 55 38.89 90 8.3 19.IAA400ppm(20S) 90 30 60 33.33 90 8.5 20.IAA500ppm(20S) 90 27 63 30.00 90 9.1 21.IBA100ppm(20s) 90 27 63 30.00 90 7.3 22.IBA200ppm(20s) 90 29 61 32.22 90 7.4 23.IBA300ppm(20s) 90 32 58 35.56 90 7.7 24.IBA400ppm(20s) 90 36 54 40.00 90 8.1 25.IBA500ppm(20s) 90 30 60 33.33 90 7.7 26.NAA100ppm(20s) 90 38 52 42.22 90 7.1 27.NAA200ppm(20s) 90 43 47 47.78 90 8.2 28,NAA300ppm(20s) 90 44 46 48.89 90 8.8 29.NAA400ppm(20s) 90 57 33 63.33 90 9.2 30.NAA500ppm(20s) 90 43 47 47.78 90 9.5 46 Control formula 90 21 69 23.33 90 Tbj 539 No rooting (stem cutting) Rate of rooting 901 Tai 1440 The root length level I 2.1 The shoot length 8.2 Standard test when squared to survival with immersion time 20s TT 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 Tai 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 2880 Tbj 539 539 539 539 539 539 539 539 539 539 539 539 539 539 539 539 901 901 901 901 901 901 901 901 901 901 901 901 901 901 901 901 23040 ft 23 24 35 30 27 27 29 32 36 30 38 43 44 57 43 21 67 66 55 60 63 63 61 58 54 60 52 47 46 33 47 69 1440 Look up table fl 33.69 33.69 33.69 33.69 33.69 33.69 33.69 33.69 33.69 33.69 33.69 33.69 33.69 33.69 33.69 33.69 56.31 56.31 56.31 56.31 56.31 56.31 56.31 56.31 56.31 56.31 56.31 56.31 56.31 56.31 56.31 56.31 cn^2 3.39 2.79 0.05 0.40 1.33 1.33 0.65 0.08 0.16 0.40 0.55 2.57 3.16 16.13 2.57 4.78 2.03 1.67 0.03 0.24 0.79 0.79 0.39 0.05 0.09 0.24 0.33 1.54 1.89 9.65 1.54 2.86 64.50 24.99579 Compare and find the experiment with the highest survival rate using the standard of U root tuber a standard distribution (immersion time 20s) ET Experimental stem cutting Number of rooting (stem cutting) 28,NAA300ppm (20s) 90 44 46 48.89 29.NAA400ppm (20s) 90 57 33 63.33 Pm1 0.49 Pm2 p 0.63 0.56 q U U 0.05 0.44 1.95 1,65 No rooting Rate of (stem rooting cutting) Analysis of Anova way (drug type, concentrate) to root length (immersion time 20s) Anova: Two-Factor With Replication SUMMARY IAA IBA NAA Total Count Sum Average Variance 2.0 0.0 6.9 2.3 0.0 7.1 2.4 0.0 20 2.2 0.0 Count Sum Average Variance 7.2 2.4 0.0 7.4 2.5 0.0 8.1 2.7 0.0 22.7 2.5 0.0 Count Sum Average Variance 7.7 2.6 0.0 7.3 2.4 0.0 9.3 3.1 0.0 24.3 2.7 0.1 Count Sum Average Variance 7.2 2.4 0.0 7.6 2.5 0.0 10.9 3.6 0.0 25.7 2.9 0.4 Count Sum Average Variance 5.9 2.0 0.0 7.2 2.4 0.0 10 3.3 0.0 23.1 2.6 0.4 Count Sum Average Variance 6.2 2.1 0.0 6.2 2.1 0.0 6.2 2.1 0.0 18.6 2.1 0.0 18 40.2 2.2 0.1 18 42.6 2.4 0.0 18 51.6 2.9 0.3 SS df 3.9 5.0 0.8 40.71 0.0 2.48 4.0 2.8 0.7 11.4 2.0 10.0 36.0 53.0 2.0 0.3 0.0 104.19 14.47 0.0 0.0 3.26 2.11 Total Source Factor A (Concentrate) Factor B (Drug) Interactive AB Ramdon Total Count Sum Average Variance ANOVA Source of Variation Sample Columns Interaction Within Total MS F P-value F crit Finding the drug and concentrates giving the best results of survival rate and root length of C javanica stem cuttings under the immersion time 20s Xmax Xmax2 Sn n1 n2 t t0.05 3.6 3.3 0.7 Random concentrate variable Sample size but averaged so this is the Reepeat -0.524890659 2.131449546 Appendix 12: The results of the survival rate and root length of C javanica stem cuttings under the drugs /drug concentrates under the drug immersion time 