Determination of suitable cultural conditions for the growth of black linhzhi mushroom (ganoderma subresinosum) (khóa luận tốt nghiệp)

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 : Dr.NGO XUAN NGHIEN DEPARTMENT: MICROBIAL BIO TECHNOLOGY : Dr 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 i ACKNOWLEDGEMENT ii LIST OF FIGURES vii CHAPTER I: INTRODUCTION 1.1 Introduction 1.2 Objective 1.3 Requirement CHAPTER II: LITERATURE REVIEW2 2.1 The situation of research in the world and Viet Nam 2.1.1 The situation of research in the world 2.1.2 The situation of research in the Vietnam 2.2 Scientific classification 2.3 Distribution 2.4 Life cycle of Ganoderma sp 2.5 Biological characteristics of Ganoderma sp 2.6 Factors effect on Ganoderma sp growth and development 10 2.6.1 Internal factors 10 2.6.2 External factors 12 2.6.3 Composition of chemical and biological compounds 14 2.6.4 Major bioactive components isolated from Ganoderma sp 15 2.6.5 Health benefits of Ganoderma sp 19 CHAPTER III: MATERIAL AND METHODS 24 3.1 Time and location 24 3.2 Research object 24 3.3 Materials 24 iii 3.4 Equipment 25 3.5 Contents of the study 25 3.6 Methods 25 3.7 Methods of material processing 26 3.7.1 Preparation of Agar media 26 3.7.2 Preparation of paddy grain substrates 26 3.7.3 Preparation of substrates for cultivation 26 3.8 Method of evaluating the effect of substrates on growth and development of Ganoderma subresinosum (Ga21) 27 3.8.1 Experiment Evaluation of different temperature effects to mycelial growth of Ganoderma subresinosum 27 3.8.2 Experiment Evaluation of different pH effects to mycelial growth of Ganoderma subresinosum 27 3.8.3 Experiment Evaluation of different pure culture media effects to mycelial growth of Ganoderma subresinosum 28 3.8.4 Experiment Evaluation of different paddy grain substrates (grain master or grade 2) effects to mycelial growth of Ganoderma subresinosum 28 3.8.5 Experiment Evaluation of different substrates effect to the growth and development of Ganoderma subresinosum 29 3.9 Data collection 29 3.10 Data analysis 30 PART IV: RESULT AND DISCUSSION 31 4.1 Effect of temperature on mycelial growth of Ganoderma subresinosum 31 4.2 Effect of pH on mycelial growth of Ganoderma subresinosum 34 4.3 Effect of spawn grain substrates (grain master or grade 2) on mycelial growth of Ganoderma subresinosum 38 iv 4.4 Evaluation of different pure culture media effects to mycelial growth of Ganoderma subresinosum 41 4.5 Effect of different substrates on primordia formation and development of Ganoderma subresinosum 45 PART V: CONCLUSION AND RECOMMENDATION 52 5.1 Conclusion 52 5.2 Recommendation 52 REFERENCES 53 v LIST OF TABLES Table 1.Ecological conditions of Ganoderma sp cultivation 14 Table Pure culture media components 28 Table 3.2 Different ratio of substrates to cultivated Ganoderma subresinosum 29 Table Effect of different temperature on mycelial growth of Ganoderma subresinosum 31 Table Effect of different pH on mycelial growth of Ganoderma subresinosum 35 