Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/31248
標題: 台灣金線連莖腐病之生物防治
Biocontrol of stem rot of Taiwan Anoectochilus caused by Fusarium oxysporum Schl.
作者: 蔡金池
Tsai, Chin-Chih
關鍵字: 生物防治
Biological control
台灣金線連
鐮胞菌
木黴菌
內轉錄區間親緣性
海藻酸鈣
羧甲基纖維素
過氧化酵素
苯丙氨酸解氨酵素
β-1,3-葡萄聚醣酵素
幾丁質分解酵素
可溶性酚化物
細胞壁鍵結酚化物
木質素
誘導抗病性
Anoectochilus formosanus Hayata
Fusarium oxysporum Schl
Trichoderma asperellum
ca-alginate
sodium carboxymethyl cellulose
CMC
peroxidase
phenylalanine ammonia lyase
chitinase
β-1,3-glucancase
free soluble phenolic compounds
cell wall incorporated esterify phenolic compounds
lignin
induced systemic resistance.
出版社: 植物病理學系所
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摘要: 由埔里地區金線連根圈栽培介質分離所得之拮抗性木黴菌 (Trichodermas spp.)菌株,經由菌株生長速度、產孢測試與鐮胞菌Fusarium oxysporum F66菌株在馬鈴薯蔗糖平板行對峙培養測試其拮抗能力,以及對金線連生長促進性評估,獲得五株木黴菌拮抗菌株,隨後在南投埔里地區進行金線連莖腐病系列田間防治試驗,TA菌株為五菌株中對植株產量提升與莖腐病防治效果表現最好。在溫室試驗中,TA菌株不同濃度分生孢子對金線連莖腐病防治效果評估,顯示菌株分生孢子製劑施用濃度越高對莖腐病防治效果越明顯。以每克栽培介質施用5.7Χ106-107 分生孢子之TA菌株處理,在處理後15週莖腐病發病率僅約10%,相對於未處理之對照組發病率則達93.3%。施用方式以分生孢子製劑行根部被覆處理或栽培介質混拌處理,明顯優於厚膜孢子製劑處理。於栽培場試驗,分生孢子混合羧甲基纖維素製劑行根部被覆處理者 (CoCMC),在其種植後九週對莖腐病防治效果幾達100%,麩皮於CoCMC中之進一步添加處理,則有助於介質中木黴菌族群菌量維持在每克107 孢子左右,伴隨之則可見莖腐病之感染明顯受到抑制,試驗結果顯示TA菌株確在金線連莖腐病生物防治上極具應用價值。TA菌株生物特性研究顯示,菌絲生長最適與最高溫度分別為30℃與37℃,能產生厚膜孢子,與利用蜜三糖作為碳源,可產生異腈類 (isonitrile) 抗生物質及明顯的椰子氣味,分生孢子梗多為對稱分枝,其上著生之瓶狀枝呈瘦長瓶壺形,分生孢子圓形或近卵圓形,表面具有明顯不規則金字塔型疣狀突起,依上述形態、生理特性,及其rDNA ITS序列與NCBI資料庫既有木黴菌菌株序列資料比對其親緣關係,皆證實TA菌株歸屬於T. asperellum Samuels, Liechfeldt & Nirenberg。探討木黴菌TA菌株防治鐮胞菌引起之金線連莖腐病是否與誘導金線連產生抗病性反應有關,在無菌培養系統中,金線連根部經TA菌株孢子懸浮液 (8.3X105 conidia/ml) 處理後,金線連之根、莖、葉組織中的過氧化酵素、苯丙氨酸解氨酵素、幾丁質分解酵素與β-1,3-葡萄聚醣水解酵素活性與細胞中累積的酚化物以及木質素等,皆較未處理TA菌株之對照處理為高,以TA菌株培養在1% (w/v) 鐮胞菌胞壁或幾丁質培養基之濾液處理也得到相似的結果,而未處理TA菌株之對照組與未加1% (w/v) 鐮胞菌細胞壁或幾丁質培養基之濾液處理組則未見有相同的結果。金線連根部處理人工合成誘導後天系統性抗病物質2,6-dichloroisonicotinic acid,在金線連之莖、葉組織皆可測得抗病物質之增加,證實金線連本身具有後天系統抗病性。TA菌株孢子懸浮液處理金線連根部,其根、莖、葉組織皆可測得較對照處理為高的抗病物質,證實T. asperellum TA菌株誘導金線連後天系統抗病性反應為TA菌株防治金線連莖腐病的重要因素。
For the biological control of stem rot of Anoectochilus formosanus Hayata caused by Fusarium oxysporum Schl., a total of 5 antagonistic Trichoderma strains were isolated from A. formosanus rhizospheres in Puli. The potential of the obtained strains as biocontrol agent were evaluated by dual culture assay against Fusarium oxysporum Schl. F66 strain on potato sucrose agar. The characteristics screened included inhibition of mycelial growth for the pathogen, the growth and sporulation of the antagonists, and the growth promoting effect on target plant. A followed field trial at Puli, Nantou, aimed for the control of Fusarium stem rot of A. formosanus indicated Trichoderma TA strain was the best among the five strains in regard to the yield and the disease control effectiveness. In a greenhouse trial, the effectiveness of stem rot control was shown dependent on the conidial concentration of the antagonist applied. The stem rot incidence of test plants grown in substrates treated by 5.7Χ106-107 cfu/gram conidial preparation 15 weeks after treatment was approximately 10%, whereas that of the compared non-treated control was about 93.3%. In a serial field trials, the effectiveness of disease control was consistently found better by root coating or preplanting substrate blending with the conidial suspension than that by the chlamydospore preparation of the antagonist. A conidial formulation with carboxymethyl cellulose (CoCMC) was most effective for the disease control in practical cultivation. By root coating during transplanting, the application