臺灣博碩士論文加值系統

秀珍菇（Pleurotus ostreatus，PO）及鮑魚菇（P. cystidiosus，PC)為食藥用菌菇，具有豐富的營養成分和保健功效，並具有生技和環境上的應用價值。本研究以秀珍菇及鮑魚菇為材料，探討(1)溫度和養分對其菌絲體生長的影響，(2)評估介質、光照、太空包口的開啟方式和菌種來源對其子實體的生長、產量和營養成分的影響及(3)探討栽培介質、乾燥方式、光照處理等栽培環境下，對PO及PC子實體抗氧化活性的反應。研究結果顯示：PO及PC的菌絲體可在溫度24~32oC範圍中生長，在28oC時表現最佳。馬鈴薯葡萄糖培養基（potato dextrose agar medium，PDA）、番薯葡萄糖培養基（sweet potato dextrose agar medium，SPDA）、山藥葡萄糖培養基（yam dextrose agar medium，YDA）和麥芽萃取物培養基（malt extract agar medium，MEA）是適合PC菌絲體生長的培養基，而PDA和YDA是適合PO菌絲體生長的培養基。添加濃度1~5%的蔗糖為碳源與添加氯化銨0.03-0.09％作為氮源，可以促進PO和PC的菌絲體生長。糙米和黃玉米為生產菌種最好之資材，而雜
木屑sawdust，SD）、玉米穗軸（corn cob，CC）和甘蔗渣（sugarcane bagasse ，SB）為適合PO和PC菌絲體生長的木質纖維素來源。在栽培介質的配方中，增加使用CC和SB的比率可減少碳氮比（C/N）與增加礦物質含量的方式，有助於增加菌蓋直徑、菌柄厚度、子實體重量、產量、生物轉換率（BE）、蛋白質、纖維，灰分和礦物質含量(鈣、鉀、鎂)，因此CC可以取代全部或部分木屑的栽培介質成份。本研究顯示光照處理影響PO與PC子實體之生長、產量與生物轉換率，但對其營養成分並無影響。以黑暗處理做為對照，藍光、白光、紅光與紅外線發光二極體(LED)環境下，明顯地縮短PO菇原體形成時間8.78到11.98天，而PC縮短6.3到8.2天。紅外線LED燈和紅光LED燈是最適合PO和PC子實體生長的環境，因其縮短了菇原體形成的天數，略增加了菇體的重量與菌柄的厚度，以及總收穫量和生物轉換率。在測試太空包開口的試驗中，開口表面積越大造成介質乾燥的速度也就越快，也減少了出菇次數和採收期，然而，卻增加了菇體的數目。PO在三潮的出菇和PC在第一潮的出菇，除了脫袋法，其他袋口不同處理方式並沒有影響到PO和PC的營養成分。不要將太空包套環移除是對PC最好的栽培方式，此方式收穫達最高產量（204.3克/包）和高BE（40.80%），而在PO方面，將太空包套環移除為最好的方式，得到了最高產量（298.5克/包）和BE（59.62%）。以糙米和雜木屑作為菌種栽培介質，PO和PC不同的菌種來源並沒有對生長、產量和BE產生明顯地變化。因此，在秀珍菇的栽培上，可以利用雜木屑取代糙米作為菌種介質，達到降低成本和高獲利的目標。
關於PO和PC在抗氧化合物及抗氧化活性的分析結果如下: 介質中若含有較高的CC和SB成份（100%和50%），子實體中則有較高的總酚含量（TPC）、總類黃酮含量（TPF）以及高效率的1,1-diphenyl-2-picrylhydrazyl（DPPH）自由基清除能力、還原能力及螯合亞鐵離子能力;然而在100% SD的介質種植下，從PO和PC的子實體萃取物中，會減少TPC和TFC含量進而降低抗氧化的能力。結果顯示配方中使用CC和SB來取代部分或是全部使用的SD來栽培秀珍菇，將可增加抗氧化物質及抗氧化的能力。以冷凍乾燥法乾燥子實體較傳統烘乾法能有效率的提高TPC以及抗氧化活性。不同光處理對PO和PC之TPC含量無顯著提升，對PO之TFC含量也無明顯的影響。PC在不同光條件的處理下，對TFC有影響然差異不顯著，一般而言，在所有光照處理條件下PO和PC具有良好的抗氧化活性、對DPPH自由基清除能力及螯合亞鐵離子能力。紅外LED燈和紅色LED燈為最適合的光照處理，它不僅有助於提高產量和生物轉換率，而且也增加DPPH自由基清除能力與PO和PC的還原力，些微增加PO亞鐵離子的螯合能力。

Pleurotus ostreatus (PO) and P. cystidiosus (PC) are edible and medicinal mushrooms, which are rich in nutrition and health benefits and have the biotechnology and environmental value used as material in this study. The objective of the study was (1) to evaluate the effects of temperature and nutrition conditions on the mycelium growth of two oyster mushrooms PO and PC for spawn production; (2) to evaluate the effects of some factors such as substrates, light treating conditions, bag opening types, spawn sources on the growth, yield, and nutritional composition of PO and PC fruiting body; (3) to determine the effects of substrate, drying method, light treating condition on antioxidant compound, antioxidant activity of mushrooms PO and PC. The results of this study showed that:
The mycelium of mushroom PO and PC can grow at 24-32oC, but at 28oC present the best. Potato dextrose agar medium (PDA), sweet potato dextrose agar medium (SPDA), yam dextrose agar medium (YDA), and malt extract agar medium (MEA) were the favorable media for mushroom PC mycelium growth while PDA and YDA were the suitable media for PO mycelium growth. Addition of sucrose at 1-5% concentration as carbon source and ammonium chloride at 0.03-0.09% concentration as nitrogen source can promote the mycelium growth of mushrooms PO and PC. Brown rice was the best grain sources to produce spawn of mushroom PO and PC while sawdust (SD), corn cob (CC), and sugarcane bagasse (SB) were suitable lignocellulose substrate sources for mycelium growth as well. Increasing the rates of using CC and SB helped decreasing C/N ratio and enhancing some mineral contents of substrate formulas resulted increasing the cap diameter, stipe thickness, mushroom weight, yield, biological efficiency (BE), protein, fiber, ash, and mineral content (Ca, K, and Mg) of mushroom PO and PC, consequently