30s ET Experimental stem cutting Number of rooting (stem cutting) No rooting (stem cutting) Rate of rooting 31.IAA100ppm(30s) 90 24 66 26.67 32.IAA200ppm(30s) 90 27 63 30.00 33.IAA300ppm(30s) 90 34 56 37.78 34.IAA400ppm(30s) 90 30 60 33.33 35.IAA500ppm(30s) 90 27 63 30.00 36.IBA100ppm(30s) 90 21 69 23.33 37.IBA200ppm(30s) 90 27 63 30.00 38.IBA300ppm(30s) 90 24 66 26.67 39.IBA400ppm(30s) 90 33 57 36.67 40.IBA500ppm(30s) 90 21 69 23.33 41.NAA100ppm(30s) 90 21 69 23.33 42.NAA200ppm(30s) 90 35 55 38.89 43.NAA300ppm(30s) 90 29 61 32.22 44.NAA400ppm(30s) 90 24 66 26.67 45.NAA500ppm(30s) 90 22 68 24.44 46.Công thức đối chứng 90 21 69 23.33 Tabj 420 1020 Standard test when squared to survival with immersion time 30s TT 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 Tai 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 90 2880 Tbj 420 420 420 420 420 420 420 420 420 420 420 420 420 420 420 420 1020 1020 1020 1020 1020 1020 1020 1020 1020 1020 1020 1020 1020 1020 1020 1020 23040 Tra bảng ft 24 27 34 30 27 21 27 24 33 21 21 35 29 24 22 21 66 63 56 60 63 69 63 66 57 69 69 55 61 66 68 69 1440 fl 26.25 26.25 26.25 26.25 26.25 26.25 26.25 26.25 26.25 26.25 26.25 26.25 26.25 26.25 26.25 26.25 63.75 63.75 63.75 63.75 63.75 63.75 63.75 63.75 63.75 63.75 63.75 63.75 63.75 63.75 63.75 63.75 n^2 0.19 0.02 2.29 0.54 0.02 1.05 0.02 0.19 1.74 1.05 1.05 2.92 0.29 0.19 0.69 1.05 0.08 0.01 0.94 0.22 0.01 0.43 0.01 0.08 0.71 0.43 0.43 1.20 0.12 0.08 0.28 0.43 18.77 24.99579 Compare and find the experiment with the highest survival rate using the standard of U root tuber a standard distribution 30s) ET Experimental stem cutting Number of rooting (stem cutting) No rooting (stem cutting) Rate of rooting 42.NAA200ppm(30s) 90 35 55 38.89 33.IAA300ppm(30s) 90 34 56 37.78 Pm1 0.388889 Pm2 0.377778 p 0.383333 q 0.616667 U -0.1533 U0.05 1,65 Analysis of Anova way (drug type, concentrate) to root length (immersion time 30s Anova: Two-Factor With Replication SUMMARY IAA IBA NAA Total Count Sum Average Variance 6.4 2.1 0.0 5.7 1.9 0.0 5.9 2.0 0.0 18 2.0 0.0 Count Sum Average Variance 7.9 2.6 0.0 6.8 2.3 0.0 7.1 2.4 0.0 21.8 2.4 0.0 Count Sum Average Variance 9.1 3.0 0.0 5.8 1.9 0.0 7.7 2.6 0.0 22.6 2.5 0.2 Count Sum Average Variance 2.7 0.0 6.5 2.2 0.0 7.3 2.4 0.0 21.8 2.4 0.1 Anova: Two-Factor With Replication SUMMARY IAA IBA NAA Total Count 3 Sum 7.2 6.6 5.7 19.5 Average 2.4 2.2 1.9 2.2 Variance 0.0 0.0 0.0 0.1 Count Sum Average Variance 6.2 2.1 0.0 6.2 2.1 0.0 6.2 2.1 0.0 18 44.8 2.5 0.1 18 37.6 2.1 0.0 18 39.9 2.2 0.1 df MS 18.6 2.1 0.0 Total Count Sum Average Variance ANOVA Source of Variation Source Factor A (Concentrate) Factor B (Drug type) Interactive AB Ramdon Sample Columns Interaction Within Total Total SS 2.1 1.5 1.4 0.4 5.0 2.0 10.0 36.0 5.323148 53 0.4 0.8 0.1 0.0 F 39.79 72.45 13.35 P-value 0.0 0.0 0.0 F crit 2.48 3.26 2.11 Finding the drug and concentrates giving the best results of survival rate and root length of C javanica stem cuttings under the immersion time 30 Xmax Xmax2 2.7 Sn 0.4 Variable Concentration Random n1 Sample size but averaged so this is the Reepeat n2 t t0.05 -0.91856 2.13145 Appendix 13 Analysis of Anova growing in diameter at different shading experimental