VIETNAM NATIONAL UNIVERSITY OF AGRICULTURE FACULTY OF BIOTECHNOLOGY UDERGRADUATE THESIS TITLE: “DETERMINATION OF SUITABLE CULTURAL CONDITIONS FOR THE GROWTH OF BLACK LINGZHI MUSHROOM (GANODERMA SUBRESINOSUM)” Supervisor : 1.NGO XUAN NGHIEN DEPARTMENT: MICROBIAL BIO TECHNOLOGY : 2. NGUYEN THI BICH THUY DEPARTMENT : MICROBIAL BIO TECHNOLOGY Student : NGUYEN THI THU TRANG Class : K61CNSHE Student ID : 610783 HÀ NỘI - 2021 COMMITMENT I hereby declare that all the data and results in this thesis are honest and have never been used in any report yet. Also, I would like to assure that all information in this report are shown exactly resource, and all effort, assistance are all mentioned. Hanoi, February 2nd 2021 Student Nguyen Thi Thu Trang i ACKNOWLEDGEMENT First and foremost, I have to express out appreciation to the directory of Vietnam National University of Agriculture, board of deans and lectures in faculty of biotechnology for creating an interesting learning environment and provided me the sufficient knowledge throughout four academic years.
Particularly, I would like to send my gratitude to Dr. Ngo Xuan Nghien belong to Department of Microbial bio technology, Faculty of Biotechnology, Vietnam National University of Agriculture, for him ongoing and suggestions through my undergraduate thesis. Without him, this thesis would not have been possible. Next, I would like to say thanks to Dr.
Nguyen Thi Bich Thuy belong to Department of Microbial bio technology, Faculty of Biotechnology, Vietnam National University of Agriculture, who directly introduced me for my whole period on doing experiments and provided me the guideline from the beginning until completing my whole thesis. I am indebted to Mrs. Nguyen Thi Luyen, the assistance of Mushroom Research and Development Center and all my friends at Mushroom Research and Development Center, who support me unconditionally during the time doing research. I would never have been able to finish my thesis without their aid and guidance.
Last but not least, I would like to thank my parents who encouraged me to try my best until the job is done, and who never ceased in providing me with the love and support I needed. Hanoi, February 2nd 2021 Student Nguyen Thi Thu Trang ii TABLE OF CONTENTS COMMITMENT. ii LIST OF FIGURES. vii CHAPTER I: INTRODUCTION.
2 CHAPTER II: LITERATURE REVIEW2 .1 The situation of research in the world and Viet Nam .1 The situation of research in the world. The situation of research in the Vietnam. Life cycle of Ganoderma sp. Biological characteristics of Ganoderma sp.
Factors effect on Ganoderma sp growth and development. Composition of chemical and biological compounds. Major bioactive components isolated from Ganoderma sp. Health benefits of Ganoderma sp.
19 CHAPTER III: MATERIAL AND METHODS. Time and location. Contents of the study. Methods of material processing.
Preparation of Agar media. Preparation of paddy grain substrates. Preparation of substrates for cultivation. Method of evaluating the effect of substrates on growth and development of Ganoderma subresinosum (Ga21).
Evaluation of different temperature effects to mycelial growth of Ganoderma subresinosum. Evaluation of different pH effects to mycelial growth of Ganoderma subresinosum. Evaluation of different pure culture media effects to mycelial growth of Ganoderma subresinosum. Evaluation of different paddy grain substrates (grain master or grade 2) effects to mycelial growth of Ganoderma subresinosum.
Evaluation of different substrates effect to the growth and development of Ganoderma subresinosum. 30 PART IV: RESULT AND DISCUSSION. Effect of temperature on mycelial growth of Ganoderma subresinosum. Effect of pH on mycelial growth of Ganoderma subresinosum.
Effect of spawn grain substrates (grain master or grade 2) on mycelial growth of Ganoderma subresinosum. Evaluation of different pure culture media effects to mycelial growth of Ganoderma subresinosum. Effect of different substrates on primordia formation and development of Ganoderma subresinosum. 45 PART V: CONCLUSION AND RECOMMENDATION.