Table The length of mycelial growth of Ganoderma subresinosumin 4,8,12,16,20 days after incubation 38 Table 4.4 Evaluation of different grain substrates for spawn development 39 Table Effect of different pure culture media on mycelial growth of Ganoderma subresinosum 41 Table The length of mycelial growth of Ganoderma subresinosumin after 4, 8, 12, 16, 20 days of incubation 45 Table Comparison of antlers appearances period and mycelial growth rate 46 Table 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 Fig Effect of different temperature on mycelial growth of Ganoderma subresinosum 32 Fig 4.2 Mycelial growth on different temperature for days after inoculation 33 Fig 4.3 Mycelial growth on different temperature for days after inoculation 33 Fig 4 Effect of different pH on mycelial growth of Ganoderma subresinosum 37 Fig Mycelial growth on different pH for days after inoculation 37 Fig 4.6 Effect of different grain substrates on mycelial growth of Ganoderma subresinosum 40 Fig 4.7 Mycelial growth for eight days 40 Fig Mycelial growth for 14 days 40 Fig 4.9 Effect of different pure culture media on mycelial growth of Ganoderma subresinosum 43 Fig 10 Mycelial growth on different media for days after inoculation 44 Fig 11 Mycelial growth on different media for days after inoculation 44 Fig 4.12 Mycelial colonized half the bags 47 Fig 4.13 Mycelial colonized fully the bags 47 Fig 4.14 Comparison of Biological efficiency in different substrates of Ganoderma subresinosum 48 Fig 4.15 Ganoderma subresinosum in the button neck 49 Fig 4.16 Ganoderma subresinosum in the body of the bag 50 Fig 4.17 Ganoderma subresinosum is eaten by worms 51 Fig 4.18 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.g., βglucans), 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.g., 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 PART V: CONCLUSION AND RECOMMENDATION 5.1 Conclusion -The optimal temperature and pH for mycelial growth of strain Ga21 were at 25 ° C ± and pH = 8.0-9.0, respectively -Formula (69% of rice grains + 30% sawdust + 1% CaCO3) was considered as the optimal spawn substrate for luxuriant mycelial growth of strain Ga21 - The highest mycelium growth rate was obtained when cultivated on Treatment (sawdust 99% + wheat bran 0% + CaCO3 1%), whereas the best yield performance was found in formula (2, 95%) 5.2 Recommendation Further studies are needed to: - Evaluate the medicinal value of strain Ga21 - Evaluate the resistant of strain Ga21 to bacteria, virus, and mould - Identify the taxonomic position of strain Ga21 52 REFERENCES BREENE;, W.M., Nutritional and Medicinal Value of Specialty Mushrooms J Food Prot 1990 53(10): p 883-894 Friedman, M., Mushroom Polysaccharides: Chemistry and Antiobesity, Antidiabetes, Anticancer, and Antibiotic Properties in Cells, Rodents, and Humans Foods, 2016 5(4) D, Z.J and Z.X Q;, Importance, distribution and taxonomy of Ganodermataceae in China Proceedings of Contributed Symposium, B 5th International Mycological Congress, Vancouver 8-1994: p 