、發明說明： 【發明所屬之技術領域】 本發明關於一種菇類栽培介質及一種新穎之 木黴菌菌株，尤指一種利用該木黴菌菌株堆積發酵 之供杏鮑菇培養之栽培介質。 【先前技術】 杏鮑菇(户―(DC.Fr.) Qu 1.)是分佈 於亞熱帶及草原地帶之蠔菇屬的一種白腐真菌。 鮑菇其菌傘及菌柄質地肉質豐厚，口感 ， 呼。㈣關乎沒有野^ 過不斷人工培育改良所長成_，主 包、歸瓶填充介f ^以栽培。杏_ 木：心^主田要原严為木屬’木屑以山黃麻、楓 發酵木屑做佳：以f鮮！經過適當堆積 及品質，出^= 可獲得較高之杏鮑菇產量 出菇也較整齊，利於採收。 法，^= 發酵大多採用自然堆肥 及通氣性等，，吏自缺為Υ:材:大小、水份含量 中，經過if I 生物滋生於木屑材料 均-、化學酵時程後’生產出物理性狀 待木屑堆積出H兹裁培介質’因此，等 需要三個月：適合的栽培介質(-般而言 免的時間成本。 B ，係為杏鮑菇培養不可避 、、田声/ΐ發酵是一個動態的過程，勺入女地 =以物的變化，尤其ΐ其以= 4長，因此’為了増進有機 效率，調配不同有機物材料用量比例，或施予適當 f微生物菌種，是堆積發酵的關鍵之一，而微生物 菌種又與整體發酵過程的溫度變化習習相關。 一積發酵過程可分為升溫期、高溫、中溫 ，後腐热等不同階段。當有機材料混合後開始堆積 製作，堆肥化條件控制適當時，微生物開始繁殖， 熱能的㈣累積使整體材料的溫度開始上升；當溫 ，上，至55-60。〇時，熱能的累積達到高峰，此刻 :溫菌主宰整個過程；隨後高溫菌的繁殖逐漸減 緩，，而代之的是中溫菌的出現；當溫度到達 = -50 C時，中溫菌的族群逐漸消失，最後即為後腐 Ξ階段’此時有機材料中的腐植質成分隨著溫度的 邊化而逐漸增加’使有機組成分更加敎與腐熟 :目J的技術而言’杏鲍菇生產以採收 ===因在於第二㈣體產量和品質 如第一周期，且出菇時間較長， =而，栽培出兹―次後的廢棄栽培 然Ϊ高，必須適當處理才能U ; 3 J研發作為有機肥料，或添加其他^$ 病"貝，或做為其他菇類之栽培介質用。作為抑 以、商雖然培養過杏鮑菇後的栽捭介- 再利用’但就杏_的栽S二貝： -人新裁都需要使用新的木屑，不符人 /，母 概念’於成本的考量上也未绩理想。二仃的環保 $業迫切需要一種新穎的杏鮑菇栽捭八哲杏鮑菇 術’以減少新木屑的用量並縮短發酵：質製造技 【發明内容】 爰是，本發明之一目的為提供一種新穎的杏鲍 菇栽培介質的製造方法，可縮短堆積發酵的時間， 降低時間成本。 本發明之另一目的為提供一種新穎的杏鮑菇 栽培介質，可減少新木屑的用量，達到環保的要求 並降低生產成本。 為達到上述目的，本發明提供一種木黴菌菌 株，該木黴菌菌株在中華民國食品工業發展研究所 菌種中心的寄存編號為BCRC 930113。 本發明另提供一種用於杏鮑菇培養的栽培介 質，其係包含：基質；及如前述之木黴菌菌株。 本發明再提供一種製備培養杏鮑菇之栽培介 質的方法，其係包含下列步驟：提供一基質；混合 一木黴菌懸浮液·於前述基質中以形成一介質材 料，該木黴菌在中華民國食品工業發展研究所菌 種中心的寄存編號為BCRC 930113 ;調整前述介 質材料的水分含量；及將前述介質材料進行堆積發 酵。 較佳地，前述木黴菌菌株係濃度為lxlO7〜lx 108 cfu/ml的木黴菌懸浮液。 較佳地，每一立方公尺的前述基質混有15〜25 公升的前述木黴菌懸浮液。 較佳地，前述基質為未使用的木屑。 較佳地，前述基質包含舊栽培介質，該舊栽培 1378143· 介質係指已培養過杏鮑菇的栽培介質。 較佳地，前述舊栽培介質佔前述基質的20〜40 wt%。 較佳地，前述介質材料的堆積高度為2.0-2.5 公尺。 較佳地，前述介質材料的水分含量係調整至 55〜65 wt% 〇 本發明又提供一種用於杏鮑菇培養的栽培介 • 質，其係包含前述未使用的乾燥木屑作為基質；以 及前述寄存編號為BCRC 930113的木黴菌菌株， 並以前述方法製得。 本發明更提供一種用於杏鮑菇培養的栽培介 質，其係包含前述未使用的乾燥木屑和前述舊栽培 介質作為基質；以及前述寄存編號為BCRC 930113 的木黴菌菌株，並以前述方法製得。 綜上述，本發明關於一種新穎的用於杏鮑菇培 養的栽培介質及其製造方法，其包含寄存編號為 • BCRC 930113的木黴菌菌株，並適當地混合使用 新木屑以及舊栽培介質，使整體堆積發酵的時間 縮短三分之一，而得以減少新木屑的用量，達到 降低成本並符合環保的目的。 【實施方式】 本發明關於一種新穎的供杏鮑菇培養的栽培 介質及其製造方法，其技術重點在於使用寄存編號 為BCRC 930113的木黴菌菌株（其分離株編號為 6 [S1 介 1°9768) ’並適當地搭配使用新木屑和舊裁培 杏飽用「介質材料」-詞，係指混合有供 積發酵:材以:質的所需材料’而尚未完成堆 杏鮑所用「栽培介質」—詞，係指混合有供 發酵的i程之栽培介f的所需材料’並已完成堆積 i酵的過’而得以❹於杏·縣培養之材料混合 或招思2可，，但不限於山黃麻、楓木、楠木 栽i立二°:述舊栽培介質係指已培養過杏飽兹的 i杏:!4:rr。係指已培養過至少一個周期 之優 以下實施例係用於進一步了解本發明 點，並非用於限制本發明之申請專利範圍。 實施例一：菌種分離與篩選 ^發明之木黴菌分離菌株(TCF09768)係由有 農場1壌中’以減半營養抽出物水瓊脂(Nutrient agar N.A.)洋菜平板法進行微生物分離。簡單地說， 先稱取1克土壤置於lOc.c.無菌水中，經振盪後， 取過濾液於半營養抽出物之水瓊脂/洋菜培養皿上 進=平板劃線分離以獲得單一菌落，每種土壤樣品 ，行5個培養皿，將完成平板劃線的培養皿移至％ C之黑暗培養箱中培養二天後，挑取單一菌落移到 營養抽出物（Nutrient agar N.A·)洋菜平板培養基 上’培養三天後觀察各菌落生長形態。 接者由其中挑取似木徽菌菌落形態之菌落，進 7 ‘ < 1378,143. 行二次純系分離後取單一菌落置於裝有5 cc無菌 水之螺旋試管中保存。各分離菌株以馬鈴薯葡g糖 培養基（potato dextrose agar，PDA)培養 7 天後，以 5 c.c.無菌水將培養皿上之菌落洗下，置入裝1〇克 稻穀培養奉之試管中’並於4(TC下培養’以<觀察各 。菌株在其中生長狀況以及其產孢情形，以挑取於4〇 °C下能快速纏繞稻榖培養基及產孢之菌株。 實施例二：木徵菌生長條件測試及菌株特徵 將分離株TCF09768 (97ID104)進行菌株特徵 及菌種學名鑑定。於含有萘乙酸鈉（s〇(jium Naphthaleneacetic acid，SNA，一種生長調節劑） 的培養基上(20°C )培養4天的菌落具有以下特徵： 菌落直控約為90mm ;菌絲白色十分稀疏；產孢很 多，其孢子呈帶灰的綠色到深綠色（grayish green-dark green ’ 26D5-26F5);產孢構造為聚集簇 生(pustulate)並且呈半球形。 在顯微鏡下觀察其微構造之結果顯示，該菌落 之分生抱子柄（conidiophores)產生短的側生分支 (primary branches)，而近基部處之二次分支 (secondary branches)很少有再分支之情形，分支大 都沿著主支單一或成對(in pairs)產生，並與主支呈 近90度的角度。 該菌落的產孢細胞（conidiogenous cell， phialides)的形態為瓶梗（phialide)且呈安瓶形 (ampulliform)，中間略為膨大，長寬約為 7.4-9.4χ2.0-3_2μηι，大部分為 2-3 輪生(whorls)，主 支頂部具瓶梗(phialide)，形態上較為細長，約為 8 m 12.47-13.38><2·08-2.50μηι。分生跑子成熟時呈暗綠 色’形癌為球形（spherical)、近球形（subSphericaI) 或卵形（ovoidal) ((2.8-)3.5-3.8(-4.0) X (2.8-)3.0-3.5 μηι，係指非生孢子之橫徑X縱徑為3.5-3.8x3.0-3.5 ; 而（2.8-)(-4.0)為橫徑差值範圍），外壁呈細刺狀 (spinulose)，其紋飾（ornamentation，又稱外飾物） 有時不易觀察。 综合鑑定結果顯示，木黴菌分離株TCF09768 為 Samuels，Lieckfeldt &[Technical Field] The present invention relates to a mushroom cultivation medium and a novel strain of Trichoderma, and more particularly to a cultivation medium for cultivation of Pleurotus eryngii by the fermentation of the Trichoderma strain. [Prior Art] Pleurotus eryngii (DC.Fr.) Qu 1.) is a white rot fungus of the genus Pleurotus ostreatus distributed in the subtropical and prairie regions. The mushroom and the handle of the mushroom are rich in fleshy texture, taste, and call. (4) It is concerned with the fact that there is no wilderness, continuous cultivation and improvement, and the main package, the bottle filling, and the cultivation. Apricot _ wood: heart ^ main field to the original strict for the wood genus 'wood chips to mountain yellow hemp, maple fermented