53 v LIST OF TABLES Table 2.Ecological conditions of Ganoderma sp. Pure culture media components. Different ratio of substrates to cultivated Ganoderma subresinosum. Effect of different temperature on mycelial growth of Ganoderma subresinosum.
Effect of different pH on mycelial growth of Ganoderma subresinosum. The length of mycelial growth of Ganoderma subresinosumin 4,8,12,16,20 days after incubation. Evaluation of different grain substrates for spawn development. Effect of different pure culture media on mycelial growth of Ganoderma subresinosum.
The length of mycelial growth of Ganoderma subresinosumin after 4, 8, 12, 16, 20 days of incubation. Comparison of antlers appearances period and mycelial growth rate. Effect of different formulas on formation fruiting body and Biological Efficiency of Ganoderma subresinosum. 47 vi LIST OF FIGURES Fig 2.1 Life cycle of Ganoderma sp.
Effect of different temperature on mycelial growth of Ganoderma subresinosum. Mycelial growth on different temperature for 4 days after inoculation. Mycelial growth on different temperature for 8 days after inoculation. Effect of different pH on mycelial growth of Ganoderma subresinosum.
5 Mycelial growth on different pH for 7 days after inoculation. Effect of different grain substrates on mycelial growth of Ganoderma subresinosum .7 Mycelial growth for eight days. 8 Mycelial growth for 14 days. Effect of different pure culture media on mycelial growth of Ganoderma subresinosum.
Mycelial growth on different media for 5 days after inoculation. Mycelial growth on different media for 9 days after inoculation. Mycelial colonized half the bags. Mycelial colonized fully the bags.
Comparison of Biological efficiency in different substrates of Ganoderma subresinosum. Ganoderma subresinosum in the button neck. Ganoderma subresinosum in the body of the bag. Ganoderma subresinosum is eaten by worms.
Ganoderma subresinosum is infected with mold. 51 vii LIST OF ABBREVIATIONS Fig Figure LSD Least Significant Difference SD Standard Deviation PGA Potata Glocuse Agar viii CHAPTER I: INTRODUCTION 1.1 Introduction As precious medical macrofungi, Ganoderma (common name: reishi mushroom or Ling Zhi) has been successfully cultivated for 4000 years ago, particularly in the Far East countries (China, Japan, Korea, etc. This genus was classified as a member of the family Ganodermataceae and the class Agaricomycetes. Extensive clinical studies, primarily in Japan, have clearly demonstrated that mushroom have medicinal and therapeutic value, by injection or oral administration, in the prevention/treatment of cancer, viral diseases (influenza, polio), hypercholesterolemia, blood platelet aggregation, and hypertension,and most of the studies are on Ganoderma sp.
many of the active substances which include polysaccharides (e., β- glucans), nucleic acid derivatives (the hypocholesterolemic eritadenine), lipids, peptides, proteins, and glycoproteins, have been isolated and identified. Some of the mechanisms of activity have been elucidated, e., antiviral activity via stimulation of interferon production in the host. Additional medical claims which were less well documented may nonetheless have some validity and merit further study. [1] Reishi has long been studied in long time by scientists because of their great use, and most studies are done on Ganoderma lucidum - the most popular type of Reishi.
Meanwhile, not much research has been done on the black Reishi or Ganoderma subresinosum , which is not difficult to cultivate, and has high practicality. Vietnam is a country of abundant raw materials and human resources, which are suitable for growing mushrooms, so it can contribute to 1 sustainable economic development. For that reason, I chose Ganoderma subresinosum as the research object for this thesis. To grow Ganoderma subresinosum or any other fungus, besides choosing varieties and farming methods, an factor is very interested and focused on what is the enviromental condition .Depending on the location and climate of each country, it is possible to determine the most suitable farming conditions,which have not been studied much.