they can replace some parts or whole SD in substrate formulas. In this study, light treating conditions had effect on the growth, yield, BE and no effect on nutrition composition of oyster mushrooms PO and PC. In comparison with dark condition, light stimulated primordial formation early, decreasing the primordial formation days, so that total harvesting period under light treating was significantly shorter from 8.78 to 11.98 days in case of mushroom PO and from 6.3 to 8.2 days in case of mushroom PC. Infrared-LED (light emitting diode) light and red-LED light were the most suitable treating conditions because of reducing total harvesting period and slight increasing stipe thickness, mushroom weight, total yield, and BE of mushrooms PO and PC. This result obtained showed bag opened with larger surface induced rapidly dried substrate and reducing the number of flushes and total harvesting period. However, it induced increasing the number of effective fruiting bodies, the yield of three flushes of mushroom PO and the first flush of mushroom PC except de-plastic culture bag method, while bag opening method did not impact on nutritional composition of mushrooms PO and PC. No removing ring was the best method to get the highest yield (204.3 g/bag) and BE (40.80%) of mushroom PC, while removing ring was identified as the best method for cultivation of mushroom PO with the highest yield (298.5 g/bag) and BE (59.62%). The result showed that spawn source from brown rice grain had no significant effect on the growth, yield, and BE of mushrooms PO and PC when compared with spawn source from sawdust substrate. In large scale of oyster mushroom cultivation, spawn source from sawdust substrate should be used to replace spawn source from brown rice grain with purpose of low cost and high benefit.
Regarding to antioxidant compound and activity of mushrooms PO and PC, the results showed that substrates containing higher contents of CC and SB (100% and 50%) resulted in higher values of total phenolic contents (TPC), total flavonoid contents (TFC) as well as high efficiency of 1,1- diphenyl-2-picrylhydrazyl (DPPH) radical scavenging ability, reducing power, and chelating ferrous ions ability. Whereas, substrate 100% SD reduced TPC and TFC which directly linked to a decreased antioxidant activity of mushroom PO and PC extracts. These results suggested that CC and SB can be used to replace some parts or all SD in substrate formulas for oyster mushroom cultivation which also improved antioxidant component and antioxidant activity of mushroom extracts. With freeze drying method, mushrooms PO and PC cultivated in almost substrates showed efficiency in improving the TPC as well as antioxidant activity in comparison with oven drying method. The results also indicated that different light treating conditions had no significant effect on TPC of PO and PC mushroom, and TFC of mushroom PO, however, it has effect on TFC of mushroom PC. In general, mushroom PO and PC had good antioxidant activity, especially DPPH radical scavenging ability and chelating ability on ferrous ions at all light treating conditions. Different light treating conditions had slight effect on DPPH radical scavenging ability, chelating ability and reducing power at 5 mg/mL concentration of extract in methanol. Red-LED light and infrared-LED light were the suitable light treating condition, it not only helps slightly increasing yield and BE but also increases DPPH radical scavenging ability, reducing power of mushroom PO and PC as well as slight increasing chelating ability on ferrous ions of mushroom PO.