wood chips do better: to f fresh! After proper accumulation and quality, ^= can obtain higher yield of Pleurotus eryngii. The mushroom is also neat and tidy, which is conducive to harvesting. Method, ^= Most of the fermentation uses natural composting and aeration, etc., and the sputum is self-deficient: material: size, moisture content, after if I organisms are grown in wood chips - and after chemical fermentation time period, 'production physics The traits are to be piled up by the wood chips. Therefore, it takes three months: the suitable cultivation medium (-the general time cost. B, the cultivation of Pleurotus eryngii is inevitable, and the field sound/ΐ fermentation is A dynamic process, scooping into women's land = changes in things, especially in the case of = 4 long, so 'in order to break into the organic efficiency, blending the proportion of different organic materials, or to apply appropriate f microbial strains, is the accumulation of fermentation One of the key points, and the microbial strains are related to the temperature change of the whole fermentation process. The accumulation process can be divided into different stages such as heating period, high temperature, medium temperature and post-corrosion. When the composting conditions are properly controlled, the microorganisms begin to multiply, and the accumulation of thermal energy (4) causes the temperature of the whole material to start to rise; when it is warm, up to 55-60. When the heat energy reaches a peak, at this moment: The warm bacteria dominate the whole process; then the reproduction of the high temperature bacteria gradually slows down, and instead the appearance of the moderate temperature bacteria; when the temperature reaches = -50 C, the population of the moderate temperature bacteria gradually disappears, and finally the post-corrosion stage 'At this time, the humus components in the organic material gradually increase with the temperature edge-making the organic component more sturdy and decomposed: in terms of the technology of the eye J, the production of the oyster mushroom is harvested === (4) The body yield and quality are as in the first cycle, and the time for mushrooming is longer, = and the cultivated cultivation after the cultivation is high, and must be properly treated to be U; 3 J R&D as an organic fertilizer, or add other ^ $病"Bei, or as a cultivation medium for other mushrooms. As a restraint, although the cultivation of Pleurotus eryngii has been cultivated - reused, but the apricot _ planted S two shells: - people new The need to use new wood chips, the non-conformity / the mother concept 'has not been satisfactory in terms of cost considerations. The environmental protection of the two industries is urgently needed for a novel Pleurotus eryngii planted with eight scented oyster mushrooms to reduce The amount of new wood chips and shorten the fermentation: quality manufacturing technology [invention content] It is an object of the present invention to provide a novel method for producing a culture medium of Pleurotus eryngii, which can shorten the time of accumulation fermentation and reduce the time cost. Another object of the present invention is to provide a novel cultivation medium for Pleurotus eryngii, which can be To reduce the amount of new wood chips, to meet environmental protection requirements and to reduce production costs. To achieve the above object, the present invention provides a Trichoderma strain, which is registered under the registration number BCRC 930113 of the Republic of China Food Industry Development Research Institute. The present invention further provides a cultivation medium for cultivating Pleurotus eryngii, comprising: a substrate; and a Trichoderma strain as described above. The present invention further provides a method for preparing a cultivation medium for cultivating Pleurotus eryngii, which comprises the following steps Providing a substrate; mixing a Trichoderma suspension in the matrix to form a dielectric material, the registration number of the Trichoderma in the Center of Culture Development of the Republic of China Food Industry Development Institute is BCRC 930113; adjusting the moisture content of the aforementioned dielectric material And the above-mentioned dielectric