14-21 Chiu, S.W., et al., Food and chemical toxicology 2000: p 173-178 Wang Chi, Z and H Y, A new kind of preparation of Ganoderma lucidum in treating 35 patients with coronary cardiac disease, Proc, 94 Inter, Sym, On Ganoderma Res, 1994: 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============================================================================= REPEAT 855022 427511 1.27 0.374 FORMULA 21.7848 10.8924 32.48 0.005 * RESIDUAL 1.34131 335329 * TOTAL (CORRECTED) 23.9812 2.99764 BALANCED ANOVA FOR VARIATE DAYS FILE TEMP 25/ 1/21 17:36 :PAGE VARIATE V004 DAYS SUMS OF MEAN F RATIO PROB ER SQUARES SQUARES LN ============================================================================= REPEAT 3.69662 1.84831 1.82 0.274 FORMULA 444.220 222.110 218.55 0.000 * RESIDUAL 4.06513 1.01628 * TOTAL (CORRECTED) 451.981 56.4977 BALANCED ANOVA FOR VARIATE DAYS FILE TEMP 25/ 1/21 17:36 :PAGE VARIATE V005 DAYS SUMS OF MEAN F RATIO PROB ER SQUARES SQUARES LN ============================================================================= REPEAT 10.7500 5.37501 1.63 0.304 FORMULA 859.282 429.641 130.25 0.001 * RESIDUAL 13.1944 3.29861 * TOTAL (CORRECTED) 883.226 110.403 BALANCED ANOVA FOR VARIATE DAYS FILE TEMP 25/ 1/21 17:36 :PAGE VARIATE V006 DAYS SUMS OF MEAN F RATIO PROB ER SQUARES SQUARES LN ============================================================================= REPEAT 3.32163 1.66081 0.99 0.448 FORMULA 628.943 314.472 188.01 0.001 * RESIDUAL 6.69064 1.67266 * TOTAL (CORRECTED) 638.956 79.8695 TABLE OF MEANS FOR FACTORIAL EFFECTS FILE TEMP 25/ 1/21 17:36 :PAGE MEANS FOR EFFECT REPEAT REPEAT NOS 3 DAYS 6.63667 7.39000 DAYS 18.2500 19.7767 58 DAYS 26.2767 28.9467 DAYS 32.6933 33.8033 3 7.05667 19.3300 27.7800 32.3900 SE(N= 3) 0.334330 0.582032 1.04859 0.746695 5%LSD 4DF 1.31050 2.28144 4.11024 2.92688 MEANS FOR EFFECT FORMULA FORMULA NOS 3 3 DAYS 5.05667 8.86000 7.16667 DAYS 9.30333 25.3600 22.6933 DAYS 13.8633 35.1100 34.0300 DAYS 21.1400 38.8867 38.8600 SE(N= 3) 0.334330 0.582032 1.04859 0.746695 5%LSD 4DF 1.31050 2.28144 4.11024 2.92688 ANALYSIS OF VARIANCE SUMMARY TABLE FILE TEMP 25/ 1/21 17:36 :PAGE F-PROBABLIITY VALUES FOR EACH EFFECT IN THE MODEL SECTION - VARIATE DAYS DAYS DAYS DAYS GRAND MEAN (N= 9) NO OBS 7.0278 19.119 27.668 32.962 STANDARD DEVIATION C OF V |REPEAT SD/MEAN | BASED ON BASED ON % | TOTAL SS RESID SS | 1.7314 0.57908 8.2 0.3738 7.5165 1.0081 5.3 0.2744 10.507 1.8162 6.6 0.3040 8.9370 1.2933 3.9 0.4480 |FORMULA | | | 0.0049 0.0005 0.0008 0.0006 | | | | 4.2: Investigation of the effect of pH on the mycelial growth of Ganoderma subresinosum BALANCED ANOVA FOR VARIATE DAYS FILE PH 25/ 1/21 16:54 :PAGE VARIATE V003 DAYS LN SOURCE OF VARIATION DF LN SOURCE OF VARIATION DF LN SOURCE OF VARIATION DF SUMS OF MEAN F RATIO PROB ER SQUARES SQUARES LN ============================================================================= REPEAT 830659 415329 2.00 0.170 FORMULA 171.919 24.5598 118.55 0.000 * RESIDUAL 14 2.90035 207168 * TOTAL (CORRECTED) 23 175.650 7.63695 