TABLE OF CONTENTS
摘 要…………………………………...………………………………………i
ABSTRACT iv
LIST OF ABBREVIATION viii
ACKNOWLEDGEMENTS ix
TABLE OF CONTENTS x
LIST OF TABLES xviii
LIST OF FIGURES…………………………………………….….......……xxi
CHAPTER 1 1
INTRODUCTION 1
CHAPTER 2 5
LITERATURE REVIEW 5
2.1. Introduction of mushroom 5
2.2. Life cycle of mushroom 6
2.3. Edible mushroom production and consumption in the world 8
2.4. Classification and Biological characteristics of oyster mushrooms PO and PC 9
2.5. Nutritional contribution of oyster mushroom 12
2.5.1. Active constituents 13
2.5.2. Proteins 13

2.5.3. Lipids 14
2.5.4. Carbohydrates and fibers 15
2.5.5. Vitamins 16
2.5.6. Mineral constituents 16
2.6. Potential development of oyster mushroom in Vietnam 17
2.7. Cultivation of oyster mushrooms 17
2.7.1. Cultivation methods 17
2.7.2. Spawn production and their effects on mushroom production 18
2.7.3. Substrates for oyster mushroom cultivation 20
2.7.4. Light requirement for mushroom 22
2.8. Antioxidants 24
2.8.1. Antioxidant and free radical introduction 24
3.8.2. Phenolic compound 27
3.8.2.1. Antiproliferative of phenolic compound 28
3.8.2.2. Phenolic acid 29
3.8.2.3. Flavonoid 31
3.9. Antioxidant of oyster mushroom 32
CHAPTER 3 35
MATERIAL AND METHODS 35
Folow chart of the reseach…………………………..………………………..….36
3.1. Materials 37
3.1.1. Oyster mushrooms Pleurotus ostreatus (PO) and P. cytidiosus (PC) 37
3.1.2. Grain and lignocellulosic substrate 37
3.1.3. Chemicals 37
3.1.4. Instruments 39
3.2. Methods 40
3.2.1. Evaluating the factors on the mycelium growth of two oyster mushrooms PO and PC for spawn production 40
3.2.1.1. Experimental design 40
3.2.1.2. Evaluating the culture media on mycelium growth 41
3.2.1.3. Evaluating the temperatures on mycelium growth 41
3.2.1.4. Evaluating the carbon sources and concentrations on mycelium growth 42
3.2.1.5. Evaluating the nitrogen sources and concentrations on mycelium growth 42
3.2.1.6. Evaluating the different grain sources on mycelium growth 43
3.2.1.7. Evaluating the lignocellulosic substrate sources on mycelium growth 43
3.2.2. Evaluating the factors on the mushroom PO and PC fruiting body production 44
3.2.2.1. Evaluating the substrate formulas on the growth, yield and nutritional composition of two oyster mushrooms PO and PC 44
3.2.2.1.1. Experimental design 44
3.2.2.1.2 Mushroom cultivation material and spawn preparation 44
3.2.2.1.3. The preparation of substrates in plastic bag and inoculation 45
3.2.2.1.4. Incubation and harvest 46
3.2.2.1.5. Substrate analysis 47
3.2.2.1.6. Fruiting body analysis 49
3.2.2.2. Evaluating the light treating conditions on the growth, yield and nutritional composition of two oyster mushrooms PO and PC 51
3.2.2.3. Evaluating the bag opening methods on the growth, yield, and nutritional composition of two oyster mushrooms PO and PC 53
3.2.2.4. Evaluating the spawn sources on growth, yield, and biological efficiency (BE) of two oyster mushrooms PO and PC 53
3.2.3. Evaluating the factors on antioxidant compound and antioxidant activity of fruiting body 54
3.2.3.1. Evaluating the substrates and drying methods on antioxidant compound and activity of fruiting body of two oyster mushrooms PO and PC 54
3.2.3.1.1. Sample preparation 55
3.2.3.1.2. Determination of total phenolic contents (TPC) 55
3.2.3.1.3. Determination of total flavonoid contents (TFC) 56
3.2.3.1.4. Scavenging ability on 1,1- dephenyl-2-picrylhydrazyl (DPPH) radical 57
3.2.3.1.5. Reducing power assay 57
3.2.3.1.6. Chelating ability on ferrous ions 58
3.2.3.2. Evaluating the light treating conditions on antioxidant compound and activity of fruiting body of two oyster mushrooms PO and PC 58
3.2.4. Data analysis 59
CHAPTER 4 60
RESULTS AND DISCUSSION 60
4.1. Effect of factors on the mycelium growth for spawn production 60
4.1.1. Effect of different culture media on the mycelium growth 60
4.1.2. Effect of temperatures on mycelium growth 63
4.1.3. Effect of carbon sources and sucrose concentration on mycelium growth 65
4.1.4. Effect of nitrogen sources and concentration on mycelium growth 70
4.1.5. Effect of different grain sources on the mycelium growth 75
4.1.6. Effect of different lignocellulosic substrate sources on the mycelium growth 78
4.1.7. Conclusion for experiment of the factors on the mycelium growth of two oyster mushrooms PO and PC for spawn production 82
4.2. Effect of factors on the mushroom PO and PC fruiting body production 83
