1306448 玖、發明說明： 【發明所屬之技術領域】 本發明係關於一種稻榖培養基及其製備微生物之方法係以 未去殼之水稻榖粒加水後經滅菌製得。利用稻榖培養基所製備之 菌種，可添加到堆肥中，促進堆肥發酵，縮短堆肥製程所費時間， 並可製成品質優良的有機質肥料，促進田間農作物栽培之使用效 益。 【先前技術】 一般農業廢棄物均兼具污染性及資源性，如妥為處理，將能 鲁轉化為農業生產系統中的養分源（氣、磷、軒）及能量源（碳）。 因此將農業廢棄物回歸於農田，不僅合乎資源再利用的自然法 則，而且也是現今消納有機廢棄物之重要方向之一。然而施用未 腐熟的有機物，容易造成土壤過度還原性及釋出毒性物質等問 題。因此有機廢棄物需經過適當的堆肥化處理以除去不良有機成 分及毒性物質等限制作物生長的因子。所謂堆肥化作用即利用廣 泛分佈於自然界之微生物，在控制的條件下，將廢棄物中不穩定 的有機成份加以分解，轉換為安定的腐植質成份，即腐熟的堆 Φ 肥。在堆肥化過程中，有機物基質中所含碳水化合物會迅速被微 生物作用而分解，同時微生物之增殖必須吸收氮、磷等營養成份 以合成微生物體質（biomass)，所以堆肥化前有機物基質中應含 有豐富的營養要素成份，並需將堆肥化前有機物基質中各種成份 調整至較適宜比例範圍内，以利於微生物進行堆肥化作用。 施用新鮮或未腐熟之有機物於土壤可能引起不良後果，如土 壤缺氮 '產生有機酸或土壤還原性阻害作物生長、傳播病原菌、 雜草種子等。因此，製作堆肥之堆肥化過程；讓有機物充分腐熟， 使它轉變為安全、穩定之高品質有機質肥料，相當重要。一般堆 肥腐熟之目的包括有： 1306448 1.調整有機物腐熟度 一般新鮮或未腐熟之有機物，其有機組成分相當不穩定，在 堆肥化過程中’有機廢棄物中碳氮比是—個非常重要的因素，堆 肥中微生物1要碳素當作生活能源，同時也需氮素來維持生命及 建以體細胞。當堆肥材料碳氮比太高時，會因氮素缺乏，致使微 生物無法大量繁瘦，堆肥化過程進行相當緩慢。如果碳氣比太 低’微生物分解出過多之氨’而易從堆肥中逸散，導致氮素損失。 堆積過程進行時’有機廢棄物中之碳氮比逐漸減少至20:1左右。 _ 2.減少有害成分 有機物77解會產生甲院、氫氣、盼酸、有機酸等有毒物質， 對作物生長有害’若經過堆積腐熟分解可減少有害成分。 3. 避免有機物分解所產生的不良因子 -般有機物含有_貞、脂肪酸等成分，當微生物分解時會產 生高溫及局部的缺氧情形，微生物大量繁殖，產生土壤還原性， 不利作物根部生長’如事先堆積分解，則可避免直接施用時影響 作物生長。 4. 消滅病菌、蟲卵、雜草種子 1 有機材料如穀殼、蒿桿 '糞尿等均附著許多病原菌、蟲印及 雜草種子。堆肥如經適當_，溫度最高可達_以上大部分 的病原菌、蟲印及雜草種子在此種溫度下可被殺滅。 5. 改良物理性 终多有機材料如蒿桿、殘枝、樹皮等材質堅硬、纖維 經過堆積分解後會變成脆細、柔軟、利於撒布，與土壤混合均勾。 6. 減少臭味，維護田間衛生 經過適當堆積腐熟的堆肥會降低令人不快的臭味，於田間施 用也可減少蚊蠅茲生，維護環境衛生。 7 ·可調配堆肥成分、增進肥效 1306448 不同有機材料所含營養成分差異頗大，如經適當調配，製成 的堆肥成分能配合不同作物生長所需，且堆積腐熟後可增加有機 材料的有效性磷與氮素’可增進作物產量及品質。 在作物永續栽培過程中使用生物製劑為現今栽培的新趨勢， 其生物製劑使用的種類包含有幾大項，（1)促進植物生長，如菌根 菌、固氮菌、根圈有益微生物群。（2)病蟲害生物防治，如蘇力菌、 木黴菌、枯草桿菌。（3)改善作物生長環境，如溶磷菌。其功用主 要在促進植物生長、增加產量、減少病蟲害，其它功能尚包含有 產生植物贺爾蒙、誘發植物抗病反應、降低土壤酸化、減低土壤 鹽類累積、誘使其它有益微生物產生，這些生物製劑近年來已有 相當多的研究人員投入。生物製劑的定義如以廣泛的生物種類區 分’則其除包含作物本身外’尚包含與作物發生反應之各種生 物。如僅以微生物的製劑而言，則生物製劑只包含微生物體或其 相關代謝產物。微生物製劑使用在作物生長上的效益可粗分為直 接利益及間接利益二大類。直接利益微生物包括與亞科共生根瘤 菌（Rhizobium)，與森林作物共生之放線菌（Frankia)，促進植物生 長之固氣細囷，囊叢枝勤根囷（vesicualar-arbuscular mycorrhizal fungi)及外生菌根菌（ectomycorrhizal fungi)等五大類，皆以能直接 促進植物營養吸收方式促進生長；而間接利益微生物則包括對植 物病原菌、植物蟲害及雜草競爭之生物控制作用，所以是屬於降 低影響作物生長不良環境因子作用的間接機制。無論其所採取的 機制為何，這些微生物在實驗過程中均能顯現促進作物生長潛 能’能取代無機肥料及化學殺蟲劑，降低生產成本及減少環境污 染，並能確保作物產量。是以目前在各項農業資源遭受威脅的今 天，微生物肥料的研發可能是一條明智之途。一般農業廢棄物均 兼具污染性及資源性，如利用微生物妥為處理，將能轉化為農業 生產系統中的養分源（氮、磷、鉀）及能量源（碳）。因此將農業廢 1306448 棄物回歸于農田，不僅合乎資源再利用的自然法則，而且也是現 今消納如此龐大量有機廢棄物之重要方向之一。 微生物在堆肥化過程中，擔任有機物分解與堆肥穩定化之重 要角色。不同的堆積材料如能接種適當的微生物菌種，可以加速 堆肥醱酵。為達到最有效率之堆肥化作用，除了添加適當的微生 物菌種外，在堆積材料環境中，維持微生物最適宜之生長條件， 使微生物充分的活動與繁殖，亦能加強堆肥材料的發酵與分解。 一般堆肥化微生物菌種的繁殖，可以採用自然堆肥法，即依照堆 肥化條件，於木屑、豆粕等堆肥材料中，調整適當的材料大小、 水份含量及通氣性等，再進行堆積腐熟，使自然界中微生物滋生 於堆肥材料中，最後在腐熟堆肥中即含有許多分解有機物的微生 物菌種，故可做為下一批堆肥製作時之微生物菌種。但為了增進 堆肥材料發酵分解效率，針對不同有機物材料特性，施予適當的 微生物菌種，將是堆肥製作過程之重要步驟之一。其中有關於利 用微生物菌種的關鍵機制，應包括有篩選出適當的微生物菌種、 建立有效率的菌種培養繁殖方法與應用於堆肥材料中的接種方 法等。 【發明内容】 本發明係針對蔗渣木屑與稻殼等纖維性高之資材，調配出最 適宜的用量比例，並接種適當的有益微生物，以使蔗渣木屑及稻 殼在堆肥化過程中，能夠迅速充分發酵及腐熟。 本發明之目的係提供一種稻榖培養基，其係由未去殼之水稻 穀粒與水混合經滅菌程序製得，其中前述未去殼之水稻穀粒與水 之體積混合比例係為3:1至5:1。 前述未去殼之水稻穀粒與水之混合比例較佳係為體積比4:1。 前述滅菌程序係將水稻穀粒與水混合後置於高溫高壓滅菌。 本發明之另一目的係提供一種利用稻穀培養基製備微生物 1306448 之方法，係包含下❹驟：提供—欲培制種之孢子 2養菌種，孢子加入稻榖培養基中培養種菌；及種菌成：時： 並以稻縠培養基進行種菌量產程序。 ’’、· 别述欲培養菌種之孢子係可由各 離與_選獲得。 讀了由各種堆肥成》知微生物分 前述菌種係為木黴菌。 培養=培養種菌係將欲培養菌種之孢子製成菌懸液加入稻榖 前述培養種菌之時間為2〜3週。 刚述1產程序係以種菌+稻縠培養 培養法進行種菌之量產。 一明/口夾層 ^發明之又—目的係提供—種利用稻榖培養 含下列步驟:提供-木黴菌菌種之孢子;= 2成囷驗加人稻穀培養Μ培養木黴8之 =取出：並以稻毅培養基進行種菌量產程序，其中前 之量Γ種國+&穀培養基+種菌之三明治夾層培養法進行種菌 前述培養種菌之時間為2〜3週。 色抱^述種g成熟料木黴㈣絲長滿水稻穀粒後開始形成綠 物技明=再—目的係提供—種利用稻榖培養基製備之微生 方法，！將利用稻穀培養基量產完成含有菌種 料！立方公"尺:^體二口釋⑽倍成㈣…堆肥材 中。 。 A升之比例，將菌懸液混入堆肥材料 前述菌種係為木黴菌。 别述堆肥材料係為高纖維性之材料，例如稻殼堆肥或嚴渣木 1306448 屑堆肥。 前述稻殼堆肥的材料包括稻殼、米糠、棕櫚粕等以乾物重用 量比例約為6 : 2 : 2混和。 前述蔗渣木屑堆肥的材料包括蔗渣、太空包廢木屑、豬糞、 菜仔粕等以乾物重用量比例約為4 : 3 : 1 : 2混和。 前述菌稀釋液混入堆肥材料後需將水分含量調整至60%，堆 積高度維持約1.5-2公尺，爾後立即進行堆積製作。 本發明所提供之稻穀培養基價格低廉，其所培養出菌種利用 接種用量比例技術施用於生物性堆肥時，不僅具有高分解能力， 而且可以加速堆肥腐熟，減少堆肥化時程，並降低堆肥製作成本。 【實施方式】 本發明提供一種由未去殼之水稻穀粒與水混合經滅菌程序 製得之稻穀培養基，其中未去殼之水稻穀粒與水之體積混合比例 係為3:1至5:1，較佳係為4:1，滅菌程序係將水稻穀粒與水混合 後置於高溫高壓下滅菌。 本發明另提供一種利用稻穀培養基繁殖微生物之方法，如第 一圖所示，其步驟如下：首先提供一欲培養菌種之孢子，接著將 孢子加入稻穀培養基中培養種菌，等到種菌成熟時取出以稻穀培 養基及三明治培養法進行種菌量產程序。 前述欲培養菌種之孢子係可由各種堆肥成分進行微生物分 離與篩選獲得，以本發明為例所篩選獲得之菌種係為木黴菌，接 著將木黴菌之孢子製成菌懸液加入稻穀培養基中培養木黴菌種 菌之時間為2〜3週，當木黴菌菌絲長滿水稻穀粒後開始形成綠 色孢子即為木黴菌種菌成熟，接著將木黴菌種菌取出，以種菌+ 稻穀培養基+種菌之三明治夾層培養法進行木黴菌之量產可製 得孢子濃度lxlO9 spore/g之固體木黴菌。 本發明另提供一種利用稻榖培養基繁殖之固體木黴菌接種 1306448 堆肥材料的方法，如第二圖所示，係將利用稻穀培養基量產完成 含有lxl09spore/g之固體木黴菌種稀釋200倍成菌懸液，再以堆 肥材料1立方公尺與菌懸液20公升之比例，將菌懸液混入堆肥 材料中。堆肥材料可為稻殼、米糠、棕櫚粕等以乾物重用量比例 約為6 : 2 : 2混和之稻殼堆肥，所接種之菌種係為木黴菌 (TCT111)，或者以蔗渣、太空包廢木屑、豬糞、菜仔粕等以乾 物重用量比例約為4 : 3 : 1 : 2混和之蔗渣木屑堆肥，所接種之 菌種係為木黴菌（TCT103)。 當菌稀釋液混入堆肥材料後需將水分含量調整至60%，並使 ® 堆積高度維持約1.5-2公尺，爾後立即進行堆積製作。 以下實施例係用於進一步了解本發明之優點，並非用於限制 本發明之申請專利範圍。 實施例1·稻穀培養基之製備 取未去殼之水稻穀粒與水，以體積比4:1之比例混合後進行 滅菌程序，滅菌程序可於高壓滅菌釜中以120°C之高溫進行滅菌 40分鐘2次達成，製得之稻穀培養基可用於後續菌種分離、篩選 與培養。 φ 實施例2.