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研究生:鍾宜穎
研究生(外文):Chung,I-Ying
論文名稱:百合Rhizoctonia病害之生物防治
論文名稱(外文):Biocontrol of Rhizoctonia disease on lily
指導教授:吳文希吳文希引用關係
指導教授(外文):Wu,Wen-Shi
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:植物病理學研究所
學門:農業科學學門
學類:植物保護學類
論文種類:學術論文
論文出版年:1999
畢業學年度:88
語文別:中文
論文頁數:102
中文關鍵詞:百合立枯絲核菌生物防治牛糞堆肥生態
外文關鍵詞:lilyRhizoctonia solanibiocontrolcattle manureetiology
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擬在本土生產健康百合種球之際,土媒病原菌-立枯絲核菌(Rhizoctonia solani)的存在卻為其生產的限制因子之一,為有效生物防治此種病原菌,乃由各地土壤、百合植株及植體內分得325株細菌,8株真菌,再經由對峙及共同培養篩選出20株細菌,根據在鱗片上競爭佔據檢定,挑出六株菌株進行多次溫室生物檢定,而得到二株有效之拮抗細菌- S26和18,經初步鑑定為Bacillus megaterium,均可顯著地(P=0.05)減少百合植株壞疽及倒伏,百合種球之鮮重及乾重亦較未處理對照組為佳。此外,混合施用低濃度的拮抗菌(104CFU/ml)和殺菌劑difenoconazole(62.5ppm),防治R. solani的效果較單獨施用高濃度殺菌劑difenoconazole(125ppm)為佳,且和健康對照組無顯著性(P=0.05)差異。當百合種植於含牛糞堆肥的泥炭土中,牛糞堆肥不論滅菌與否,其出土率及莖長皆明顯地(P=0.05)較加或不加台糖有機肥料之泥炭土者為佳,其最佳調配比例為2或3份牛糞堆肥混合1份泥炭土,將B. megaterium懸浮液浸泡百合種球而後種植於此比例含病原菌之牛糞堆肥栽培介質中,則百合不論在莖長、根數和病害級數方面,皆明顯地(P=0.05)較未防治之對照組佳。探討R. solani在不同土壤深度(5, 10, 20, 30 cm)、水份潛勢(-1.37, -0.51, -0.23, -0.12 bars)和溫度(20, 25, 30, 35℃)等情況下於百合莖段上腐生存活的能力,每隔10天回收各處理之百合莖段,50天內土壤水份潛勢和R. solani埋在土壤中的天數,對R. solani存活有顯著性地(P=0.05)影響,且兩者互有交感作用存在,而土壤溫度和深度對R. solani之存活並無顯著性地(P=0.05)影響。以FDA的水解程度測量添加不同濃度之B. megaterium懸浮液於土壤後之微生物活性,當土壤微生物活性愈高,相對地R. solani的存活量則愈低。應用B. megaterium可防治R. solani的原因,部份係由於B. megaterium對病原菌生長有明顯的抗生抑制效果,因當拮抗細菌培養六天時,1%濃度代謝液即可有效抑制R. solani的菌絲生長;在掃描式電子顯微鏡下觀察,B. megaterium能附著或定殖於R. solani菌絲上,引起菌絲之變形、膨大和凹陷,甚至會造成菌絲崩潰溶解的現象,嚴重影響該病原菌之正常生長。