of CoCMC resulted in nearly 100% protection of the test plants for 9 weeks, whereas the disease incidence of non-treated control at the same time reached about 82 %. The protective effect of CoCMC appeared to be further strengthened by the addition of wheat bran as a food base. The Trichoderma/ Fusarium population dynamics contained in the growth substrates indicated a well sustained Trichoderma population at approximately 107 cfu/g throughout the test period. And in accompany to that was the decrease of disease incidence. These results suggest T. asperellum TA strain have an excellent potential to be used as biocontrol agents for the control of stem rot disease of A. formosanus. Investisation of the biological characteristics of TA strain indicated that the fungus produced chlamydospores on culture and showed optimum and maximum mycelial growth temperature at 30 and 37℃, respectively. It may utilize melezitose as sole carbon source and produce antifungal isonitrinic acids and unique coconut smells. Also, upon conidiation, it produced conidiophores predominantly in paired branching, and among them bore slender-ampulliform phialides with characteristic viride-type morphology. The conidia were ovoid to subglobose structure covered with conspicuous irregular pyramidal warts. The morphological characteristics indicated TA strain a member of T. asperellum (Samuels, Liechfeldt & Nirenberg). By polymerase chain reaction, the rDNA internal transcribed spacer (ITS) sequence of the TA strains was amplified and sequenced. Results obtained from the pair-wise comparison of the sequence data of comparative strains available from NCBI and the phylogenetic analysis supported fully the above species identification based on the morphological characteristics. The involvement of induced resistance in the effectiveness of T. asperellum TA strain as a biological agent for controlling stem rot of A. formosanus caused by F. oxysporum Schl. was explored. For A. formosanus grown in a gnotobiotic culture system, inoculation of a TA strain germinating conidial suspension (8.3X105 conidia/ml) was shown leading to transcient but significant increases of activities of phenylalanine ammonia lyase, peroxidase, chitinase, and β-1,3-glucancase, and as well the accumulation in cell wall free soluble phenolics, and lignin among leaves, stem and root tissues of treated A. formosanus. Similar stimulatory effects were also observed on test plants treated with culture filtrate of TA strain grown in 1% (w/v) cell wall of F. oxysporum or chitin. Whereas that among control plants treated with non-cultured broth medium were not detected. In the case of changes of phenylalanine ammonia lyase and peroxidase activities, the stimulatory effect was also detected by treatment with 2,6-dichloroisonicotinic acid- the chemical known to provoke systemic acquired resistance on various crop species. The stimulatory effects detected on stem and leaf tissues indicated the systemic effect of the treatment which was applied primarily on the roots. The accumulated data indicated the importance of systemic aquired resistance as a major factor contributing to the effectiveness of biological control of stem rot disease by T. asperellum TA strain.
URI: http://hdl.handle.net/11455/31248
其他識別: U0005-2108200814463800
文章連結: http://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-2108200814463800
Appears in Collections:植物病理學系

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