利用稻穀培養基進行木黴菌培養繁殖 菌種分離與篩選： 由各種堆肥成份，如菇類太空包、廢木屑、稻穀、蔗渣中， 以2%水瓊脂（Water agar W.A.)洋茶平板法進行微生物分離，各資 材先切成0.5公分小段後取5小段置於WA中，每種類資材5個 培養皿，各處理於28°C培養箱中不照光培養三天後，挑取片段菌 絲尖端移到馬鈴薯葡萄糖培養基上，培養7天後觀察各菌絲生長 形態，由其中挑取似木黴菌菌絲形態之菌株，進行二次純系分離 後切取0.5平方公分之菌絲塊置於裝5毫升無菌水之螺旋試管中 保存。各分離菌株以PDA培養7天後，切取菌絲塊，置入裝200 11 1306448 克稻穀培養基之三角瓶中培養，觀察各菌株在其中生長情形，以 挑取能快速纏繞稻穀培養基之菌株。 木黴菌培養繁殖： 木黴菌（TWc/zoi/erma TCT103、TCT111)原菌種培養於馬铃 薯葡萄糖洋菜平板培養基，於培養七天後，以無菌水製成孢子懸 浮液1X1 〇3 spore/ml，加入實施例1所製備之稻穀培養基中培養兩 ~三週，於木黴菌菌絲長滿水稻榖粒後開使形成綠色孢子此即為 量產之種菌，種菌培養成熟時取出與消毒完全之稻榖培養基以三 明治夾層培養法量產接種菌種，三明治夾層培養法係以種菌+消 毒完全之稻穀+種菌方式培養，以此法進行接種源之後發酵，稻 穀培養基後發酵成固體菌種其孢子濃度可達lxl〇9 spore/g。 實施例3.生物性蔗渣木屑堆肥之製作 生物性蔗渣木屑堆肥係以蔗渣及廢木屑（例如太空包廢木 屑）為主要原料，配合豬糞、油粕類等次要材料。其中蔗渣氮含 量約0.62%、磷含量約0.11%、鉀含量約0.85%、鈣含量約0.54%、 鎮含量約0.36%、辞含量約18ppm、銅含量約3ppm。太空包廢木 屑氮含量約1.12%、磷含量約0.32%、鉀含量約1.11%、鈣含量約 0.42%、鎂含量約1.29%、鋅含量約73ppm、銅含量約8ppm。豬 糞氮含量約2.23%、磷含量約0.69%、鉀含量約1.18%、鈣含量約 0.41%、鎂含量約0.38%、辞含量約190ppm、銅含量約33ppm。 菜仔粕氮含量約4.14%、磷含量約1.16%、鉀含量約3.57%、鈣含 量約1.65%、鎂含量約1.21%、辞含量約28口卩111、銅含量約11^卩111。 經由以上材料化學成分含量與乾物重量相互換算，當蔗渣木 屑堆肥的材料包括蔗渣：太空包廢木屑：豬糞：菜仔粕等乾物重 用量比例依序為40%、30%、10%及20%，堆肥材料中碳氮比為 30。堆肥試作用量分別為蔗渣2,000公斤、太空包廢木屑1,500 公斤、豬糞500公斤、菜仔粕1，000公斤。取用適量含有約109 12 1306448 spore/g木黴菌（TCT103)之稻穀培養基，先加水稀釋200倍成 菌懸液，再以1立方公尺堆肥材料與菌懸液20公升之比率，將 菌稀釋液混入堆肥材料中，最後將堆肥材料水份含量調整至 60%，堆積高度維持約1.5-2.0公尺，爾後立即進行堆積製作，堆 肥化期間約5-7曰利用鏟裝機翻堆一次，一直持續到堆肥腐熟為 止，堆肥化期間溫度變化如表一，使用木黴菌堆肥初期高溫上升 較快且略高，且較快降至低溫期（<50°C )而漸趨穩定，所以約 比未使用木黴菌堆肥之對照提早3-6日腐熟。 天數 組別 溫度 0-5 6-10 11-15 16-20 21-25 26-30 31-35 36-45 生物性 蔗渣木屑堆肥 62 71 63 58 56 52 49 42 一般 蔗渣木屑堆肥 61 68 64 57 53 54 50 43 實施例4·生物性稻殼堆肥材料之製作 稻殼堆肥係以稻殼為主要原料，配合米糠、油粕類等次要材 料。由堆肥材料之養分含量分析結果顯示，稻殼氮含量約0.39%、 磷含量約0.05%、鉀含量約0.28%、鈣含量約0.07%、鎂含量約 0.04%、鋅含量約26ppm、銅含量約3ppm，其次米糠氮含量約 1.02%、構含量約0.34% '卸含量約1.52%、釣含量約0.38%、鎂 含量約1.21 %、辞含量約84ppm、銅含量約15ppm，棕搁粕氮含 量約2.38%、磷含量約0.37%、鉀含量約1.03%、鈣含量約0.42%、 鎮含量約0.78%、辞含量約23ppm、銅含量約8ppm。 經由以上材料化學成分含量與乾物重量相互換算，當稻殼堆 肥的材料包括稻殼：米糠：棕櫚粕等乾物重用量比例約為6 : 2 : 2，此堆肥材料中碳氮比約為30左右。堆肥試作用量分別為稻殼 13 13064481306448 发明Invention Description: [Technical Field] The present invention relates to a rice blast medium and a method for preparing the same, which are obtained by sterilizing water without unshelled rice granules. The strain prepared by the rice blast medium can be added to the compost to promote compost fermentation, shorten the time required for the composting process, and can be made into high quality organic fertilizer to promote the use of field crop cultivation. [Prior Art] General agricultural wastes are both polluting and resource-intensive. If properly disposed of, they can be converted into nutrient sources (gas, phosphorus, xenon) and energy sources (carbon) in agricultural production systems. Therefore, returning agricultural waste to farmland not only conforms to the natural laws of resource reuse, but is also one of the important directions for the elimination of organic waste. However, the application of unfertilized organic matter is liable to cause problems such as excessive reduction of soil and release of toxic substances. Therefore, organic waste needs to be properly composted to remove factors such as undesirable organic components and toxic substances that limit crop growth. The so-called composting effect is to use the microorganisms widely distributed in nature to decompose the unstable organic components in the waste under controlled conditions and convert them into stable humic components, that is, decomposed piles of Φ fertilizer. During the composting process, the carbohydrates contained in the organic matrix are rapidly decomposed by the action of microorganisms. At the same time, the proliferation of microorganisms must absorb nutrients such as nitrogen and phosphorus to synthesize micromass. Therefore, the organic matter matrix should be contained before composting. Rich in nutrient elements, and the various components in the organic matrix before composting should be adjusted to a suitable ratio to facilitate the composting of microorganisms. The application of fresh or unfermented organic matter to the soil may cause adverse consequences, such as soil nitrogen deficiency, which produces organic acids or soil reduction, which hinders crop growth, spreads pathogens, and weed seeds. Therefore, it is very important to make the composting process of composting; it is very important to make the organic matter fully fermented and turn it into a safe and stable high quality organic fertilizer. The general purpose of composting includes: 1306448 1. Adjusting the organic maturity of fresh or undecomposed organic matter, the organic composition is quite unstable. In the process of composting, the carbon-nitrogen