Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/31906
標題: 具有防治十字花科蔬菜根瘤病潛力的技術開發
Development of the Potential Techniques for Control of Cruciferous Clubroot Disease
作者: 鄭倩芸
Cheng, Chien-Yun
關鍵字: Plasmodiophora brassicae
Clubroot
十字花科根瘤病
中興一百衍生物
生物防治
蕈狀芽孢桿菌
Plasmodiophora brassicae
crucifer
fast analysis technique
biological control
Bacillus mycoides
出版社: 植物病理學系所
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摘要: 西元1933年,台灣新竹與桃園一帶發現十字花科蔬菜發生葉片失水萎凋與根部畸形腫大之病徵,研究證實係由Plasmodiophora brassicae Woronin引起之根瘤病。近年來,十字花科根瘤病廣泛發生於台灣新竹、桃園、台中及宜蘭等地區,造成十字花科蔬菜的收益鉅減。田間P. brassicae從感染甘藍植株直到出現顯著病徵約需二至三個月,若要調查其各種防治成效卻頗為耗時,因此本文的主要目的即在建立一套快速且能精準評估十字花科蔬菜根瘤病危害的分析技術與標準流程,並利用此技術篩選拮抗微生物與土壤添加物之防病潛力。首先利用甘藍種子播種於病土中8天後進行根部染色,並調查根毛受感染之百分率,結果發現在24 oC接種病原菌濃度108 spores/g dry cultural medium時,主根在2-4公分有最高的感染率;將此區間分成4等分所計算出之數據,證實其與主根2-4公分間所有根毛的感染率相近。利用主根2-4公分間所調查的結果,進行統計分析發現接種濃度與感染率 (Y = 1.4773 + 12.3707 X – 0.4161 X2 , r2 = 0.98)、罹病度 (Y = 2.9764 + 2.3577 X + 1.3008 X2 , r2 = 0.97) 及植株鮮重 (Y = 8.2314 – 0.6148 X , r2 = 0.91 ; Y = 2.2850 – 0.1840 X , r2 = 0.95) 間均具有高度的相關性;至於感染率與罹病度 (Y = 3.0320 – 0.1336 X + 0.0193 X2 , r2 = 0.94) 間亦具有頗高的相關性。進一步採取此評估方法篩選有效防治十字花科根瘤病的微生物和土壤添加物,結果發現拮抗微生物Wt15及Wt16兩菌株可有效降低64.0 %和56.4 %的根毛感染率,並可使罹病度分別減少44.4 %與40.8 %,兩菌株經由生理生化及分子鑑定等結果證明均為蕈狀芽孢桿菌Bacillus mycoides。將兩拮抗菌株以種子粉衣、混拌入介質及澆灌等三種不同施用方式處理甘藍幼苗,結果以B. mycoides Wt15菌株粉衣種子之處理效果最為顯著,不僅較接種根瘤病菌之對照組延遲兩週產生病徵,罹病度亦由100 %降至48.15 %,且分別可提高甘藍地上部與地下部植體的鮮重達5.4與2.3倍;進一步分析整體根系結瘤比例後,結果顯示B. mycoides Wt15菌株粉衣種子之處理組植株的結瘤比為22.57 %,而對照組則為69.53 %。利用B. mycoides Wt15菌株處理本病原菌的休眠孢子36小時後,結果顯示孢子發生縊縮變形的現象,孢子處理48小時後活性由66.3 %降至44.7 %。至於土壤添加物方面,中興一百衍生物雖亦可有效降低53 %的根毛感染率,惟其防治根瘤病的效果與拮抗菌B. mycoides Wt15菌株的處理間並無顯著的協力加成效果。
The clubroot disease of cruciferous crops caused by Plasmodiophora brassicae, is one of the most important soilborne diseases. Infected plants typically exhibit abnormal growth, foliar wilting and swollen roots. In Taiwan, the disease was seriously found in Taoyuan, Hsinshu, Taichung and Ilan, causing huge ecomonic losses in agricultural production. For surveying the clubroot disease severity of cabbage plants in the field, it takes long time and heavy work. Therefore, the main purpose of this study is to establish a fast analysis technique for developing and evaluating the control measures of cruciferous clubroot disease in early stage of the crop. In this study, we found the highest infection occurred within 2-4 cm of the main root at the 8th day of inoculation under 24℃ . Besides, examining the infection of 40 root hairs within 2-4 cm region could stand for the whole infection. On the other hand, we also found that relationships among inoculum density, infection, fresh wieght, root wieght and disease severity were highly correlative. By using the fast analysis technique, we screened the antagonistic bacteria and soil amendments for the disease control. The results showed that Bacillus mycoides Wt15 delayed the disease development over two weeks which could reduce 52 % and 47 % of disease severity and clubbed root ratio. Moreover, B. mycoides Wt15 could also increase respectively 5.4 and 2.3 fold of fresh weight and root weight by seed coating method. The observation under the fluorescence microscopy indicated that B. mycoides Wt15 could reduce 21 % viability of resting spores compared with the control and result in shrinking shape of germinated resting spores. The CH100 derivative (250X) was able to reduce 53 % of root hairs infection, however, it did not show the synergistic effect with B. mycoides Wt15 on reducing disease severity.
URI: http://hdl.handle.net/11455/31906
其他識別: U0005-1408201204461400
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