The presence of soilborne Rhizoctonia solani is one of the limiting factors for producing healthy lily bulbs. For effective controlling this pathogen thru bioagents, microorganisms were isolated from various soil samples and lily tissue. 325 isolates of bacteria and 8 isolates of fungi were obtained. Among them, 20 isolates of bacteria expressed significantly (P=0.05) inhibitory capability to suppress the growth of R. solani on cultural media by means of dual and concomitant cultures. Six isolates among these 20 isolates had competitive saprophytic ability to colonize lily bulbs and exclude the colonization of R. solani. Among these six isolates, antagonistic isolates S26 and #18 were able to reduce significantly (P=0.05) the disease severity and damping-off. They were identified to be Bacillus megaterium by Biolog system. Besides reducing disease severity, fresh and dry weights of lilies treated with B. megaterium S26 or #18 were significantly (P=0.05) healthier than untreated control. Lily bulbs applied with the mixture of low concentration of B. megaterium (104 CFU/ml) and difenoconazole (62.5 ppm) grew significantly (P=0.05) healthier than bulbs treated with 125ppm difenoconazole but not significantly (P=0.05) different from healthy control. The emergence rate of lilies which were planted in the potting media containing sterilized or unsterilized composted cattle manure was significantly (P=0.05) higher than planting in the potting medium containing Taiwan Sugar Company organic amendment and peat. The best ratio of the components of this potting medium is composted cattle manure:peat = 2:1 or 3:1. Lily bulbs treated with B. megaterium suspension had significantly (P=0.05) longer stem length, more root numbers and less disease index than non-treated control in R. solani-infested potting medium. R. solani cultured on lily stem fragments was placed at different soil depths (5, 10, 20, 30cm), water potentials (-1.37, -0.51, -0.12 bars) and temperatures (20, 25, 30, 35℃) for studying its survivability. Every 10 days, lily stem fragments were removed. This study was last for 50 days. Soil water potentials and number of days that lily stem fragments were buried had the significantly (P=0.05) effect on the survival of R. solani, and there were interactions between them. Soil temperatures and depths had no significantly (P=0.05) effect on R. solani survival. The concentration of B. megaterium was positively correlated with fluorescein diacetate (FDA) concentration. This parameter could reflect the microbial activity in soil. The survivability of R. solani was decreasing with the increasing of microbial activity in soil. 1% of the metabolites of B. megaterium which was cultured in PDB for six days was effective to inhibit the growth of R. solani. Besides, B. megaterium could colonize on the hyphae of R. solani. The B. megaterium-colonized hyphae malformed and lysed eventually.

壹、前言………………………………………………….…….1
貳、前人研究
一、百合病害…………………………………………………………3
二、立枯絲核菌之特性與生態………………………………………9
三、生物防治……………………………………………………….13
四、有機廢棄物在防治植物病害上之應用……………………….15
參、材料與方法
一、百合病害調查………………………………………………….19
二、立枯絲核菌對百合半致死量的測定………………………….19
三、拮抗微生物的分離…………………………………………….20
四、拮抗微生物之篩選…………………………………………….20
五、溫室生物及化學防治效力評估……………………………….21
六、田間生物及化學防治試驗…………………………………….22
七、栽培介質的調配……………………………………………….23
八、拮抗微生物在牛糞堆肥中之防治效果……………………….24
九、立枯絲核菌之生態探討……………………………………….24
十、拮抗微生物活性對立枯絲核菌存活之影響………………….27
十一、拮抗微生物之鑑定………………………………………….28
十二、初步拮抗機制的探討……………………………………….29
肆、 結果
一、百合病害調查………………………………………………….31
二、立枯絲核菌對百合半致死量的測定………………………….31
三、拮抗微生物的分離…………………………………………….31
四、拮抗微生物之篩選…………………………………………….32
五、溫室生物及化學防治效力評估……………………………….32
六、田間生物及化學防治試驗…………………………………….33
七、栽培介質的調配……………………………………………….34
八、拮抗微生物在牛糞堆肥中之防治效果……………………….35
九、立枯絲核菌之生態探討……………………………………….35
十、拮抗微生物活性對立枯絲核菌存活之影響………………….36
十一、拮抗微生物之鑑定………………………………………….37
十二、初步拮抗機制的探討……………………………………….38
伍、討論………………………………………………………74
陸、中文摘要…………………………………………………82
柒、英文摘要…………………………………………………84
捌、參考文獻…………………………………………………86

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