ratio in organic waste is very important. Factors, microbes in composting require carbon as a source of energy, and also need nitrogen to sustain life and build somatic cells. When the carbon-nitrogen ratio of the compost material is too high, the nitrogen deficiency is caused, so that the microorganisms cannot be bulky, and the composting process is rather slow. If the carbon to gas ratio is too low, 'the microorganism decomposes too much ammonia' and easily escapes from the compost, resulting in nitrogen loss. When the accumulation process is carried out, the carbon to nitrogen ratio in organic waste is gradually reduced to about 20:1. _ 2. Reducing harmful ingredients The organic matter 77 will produce toxic substances such as a hospital, hydrogen, acid, organic acids, etc., which are harmful to crop growth. 3. Avoid the bad factors caused by the decomposition of organic matter - organic matter contains ingredients such as 贞, fatty acids, etc. When the microorganism decomposes, it will produce high temperature and local anoxic conditions. The microorganisms will multiply and produce soil reducing, which will be unfavorable for crop root growth. Accumulation and decomposition in advance can avoid affecting crop growth when applied directly. 4. Elimination of germs, eggs, and weed seeds 1 Organic materials such as chaff, artemisia, and feces have many pathogens, insect prints, and weed seeds. If the compost is properly _, the highest temperature, up to _, most of the pathogens, insect prints and weed seeds can be killed at this temperature. 5. Improved physical properties The final multi-organic materials such as artemisia, rods, bark and other materials are hard, and the fibers will become brittle, soft, and easy to spread after being piled up and decomposed. 6. Reduce odor and maintain field hygiene. Proper accumulation of decomposed compost will reduce unpleasant odors, and application in the field can also reduce mosquitoes and maintain environmental hygiene. 7 · Adjustable composting composition and improve fertilizer efficiency 1306448 Different organic materials contain different nutrients. If properly formulated, the compost composition can be matched with the growth of different crops, and the effectiveness of organic materials can be increased after stacking and decomposing. Phosphorus and nitrogen' can increase crop yield and quality. The use of biological agents in the continuous cultivation of crops is a new trend in today's cultivation. The types of biological agents used include several major items. (1) Promote plant growth, such as mycorrhizal fungi, nitrogen-fixing bacteria, and beneficial microflora of the root ring. (2) Biological control of pests and diseases, such as Suri, Trichoderma, and Bacillus subtilis. (3) Improve crop growth environment, such as phosphate solubilizing bacteria. Its function is mainly to promote plant growth, increase yield, reduce pests and diseases, and other functions include plant hormone production, induce plant disease resistance, reduce soil acidification, reduce soil salt accumulation, and induce other beneficial microorganisms. Formulations have been put into considerable research by researchers in recent years. The definition of a biological agent, if it is distinguished by a wide range of biological species, then contains various organisms that react with the crop, in addition to the crop itself. If only in the case of a microbial preparation, the biological preparation contains only the microorganism or its associated metabolite. The benefits of using microbial preparations in crop growth can be broadly divided into two categories: direct benefits and indirect benefits. Direct interest microbes include Rhizobium, a symbiotic actinomycete (Frankia) that promotes plant growth, and vesicualar-arbuscular mycorrhizal fungi and ectomycorrhizal fungi. The five major types of bacteria (ectomycorrhizal fungi) promote growth by directly promoting plant nutrient absorption; while indirect microbes include biological control of plant pathogens, plant pests and weed competition, so it is a reduction in adverse growth of crops. An indirect mechanism of the role of environmental factors. Regardless of the mechanism they are employed, these microorganisms can demonstrate the potential to promote crop growth during the experiment. They can replace inorganic fertilizers and chemical pesticides, reduce production costs and reduce environmental pollution, and ensure crop yield. With the current threat to various