臺灣博碩士論文加值系統

為因應人口增加所帶來可能的糧食不足問題，世界各國無不在為糧食增產做努力。然如何在糧食生產又不破壞環境與生態體系間取得平衡，則是各國農業政策的努力目標，推廣有機農業則是其政策之一。生物防治則是被公認是未來替代或減少化學農藥的重要策略之一，因此本論文擬從有機茶園中分離出拮抗微生物，並測試其病害防治效果，以期未來能開發成生物農藥。
本試驗地點選在臺大實驗林鳳凰自然教育園區，探討在有機農法和慣行農法的管理方式下，對於茶園土壤微生物相結構組成有何差異，並從其土壤中篩選有益微生物，並檢測對炭疽病菌(Colletotrichum higginsianum)生長之影響及其引起的病害是否有防治效果。試驗自2017年7月開始，每隔3月採樣一次，為期一年，土壤採樣區包括一處慣行茶園及兩處有機茶園。經土壤微生物相調查，結果顯示有機茶園的真菌平均數量(59.82×10^3cfu/g)普遍較慣行茶園(18.86×10^3cfu/g)數量要高；同樣的，具有指標性的木黴菌平均數量在有機茶園(23.72×10^3cfu/g)也較慣行茶園(5.78×10^3cfu/g)約高出4倍量。將各個微生物族群數量與全年雨量及溫度比對後，顯示真菌及放線菌數量與該地區的雨量多寡呈正相關，但各微生物數量與溫度則無顯著相關。經培養基做對峙試驗，從中篩選出8株對炭疽病菌生長抑制較佳的菌株，並於溫室中檢測對炭疽病的防治效果。測試結果顯示，篩選出的8株拮抗真菌對白菜炭疽病菌感染小白菜的罹病度，與對照組比較分別降低32~53%，其中以菌株O1-5-3最佳。此菌株O1-5-3經基因定序比對，可確定為木黴菌屬之綠木黴菌(Trichoderma virens)。

In order to cope with the possible shortage of food caused by the increase in population, all countries in the world are working hard to increase food production. However, how to achieve a balance between food production and environmental and ecological systems is the goal of national agricultural policy efforts, such as promoting organic agriculture is one of its policies. Biological control is recognized as one of the important means to replace or reduce chemical pesticide use in the future. Therefore, this thesis is conducted to isolate antagonistic microorganisms from organic tea garden and test their disease control ability, in order to develop bio-pesticide in the future.
The experimental fields were selected in the Fenghuang Nature Education Areas of the Experimental Forest National Taiwan University to explore the differences in the composition of soil microbial phase structure in tea gardens under the management of organic farming methods and conventional farming methods, and to screen for beneficial microorganisms from their soils and to detect anthracnose disease control of chinese cabbage caused by Colletotrichum higginsianum and the pathogen growth. According to soil microbial survey, the average number of fungi in organic tea gardens (59.82×103 cfu/g) is generally higher than that of conventional tea gardens (18.86×10^3 cfu/g); similarly, the average number of Trichoderma spp. is in organic tea gardens (23.72×10^3 cfu/g) is also about 4 times higher than the conventional tea garden (5.78×10^3 cfu/g). For bacteria and actinomycetes number investigations, the results showed no significant difference between organic tea gardens and conventional tea gardens. Comparing the number of microbial populations with the annual rainfall and temperature, it was found that the number of fungi and actinomycetes was positively correlated with the amount of rainfall in the area, but the number of all microorganisms was not significantly correlated with temperature. For dual culture of pathogen and antagonists on medium, 8 strains of Trichoderma spp. which had better growth inhibition of C. higginsianum were screened out, and to control anthracnose disease of chinese cabbage was detected in the greenhouse. The results showed that the 8 selected strains of antagonists were effective to reduce the anthracnose disease of chinese cabbage about 32~53% compared with the untreated control, among which strain O1-5-3 was the best. This strain O1-5-3 was identified by gene sequencing and assigned as Trichoderma virens.

摘要........................................................i
Abstract...................................................ii
誌謝.......................................................iv
目錄........................................................v
表目錄....................................................viii
圖目錄......................................................ix
第一章 前言..................................................1
第二章 文獻回顧..............................................2
2.1 土壤微生物的重要性........................................2
2.2土壤微生物的種類及作用.....................................2
2.2.1 病毒...................................................2
2.2.2 細菌...................................................3
2.2.3 放線菌.................................................4
2.2.4 真菌...................................................4
2.2.5 藻類...................................................5
2.2.6 原生動物................................................5
2.3 有機農業..................................................6
2.3.1 有機農業之定義...........................................6
2.3.2 有機農業的發展現況.......................................6
2.3.3 生物應用與有害物之防治....................................7
2.4 氣候條件對土壤生物相的影響及研究方法.........................7
2.5 試驗地區環境及背景介紹......................................8
2.5.1 鳳凰自然教育園區有機茶園之環境介紹.........................8
2.5.2 有機茶園的栽培史..........................................8
2.5.3 氣象.....................................................8
2.5.3.1 氣溫...................................................9
2.5.3.2 雨量...................................................10
2.5.3.3 風向、相對溼度..........................................10
2.5.4 茶園管理方式..............................................11
2.6 不結球白菜簡介..............................................11
2.6.1 簡介......................................................11
2.6.2 經濟價值..................................................11
2.7 Colletotrichum higginsianum介紹.............................12
2.7.1 Colletotrichum higginsianum分類...........................12
2.7.2 Colletotrichum higginsianum之型態及生長特性................12
2.7.3 Colletotrichum higginsianum之病癥及傳播方式................12
2.7.4 Colletotrichum higginsianum之防治方式......................13
第三章 材料與方法................................................14
3.1 茶園採樣及菌相調查...........................................14
3.1.1 土壤採樣時間...............................................14
3.1.2 土壤採樣地點...............................................14
3.1.3 土壤樣本採樣...............................................15
3.1.4 培養基配制.................................................15
3.1.5 土壤處理及分析方法..........................................15
3.2 從土壤中篩選可抑制小白菜炭疽病菌生長的真菌菌株..................16
3.2.1真菌菌株來源................................................16
3.2.2 病原菌菌種來源..............................................16
3.2.3 拮抗真菌與白菜炭疽病菌之培養與保存............................16
3.2.4 土壤分離之菌株與白菜炭疽病菌之對峙試驗........................16
3.3 溫室試驗.....................................................17
3.3.1 供試植株...................................................17
3.3.2 拮抗真菌懸浮液之制備........................................17
3.3.3 白菜炭疽病菌孢子懸浮液之製備.................................17
3.3.4 計算葉片病害面積方法及病害調查法.............................17
3.3.5 拮抗真菌對小白菜生長影響測試.................................18
3.3.6 測試白菜炭疽病菌濃度對小白菜罹病度之影響......................18
3.3.7測試拮抗真菌對白菜炭疽病菌在小白菜上的抑制能力..................19
3.3.8不同濃度之拮抗真菌對白菜炭疽病菌在小白菜上的抑制能力影響.........19
3.4數據處理與分析.................................................19
3.5 拮抗真菌鑑定..................................................20
第四章 結果.......................................................21
4.1 茶園微生物相之調查.............................................21
4.1.1 有機茶園與慣行茶園微生物相之調查..............................21
4.1.2 不同栽培區菌落數在不同季節下的消長............................22
4.2 抑制炭疽病菌生長能力之拮抗真菌株篩選.............................26
4.3 拮抗真菌對於溫室小白菜生長之影響................................28
4.4白菜炭疽病菌濃度對於溫室小白菜罹病率之影響........................30
4.5 測試拮抗真菌對白菜炭疽病菌的抑制能力.............................32
4.6 測試不同濃度拮抗真菌對白菜炭疽病菌在小白菜的抑制能力..............33
4.7菌種鑑定結果...................................................34
第五章 討論.......................................................35
第六章 結論.......................................................38
參考文獻..........................................................39
附錄1真菌菌種鑑定報告..............................................43
Extended Abstract................................................48

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