BALANCED ANOVA FOR VARIATE DAYS FILE PH 25/ 1/21 16:54 :PAGE VARIATE V004 DAYS SUMS OF MEAN F RATIO PROB ER SQUARES SQUARES LN ============================================================================= REPEAT 441758 220879 1.63 0.230 FORMULA 1744.28 249.182 ****** 0.000 * RESIDUAL 14 1.89872 135623 * TOTAL (CORRECTED) 23 1746.62 75.9399 BALANCED ANOVA FOR VARIATE DAYS FILE PH 25/ 1/21 16:54 :PAGE VARIATE V005 DAYS SUMS OF MEAN F RATIO PROB ER SQUARES SQUARES LN ============================================================================= 59 REPEAT 2.80030 1.40015 1.15 0.345 FORMULA 2955.03 422.147 347.20 0.000 * RESIDUAL 14 17.0222 1.21587 * TOTAL (CORRECTED) 23 2974.85 129.341 BALANCED ANOVA FOR VARIATE DAYS FILE PH 25/ 1/21 16:54 :PAGE VARIATE V006 DAYS LN SOURCE OF VARIATION SUMS OF MEAN F RATIO PROB ER SQUARES SQUARES LN ============================================================================= REPEAT 2.18628 1.09314 3.50 0.057 FORMULA 3083.41 440.488 ****** 0.000 * RESIDUAL 14 4.36769 311978 * TOTAL (CORRECTED) 23 3089.97 134.346 TABLE OF MEANS FOR FACTORIAL EFFECTS FILE PH 25/ 1/21 16:54 :PAGE MEANS FOR EFFECT REPEAT REPEAT NOS 8 DF DAYS 5.80875 5.76125 6.17750 DAYS 15.8438 15.7700 16.0875 DAYS 24.8075 24.3438 25.1787 DAYS 31.1175 31.4275 31.8538 SE(N= 8) 0.160922 0.130203 0.389851 0.197477 5%LSD 14DF 0.488113 0.394935 1.18250 0.598992 MEANS FOR EFFECT FORMULA FORMULA NOS 3 3 3 3 DAYS 0.836667 3.72333 4.21333 6.19333 7.16667 9.25000 9.11000 6.83333 DAYS 1.30333 6.97333 8.58333 17.4267 21.6100 25.4700 25.1667 20.6700 DAYS 1.91667 14.8600 18.8167 28.8233 31.8333 35.2600 35.0000 31.7033 DAYS 3.74667 26.5567 30.2200 36.4533 38.5300 38.8700 39.0000 38.3533 SE(N= 3) 0.262785 0.212621 0.636624 0.322479 5%LSD 14DF 0.797085 0.644927 1.93102 0.978150 ANALYSIS OF VARIANCE SUMMARY TABLE FILE PH 25/ 1/21 16:54 :PAGE F-PROBABLIITY VALUES FOR EACH EFFECT IN THE MODEL SECTION - VARIATE DAYS DAYS DAYS DAYS GRAND MEAN (N= 24) NO OBS 24 5.9158 24 15.900 24 24.777 24 31.466 STANDARD DEVIATION C OF V |REPEAT SD/MEAN | BASED ON BASED ON % | TOTAL SS RESID SS | 2.7635 0.45516 7.7 0.1702 8.7143 0.36827 2.3 0.2305 11.373 1.1027 4.5 0.3454 11.591 0.55855 1.8 0.0575 |FORMULA | | | 0.0000 0.0000 0.0000 0.0000 | | | | 4.3: Investigation of the growth of mycelial on paddy grain substrates (grain master or grade 2) BALANCED ANOVA FOR VARIATE DAYS FILE PADDY 25/ 1/21 18:13 :PAGE VARIATE V003 DAYS 60 LN SOURCE OF VARIATION DF SUMS OF MEAN F RATIO PROB ER SQUARES SQUARES LN ============================================================================= REPEAT 1.18820 594100 0.57 0.606 FORMULA 38.0000 19.0000 18.33 0.012 * RESIDUAL 4.14520 1.03630 * TOTAL (CORRECTED) 43.3334 5.41668 BALANCED ANOVA FOR VARIATE DAYS FILE PADDY 25/ 1/21 18:13 :PAGE VARIATE V004 DAYS LN SOURCE OF VARIATION DF LN SOURCE OF VARIATION DF LN SOURCE OF VARIATION DF LN SOURCE OF VARIATION DF SUMS OF MEAN F RATIO PROB ER SQUARES SQUARES LN ============================================================================= REPEAT 4.31976 