agricultural resources, the development of microbial fertilizers may be a wise way. General agricultural wastes are both polluting and resource-intensive. If properly treated with microorganisms, they can be converted into nutrient sources (nitrogen, phosphorus, potassium) and energy sources (carbon) in agricultural production systems. Therefore, the return of agricultural waste 1306448 to farmland not only conforms to the natural law of resource reuse, but also is one of the important directions for the elimination of such a large amount of organic waste. During the composting process, microorganisms play an important role in the decomposition of organic matter and the stabilization of compost. Different accumulation materials, such as the ability to inoculate appropriate microbial strains, can accelerate composting. In order to achieve the most efficient composting, in addition to the addition of appropriate microbial strains, in the environment of the stacked materials, maintaining the optimal growth conditions of the microorganisms, allowing the microorganisms to fully move and reproduce, can also enhance the fermentation and decomposition of the compost materials. . In general, the composting of microbial strains can be carried out by natural composting method, that is, according to the composting conditions, in the composting materials such as wood chips and soybean meal, the appropriate material size, moisture content and air permeability are adjusted, and then piled and decomposed. Microorganisms in nature are born in composting materials. Finally, in the composting compost, there are many microbial strains that decompose organic matter, so it can be used as the microbial strain in the next batch of composting. However, in order to improve the fermentation efficiency of composting materials, it is one of the important steps in the composting process to apply appropriate microbial strains for different organic material properties. Among them, there are key mechanisms for the use of microbial strains, including the selection of appropriate microbial strains, the establishment of efficient strain culture and propagation methods, and the inoculation methods used in composting materials. SUMMARY OF THE INVENTION The present invention relates to a material having high fiber content such as bagasse wood chips and rice hulls, and formulates an optimum dosage ratio, and inoculates appropriate beneficial microorganisms, so that bagasse wood chips and rice husks can be quickly formed during composting. Fully fermented and decomposed. The object of the present invention is to provide a rice bran culture medium which is prepared by mixing a non-hulled rice grain and water through a sterilization process, wherein the volume ratio of the unhulled rice grain to water is 3:1. To 5:1. The mixing ratio of the aforementioned unhulled rice grains to water is preferably 4:1 by volume. The aforementioned sterilization procedure is to mix rice grains with water and then to be autoclaved. Another object of the present invention is to provide a method for preparing microorganism 1306448 by using a rice medium, comprising the steps of: providing a spore-producing strain of the seed to be cultured, adding the spore to the rice bran medium for cultivating the inoculum; and growing the inoculum: Time: The inoculum mass production procedure is carried out in rice blast medium. The spore line of the species to be cultured may be obtained by separate and _ selection. Read the various microorganisms into the "microorganisms", the aforementioned strain is Trichoderma. Culture = culture of the inoculum The suspension of the spores to be cultured is added to the rice blast. The time for cultivating the above-mentioned inoculum is 2 to 3 weeks. Just mentioned the 1 production procedure is the mass production of the inoculum by the inoculum + rice bran culture method. A Ming / mouth sandwich layer ^ Invented again - the purpose is to provide - the use of rice cultivating culture with the following steps: provide - spores of Trichoderma species; = 2 into the test of the addition of rice cultivation Μ culture of Trichoderma 8 = Take out: The inoculum mass production procedure is carried out using the rice cultivar medium, wherein the pre-measuring amount of the cultivar + & gluten medium + the inoculum sandwich sandwich culture method is carried out for the cultivating the inoculum for 2 to 3 weeks. Color hug ^ described species of mature plant Trichoderma (four) silk overgrown