material is subjected to stack fermentation. Preferably, the Trichoderma strain is in a concentration of 1×10 7×1×108 cfu/ml of the Trichoderma suspension. Preferably, the aforementioned substrate per cubic meter is mixed with 15 to 25 liters of the aforementioned Trichoderma suspension. Preferably, the aforementioned substrate is unused wood chips. Preferably, the matrix comprises an old cultivation medium, and the old cultivation 1378143· medium refers to a cultivation medium in which Pleurotus eryngii has been cultured. Preferably, the aforementioned old cultivation medium accounts for 20 to 40% by weight of the aforementioned substrate. Preferably, the aforementioned dielectric material has a stack height of 2.0 to 2.5 meters. Preferably, the moisture content of the dielectric material is adjusted to 55 to 65 wt%. The present invention further provides a cultivation medium for cultivating Pleurotus eryngii, which comprises the aforementioned unused dry wood chips as a substrate; The Trichoderma strain numbered BCRC 930113 was deposited and prepared as described above. The present invention further provides a cultivation medium for cultivating Pleurotus eryngii, comprising the aforementioned unused dried wood chips and the aforementioned old cultivation medium as a substrate; and the aforementioned Trichoderma strain having the accession number BCRC 930113, which is obtained by the aforementioned method. . In view of the above, the present invention relates to a novel cultivation medium for cultivating Pleurotus eryngii and a method for producing the same, which comprises a Trichoderma strain having the registration number BCRC 930113, and appropriately mixing new wood chips and old cultivation medium to make the whole The time to accumulate fermentation is shortened by one-third, and the amount of new wood chips can be reduced to achieve cost reduction and environmental protection. [Embodiment] The present invention relates to a novel cultivation medium for cultivating Pleurotus eryngii and a method for producing the same, the technical focus of which is to use a Trichoderma strain having the accession number BCRC 930113 (the isolate number is 6 [S1 介1°9768] ) 'And use the new wood chips and the old cut apricots with the "media material" - the word refers to the mixed fermentation: the material is: the required material of the quality' and has not completed the "cultivation medium" used in the apricot "-" refers to the material required to mix the fermented i-process and has completed the accumulation of i-fermentation and can be mixed with the materials cultivated in the apricot and county. It is not limited to mountain jute, maple, and nanmu planting. Two degrees: The old cultivation medium refers to the apricot that has been cultivated with apricots: 4: rr. It is preferred that the following examples have been cultivated for the purpose of further understanding of the present invention and are not intended to limit the scope of the invention. Example 1: Isolation and Screening of Strains The isolated strain of Trichoderma (TCF09768) was isolated by microbial isolation in a farm with a nutrient agar (Nutrient agar N.A.). Briefly, first weigh 1 gram of soil and put it in lOc.c. sterile water. After shaking, take the filtrate on the water agar/aquaculture dish of semi-nutrient extracts to form a single colony. For each soil sample, take 5 culture dishes, transfer the plate-dished dishes to the %C dark incubator for 2 days, pick a single colony and transfer to the nutrient extract (Nutrient agar NA·). The growth morphology of each colony was observed after three days of culture on the plate medium. The colony was picked from the colony of the colony of the Phytophthora, and was subjected to secondary detachment. A single colony was taken and placed in a spiral tube containing 5 cc of sterile water. After the isolated strains were cultured for 7 days in potato dextrose agar (PDA), the colonies on the culture dishes were washed with 5 cc of sterile water and placed in a test tube containing 1 gram of rice culture. (Cultivate under TC' to observe the growth status of the strain and its sporulation in order to pick up the strain of rice blast medium and sporulation at 4 ° C. Example 2: Growth condition test and strain