2.15988 1.89 0.264 FORMULA 234.337 117.168 102.52 0.001 * RESIDUAL 4.57140 1.14285 * TOTAL (CORRECTED) 243.228 30.4035 BALANCED ANOVA FOR VARIATE 12 DAYS FILE PADDY 25/ 1/21 18:13 :PAGE VARIATE V005 12 DAYS SUMS OF MEAN F RATIO PROB ER SQUARES SQUARES LN ============================================================================= REPEAT 2.26646 1.13323 9.07 0.034 FORMULA 295.584 147.792 ****** 0.000 * RESIDUAL 499566 124891 * TOTAL (CORRECTED) 298.350 37.2938 BALANCED ANOVA FOR VARIATE 16 DAYS FILE PADDY 25/ 1/21 18:13 :PAGE VARIATE V006 16 DAYS SUMS OF MEAN F RATIO PROB ER SQUARES SQUARES LN ============================================================================= REPEAT 2.90002 1.45001 3.39 0.138 FORMULA 188.068 94.0341 220.01 0.000 * RESIDUAL 1.70965 427413 * TOTAL (CORRECTED) 192.678 24.0847 BALANCED ANOVA FOR VARIATE 20 DAYS FILE PADDY 25/ 1/21 18:13 :PAGE VARIATE V007 20 DAYS SUMS OF MEAN F RATIO PROB ER SQUARES SQUARES LN ============================================================================= REPEAT 5.95902 2.97951 1.64 0.302 FORMULA 101.446 50.7232 27.98 0.006 * RESIDUAL 7.25138 1.81285 * TOTAL (CORRECTED) 114.657 14.3321 TABLE OF MEANS FOR FACTORIAL EFFECTS FILE PADDY 25/ 1/21 18:13 :PAGE MEANS FOR EFFECT REPEAT REPEAT NOS 3 3 SE(N= 3) DAYS 15.7800 16.6700 16.2200 DAYS 35.6667 36.7767 37.3333 12 DAYS 60.9967 62.0000 62.1133 16 DAYS 90.7767 91.7767 92.1133 0.587735 0.617211 0.204036 0.377453 61 5%LSD 4DF 2.30380 REPEAT NOS 3 3 2.41934 0.799775 1.47954 20 DAYS 118.777 120.443 120.557 SE(N= 3) 0.777356 5%LSD 4DF 3.04707 MEANS FOR EFFECT FORMULA FORMULA NOS 3 3 SE(N= 5%LSD 3) 4DF FORMULA NOS 3 3 DAYS 15.8900 18.8900 13.8900 DAYS 33.5533 43.7800 32.4433 12 DAYS 55.0000 69.0000 61.1100 16 DAYS 87.8900 98.0000 88.7767 0.587735 2.30380 0.617211 2.41934 0.204036 0.799775 0.377453 1.47954 20 DAYS 117.333 124.667 117.777 SE(N= 3) 0.777356 5%LSD 4DF 3.04707 ANALYSIS OF VARIANCE SUMMARY TABLE FILE PADDY 25/ 1/21 18:13 :PAGE F-PROBABLIITY VALUES FOR EACH EFFECT IN THE MODEL SECTION - VARIATE 12 16 20 DAYS DAYS DAYS DAYS DAYS GRAND MEAN (N= 9) NO OBS 16.223 36.592 61.703 91.556 119.93 STANDARD DEVIATION C OF V |REPEAT SD/MEAN | BASED ON BASED ON % | TOTAL SS RESID SS | 2.3274 1.0180 6.3 0.6064 5.5139 1.0690 2.9 0.2645 6.1069 0.35340 0.6 0.0344 4.9076 0.65377 0.7 0.1378 3.7858 1.3464 1.1 0.3016 |FORMULA | | | 0.0116 0.0011 0.0001 0.0005 0.0061 | | | | 4.4: Investigation of the growth of Ganoderma subresinosum mycelial on culture media BALANCED ANOVA FOR VARIATE DAYS FILE AGAR 25/ 1/21 22:11 :PAGE VARIATE V003 DAYS LN SOURCE OF VARIATION DF LN SOURCE OF VARIATION DF SUMS OF MEAN F RATIO PROB ER SQUARES SQUARES LN ============================================================================= REPEAT 164433 822167E-01 4.20 0.047 FORMULA 182183 364367E-01 1.86 0.188 * RESIDUAL 10 195633 195633E-01 * TOTAL (CORRECTED) 17 542250 318971E-01 BALANCED ANOVA FOR VARIATE DAYS FILE AGAR 25/ 1/21 22:11 :PAGE VARIATE V004 DAYS SUMS OF MEAN F RATIO PROB ER SQUARES SQUARES LN ============================================================================= 62 REPEAT 3.33941 1.66971 1.61 0.247 FORMULA 22.4026 4.48051 4.32 0.024 * RESIDUAL 10 10.3783 1.03783 * TOTAL (CORRECTED) 17 36.1202 2.12472 BALANCED ANOVA FOR VARIATE DAYS FILE AGAR 25/ 1/21 22:11 :PAGE VARIATE V005 DAYS LN SOURCE OF VARIATION DF SUMS OF MEAN F RATIO PROB ER SQUARES SQUARES LN ============================================================================= REPEAT 2.35414 1.17707 0.61 0.568 FORMULA 325.129 65.0258 33.51 0.000 * RESIDUAL 10 19.4045 1.94045 * TOTAL (CORRECTED) 17 346.887 20.4051 BALANCED ANOVA FOR VARIATE DAYS FILE AGAR 25/ 1/21 22:11 :PAGE VARIATE V006 DAYS LN SOURCE OF VARIATION DF LN SOURCE OF VARIATION DF SUMS OF MEAN F RATIO PROB ER SQUARES SQUARES LN ============================================================================= REPEAT 4.13230 2.06615 1.23 0.333 FORMULA 502.316 100.463 59.89 0.000 * RESIDUAL 10 16.7760 1.67760 * TOTAL (CORRECTED) 17 523.224 30.7779 BALANCED ANOVA FOR VARIATE 10 DAYS FILE AGAR 25/ 1/21 22:11 :PAGE VARIATE V007 10 DAYS SUMS OF MEAN F RATIO PROB ER SQUARES SQUARES LN ============================================================================= REPEAT 258534 129267 0.22 0.804 FORMULA 440.797 88.1593 153.24 0.000 * RESIDUAL 10 5.75288 575288 * TOTAL (CORRECTED) 17 446.808 26.2828 TABLE OF MEANS FOR FACTORIAL EFFECTS FILE AGAR 25/ 1/21 22:11 :PAGE MEANS FOR EFFECT REPEAT REPEAT SE(N= 5%LSD NOS 6 6) 10DF REPEAT NOS 6 DAYS 1.43333 1.44500 1.23667 DAYS 8.41667 7.36167 7.88000 DAYS 18.3333 19.1650 19.0133 DAYS 25.6667 26.7517 26.5967 0.571013E-01 0.415898 0.179928 1.31051 0.568689 1.79196 0.528772 1.66618 10 DAYS 31.9583 31.9050 32.1817 SE(N= 6) 0.309647 5%LSD 10DF 0.975710 MEANS FOR EFFECT FORMULA FORMULA NOS 3 DAYS 1.41667 1.47333 DAYS 7.39000 7.22333 63 DAYS 14.8033 16.7467 DAYS 19.2533 24.3067 3 3 SE(N= 5%LSD 3) 10DF FORMULA NOS 3 3 3 1.28000 1.19333 1.45000 1.41667 7.27667 6.83333 8.48333 10.1100 16.6667 15.5833 22.9167 26.3067 24.1367 23.6133 32.3033 34.4167 0.807534E-01 0.588168 0.254457 1.85334 0.804248 2.53422 0.747796 2.35633 10 DAYS 24.3900 29.8900 30.9200 30.2233 38.2233 38.4433 SE(N= 3) 0.437907 5%LSD 10DF 1.37986 ANALYSIS OF VARIANCE SUMMARY TABLE FILE AGAR 25/ 1/21 22:11 :PAGE F-PROBABLIITY VALUES FOR EACH EFFECT IN THE MODEL SECTION - VARIATE 10 4.5: DAYS DAYS DAYS DAYS DAYS GRAND MEAN (N= 18) NO OBS 18 1.3717 18 7.8861 18 18.837 18 26.338 18 32.015 STANDARD DEVIATION C OF V |REPEAT SD/MEAN | BASED ON BASED ON % | TOTAL SS RESID SS | 0.17860 0.13987 10.2 0.0469 1.4576 1.0187 12.9 0.2475 4.5172 1.3930 7.4 0.5682 5.5478 1.2952 4.9 0.3334 5.1267 0.75848 2.4 0.8043 |FORMULA | | | 0.1879 0.0239 0.0000 0.0000 0.0000 | | | | Investigation of the growth and development of Ganoderma subresinosum cultivated on different substrates BALANCED ANOVA FOR VARIATE DAYS FILE GROWTH 25/ 1/21 21:17 :PAGE VARIATE V003 DAYS LN SOURCE OF VARIATION DF LN SOURCE OF VARIATION DF SUMS OF MEAN F RATIO PROB ER SQUARES SQUARES LN ============================================================================= REPEAT 