with rice grains began to form green matter skill = re-purpose to provide a species of micro-methods prepared using rice blast medium,! The rice seed culture medium will be used to mass-produce the bacteria-containing seeds! Cubic public " ruler: ^ body two mouth release (10) times into (four) ... compost material. . The proportion of A is increased, and the bacterial suspension is mixed into the compost material. The aforementioned strain is Trichoderma. The composting materials are high-fibrous materials, such as rice husk compost or slag wood 1306448. The above-mentioned rice husk composting materials include rice husks, rice bran, palm sorghum, etc., which are mixed in a dry matter ratio of about 6:2:2. The materials of the aforementioned bagasse wood chip compost include bagasse, space waste wood chips, pig manure, rapeseed meal, etc., and the dry matter weight ratio is about 4:3:1:2. After the above-mentioned bacterial dilution liquid is mixed into the composting material, the moisture content is adjusted to 60%, the stacking height is maintained at about 1.5-2 meters, and the stacking is immediately performed. The rice medium provided by the invention is low in price, and the cultured strain is applied to the biological compost by the technique of inoculation ratio, which not only has high decomposition ability, but also accelerates compost maturity, reduces composting time course, and reduces compost production. cost. [Embodiment] The present invention provides a rice culture medium prepared by mixing a non-shelled rice grain and water through a sterilization process, wherein the volume ratio of the unhulled rice grain to the water is 3:1 to 5: 1, preferably 4:1, the sterilization process is to mix the rice grains with water and then sterilize under high temperature and high pressure. The invention further provides a method for propagating microorganisms by using rice medium, as shown in the first figure, the steps are as follows: firstly, a spore of a strain to be cultured is provided, then the spore is added to the rice medium to culture the inoculum, and when the inoculum matures, the extract is taken out. The rice culture medium and the sandwich culture method are used for the inoculum production process. The spores of the above-mentioned cultured species can be obtained by microbial separation and screening of various composting components. The strain obtained by the present invention is a Trichoderma strain, and then the spores of Trichoderma sp. are added to the rice culture medium. The time for cultivating the Trichoderma strain is 2 to 3 weeks. When the Trichoderma mycelium grows over the rice grain, the green spore begins to form, and the Trichoderma species is matured. Then the Trichoderma strain is taken out, and the inoculum + rice medium + inoculum sandwich is taken. The Trichoderma spp. can produce a solid Trichoderma spore with a spore concentration of lxlO9 spore/g. The invention further provides a method for inoculating 1306448 compost material by using solid Trichoderma sp. which is propagated by rice blast culture medium. As shown in the second figure, the solid Trichoderma species containing lxl09spore/g is diluted by the rice medium to produce 200 times of bacteria. The suspension is mixed with the bacterial suspension into the compost material at a ratio of 1 cubic meter of compost material to 20 liters of the bacterial suspension. The compost material may be rice husk, rice bran, palm sorghum, etc., which is about 6:2:2 mixed rice husk compost, and the inoculated strain is Trichoderma (TCT111), or waste in bagasse and space. Wood chips, pig manure, rapeseed meal, etc. are about 4:3 : 1 : 2 mixed bagasse wood chips composted with the dry matter weight ratio, and the inoculated strain is Trichoderma (TCT103). When the bacteria dilution is mixed into the compost material, the moisture content should be adjusted to 60%, and the stack height should be maintained at about 1.5-2 meters, and then stacked immediately. The following examples are intended to further understand the advantages of the present invention and are not intended to limit the scope of the invention. Example 1 Preparation of Rice Medium The un-shelled rice grains and water were mixed at a volume ratio of 4:1 and then subjected to a sterilization procedure, and the sterilization procedure was carried out in an autoclave at a high temperature of 120 ° C. The rice culture medium can be used for the separation, screening and culture of the subsequent strains. φ Example 2. Separation and screening of Trichoderma culture and propagation strains using rice culture medium: 2% water agar (Water agar WA) tea from various composting ingredients such as mushroom space bag, waste wood chips, rice, bagasse The plate method was used for microbial separation. Each material was first cut into 0.5 cm segments and then placed in WA for 5 small sections. Each culture material was 5 culture dishes. Each treatment was carried out in a 28 °C incubator for 3 days without light culture. The mycelial tip was moved to the potato dextrose medium, and the growth morphology of each mycelium was observed after 7 days of culture. The strains of the mycelium-like mycelium were picked and subjected to secondary pure separation, and then 0.5 square centimeter of hyphae was cut and placed. Store in 5 ml sterile water in a spiral tube. After the isolated strains were cultured for 7 days in PDA, the mycelial masses were excised, placed in a triangular flask containing 200 11 1306448 g of rice medium, and the growth of each strain was observed to pick up a strain capable of rapidly winding the rice medium. Trichoderma culture and propagation: The original strain of Trichoderma (TWc/zoi/erma TCT103, TCT111) was cultured in potato dextrose plate culture medium. After 7 days of culture, spore suspension 1X1 〇3 spore/ml was made with sterile water. Adding to the rice medium prepared in Example 1 for two to three weeks, after the mycelium of the Trichoderma is overgrown with rice, the green spores are formed, which is a mass-produced inoculum, and the inoculum is completely taken out and disinfected when the culture is matured. The rice bran culture medium is produced by sandwich culture method, and the sandwich sandwich culture method is cultured by inoculum + disinfected rice + inoculum. After the inoculation source is fermented by this method, the rice medium is fermented into solid strains and spores. Concentration up to lxl〇9 spore/g. Example 3. Preparation of Biological Bagasse Saw Compost The biological bagasse wood chip composting is mainly composed of bagasse and waste wood chips (such as space debris), and is matched with secondary materials such as pig manure and oil mites. The bagasse has a nitrogen content of about 0.62%, a phosphorus content of about 0.11%, a potassium content of about 0.85%, a calcium content of about 0.54%, a town content of about 0.36%, a rhodium content of about 18 ppm, and a copper content of about 3 ppm. The space waste wood has a nitrogen content of about 1.12%, a phosphorus content of about 0.32%, a potassium content of about 1.11%, a calcium content of about 0.42%, a magnesium content of about 1.29%, a zinc content of about 73 ppm, and a copper content of about 8 ppm. The pig has a fecal nitrogen content of about 2.23%, a phosphorus content of about 0.69%, a potassium content of about 1.18%, a calcium content of about 0.41%, a magnesium content of about 0.38%, a rhodium content of about 190 ppm, and a copper content of about 33 ppm. The cultivar has a nitrogen content of about 4.14%, a phosphorus content of about 1.16%, a potassium content of about 3.57%, a calcium content of about 1.65%, a magnesium content of about 1.21%, a reciprocal content of about 28 卩111, and a copper content of about 11^卩111. Through the conversion of the chemical composition content of the above materials and the dry matter weight, when the bagasse wood chip composting material includes bagasse: space bag waste wood chips: pig manure: rapeseed meal and other dry matter weight ratios are 40%, 30%, 10% and 20, respectively. %, the carbon to nitrogen ratio in the compost material is 30. The amount of composting test was 2,000 kg of bagasse, 1,500 kg of waste wood chips, 500 kg of pig manure, and 1,000 kg of rapeseed meal. Take an appropriate amount of rice medium containing about 109 12 1306448 spore/g Trichoderma (TCT103), first dilute with water to make a bacterial suspension 200 times, and then dilute the bacteria with a ratio of 1 cubic meter of compost material to 20 liters of bacterial suspension. The liquid is mixed into the compost material, and finally the moisture content of the compost material is adjusted to 60%, the stacking height is maintained at about 1.5-2.0 meters, and the stacking is immediately performed. During the composting period, about 5-7 翻 is piled up by the shovel loader. Continue until the compost is ripe, the temperature change during composting is shown in Table 1. The high temperature rise in the