characteristics The isolate strain TCF09768 (97ID104) was identified for strain characteristics and strain name. It was cultured on a medium containing sodium naphthaleneacetic acid (SNA) (20 ° C). The colonies cultured for 4 days have the following characteristics: The colony is directly controlled to be about 90 mm; the hyphae are very sparse white; the spores are many, and the spores are grayish green to dark green (grayish green-dark green '26D5-26F5); sporulation Constructed as a cluster of pustulate and hemispherical. The results of observing the microstructurals under the microscope show that the colony's conidiophores produce short lateral branches. The secondary branches at the base are rarely re-branched, and the branches are mostly produced along the main branch in pairs or in pairs and at an angle of nearly 90 degrees to the main branch. The form of the conidiogenous cell (phialides) is phialide and is ampulliform, slightly enlarged in the middle, and the length and width are about 7.4-9.4χ2.0-3_2μηι, most of which are 2-3 rounds ( Whorls), the main branch has a phialide at the top, which is relatively slender in shape, about 8 m 12.47-13.38><2·08-2.50μηι. The split running is dark green when it is mature. (spherical), near-spherical (subSphericaI) or oval (ovoidal) ((2.8-)3.5-3.8(-4.0) X (2.8-)3.0-3.5 μηι, refers to the transverse diameter of non-spores X longitudinal diameter is 3.5 -3.8x3.0-3.5 ; (2.8-) (-4.0) is the range of the transverse diameter difference), the outer wall is spinulose, and its ornamentation (also known as exterior decoration) is sometimes difficult to observe. The identification results showed that the Trichoderma isolate TCF09768 was Samuels, Lieckfeldt &
Nirenberg。將此木黴菌菌株TCF09768於民國97 年9月24日寄存於中華民國食品工業發展研究所 菌種中心，其寄存編號為：BCRC 930113。Nirenberg. The Trichoderma strain TCF09768 was deposited in the Center of Fungi of the Republic of China Food Industry Development Research Institute on September 24, 1997. The registration number is BCRC 930113.
實施例三：本發明栽培介質之效率評估I 欲了解添加本發明之木黴菌（TCF09768)和舊 栽培介質與否對於介質材料之堆積發酵的效率評 估，首先設計不同之介質材料的配方如下表一： 表一：實驗設計（表中「+」表示有添加；「-」表示無添加） 組別 ABC D 新木屑（相思木） + + + + 舊栽培介質 + 木黴菌懸浮液(TCF09768) + 其中前述新木屑及/或前述舊栽培介質組成本 實施例之介質材料的基質：A組和B組的基質完全 使用新木屑丨C組和D組的基質中，新木屑和舊栽 培介質各占整體基質的75 wt%和25 wt%。 依據前述表一的設計，製作前述A〜D組栽培 介質的步驟為··取得未使用的相思木木屑及杏鲍菇 类負舊栽培介質’將未使用的相思木木屑及杏鲍菇類 舊^培介質依據表一混合成介質材料的基質，於本 實施例中9，該基質總重量為80公噸。接著取菌數 約lxl09cfu/ml的木黴菌菌株（TCF〇9768)加水稀 釋7 1〇〇倍以製備本發明之木黴菌懸浮液（1 χ 二cfu/nU)。然後以每lm3之前述基質：2。公升之 木，㈣浮稀釋液之比例，將前述木黴菌懸浮 ，入=34基質巾’朗整整體水分含量調整至60 ^，以氯成本實施例A〜D組的介質材料，苴中α 加前述木徽菌懸浮液’僅加“替以 5周整削4水分含量。 、曰 最後將前述介質材料堆積為高 ί以積發酵。於堆積發酵 酵完m堆材料一次’直到堆肥腐熟（堆積發 由於溫度可反應出介質材料 中有機物之分解階段和微生 Y的H 堆積發酵過程 堆積發酵過程，初期溫度急速C’予常進行的 後逐漸下降至周圍環境溫度並以上’然 機材料的腐熟與穩定。因此藉著慧疋，代表著有 酵過程中溫度對應發酵天數的變彳質材料於發 過程的進度。 可以窺知發酵 請參下表二，首先觀察A_D 期的天數·· A組於第45天到組別進入高溫Example 3: Evaluation of the efficiency of the cultivation medium of the present invention I For understanding the efficiency of the addition of the Trichoderma sp. (TCF09768) of the present invention and the old cultivation medium for the bulk fermentation of the medium material, firstly, the formulation of the different dielectric materials is as follows: : Table 1: Experimental design ("+" indicates that there is added; "-" indicates no addition) Group ABC D New sawdust (Acacia) + + + + Old cultivation medium + Trichoderma suspension (TCF09768) + The aforementioned new wood chips and/or the aforementioned old cultivation medium constitute the matrix of the medium material of the present embodiment: the matrix of the group A and the group B completely use the matrix of the new wood chips C group and the D group, and the new wood chips and the old cultivation medium each occupy the whole. 75 wt% and 25 wt% of the matrix. According to the design of the above Table 1, the steps of preparing the above-mentioned Group A to D cultivation medium are: obtaining unused Acacia wood chips and Pleurotus eryngii negative old cultivation medium 'The unused Acacia wood chips and Pleurotus eryngii The medium was mixed into a matrix of the dielectric material according to Table 1. In this example, the total weight of the substrate was 80 metric tons. Then, the Trichoderma strain (TCF〇9768) having a bacterial count of about lxl09 cfu/ml was diluted with water to a ratio of 7 1〇〇 to prepare a Trichoderma suspension of the present invention (1 χ 2 cfu/nU). Then with each of the aforementioned substrates per lm3: 2. The ratio of the liter wood, (4) floating diluent, the above-mentioned Trichoderma suspension, into the = 34 substrate towel 'rounding the whole moisture content is adjusted to 60 ^, the cost of chlorine in the dielectric materials of the groups A to D, 苴中α加The above-mentioned Mucor suspension is 'added only' to 5 times to cut 4 moisture content. 