芽孢桿菌 Bacillus subtilis B34 and Bacillus thuringiensis B163 接種於菜籽粕與蓖麻粕粉對水溶性作物養分濃度影響之探討

The effect of inoculating Bacillus subtilis B34 and Bacillus thuringiensis to Reapseed meal and Castor meal on the concentrations of water soluble nutrients for crop growth

陽鎮宇(Chen-Yu Yang)

指導教授 ： 簡宣裕 ； 陳齊聖

朝陽科技大學/理工學院/應用化學系碩士在職專班/碩士(2011年)

摘要

從工業革命以來化學產業不斷進步，不管在人事物對於反應時間也越來越重視，這觀念也開始影響人類對於傳統產業經營觀念與作業方式，早期農業主要是以堆肥做為作物生長所需的養分來源，轉而被化學合成肥料取代，做為作物所需的養分，因化學肥料可以快速滿足作物需求減少生長時間與增加品質達到經濟效益，但是長期不合理化施用化學肥料的結果是對於土壤、生態、作物及人類的健康都有相當的傷害，且化學合成肥料於製造過程需耗費大量能源及石化原料對於地球資源損耗與汙染影響甚大。而有機液肥肥效反應與化學肥料相近，故本研究將農業廢棄物之菜籽粕與蓖麻粕作為醱酵基質，接種入由土壤、作物根部、及分解中有機物分離、純化、篩選出之分解蛋白質能力強的菌株 B. subtilis B 34 與 B. thuringiensis B 163，進行液態醱酵，以生物降解作用快速有效的提升有機液肥之有效性養分，提供葉菜類作物吸收利用，以減少對化學肥料依賴性。研究中分別探討對於不同溫度、通氣量、菌液比例、時間、有無磨粉及水粕比例環境變因對此兩株蛋白分解菌降解菜籽粕與蓖麻粕產出水溶性氮濃度之影響並觀測水溶性磷、鉀、 pH 及 EC 量之變化，並由實驗結果選取各變因中較適合蛋白分解菌降解菜籽粕與蓖麻粕產出水溶性氮之條件，以符合液肥製作過程中節能降低成本效益，將之組合起來做為液態醱酵之條件，以探討是否為此兩株蛋白分解菌降解菜籽粕與蓖麻粕產出較高有效性氮濃度之最適條件。 B. subtilis B 34 降解菜籽粕，進行液態醱酵之較合是條件設定為 溫度 30 ℃ 、轉速 125 rpm 、 菜籽粕與水比例為 10 g ： 100 mL 、菌液接種量為 1 mL 、醱酵時間 4 週；B. thuringiensis B 163 降解蓖麻粕，進行液態醱酵之較合適條件設定為 溫度 30 ℃ 、轉速 150 rpm 、 蓖麻粕與水比例為 15 g ： 100 mL 、菌液接種量為 1 mL 、醱酵時間 4 週，由實驗結果得知 B. subtilis B 34 降解菜籽粕，水溶性氮含量可達到 1.92 mg‧mL-1 ，比對照組高出 16167 ％ ，並在實驗中發現B. subtilis B34 降解菜籽粕時對於增加水溶性磷也具有一定能力。 B. thuringiensis B 163 在設定的較適條件下降解蓖麻粕，所產出水溶性氮含量可達到 2.05 mg‧mL-1 ，比對照組高出 2344 ％ 。 B. subtilis B34 與 B. thuringiensis B 163 分別降解菜籽粕與蓖麻粕，增加有機液肥中的水溶性氮量具有一定能力。此外蓖麻粕與菜籽粕以煮沸30分鐘替代高溫高壓滅菌，然後進行液態醱酵實驗中，也發現蓖麻粕與菜籽粕本身含有蛋白分解能力菌株，在增加水溶性氮量也具有一定能力，這將是未來進一步研究探討之方向。

關鍵字

枯草桿菌 ； 蓖麻粕 ； 菜籽粕 ； 葉菜類植物 ； 有機液肥 ； 蘇力菌

並列摘要

Since 19th century industrial revolution, the chemical industry had also taken a big step. The concept of efficiency and efficacy become an important element in the chemical industry, and it also made a revolution in the development of traditional agriculture. In ancient times, agriculture is limited in given natural organic component, waiting long time for the crops to absorb these nutrition by themselves. As the social structure changed, to gain the best profit, making fast-growing and good-quality crops becomes the new direction of agriculture . Chemical-synthesis components that can be absorbed directly and easily by the crops become the new land of agriculture. But as times goes by, people start to realize that those chemical-synthesis components are harmful to land, cops, ecology, and even our own health,if it is not used reasonaldy. It not only consumes our nature source but also make irreversible pollution to our land.
Our research goal is to produce efficient microorganism-made organic liquid fertilizer that provides abundant nitrogen for leafy vegetables to grow.
First, we used agriculture waste, rape seed meal and castor seed meal, as our fermented component. Second, we choose bacteria, B. subtilis B 34 and B. thuringiensis B 163 isolated form soil and the decaying organic material. Both of them have the ability to break down protein to release large quantity of water-soluble nitrogen form rape seed meal and castor seed meal. Our research focus on how different environmental elements, including temperature, rotationg speed, the quantity of inoculam, and the ratio of water and meal, affect these bacteria to ferment rape seed meal and castor seed meal containing solution to produce aridble nutrients. We evaluate their protein degeneration ability and the production of water-soluble nitrogen. We also observe the change of water-soluble phosphate, potassium, pH value and EC.
We look in the fermented soluions proper for the conditions of each environmental elements separately, and try to make the lowest cost at the same time. Then we put all those proper conditions together to figure whether this condition is the best for protein-degrading bacterium to degenerate rape seed meal and castor seed meal to produce available nutrients effectively. Our experiment result showed that the fermcntation condition for B. subtilis B 34 and B. thuringiensis B 163 to work effictiuely is 30 ℃ rotation rate with 125 rpm, rape seed meal 10 g accompanied with 1 mL water for fermenting four weeks and 30 ℃,rotation rate with 150 rpm ,castor seed meal 15 g accompanied with 1 mL water for fermenting four weeks respectiuely . Under best fermentation condition, B. subtilis B 34 fermented rape seed meal containg solution to produce water-soluble nitrogen up to 1.92 mg‧mL-1 which is 1617 ％ higher than that of control group. At the same time, B. subtilis B 34 shows its good ability to degenerate water-soluble phosphate form rape seed meal. Under the best fermentation condition,B. thuringiensis B 163 fermented castor seed meal containing solution to produce water-soluble inorgomic nitrogen up to 2.05 mg‧mL-1 which is 2344 ％ high than that of control group. Our results indicated that by degrading rape seed meal and castor seed meal, B. subtilis B34 and B. thuringiensis B 163 show very good ability in increasing water-soluble inorgornic nitrogen contents in the fermented organic liquid fertilizers. We also found that after boiling for 30 minutes only or without sterilization,poteolytic bacteria inside rape seed meal and castor seed meal show their ability in producing water-soluble inorganic nitrogen. This finding is our next step study in the future for producing liquid fertilizers using rape seed and castor seed meal as fermented materials.

並列關鍵字

Bacillus thuringiensis ； organic liquid fertilizer ； Bacillus subtilis ； Castor seed meal ； Rape seed meal ； Leaf vegetables

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