formulas (A.longiligulare) Anova: Single Factor SUMMARY Groups 0.25 0.5 0.75 ANOVA Source of Variation Between Groups Within Groups Total Count 58 74 71 48 Sum 321.6 433.8 395.9 221.6 Average 5.544828 5.862162 5.576056 4.616667 Variance 1.046025 0.581014 1.583276 0.875887 SS 47.45642 254.0335 301.4899 df MS 15.81881 1.028476 F 15.38 247 250 P-value 0.00 F crit 2.64 Appendix 14 Analysis of Anova's height growth at different shading experimental formulas (A.longiligulare) Anova: Single Factor SUMMARY Groups Count Sum Average Variance 0.25 58 1678 28.93103 75.32849 0.5 74 2455 33.17568 64.1194 0.75 71 2284 32.16901 78.97103 48 1249 26.02083 112.0208 ANOVA Source of Variation SS df MS Between Groups 1832.91 610.9731 Within Groups 19767.3 247 80.02992 Total 21600.3 250 F P-value 7.63 0.00 F crit 2.64 Appendix 15 Analysis of Anova growth in diameter at different potting formulas (A.longiligulare) Anova: Single Factor SUMMARY Groups Count Sum Average Variance ET 68 370.7 5.451470588 1.030893 ET 80 476.1 5.95125 0.71696 ET 74 412.5 5.574324324 1.522208 ANOVA Source of Variation SS df MS Between Groups 10.25144 5.125721615 Within Groups 236.8309 219 1.081419836 Total 247.0824 221 F P-value 4.739807 0.009657 F crit 3.037088 Appendix 16 Analysis of Anova's height growth at different potting formulas (A.longiligulare) Anova: Single Factor SUMMARY Groups Count Sum Average Variance ET 68 1961 28.83823529 74.97344 ET 80 2827 35.3375 54.22642 ET 74 2440 32.97297297 67.25953 ANOVA Source of Variation SS df MS Between Groups 1571.685 Within Groups 14217.05 219 Total 15788.74 221 F 785.8423523 12.10514 64.91805495 P-value 1.03E-05 F crit 3.037088 PHOTOS Plate 1: Codonopsis javanica (Blume) Hook F et Thomson (a) fruit and flower ; (b) Freshly harvested ripe fruit Plate 2: the seeds in test samples from batches of seeds Plate3: C javanica seeds germinating in the temperature experiment Plate 4: Codonopsis javanica sprouts at ET2 of the effect of potting soil on the rate of seedling and sprouting from seed Plate5: Sizes of experimental fruit in the Experiment of the effect of root tuber size on the growth of C.janvanica seedlings in the nursery stage ET1 ET2 ET3 Plate 6: Illustration of the new shoot of Codonopsis javanica (Blume) Hook F et Thomson in the experiments of the effect of he size of the tuber on seedling Plate 7: Harvesting Codonopsis javanica (Blume) Hook F et Thomson cuttings from the forest (left) and cutting cuttings (right) Plate 8: Codonopsis stem cuttings and budding in the experiment A Longiligular plant B Longiligular flowers Plate 9: Amomum longiligulare T.L.Wu natural distribution in the wild A Clusters of spiky capsules on a long B The spiky fruit clusters peduncle arising from the horizontal rootstock Plate 10: the cluster of Amomum longiligulare fruits Plate 11: Seedlings in the experiment the effect of shading Plate 12: Measurement of growth of Amomum longiligulare T.L.Wu (left), the experiment on the effect of potting intestine on growth of Amomum longiligulare T.L.Wu after months (right) Plate 13: Model of planting Amomum longiligulare T.L.Wu under the forest canopy (at Pha Din Top survey site, Phong Lai commune, Thuan Chau district, Son La province)