曰 Finally, the above-mentioned medium materials are piled up to be high-yield to ferment. After stacking the fermented yeast, the pile material is once until the compost is decomposed (stacked) The temperature can reflect the decomposition stage of the organic matter in the dielectric material and the H accumulation fermentation process of the micro-Y. The initial temperature rapid C' is gradually decreased to the ambient temperature and above. And it is stable. Therefore, by means of Huiyi, it represents the progress of the process of transforming the enamel material in the fermentation process with the temperature corresponding to the fermentation days. It can be seen that the fermentation is as follows in Table 2, first observe the number of days in the A_D period·· Group A On the 45th day, the group entered the high temperature.
運〆、最高溫60。(： ； B 組在第30天即達到60°C，而其最高溫可達62°C (第 45天）；C和D組的溫度則於第14天就已經分別到 達66°C和67t：，其最高溫分別為71°C和73°C(皆 在第30天）。 接著觀察各組降溫的趨勢：A組的溫度於第74 天將至46°C ; B組在第74天已降至更低的45°C ; C和D組的溫度則分別於第60天將至45°C和46 r。 表二：A〜D組之介質材料於堆積發酵過程中的溫度變化 (°C) 組別 第14天 第30天 第45天 第53天 第60天 第74天 A 53 58 60 57 52 46 B 55 60 62 58 51 45 C 66 71 62 55 45 43 D 67 73 63 53 46 42 總結來說，添加舊栽培基質及/或本發明之木黴 菌懸浮液的B、C和D組較僅含有新木屑的A組更 快進入高溫，也更快降至周圍環境溫度，尤其是添 加有木黴菌懸浮液和舊栽培基質的D組，是所有實 驗組別令，最快進入高溫也最快降至周圍環境溫度 的組別。特別的是，根據實驗數據可以發現，B、C 和D組的最南溫都比A組來得南（D組最南來到7 3 °C)，顯示加入木黴菌懸浮液及/或舊栽培基質增進 了介質材料的發酵反應。 維持生今及逮、止妈二田作月匕量來源，並需要氮素來 ==的?;中，碳素會代謝產生二氧化 機紐点八素則轉化合成微生物體内的有 會隨荖二籍获缺正體介質材料中的碳氮比值（c/n) 的過程，逐漸降低至某-穩定數 瘫1盔”因此A和B組之碳氮比值（C/Ν)的起始值 以丨：’而組之碳氮比值(⑽)的起始 的參下表三，比較第】4天的時候六和18兩組 ^致據，顯不Β組之碳氮比值（C/N)的下降幅度大 二土組。再根據第74天所得之數據，可知B組之 =氮比值(C/N)的下降幅度穩定的大於A組。再比 組和D組的數據，在第14天的時候，D組之 灭氮比值(C/N)的下降幅度大於c組，且D組在第 〇天時，碳氮比值(C/N)便來到68.1的數字，相較 於C組的碳氮比值（C/N)則在第％天才到達68 4。 並且，從D組第60天和第74天的數據可知，D& 的碳氮比值(C/N)已來到穩定值。 表二：A〜D組之介質材料於堆積發酵過程中的碳氮比值 變化（C/N)The highest temperature is 60. (: ; Group B reached 60 °C on the 30th day, and its highest temperature reached 62 °C (Day 45); the temperature of Groups C and D reached 66 °C and 67t on the 14th day, respectively. The highest temperatures were 71 ° C and 73 ° C (both on day 30). Then observe the trend of cooling in each group: the temperature of group A will reach 46 ° C on the 74th day; the group B will have fallen on the 74th day. To a lower 45 ° C; the temperature of the C and D groups will be 45 ° C and 46 r on the 60th day respectively. Table 2: Temperature changes of the dielectric materials of the groups A to D during the accumulation fermentation (°C ) Group 14 Day 30 Day 45 Day 53 Day 60 Day 74 Day A 53 58 60 57 52 46 B 55 60 62 58 51 45 C 66 71 62 55 45 43 D 67 73 63 53 46 42 Summary In contrast, Groups B, C, and D added with the old culture substrate and/or the Trichoderma suspension of the present invention enter the high temperature faster than the Group A containing only the new wood chips, and also fall to the ambient temperature more quickly, especially with the addition. Group D of Trichoderma suspension and old culture substrate is the group of all experimental groups, the fastest to enter the high temperature and the lowest to the ambient temperature. In particular, according to the experimental data, B, C and D can be found. The most southern temperature of the group was higher than that of the A group (the most south of the D group came to 7 3 °C), indicating that the addition of the Trichoderma suspension and/or the old cultivation substrate promoted the fermentation reaction of the medium material. Stop the mother's second crop as the source of the monthly sputum, and need nitrogen to ==?, in which carbon will be metabolized to produce a new machine, the hexaphorin will be transformed into a synthetic microbe, and there will be a deficiency in the body. The process of carbon-nitrogen ratio (c/n) in the dielectric material is gradually reduced to a certain - stable number 盔1 helmet. Therefore, the initial