initial stage of Trichoderma composting is faster and slightly higher, and it is gradually lowered to the low temperature period (<50 °C) and gradually stabilizes. Controls that did not use Trichoderma compost were degraded 3-6 days earlier. Day array temperature 0-5 6-10 11-15 16-20 21-25 26-30 31-35 36-45 Biological bagasse wood chip composting 62 71 63 58 56 52 49 42 General bagasse wood chip composting 61 68 64 57 53 54 50 43 Example 4 Preparation of Biological Rice Hull Composting Material The rice husk composting system uses rice husk as the main raw material and is matched with secondary materials such as rice bran and oil glutinous rice. The nutrient content analysis results of the compost material showed that the rice shell nitrogen content was about 0.39%, the phosphorus content was about 0.05%, the potassium content was about 0.28%, the calcium content was about 0.07%, the magnesium content was about 0.04%, the zinc content was about 26 ppm, and the copper content was about 3ppm, followed by rice nitrogen content of about 1.02%, structure content of about 0.34% 'unloading content of about 1.52%, fishing content of about 0.38%, magnesium content of about 1.21%, content of about 84ppm, copper content of about 15ppm, brown shelf nitrogen content of about 3ppm 2.38%, phosphorus content of about 0.37%, potassium content of about 1.03%, calcium content of about 0.42%, town content of about 0.78%, content of about 23 ppm, and copper content of about 8 ppm. Through the conversion of the chemical composition content of the above materials and the dry matter weight, when the rice husk composting material includes the rice husk: rice bran: palm sorghum and the like, the dry matter weight ratio is about 6:2:2, and the carbon-nitrogen ratio in the composting material is about 30 or so. . The amount of composting test is rice husk 13 1306448
6，_9公斤、米糠2，_公斤、掠櫚相2，咖公斤。取用適量含有 約10 spore/g木黴菌（TCT103)之稻穀培養基，先加水稀釋200 倍減懸液’再m立方公尺堆肥材料與菌懸液2G公升之比率， 將囷稀釋液混入堆肥材料中，最接收A -.,5i〇 肥化期間約5-7日利用鏟裝機翻堆—次，一直堆積製作，堆 止，堆肥化期間溫度變化如表二’使用木黴菌堆= = 較快且略高，錢快降至低溫期（<5代）上升 比未使用木黴菌堆肥之對照提早3-6日腐熟。 ^ 所以約 表 天數 組別 溫度 0-5 6-10 11-15 16-20 21-25 26- 生物性 稻殼堆肥 62 70 66 57 54 5 一般稻殼堆肥 60 67 65 55 56 5( 實施例5.生物性蔗渣木屑堆肥之效益 φ 生物性蔗渣木屑堆肥之養分含量分析結果如表三， 、 ~~矛彳用本發 明稻榖培養基培養之木黴菌（TCT103)接種之嚴逢木屑堆肥直 氮含量約2.13%、磷含量約0.98%、鉀含量約！ 8ΐς7 Α ’、 、鈣含量約 1.03%、鎂含量約0.78%、有機質含量約65.5%'鋅令晷认 〒3重約98ppm、 銅含量約27ppm。未接種木黴菌之蔗渣木屑堆人旦 氧θ 約 1.87%、磷含量約0.85%、鉀含量約1.74%、鈣含量約〇 92^ , 含量約0.67%、有機質含量約63.1%、鋅含量約9〇ppm、銅八旦 約26ppm。顯然有接種木黴菌之蔗渣木屑堆肥的氮、碟、卸里 錢、鋅及銅等含量均略高於一般蔗渣木屑堆肥。 14 1306448 表三 元素 N P K Ca Mg OM Zn Cu 組別 (%) (mg/kg) 生物性 蔗渣木屑堆肥 2.13 0.98 1.81 1.03 0.78 65.5 98 27 一般 蔗渣木屑堆肥 1.87 0.85 1.74 0.92 0.67 63.1 90 26 實施例6.生物性稻殼堆肥之效益 生物性稻殼堆肥之養分含量分析結果如表四，利用本發明稻 穀培養基培養之木黴菌（TCT111)接種之生物性稻殼堆肥，其氮含 量約1.63%、磷含量約0.44%、鉀含量約1.16%、鈣含量約1.89%、 鎂含量約0_88%、有機質含量約67.4%、辞含量約55ppm、銅含 量約17ppm。未接種木黴菌之稻殼堆肥氮含量約1.56%、磷含量 約0·38%、鉀含量約1.13%、鈣含量約1.92%、鎂含量約0.87%、 有機質含量約68.1 %、辞含量約49ppm、銅含量約16ppm。顯然 有接種木黴菌之稻殼堆肥的氮、磷、鉀、鋅及銅等含量均略高於 未接種木黴菌之一般稻殼堆肥。 表四 元素 組別 N P K Ca (%) Mg OM Zn Cu (mg/kg) 生物性 稻殼堆肥 1.63 0.44 1.16 1.89 0.88 67.4 55 17 一般稻殼堆肥 1.56 0.38 1.13 1.92 0.87 68.1 49 16 實施例7.木黴菌之存活判定 從實施例5及6有接種本發明之稻穀培養基培養之木黴菌 (TCT103、TCT111)菌種於堆肥製作過程中，以及生物性蔗渣 15 1306448 木屑堆肥、稻殼堆肥等製成品中，均可分離出所添加之菌種，分 離率約為104至105 spore/g，顯示本發明所添加之木黴菌菌種可 在堆肥化過程及堆肥成品中存活。 綜上所述，本發明利用「稻殼培養基」培養法，培養木黴菌 (TWc/iWerma π. ; TCT103、TCT111 )，兩~三週後，可以培養發 酵成固體菌種可達lxio9 spore/g。不僅操作方法簡易，且材料成 本低廉，以此法培養之菌種可在室溫或低溫條件下儲存一年以 上，活性不受影響。利用木黴菌接種用量比例技術，可以快速促 進木黴菌在堆肥材料中生長繁殖，且在堆肥化過程中及堆肥成 品，能使木黴菌菌數維持穩定成長，增進及穩定堆肥成分含量及 品質。 雖然本發明已以較佳實施例揭露如上，然其並非用以限定本 發明，任何熟悉此技藝者，在不脫離本發明之精神和範圍内，當 可作各種之更動與潤飾，因此，本發明之保護範圍，當視後附之 申請專利範圍所界定者為準。6, _9 kg, rice bran 2, _ kg, grazing palm phase 2, coffee kg. Take appropriate amount of rice medium containing about 10 spore/g Trichoderma (TCT103), first dilute with water to reduce 200 times of suspension 're-m cubic meter compost material and 2G liter of bacterial suspension, mix the sputum dilution into compost material In the middle, the most received A -., 5i 〇 〇 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约And slightly higher, the money quickly fell to the low temperature period (<5 generations) and the prevalence was 3-6 days earlier than the control without the Trichoderma compost. ^ Therefore, the temperature of the array is about 0-5 6-10 11-15 16-20 21-25 26- Biological rice husk compost 62 70 66 57 54 5 General rice husk compost 60 67 65 55 56 5 (Example 5 Benefits of bio-bagasse wood chip composting φ The results of nutrient content analysis of bio-bagasse wood chip composting are shown in Table 3, and ~~ spears are inoculated with Trichoderma viride (TCT103) cultured in the rice bran medium of the present invention. About 2.13%, phosphorus content is about 0.98%, potassium content is about! 