臺灣博碩士論文加值系統

對於全球的農業而言，豆科根瘤菌生物性固氮作用是很重要的氮肥來源。敏豆是全世界產量僅次於黃豆的豆類作物，也是台灣夏季常見蔬菜。土壤酸化是豆科植物生長的限制因子，影響根瘤菌與植物根的共生效益及宿主植物的生長，降低豆科作物的產量。根瘤菌菌種的不同，對酸性的耐受性有相當大的差異。篩選出耐酸性根瘤菌，對於豆科作物根圈的固氮共生效益，將會有提升的作用。我們假設在實驗室酸性條件培養下，存活率較高的菌株在酸性土壤中可獲得更高的存活力。本研究從敏豆根瘤篩選出16株Rhizobium spp. 以及一株耐酸參考菌株CIAT899，利用homopipes調整培養基的酸鹼值 (pH = 4、5、及6)、以Al2(SO4)3 調整不同的鋁離子濃度 (1, 2及3 mmol L-1)，並以砂耕環境下 (pH 5) 評估固氮活性和共生效益，篩選耐酸性根瘤菌。本研究篩選出耐酸耐鋁菌株B412，耐酸不耐鋁菌株B379、耐鋁不耐酸菌株B378和不耐鋁不耐酸菌株B374進行酸性土壤盆栽試驗。本實驗使用兩種酸性土壤 (pH 5.1)，分別是仁愛土壤 ([Al3+] = 2.01 mg kg -1) 及通霄土壤 ([Al3+] = 3.72 mg kg -1)。酸性土壤盆栽試驗下，在仁愛土壤中種植敏豆，以接種B379菌株獲得最高的固氮活性，在鋁含量較高的通霄土壤中，耐鋁毒性菌株B378獲得最高之結瘤數，而耐酸耐鋁菌株B412則是在兩種土壤中皆獲得高的豆莢乾重，特別是在仁愛土壤中接種B412菌株之產量較化肥處理者顯著增加50 % 。本研究之篩選方法可篩選出耐酸耐鋁的根瘤菌，促進豆科作物在酸性土壤中的生長，並增加根部結瘤數、共生效益和產量。除了提升敏豆在酸性土壤中的產量，也可降低化學氮肥之施用量，節省生產成本，並有利於永續農業之發展。

Biological nitrogen fixation mediated by the legume-rhizobia symbioses is important for world agriculture. The yield of common bean (Phaseolus vulgaris L.) ranks second only to soybeans, and it is a common summer vegetable in Taiwan. Soil acidity is a limiting factor for legume growth because it affects the symbiotic efficiency of Rhizobium, host plant growth, and the legume production. Species of rhizobia vary significantly in their tolerance to low pH. Selecting acid-tolerant rhizobia may increase their symbiotic efficiency in acid soil. This study aimed to examine whether strains of Rhizobium spp. selected for growth on acid culture in vitro would also survive and grow better in acid soils. Sixteen Rhizobium spp. were isolated from commom bean nodules, and one commercial strain CIAT899 known for its acid-tolerance was included as a standard. Selective media with different pH levels (4, 5, and 6) and Al3+ concentrations (1, 2, and 3 mmol L-1) were used for selecting acid-tolerant rhizobia, and the nitrogen fixation and symbiotic efficiency of these rhizobia with common bean were evaluated in sandy medium with pH5. Four Rhizobium strains were selected for pot experiments including the acid- and Al3+-tolerant strain B412, acid-tolerant and Al3+-sensitive strain B379, acid-sensitive and Al3+-tolerant strain B378, and acid- and Al3+-sensitive strain B374. In addition, two soils with various Al3+ concentrations were used for pot experiments. B379 showed the highest nitrogenase activity in Renai soil with low Al3+ concentration (2.01 mg kg -1), but B378 resulted in the best nodule numbers in Tongxiao soil with is high Al3+ concentration (3.72 mg kg -1). However, B412 consistently showed a significant increase in bean yields in the two soils. Interestingly, the strain significantly increased the yield of bean by 50 % compared to that of the chemical fertilization treatment in Renai soil. Our study provides a strategy to select acid- and Al3+-tolerant rhizobia for increasing the yield of common bean grown in acid soils, increasing the nodulation and symbiotic effectiveness by Rhizobium. As a result, inoculation of common bean with acid-tolerant rhizobia may considerably reduce the use of chemical fertilizers for acid soils.

摘要 i
Abstract ii
目錄 iii
表次 iv
圖次 v
壹、前言 1
貳、前人研究 3
一、共生型固氮根瘤菌 3
二、敏豆生長特性 11
三、酸性土壤 17
參、材料與方法 22
一、研究架構 23
二、試驗根瘤菌之來源、分離與保存 24
三、根瘤菌之鑑定 26
四、生理特性測試 30
五、胞外酵素活性測試 33
六、耐酸、耐鋁毒性性測試 36
七、菌株固氮共生效益盆栽試驗 37
八、酸性土壤盆栽試驗 43
九、統計方法 49
肆、結果與討論 50
一、菌株來源及鑑定結果 50
二、菌株生理特性結果 55
三、菌株耐酸性結果 60
四、菌株耐鋁毒性結果 66
五、菌株固氮共生效益結果 71
六、酸性土壤盆栽試驗結果 79
伍、結論 92
陸、參考文獻 93
柒、附錄 105

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