value of the carbon-nitrogen ratio (C/Ν) of groups A and B is 丨:' The initial value of the carbon-nitrogen ratio ((10)) is shown in Table 3 below. When comparing the 4th and 6th groups of the 4th day, the difference between the carbon and nitrogen ratio (C/N) of the significant group is the second largest. According to the data obtained on the 74th day, it can be seen that the decrease of the ratio of nitrogen/nitrogen ratio (C/N) in group B is more stable than that in group A. The data of group and group D are again, on the 14th day, D The nitrogen loss ratio (C/N) of the group decreased more than the group c, and in the third group, the carbon-nitrogen ratio (C/N) reached the number of 68.1, compared with the carbon-nitrogen ratio of the group C. (C/N) is in the first day of the genius Arrived at 68 4. Also, from the data of the 60th day and the 74th day of the D group, the carbon-nitrogen ratio (C/N) of D& has come to a stable value. Table 2: The dielectric materials of the groups A to D are deposited and fermented. Carbon-nitrogen ratio change in process (C/N)
A B C D 2別第第14天第30天第45天第53天第60天第74天 134 126 109 99.5 93.2 88.6 87.6 119 104 93.8 90.8 83.5 78.1 99.5 89.4 80.6 75.3 70.9 68.4 93.1 84.7 75.7 71.6 68.1 68.2 134 104 104 另’由於A和B組與C和D組的起始數值並 =相同，所以計算各組於各天數的碳氮比值（C/N) 後降幅度（該天數之碳氮比值/原碳氮比值， 所侍數宇越小表示下降幅度越大）呈現，請參下表 四· 表四· A〜D組之碳氮比值（C/N)的下降幅度（％)__ 」且另1第丨4天第30天第45天第53天第6。天第74天 A 94 03 84·34 74-25 69.55 66.12 65.37 B 8880 77 61 70 67.76 62.31 58.28 C 9567 85 96 77·5 72.40 68.17 65.77 ° 89 52 81 44 72 78 68.85 65.48 65.58 總法 來說，添加本發明之木黴菌懸浮液的B組 的碳氮比值（C/N)穩定地維持最大的下降幅度。d 組的碳II比值的下降幅度雖然不如B组 和表四卜第60天和帛74天的數據，可知-〇組： 率先進入穩定期0顯示本發明之木黴菌懸浮液有助 於發酵反應的進行，在同時添加舊栽培介質的情況 中’更達到最快速進入穩定期的實驗結果’不僅縮 短了發酵所需時間，也達到減少新木屑使用量的目 的。 最後，分析本實施例之各組栽培介質中的主要 化學特性，其結果係顯示於表五。 表五：A~D組栽培介質的主要化學特性 組別 氮(％) 磷(％) 鉀(％) 鈣(％) 鎂(％) 有機質（％) PH值 A 0.50 0.03 0.26 0.73 0.17 74.4 6.5 B 0.52 0.03 0.28 0.77 0.19 73.0 6.7 C 0.60 0.15 0.39 1.34 0.31 71.5 6.9 D 0.62 0.15 0.38 1.35 0.32 70.9 6.8 前述表五顯示A〜D組介質材料經60天的堆積 發酵後所完成的栽培介質中的主要化學特性。由表 中數據可知’添加有舊栽培介質的C和E)組較未 添加舊栽培介質的A和B組有較高的氮、碟、卸、 鈣、鎂含量和pH值。在相同材料配方下，有接種 木黴菌分離菌株(TCF09768)之B及D組之介質， 其主要化學特性與相同材料配方未接種木黴^之 A及C組則無明顯差異，顯示本發明之木黴菌僅加 速發酵反應的進行，並不影響生成之栽培介質 化學特性。 實施例四.本發明栽培介質之效率評估π 本實施例中主要欲了解舊栽培介質占基質之 變，於本發β明之介質材料之堆積發酵效率的影 二二並納入是否添加本發明之木黴菌菌株之變因。 首先設计不同之介質材料的配方如下表六： ·實U(表中「+」表示有添加；「_」表示無添加； 例為重量百分比例） 組別 _______ t F G Η 80% 80% 60% 60% 20% 20% 40% 40% Μ木屑（相思木）的比例 舊栽培介質的比例 木黴菌懸浮液(TCF09768) 1述Ε〜Η組栽培介質係依據前述表六的設計 相=施例三中所狀步驟製備。本實施例中ε〜Η 夺、土質皆包含相思木木屑和舊栽培介質，其中Ε 和G組未添加本發明之木黴菌 和、 則有添加本發日狀賴❿F#°H組 由於藉著觀察介質材料於發酵過程中溫 應發酵天數的變化可以窺知發酵過程的，^此 程中的溫度變化，並記錄如下表七。㈣^酵過 1378143 表七：E〜Η組之介質材料於堆積發酵過程中的溫度變化 (°C) 组別 第15天 第30天 第60天 第90天 Ε 60 68 52 43 F 65 72 48 4】 G 62 69 50 42 Η 68 73 47 40 由表七的數據可知，所有組別的溫度皆於第15 天就到達60°C以上。添加有本發明之木黴菌懸浮液 的F和Η組表現出較未添加前述木黴菌懸浮液之E 和G組更高的最高溫度（分別為72°C和73°C)，且 更早降至50°C以下（第60天分別為48°C和47°C)， 顯見添加本發明之木黴菌懸浮液有助於介質材料 的堆積發酵效率，此結果與實施例三所獲得的結果 一致。 • 至於比較分別含有20%和40%之舊栽培介質 的E和G組或F和Η組於溫度變化的趨勢並沒有 顯著的差別，顯示舊栽培介質的量並不影響整體介 質材料於堆積發酵的效率，因此根據本發明之配 方，在減少新木屑用量的考量下，可以部分舊栽培 介質代替新木屑的使用，而無損堆積發酵的效率。 同樣的，本實施例也藉由觀察介質材料中碳氮 比值（C/N)的變化趨勢來了解發酵反應的進行程 度。以本實施例而言，Ε和F組的碳氮比值（C/N) 的起始值應該為106，而G和Η的碳氮比值（C/N) 16 [ S1 1378143 的起始值應該為96.6。 請參下表八和表九，分別顯示E〜Η組之介質 材料於堆積發酵的過程中碳氮比值（C/N)的變化與 下降幅度（該天數之碳氮比值/原碳氮比值，所得數 字越小表示下降幅度越大）。 表八：Ε〜Η組之介質材料於堆積發酵過程中的碳氮比值 變化（C/N) 組別 第0天 第15天 第30天 第60天 第90天 Ε 106 98.2 94.6 79.9 75.9 F 106 97.5 90.5 75.0 73.5 G 96.6 91.3 84.0 70.9 69.7 Η 96.6 90.6 80.5 67.5 64.7 表九：A-D組之碳氮比值（C/N)的下降幅度（％) 組另U 第14天 第30天 第60天 第90天 Ε 92.64 89.25 75.38 .71.60 F 91.98 85.38 70.75 69.34 G 94.51 86.96 73.40 72.15 Η 93.79 83.33 69.88 66.98ABCD 2 Bid Day 14 Day 30 Day 45 Day 53 Day 60 Day 74 134 126 109 99.5 93.2 88.6 87.6 119 104 93.8 90.8 83.5 78.1 99.5 89.4 80.6 75.3 70.9 68.4 93.1 84.7 75.7 71.6 68.1 68.2 134 104 104 In addition, since the starting values of Groups A and B are the same as those of Groups C and D, the carbon-nitrogen ratio (C/N) of each group is calculated. The carbon-nitrogen ratio of the number of days/carbon nitrogen The ratio, the smaller the number of servants, the greater the decrease, the presentation, please refer to the following table. Table 4· The decrease of the carbon-nitrogen ratio (C/N) of the groups A to D (%) __ ” and the other 1丨 4th day 30th day 45th day 53rd day 6th. Day 74 Day A 94 03 84·34 74-25 69.55 66.12 65.37 B 8880 77 61 70 67.76 62.31 58.28 C 9567 85 