臺灣博碩士論文加值系統

立枯絲核菌（Rhizoctonia solani kühn AG-4）可以引起甘藍幼苗猝倒、萎凋與死亡的現象，是甘藍苗床栽培的主要病原菌之一。使用種子誘釣法篩選具有抑制R. solani纏據甘藍種子效果的有機添加物，結果發現BVB No.4 泥炭苔中添加2％（w/v）苦茶粕處理當天即可降低種子纏據百分率達68％，處理一星期後，也可得到類似的效果。添加1％（w/v）苦茶粕可降低甘藍苗立枯病的發病率達40％左右。進一步探討苦茶粕防病的原理，發現添加1％（w/v）苦茶粕於BVB No.4 泥炭苔中，會提高微生物族群量，其中細菌菌量上升9倍，真菌及放線菌則分別增加2~3倍，但對BVB No.4 泥炭苔pH值影響不大。去除BVB No.4 泥炭苔中的微生物，發現苦茶粕仍能降低R. solani纏據種子百分率30％以上。以5倍體積甲醇在室溫下萃取苦茶粕，進一步使用管柱純化後，依Liebermann-Burchard反應確定其為三萜類皂素。在馬鈴薯葡萄糖瓊脂培養基中添加1％（w/v）苦茶粕粗萃取液，其抑菌百分率即可達50％。當無菌水中添加1％（w/v）苦茶粕粗萃取液時，將菌絲團塊置入後，可發現細胞通透性改變，到第6天導電度可以上升60μs，惟以無菌水處理菌絲團塊其導電度未明顯沒有上升。在影響苦茶粕抑菌性之因子方面，基質的pH值改變並不影響苦茶粕有效性：然而添加氮肥卻會降低苦茶粕防治甘藍立枯病的效果。

Damping-off disease caused by Rhizoctonia solani kühn AG-4 is a serious disease of cabbage seedlings especially in culture medium. Tea seed pomace was added（2％,w/v）in BVB NO.4 peat moss medium infested with R. solani AG-4. It was significantly effective in inhibiting colonization of cabbage seed caused by R. solani AG-4 at 0 and 7th days after treatment. Tea seed pomace at the rate of 1％（w/v）was able to markedly reduce the disease incidence of cabbage seedling damping-off by 40％ compared to non-treatment as a control. Tea seed pomace could stimulate proliferation of beneficial microorganisms including fungi, bacteria and actinomycetes. The pH values did not change significantly after tea seed pomace was added（1％,w/v）in BVB NO.4 peat moss medium. The methnol-extractive compound was obtained from purification procedure of Diaion HP-20column chromatography and acetone fraction. The compound was characterized by Liebermann-Burchard reaction as triterpenoid saponin. The methnol-extractives were added（1％,w/v）in potato dextrose agar for evaluating their effect on mycelial growth of R. solani AG-4. The mycelial growth was markedly inhibited higher than non-treatment about 50％. Amendment of NH4NO3 or Urea was able to nullify tea seed pomace to reduce damping-off disease incidence of cabbage seedlings caused by R. solani AG-4.

中文摘要......................................i
英文摘要.....................................ii
目錄........................................iii
表目錄.......................................vi
圖目錄......................................vii
前言..........................................1
材料與方法....................................4
一、供試菌株與植物............................4
二、接種源製備................................4
三、篩選具有抑制Rizoctonia solani AG-4纏據甘藍種子效果之有機添加物......................................4
四、不同濃度苦茶粕對Rhizoctonia solani AG-4纏據甘藍種子效果的影響........................................5
五、不同濃度苦茶粕對甘藍立枯病發生的影響......6
六、不同濃度苦茶粕對甘藍植株生長的影響........6
七、苦茶粕的防病原理探討......................6
（一）苦茶粕對栽培介質酸鹼度的影響............6
（二）苦茶粕對栽培介質中微生物族群量的影響....7
（三）在滅菌與未滅菌BVB NO.4泥炭土中添加苦茶粕對於Rhizoctonia solani AG-4纏據甘藍種子的影響.....7
八、不同公司的苦茶粕對甘藍立枯病發生的影響....8
九、有效成分的鑑定............................8
（一）萃取條件測試............................8
1. 萃取溶劑之選擇.............................8
2. 萃取時間之探討.............................9
3. 萃取溫度之探討.............................9
4. 最佳萃取溶劑比之探討.......................9
（二）苦茶粕萃取物抑菌成分之純化.............10
1. Diaion HP-20管柱層析......................10
2. 使用丙酮純化法............................11
（三）抑菌成分之鑑定.........................11
十、有效成分對Rhizoctonia solani AG-4菌絲
的影響.......................................11
（一）不同濃度苦茶粕粗萃取液對Rhizoctonia solani AG-4菌絲
生長的影響...................................11
（二）苦茶粕粗萃取液對Rhizoctonia solani AG-4菌絲細胞膜通透性的影響.....................................12
（三）苦茶粕粗萃取液對Rhizoctonia solani AG-4 isolate RST-04在甘藍莖表面形成侵入褥的影響...............12
十一、影響苦茶粕萃取物之有效性的因子.........13
（一）酸鹼值對苦茶粕萃取物有效性的影響.......13
（二）不同氮素源對苦茶粕萃取物有效性的影響...13
十二、添加不同氮肥對苦茶粕防治甘藍立枯病效果
的影響.......................................14
結果.........................................15
一、篩選具有抑制Rizoctonia solani AG-4纏據甘藍種子效果之有機添加物.....................................15
二、不同濃度苦茶粕對Rhizoctonia solani AG-4纏據甘藍種子效果的影響.......................................15
三、不同濃度苦茶粕對甘藍立枯病發生的影響.....15
四、不同濃度苦茶粕對甘藍植株生長的影響.......16
五、苦茶粕的防病原理探討.....................16
（一）苦茶粕對栽培介質酸鹼度的影響...........16
（二）苦茶粕對栽培介質中微生物族群量的影響...16
（三）在滅菌與未滅菌BVB NO.4泥炭土中添加苦茶粕對於Rhizoctonia solani AG-4纏據甘藍種子的影響....17
六、不同公司的苦茶粕對甘藍立枯病發生的影響...17
七、有效成分的鑑定...........................17
（一）萃取條件測試...........................17
（二）苦茶粕萃取物抑菌成分之純化與鑑定.......18
八、有效成分對Rhizoctonia solani AG-4菌絲
的影響.......................................18
（一）不同濃度苦茶粕粗萃取液對Rhizoctonia solani AG-4菌絲
生長的影響...................................19
（二）苦茶粕粗萃取液對Rhizoctonia solani AG-4菌絲細胞膜通透性
的影響.......................................19
（三）苦茶粕粗萃取液對Rhizoctonia solani AG-4 isolate RST-04在
甘藍莖表面形成侵入褥的影響...................19
九、影響苦茶粕萃取物之有效性的因子...........19
（一）酸鹼值對苦茶粕萃取物有效性的影響.......19
（二）不同氮素源對苦茶粕萃取物有效性的影響...20
十、添加不同氮肥對苦茶粕防治甘藍立枯病效果
的影響.......................................20
討論.........................................21
引用文獻.....................................24
圖表.........................................28

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