4.2.1. Effects of different substrate formulas on the growth, yield and nutritional composition of two oyster mushrooms PO and PC 83
4.2.1.1. Substrate analysis 83
4.2.1.2. Effect of different substrate formulas on morphological parameters 86
4.2.1.3. Effect of different substrate formulas on characteristics of fruiting body 89
4.2.1.4. Effect of different substrate formulas on yield and biological efficiency 92
4.2.1.5. Effect of different substrate formulas on nutritional composition of two oyster mushrooms PO and PC fruiting body 97
4.2.1.6. Effect of different substrate formulas on mineral contents of two oyster mushrooms PO and PC fruiting body 102
4.2.1.7. Conclusion for experiment of different substrate formulas on the growth, yield, nutritional composition of two oyster mushrooms PO and PC 105
4.2.2. Effects of light treating conditions on the growth, yield and nutritional composition of two oyster mushrooms PO and PC 106
4.2.2.1. Effect of different light treating conditions on total harvesting period and fruiting body characteristic of two oyster mushrooms PO and PC 106
4.2.2.2. Effect of different light treating conditions on the yield and biological efficiency of two oyster mushrooms PO and PC 109
4.2.2.3. Effect of different light treating conditions on nutritional composition of two oyster mushrooms PO and PC 112
4.2.2.4. Conclusion for experiment of light treating conditions on the growth, yield and nutritional composition of two oyster mushrooms PO and PC 114
4.2.3. Effect of different bag opening methods on the growth, yield, and nutritional composition of two oyster mushrooms PO and PC 114
4.2.3.1. Effect of different bag opening methods on the growth parameters and fruiting body characteristic of two oyster mushrooms PO and PC 114
4.2.3.2. Effect of different bag opening methods on the yield and biological efficiency of two oyster mushrooms PO and PC 118
4.2.3.3. Effects of different bag opening methods on nutritional composition of two oyster mushrooms PO and PC fruiting body 121
4.2.3.4. Conclusion for experiment of different bag opening methods on the growth, yield, and nutritional composition of mushrooms PO and PC 123
4.2.4. Effect of different spawn sources on the growth, yield, and biological efficiency of two oyster mushrooms PO and PC 124
4.2.4.1. Effect of different spawn sources on the harvesting time and fruiting body characteristics of two oyster mushrooms PO and PC 124
4.2.4.2. Effect of different spawn sources yield, and biological efficiency of two oyster mushrooms PO and PC 126
4.2.4.3. Conclusion for experiment of different spawn sources on the growth, yield, and biological efficiency of mushrooms PO and PC 129
4.3. Effect of factors on antioxidant compound and antioxidant activity of two oyster mushroom PO and PC fruiting body 130
4.3.1. Effect of different substrate formulas and drying methods on antioxidant compound and activity of two oyster mushrooms PO and PC 130
4.3.1.1. Effect of different substrate formulas and drying method on TPC and TFC 130
4.3.1.2. Effect of different substrate formulas and drying method on DPPH radical scavenging ability of two oyster mushrooms PO and PC 136
4.3.1.3. Effect of different substrate formulas and drying method on reducing power ability of two oyster mushrooms PO and PC 142
4.3.1.4. Effect of different substrate formulas and drying method on ferrous ions chelating ability of two oyster mushrooms PO and PC 145
4.3.1.5. Conclusion for experiment of different substrates and drying methods on antioxidant compound and antioxidant activity of mushroom PO and PC 150
4.3.2. Effect of different light treating conditions on antioxidant compound and activity of two oyster mushrooms PO and PC 151
4.3.2.1. Effect of different light treating conditions on TPC and TFC of two oyster mushrooms PO and PC 151
4.3.2.2. Effect of different light treating conditions on DPPH radical scavenging ability of two oyster mushrooms PO and PC 154
4.3.2.3. Effect of different light treating conditions on reducing power ability of two oyster mushrooms PO and PC 158
4.3.2.4. Effect of different light treating conditions on ferrous ions chelating ability of two oyster mushrooms PO and PC 161
4.3.2.5. Conclusion for experiment of different light treating conditions on antioxidant compound and activity of two oyster mushrooms PO and PC 164
CHAPTER 5 165
GENERAL CONCLUSION 165
REFERENCES 168
BIOGRAPHICAL SKETCH OF AUTHOR 196

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