96 77·5 72.40 68.17 65.77 ° 89 52 81 44 72 78 68.85 65.48 65.58 In general, add The carbon-nitrogen ratio (C/N) of Group B of the Trichoderma suspension of the present invention stably maintained the maximum decrease. Although the decrease of the carbon II ratio in group d is not as good as the data on the 60th day and the 74th day of group B and table IV, it can be seen that the group - the first to enter the stable period 0 shows that the Trichoderma suspension of the present invention contributes to the fermentation reaction. In the case of adding the old cultivation medium at the same time, the 'experimental result of the fastest entering the stable period' not only shortens the time required for fermentation, but also achieves the purpose of reducing the amount of new wood chips used. Finally, the main chemical properties in each of the cultivation media of the present examples were analyzed, and the results are shown in Table 5. Table 5: Main chemical characteristics of culture medium in group A~D Nitrogen (%) Phosphorus (%) Potassium (%) Calcium (%) Magnesium (%) Organic matter (%) PH value A 0.50 0.03 0.26 0.73 0.17 74.4 6.5 B 0.52 0.03 0.28 0.77 0.19 73.0 6.7 C 0.60 0.15 0.39 1.34 0.31 71.5 6.9 D 0.62 0.15 0.38 1.35 0.32 70.9 6.8 The above Table 5 shows the main chemical characteristics of the culture medium completed after 60 days of stacking fermentation of the A~D group of dielectric materials. . It can be seen from the data in the table that the groups C and E added with the old cultivation medium have higher nitrogen, dish, unloading, calcium, magnesium content and pH value than the groups A and B without the old cultivation medium. Under the same material formulation, there were mediums in groups B and D inoculated with Trichoderma isolates (TCF09768). The main chemical characteristics were not significantly different from the A and C groups in which the same material formulation was not inoculated with Trichoderma, indicating that the present invention Trichoderma only accelerates the fermentation reaction and does not affect the chemical properties of the culture medium produced. Example 4: Evaluation of the efficiency of the cultivation medium of the present invention π In this embodiment, it is mainly to understand that the old cultivation medium accounts for the change of the matrix, and the accumulation fermentation efficiency of the medium material of the present invention is included in the addition of the wood of the present invention. The cause of the mold strain. The formula for designing different dielectric materials is as follows: Table 6: • Real U (“+” in the table indicates addition; “_” indicates no addition; example is percentage by weight) Group _______ t FG Η 80% 80% 60% 60% 20% 20% 40% 40% proportion of eucalyptus (Acacia) proportion of old cultivation medium Trichoderma suspension (TCF09768) 1 Ε Η Η group culture medium according to the design phase of the above table 6 = Prepared in the procedure described in Example 3. In the present embodiment, ε~Η, soil contains Acacia wood chips and old cultivation medium, wherein the Ε and G groups are not added with the Trichoderma of the present invention, and the addition of the hair of the Japanese-style Lai ❿F#°H group Observing the change of the temperature of the medium in the fermentation process during the fermentation process can be used to understand the temperature change in the fermentation process, and record the following Table 7. (4) Fermentation 1378143 Table 7: Temperature change of the dielectric material of the E~Η group during the accumulation fermentation (°C) Group 15 Day 30 Day 60 Day 90 Day Ε 60 68 52 43 F 65 72 48 4] G 62 69 50 42 Η 68 73 47 40 According to the data in Table 7, the temperature of all groups reached above 60 °C on the 15th day. The F and hydrazine groups to which the Trichoderma suspension of the present invention was added exhibited higher maximum temperatures (72 ° C and 73 ° C, respectively) than the E and G groups to which the aforementioned Trichoderma suspension was not added, and dropped earlier. Up to 50 ° C or less (48 ° C and 47 ° C on the 60th day), it is obvious that the addition of the Trichoderma suspension of the present invention contributes to the stacking fermentation efficiency of the