8ΐς7 Α ', calcium content is about 1.03%, magnesium content is about 0.78%, organic matter content is about 65.5% 'zinc 晷 晷 〒 3 weight about 98ppm, copper content about 27ppm. The bagasse chips that are not inoculated with Trichoderma have a human oxygen θ of about 1.87%, a phosphorus content of about 0.85%, a potassium content of about 1.74%, a calcium content of about ^92^, a content of about 0.67%, an organic matter content of about 63.1%, and a zinc content. About 9 〇 ppm and copper 8 dan are about 26 ppm. Apparently, the contents of nitrogen, dish, unloading money, zinc and copper of the bagasse wood inoculated with Trichoderma are slightly higher than that of common bagasse wood chips. 14 1306448 Table III Element NPK Ca Mg OM Zn Cu Group (%) (mg/kg) Biological Bagasse Saw Compost 2.1 3 0.98 1.81 1.03 0.78 65.5 98 27 General bagasse wood chip compost 1.87 0.85 1.74 0.92 0.67 63.1 90 26 Example 6. Benefits of biological rice husk composting The results of nutrient content analysis of biological rice husk compost are shown in Table 4, using the rice medium of the present invention. The cultivated Trichoderma (TCT111) inoculated biological rice husk compost has a nitrogen content of about 1.63%, a phosphorus content of about 0.44%, a potassium content of about 1.16%, a calcium content of about 1.89%, a magnesium content of about 0_88%, and an organic matter content of about 67.4. %, content of about 55ppm, copper content of about 17ppm. The rice husk compost without inoculation of Trichoderma has a nitrogen content of about 1.56%, a phosphorus content of about 0.38%, a potassium content of about 1.13%, a calcium content of about 1.92%, and a magnesium content of about 0.87. %, organic matter content of about 68.1%, content of about 49ppm, copper content of about 16ppm. Obviously, the content of nitrogen, phosphorus, potassium, zinc and copper in the rice husk composted with Trichoderma is slightly higher than that of the uninoculated Trichoderma Shell compost. Table IV element group NPK Ca (%) Mg OM Zn Cu (mg/kg) Biological rice husk compost 1.63 0.44 1.16 1.89 0.88 67.4 55 17 General rice husk compost 1.56 0.38 1.13 1.92 0.87 68.1 49 16 Example 7 Trichoderma survival It is determined that the strains of Trichoderma (TCT103, TCT111) cultivated in the rice medium of the present invention are in the compost preparation process, and the biological bagasse 15 1306448 wood chip compost, rice husk compost, etc., can be used in Examples 5 and 6. The added strain was isolated, and the separation rate was about 104 to 105 spore/g, indicating that the Trichoderma strain added by the present invention can survive in the composting process and the finished compost product. In summary, the present invention utilizes the "rice-shell culture medium" culture method to culture Trichoderma (TWc/iWerma π.; TCT103, TCT111), and after two to three weeks, it can be cultured and fermented into solid strains up to lxio9 spore/g. . Not only is the method of operation simple, but the material cost is low, and the strain cultured by this method can be stored at room temperature or low temperature for more than one year, and the activity is not affected. The use of Trichoderma inoculation ratio technology can quickly promote the growth and reproduction of Trichoderma in composting materials, and in the composting process and composting products, the number of Trichoderma bacteria can be maintained stably, and the content and quality of composting components can be improved and stabilized. While the present invention has been described above in terms of the preferred embodiments thereof, it is not intended to limit the invention, and the invention may be modified and modified without departing from the spirit and scope of the invention. The scope of protection of the invention is defined by the scope of the appended claims.
16 1306448 【圖式簡單說明】 第一圖係為本發明之利用稻穀培養基製備微生物方法流程 圖。 第二圖係為利用本發明之稻榖培養基製備之微生物接種堆肥 材料的方法流程圖。16 1306448 [Simple description of the drawings] The first figure is a flow chart of the method for preparing microorganisms using rice medium according to the present invention. The second figure is a flow chart of a method for inoculating compost material using the microorganism prepared by the rice bran medium of the present invention.
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