2.11111 1.05556 0.82 0.472 FORMULA 4.14111 828222 0.64 0.675 * RESIDUAL 10 12.8972 1.28972 * TOTAL (CORRECTED) 17 19.1494 1.12644 BALANCED ANOVA FOR VARIATE DAYS FILE GROWTH 25/ 1/21 21:17 :PAGE VARIATE V004 DAYS SUMS OF MEAN F RATIO PROB ER SQUARES SQUARES LN ============================================================================= REPEAT 2.20121 1.10061 0.71 0.519 FORMULA 23.7001 4.74002 3.06 0.063 * RESIDUAL 10 15.5097 1.55097 * TOTAL (CORRECTED) 17 41.4110 2.43594 BALANCED ANOVA FOR VARIATE 12 DAYS FILE GROWTH 25/ 1/21 21:17 :PAGE 64 VARIATE V005 12 DAYS LN SOURCE OF VARIATION DF SUMS OF MEAN F RATIO PROB ER SQUARES SQUARES LN ============================================================================= REPEAT 40.8353 20.4176 1.76 0.221 FORMULA 191.643 38.3286 3.30 0.051 * RESIDUAL 10 116.308 11.6308 * TOTAL (CORRECTED) 17 348.786 20.5168 BALANCED ANOVA FOR VARIATE 16 DAYS FILE GROWTH 25/ 1/21 21:17 :PAGE VARIATE V006 16 DAYS LN SOURCE OF VARIATION DF LN SOURCE OF VARIATION DF SUMS OF MEAN F RATIO PROB ER SQUARES SQUARES LN ============================================================================= REPEAT 34.3916 17.1958 1.73 0.225 FORMULA 105.955 21.1910 2.14 0.143 * RESIDUAL 10 99.1777 9.91777 * TOTAL (CORRECTED) 17 239.524 14.0896 BALANCED ANOVA FOR VARIATE 20 DAYS FILE GROWTH 25/ 1/21 21:17 :PAGE VARIATE V007 20 DAYS SUMS OF MEAN F RATIO PROB ER SQUARES SQUARES LN ============================================================================= REPEAT 7.37029 3.68514 0.31 0.745 FORMULA 522.292 104.458 8.71 0.002 * RESIDUAL 10 119.871 11.9871 * TOTAL (CORRECTED) 17 649.534 38.2079 TABLE OF MEANS FOR FACTORIAL EFFECTS FILE GROWTH 25/ 1/21 21:17 :PAGE MEANS FOR EFFECT REPEAT REPEAT SE(N= 5%LSD NOS 6 6) 10DF REPEAT NOS 6 DAYS 5.95000 6.78333 6.45000 DAYS 11.6083 10.7750 11.0200 12 DAYS 26.4500 28.4750 24.7917 16 DAYS 47.2667 50.3217 50.0583 0.463631 1.46092 0.508424 1.60206 1.39229 4.38715 1.28568 4.05121 20 DAYS 67.2083 68.0333 66.4667 SE(N= 6) 1.41346 5%LSD 10DF 4.45385 MEANS FOR EFFECT FORMULA FORMULA SE(N= 5%LSD 3) 10DF NOS 3 3 3 DAYS 6.75000 5.70000 6.35000 6.80000 5.83333 6.93333 DAYS 10.0667 10.5000 11.1667 12.9733 9.83333 12.2667 12 DAYS 29.6833 23.6500 21.4833 28.2333 25.7667 30.6167 16 DAYS 52.0500 48.0500 47.6000 51.3333 45.2600 51.0000 0.655673 2.06605 0.719021 2.26566 1.96899 6.20437 1.81822 5.72928 65 FORMULA NOS 3 3 3 20 DAYS 72.9500 69.3833 59.1000 71.1667 60.4833 70.3333 SE(N= 3) 1.99893 5%LSD 10DF 6.29869 ANALYSIS OF VARIANCE SUMMARY TABLE FILE GROWTH 25/ 1/21 21:17 :PAGE F-PROBABLIITY VALUES FOR EACH EFFECT IN THE MODEL SECTION - VARIATE 12 16 20 DAYS DAYS DAYS DAYS DAYS GRAND MEAN (N= 18) NO OBS 18 6.3944 18 11.134 18 26.572 18 49.216 18 67.236 STANDARD DEVIATION C OF V |REPEAT SD/MEAN | BASED ON BASED ON % | TOTAL SS RESID SS | 1.0613 1.1357 17.8 0.4716 1.5608 1.2454 11.2 0.5186 4.5296 3.4104 12.8 0.2214 3.7536 3.1492 6.4 0.2251 6.1813 3.4622 5.1 0.7450 66 |FORMULA | | | 0.6749 0.0625 0.0513 0.1433 0.0023 | | | |