臺灣博碩士論文加值系統

隨著農業的發展，肥料的用量也逐年提升。尤其是磷肥，不像氮肥能夠從大氣中直接獲得，只能靠磷礦石的開採提煉而得。但是世界上的磷礦石存量大約只夠我們再使用90年，故尋求可用的替代品以讓農業能夠永續經營是當務之急。骨頭為常見的農業及食品加工業廢棄物，其成分主要為磷酸三鈣Ca3(PO4)2與氫氧基磷灰石Ca10(PO4)6(OH)2，具有與過磷酸鈣Ca(H2PO4)含量相當的磷，但骨頭於自然環境中的溶解度極低，因此不易當作肥料使用。因此若能找出將骨頭中所含的磷利用於農業生產上的方法，將能夠為農業省下可觀的成本，也使農業能夠永續發展。本研究乃利用骨粉作為養分來源以培養出能夠利用並溶解骨粉的微生物，並從中篩選出可能能夠溶出骨粉磷的菌株共 62株。經過骨粉孵育及溶骨粉磷能力測定後得到菌株Aspergillus niger為環境中最常見之黑麴菌，常被用來進行工業發酵以取得檸檬酸，對於人體與植物的致病力皆不強，其對於磷的溶出有很強的能力，在純化環境中A. niger對於骨粉磷比起其他篩選所得到菌株也有最強的溶出能力。而在液體培養基環境中能看出A. niger對羊骨粉與豬骨粉之溶磷情形不同，羊骨粉由於具有較高的碳氮比，使得磷的溶出量在初期較快速，但隨著孵育時間變長卻有減慢的情形；而豬骨粉碳氮比較低，磷溶出量緩慢上升，但經過C/N的調整試驗後發現羊骨粉磷的溶出速度在較低碳氮比時會有較大的溶出量，但豬骨則幾乎不受影響。最後將A. niger與骨粉同時或孵育後施用於盆栽中，發現無論是何種處理對於盆栽植體的磷含量影響皆不大，但會稍微提高土壤的有效性磷含量，添加羊骨粉的處理則會造成土壤裡的氮含量下降。 A. niger在土壤中雖然能夠將骨粉裡的磷溶出，但也會與植物競爭土壤裡的養分(如氮肌餓)，造成植物對於土壤養分吸收上的阻礙。

Phosphate fertilizer has been recognized as the most limiting element in agriculture production. Its natural resource, unfortunately, is also limited. The recycle of animal bone meal will be increased its importance for most country prohibiting it used as animal feed. The aim of this thesis was to screen microorganisms which can directly dissolve phosphate from the raw bone meal with no fat removal. The fat contained in raw bone meal is assumed to be the carbon source for the phosphate solubilizing microorganism. Aspergillus niger is evaluated as the only microorganism which meets the requiring that the microorganism can directly release the phosphorus from the raw bone meal of pig and goat, after screened 62 calcium phosphate solubilizing microorganisms from different resources. The main mechanism of A. niger dissolves the raw bone meal is the release of oxalic acid. In liquid media, A. niger dissolved more phosphate from the goat bone than from pig bone. The addition of extra nitrogen to lower the C/N ratio can improve the release of phosphate from goat bone, but no significant increase from pig bone. In pot experiment, although, the inoculation of A. niger increased soil available phosphate concentration of addition of all bone meal treatments, no positive effect of maize dry weight and phosphate uptake in maize shoot. These results are due to the high C/N ratio of both raw bone meals and there was no fertilizers applied except the addition of bone meals. The lower ammonium and nitrate concentrations of the bone meal addition treatments evidenced the assumption.

目錄
摘要 I
英文摘要 III
目錄 V
表次 VIII
圖次 IX
附錄 XII

壹、前言 1
貳、前人研究 3
一、磷肥 3
二、磷礦石的存量 4
三、動物骨粉 4
四、土壤微生物 5
五、土壤真菌 7
六、Aspergillus niger 8
七、微生物的生物代謝作用 9
參、材料與方法 12
一、骨粉製備 12
二、骨粉與標準磷化合物構造鑑別 12
三、骨粉化學成分測定 12
四、溶骨磷菌之分離 13
(一) 豬油分解菌的馴養 13
(二) 豬油分解菌的加強培養 (enrichment) 13
(三) 骨粉分解菌之馴養及加強培養 (enrichment) 14
(四) 骨粉分解菌之篩選 14
(五) 溶磷菌之篩選 14
五、溶骨磷菌溶磷能力之篩選 15
(一) 分離菌株對於骨粉磷的溶出試驗 15
(二) 篩選所得菌株 16
六、骨粉溶磷菌之能力測定 19
(一) 菌株於液態環境下對骨粉磷的溶出能力 19
(二) 菌株於固態環境下對骨磷的溶出能力 19
(三) 菌株於代謝時有機酸產出情形 20
七、骨粉溶磷菌的盆栽試驗 20
八、盆栽植體性質分析項目及方法 21
九、盆栽土壤性質分析項目及方法 22
肆、結果與討論 26
一、 骨粉基本性質 26
二、 菌株篩選試驗 32
(一) 菌株篩選 32
(二) 分離菌株在液體環境中對骨粉的溶解情形 38
(三) 孵育過程中有機酸的產出 45
(四) 分離菌株在固體環境中對骨粉的溶解 54
三、 菌株之實際盆栽試驗 59
四、 添加氮源對菌株溶磷能力的影響 70
伍、結論 73
陸、參考文獻 74
柒、附錄 80

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