dielectric material, and the result is consistent with the results obtained in the third embodiment. . • There is no significant difference in the trend of temperature change between the E and G groups or the F and Η groups, which contain 20% and 40% of the old cultivation medium, respectively. It shows that the amount of the old cultivation medium does not affect the overall medium material in the accumulation fermentation. The efficiency, therefore, according to the formulation of the present invention, in consideration of reducing the amount of new wood chips, some old cultivation medium can be used instead of the new wood chips without compromising the efficiency of the accumulation fermentation. Similarly, this embodiment also understands the progress of the fermentation reaction by observing the change trend of the carbon-nitrogen ratio (C/N) in the dielectric material. For the present example, the starting value of the carbon-nitrogen ratio (C/N) of the lanthanum and F groups should be 106, and the carbon-nitrogen ratio (C/N) of G and Η 16 [the starting value of S1 1378143 should be It is 96.6. Please refer to Table 8 and Table 9 below to show the change and decrease of carbon-nitrogen ratio (C/N) of the dielectric materials of E~Η group during the process of stacking fermentation (the ratio of carbon to nitrogen/the ratio of carbon to nitrogen of the days, The smaller the number obtained, the greater the decrease. Table 8: Carbon-nitrogen ratio change of the dielectric material of the Ε~Η group during the accumulation fermentation (C/N) Group 0 Day 15 Day 30 Day 60 Day 90 Day Ε 106 98.2 94.6 79.9 75.9 F 106 97.5 90.5 75.0 73.5 G 96.6 91.3 84.0 70.9 69.7 Η 96.6 90.6 80.5 67.5 64.7 Table 9: Decrease in the carbon-nitrogen ratio (C/N) of the AD group (%) Group U Day 14 Day 30 Day 60 Day 90 Scorpio 92.64 89.25 75.38 .71.60 F 91.98 85.38 70.75 69.34 G 94.51 86.96 73.40 72.15 Η 93.79 83.33 69.88 66.98
由上述表八和表九之數據可知，添加有本發明 之木黴菌懸浮液的F和Η組的碳氮比值(C/N)皆穩 定地低於其分別之未添加前述木黴菌懸浮液的組 別（Ε和G組）。顯示本發明之木黴菌懸浮液有助於 [S] 17 1378143 =質材料發酵反應的進行，此結果於實施例三_ ^照比較含有2G%和4G%之舊栽培介質的£ 和·，且或F和Η組，則顯示在此比例 之 數據可知’除了 E ·组以外，所有組別的碳氮比^ (C/N)皆來到75以下，意味著已到達分解穩定階段。 最後，分析本實施例之各組栽培介質 化學特性，其結果係顯示於表十。 表十.E〜Η組栽培介質的主要化學特性 )鎂（％) 有機質（％) pH值 0.18 76.5 6.60 0.17 74.4 6.66 0.20 72.7 6.72 0.24 72.6 6.81 材料經 60天的堆積發酵From the data of Tables 8 and 9 above, the ratio of carbon to nitrogen (C/N) of the F and lanthanum groups to which the Trichoderma suspension of the present invention is added is stably lower than that of the above-mentioned Trichoderma suspension. Groups (Ε and G groups). It is shown that the Trichoderma suspension of the present invention contributes to the progress of [S] 17 1378143 = fermentation reaction of the plastid material, and the result is compared with £ and · of the old cultivation medium containing 2G% and 4G% in the third embodiment. Or F and Η groups, the data shown in this ratio shows that except for the E · group, the carbon-nitrogen ratio (C/N) of all groups comes below 75, which means that the decomposition stabilization stage has been reached. Finally, the chemical properties of the cultivation medium of each group of this example were analyzed, and the results are shown in Table 10. Table 10. Main chemical characteristics of the culture medium of E~Η group) Magnesium (%) Organic matter (%) pH 0.18 76.5 6.60 0.17 74.4 6.66 0.20 72.7 6.72 0.24 72.6 6.81 Material 60-day accumulation fermentation
F 0-59 0.16 0.36 1. G °·61 019 〇.4〇 1.28 0.20 Η 0.64 0.21 0.43 1.28 〇.24 〜丨貝丁叼土罟化學特性。由表中數 據可知，添加本發明之木黴菌懸浮液並不影響生 之栽培介質中的化學特性，此結果與實施三一 致〇 一 若比對比較含有20%和40%之舊栽培介質的Ε =G組或F和Η組，則顯示含有4〇%之舊栽培介 質的F，Η組較含有2G%之舊栽培介質的£和g 、、且有較高的氮、磷、鉀、鈣、鎂含量和pH值，但 [S]F 0-59 0.16 0.36 1. G °·61 019 〇.4〇 1.28 0.20 Η 0.64 0.21 0.43 1.28 〇.24 丨 丨 叼 叼 叼 罟 罟 罟 罟. It can be seen from the data in the table that the addition of the Trichoderma suspension of the present invention does not affect the chemical properties in the growing cultivation medium, and the result is consistent with the implementation of the third, compared with the old cultivation medium containing 20% and 40%. Ε=G group or F and Η group, it shows F with 4〇% old cultivation medium, Η group has more than 2G% of old cultivation medium, £ and g, and has higher nitrogen, phosphorus, potassium, Calcium, magnesium content and pH, but [S]