Castor cake, a by-product generated from castor oil extraction and with over 1,383, 000 tons of annual production, has high nitrogen content, and therefore, it has been widely used as organic fertilizer for supporting plant growth and reducing the cost of waste disposal. Additionally, castor cake contains active ingredients/toxins such as ricin and ricinine, both are insecticidal, bactericidal, and nematicidal, which makes castor cake used as a pesticide as well. However, there is a few studies of the impact of the toxins to mollusks, microorganisms, or even aquatic organisms as well as crop and human health via food chain. To evaluate the effects of castor cake to soil environment and crop growth, in this study, we first conducted a release, leaching, and plant uptake experiments using pot and hydroponic systems to evluate the distribution and accumulation of toxin ricinine in soil and crops, respectively. Ricinine in solid samples is extracted by QuEChERS (quick, easy, cheap, effective, rugged and safe) method, while in liquid samples, it is extracted and concentrated through solid-phase extraction procedure followed by liquid chromatography quadrupole time of flight mass spectrometry (LC-QTOF/MS) analysis. Second, we planted lettuce in soil pots added with castor cake to determine the effect of castor cake to soil inorganic nitrogen content and lettuce growth. Last, we used castor cake extracts and different ricinine concentration to understand their effects to egg hatching rate and nematode lethality of M. incogita. Moreover, we discussed the nodulation ability of M. incognita through a pot setting. We also used a culture medium to test the inhibitory ability of ricinine to the growth of pathogen R. solani and specific beneficial bacteria as well as Biolog Ecoplate analysis and 16S rDNA analysis to evaluate the effect of castor cake on soil bacterial metabolic and species diversity. The average ricinine concentration of the castor cake used in this study was around 1093 mg kg-1. After 1 day of castor cake application, ricinine was with the highest concentration comparing to the days of 7 and 14 in soil, soil pore water and leachates, approximately 70% of the ricininie content in castor cake. The plant uptake experiment showed ricinine can be accumulated in lettuce shoots, posing risk to human health. The inorganic nitrogen concentration in the soil increased apparently within 7 days but slightly decreased from 7 to 14 days after castor application. Castor cake did inhibit the growth of M. incogita while it was with no effect on specific beneficial bacteria and enhance the metabolic and species diversity of soil bacteria. However, castor cake changed the function and dominant species of soil bacteria. Additionally, castor cake promoted the growth of Burkholderiaceae spp. bacteria in the soil. Overall, according to the results obtained from this study, we recommend farmers to apply appropriate rate for castor cake and to start planting after 14 days of castor cake application. This could not only reduce the risk of human exposed to ricinine through consumption of crops containing ricinine, but also alleviate adverse effects on crop growth and soil environment.
摘要 i
Abstract iii
目次 v
表目次 ix
圖目次 x
第一章、前人研究 1
1.1 近代農業型態之轉變 1
1.2 蓖麻粕 1
1.2.1 蓖麻粕簡介 1
1.2.2 蓖麻粕對土壤與作物之影響 2
1.2.3 蓖麻粕防治病蟲害之能力 4
1.2.4 蓖麻粕對環境微生物之影響 11
1.2.5 蓖麻粕中蓖麻毒蛋白與蓖麻鹼之毒性與致毒機制 12
1.3 蓖麻鹼之萃取與分析方法 14
1.4 土壤細菌多樣性分析方法 17
1.5. 擬解決問題 18
1.6 目的 18
第二章、材料與方法 19
2.1試驗架構 19
2.2試驗材料與化學品 21
2.2.1 試驗材料 21
2.2.2 試驗藥品 21
2.2.3 試驗土壤 22
2.2.4 蓖麻粕 24
2.3蓖麻鹼分析方法建立及應用 25
2.3.1 方法之建立 25
2.3.2 方法應用 30
2.4評估蓖麻粕中ricinine在土壤中的流佈 32
2.4.1 釋放試驗 32
2.4.2 淋洗試驗 34
2.4.3 吸附試驗 34
2.5評估ricinine在萵苣中的累積及分佈 36
2.5.1 水耕試驗 36
2.5.2 盆栽試驗 39
2.6評估施用蓖麻粕對土壤及作物中植物生長必需元素之影響 40
2.6.1 土壤分析 40
2.6.2 植體分析 41
2.7評估蓖麻粕防治南方根瘤線蟲(Meloidogyne incognita)之能力 42
2.7.1 南方根瘤線蟲之培養 42
2.7.2 蓖麻粕浸泡液對南方根瘤線蟲卵孵化率之影響 42
2.7.3 Ricinine對南方根瘤線蟲致死率之影響 43
2.7.4 蓖麻粕對南方根瘤線蟲結根瘤之影響 43
2.8 評估蓖麻粕對土壤微生物族群之影響 44
2.8.1 Ricinine對特定微生物之抑制能力 44
2.8.2 蓖麻粕對土壤細菌功能多樣性之影響 47
2.8.3 蓖麻粕對土壤細菌物種多樣性之影響 47
2.9 統計分析 49
第三章、結果與討論 50
3.1 蓖麻鹼(Ricinine)萃取與分析方法確效 50
3.1.1 SPE方法 50
3.1.2 QuEChERS方法 50
3.2 市售蓖麻粕產品ricinine含量 53
3.2.1 供試蓖麻粕的基本性質 53
3.2.2 市售蓖麻粕產品ricinine含量 54
3.3 蓖麻粕中ricinine在土壤的流佈 56
3.3.1 Ricinine自蓖麻粕釋放至土壤之情形 56
3.3.2 土壤施用蓖麻粕後ricinine在土壤中之流佈 68
3.3.3 土壤對ricinine之吸附 73
3.4 植物吸收與累積ricinine之能力 75
3.4.1 水耕萵苣對ricinine之吸收與累積 75
3.4.2 土壤中萵苣對ricinine之吸收與累積 80
3.5 蓖麻粕對土壤與植體中植物生長必需氮元素濃度之影響 82
3.5.1 對土壤無機氮元素濃度之影響 82
3.5.2 對植體內植物全氮含量之影響 85
3.5.3 土壤施用蓖麻粕對植物生長之影響 86
3.6 蓖麻粕對南方根瘤線蟲之影響 88
3.6.1 蓖麻粕浸泡液對線蟲卵孵化率之影響 88
3.6.2 Ricinine對線蟲致死率之影響 90
3.6.3 蓖麻粕對空心菜結根瘤數之影響 92
3.7 Ricinine對特定微生物之影響 94
3.8 蓖麻粕對土壤細菌功能多樣性之影響 100
3.8.1 不同蓖麻粕施用量對土壤細菌群落代謝潛勢變化 100
3.8.2 不同蓖麻粕施用量對功能多樣性指數分析 103
3.8.3 不同蓖麻粕用量對細菌碳源利用程度分析 107
3.9 蓖麻粕對土壤細菌物種多樣性之影響 114
3.9.1 不同蓖麻粕施用量對物種多樣性指數分析 114
3.9.2 不同蓖麻粕施用量對土壤物種群落變化 116
第四章、結論 120
第五章、參考資料 121
附錄 138
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