If the best growing conditions are determined, the best black Reishi mushroom strain will be produced, so this is considered a prerequisite step for the isolation and cultivation of black Reishi strains collected from heaven. As a result we do the topic “Determination of suitable cultural conditions for the growth of black lingzhi mushroom (Ganoderma subresinosum)” 1.2 Objective Research and determine the most suitable growing conditions for Ganoderma subresinosum strain, including temperature, pH, agar media and different nutritional supplements.3 Requirement Evaluate the growth of Ganoderma subresinosum on different levels of temperature, pH and agar environment to determine the optimal environment for Ganoderma subresinosum strains. Evaluate the growth and development of Ga21 strain on different sawdust substrates and then select the most potential substrate for production. 2 CHAPTER II: LITERATURE REVIEW2 2.1 The situation of research in the world and Viet Nam 2.1 The situation of research in the world Nowadays, more than 2000 species of mushrooms have been identified [2].
Among known mushroom species, approximate 80 edible and medicinal mushrooms such as oyster mushroom, straw mushroom, wood ear mushroom, and shiitake mushrooms have been successfully cultivated. Due to scarcity in nature, Ganoderma sp. are artificially cultivated in solid culture to meet demands for medicinal herbs and international markets. In Taiwan, Peng (1990) and Hseu (1992) collected and cultivated more than 10 different Ganoderma species.
In Korea, Ganoderma sp. also occupies a significant market share and pays special attention to the Ancient Lingzhi hay Artists conk species with its high anti-tumor effect. Yet China is still recognized as the world's largest center for growing and producing Reishi mushrooms [3]. According to the statistics of Moore and Chiu (2000)[4], the total global output of Reishi in 1997 reached 4300 tons, the output of China was 3000 tons.
According to Wang X. has been cultivated in China since 1621. Ganoderma sp was first reported from China by Teng (1934)[6], with four species including G. lucidum and one variety.
Later, many researchers extensively studied Chinese Ganodermataceae and introduced more than 100 species in four genera: Ganoderma, Amauroderma, Haddowia and Humphreya (Zhao & Zhang 2000, Wu & Dai 2005, Cao et al. Ganoderma was accepted as the scientific binomial of “Lingzhi” in many reports on Chinese edible and medicinal mushrooms, but it was recently described as Lingzhi based on strong molecular and morphological evidence (Cao et al. 2012, Dai et al. Mycochemical and other studies reported different species numbers in the genus Ganoderma (Peng et al.
2014, Yan et al. However, the majority 3 of Ganoderma species reported from China have not been subjected to systematic studies (Baby et al. By 1936, Zenzaburo Kasai and Yukio Naoi successfully cultivated Ganoderma sp. at Tokyo Agricultural University, Japan.
Since then, the technology of cultivating Lingzhi in Japan has developed strongly, the output from 1979 - 1995 has increased 40 times [14]. Japan is the first country to successful cultivation of Ganoderma sp. and the scientific research and Preparation of medicinal products are ranked among the top in the world. Korea is also a country with highly developed Reishi mushroom production technology.
The situation of research in the Vietnam In Vietnam, the Institute of Medicinal Materials - Hanoi successfully cultivated Ganoderma (Chinese variety) in 1978. Nine years later, in 1987, scientists from the University of Natural Sciences chose Lingzhi variety growing wild in Lam Dong mountain forest to propagate and put into production in Ganoderma farm of Central Pharmaceutical Enterprise 24, achieving good results in 1988. The Vietnamese Medicine and Pharmacy who succeeded Ly Thoi Tran discovered the use of Lingzhi mushroom such as: Le Quy Don pointed out that Lingzhi is: "The source of rare products of Dai Nam land", 200 years ago, such as Hai Thuong Lan Ong, Le Huu Trac proposed the motto "Men pharmacy to cure the Men" using Linh Chi in a number of remedies (according to Dinh Xuan Linh et al, 2012).[15] Several studies have been reported regarding the taxonomy and distribution of Vietnamese Ganodermataceae. Vietnam possesses 27 species of the Ganoderma genus; however, the classification and distribution of the Ganoderma genus in Vietnam continues to be studied.
In a 2013 report by Nguyen et al.,[16] 43 species of the Ganoderma genus were identified in the 4 Highlands of Vietnam. Five of these have been successfully cultivated in Vietnam including G. subresinosum, and one of these the G.lucidum species is most commonly cultivated.