Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/30798
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dc.contributor.advisor林益昇zh_TW
dc.contributor.advisorYi-Sheng Linen_US
dc.contributor.author許加宏zh_TW
dc.contributor.authorHsu, Chia-Hungen_US
dc.date2000zh_TW
dc.date.accessioned2014-06-06T07:40:19Z-
dc.date.available2014-06-06T07:40:19Z-
dc.identifier.urihttp://hdl.handle.net/11455/30798-
dc.description.abstract本論文之目的在於鑑定百合鱗片球根腐病之病原,和篩選拮抗細菌作為生物防治。從田間百合罹病株上分離得到之腐霉菌(Pythium spp.),計有Pythium aphanidermatum,P. ultimum,P. spinosum 和 P. splendens等四種。經完成柯霍氏法則證實上述腐霉菌主要對百合鱗片球植株具有強致病力,但對公斤球與開花球植株僅造成根尖壞疽、植株生長矮化等潛隱性病徵(subclinical symptoms)。此外從人工接種可知 P. ultimum 和 P. spinosum 最適發病溫度為16- 20 ℃ 而 P. aphanidermatum 則為 28 ℃,但這三種腐霉菌於田間百合罹病株分離率並不高。然而 P. splendens 在供試溫度16-28 ℃ 下皆對鱗片球植株具有強致病力且田間分離率高,故為百合鱗片球根腐病之主要病原。將百合罹病根部、其根圈土壤、綠力有機肥及蚯蚓排遺物等材料,加入1 % 蔗糖或1000 ppm水楊酸溶液作為處理,能夠獲得較高比率之拮抗細菌。在溫室將百合鱗片球粉衣處理拮抗細菌,並種於含腐霉菌的萬興自然病土中,以分離自百合罹病株根部代號為ADR-s-16之菌株能夠達到保護效果,但在萬興及大坪頂農場的田間試驗中並無特別顯著差異。其原因可能為台糖公司提供之鱗片球遭受污染帶有苗枯病菌(Fusarium moniliforme),或者鱗片球低溫儲藏時間不足,尚未完全打破休眠而導致田間發芽率偏低。zh_TW
dc.description.abstractThe purpose of this study was to determine the cause of root rot of lily plants from scale bulblets, and screening the antagonistic bacteria for biological control. Based on completion of Koch's Postulate, the four species of Pythium including P. aphanidermatum, P. utlimum, P. spinosum and P. splendens isolated from diseased lily plants collected from fields were proved to be highly pathogenic mainly on lily plants from scale bulblets, but only produced subclinical symptoms such as root tip necrosis and stunting on lily plants from yearling and commercial bulbs. Inoculation tests also revealed that the optimum infection temperatures were 16-20 ℃ for P. ultimum and P. spinosum, and 28 ℃ for P. aphanidermatum. These three Pythium spp. were not found frequently on lily plants in fields. However, P. splendens were quite pathogenic to lily plants from scale bulblets among the tested temperatures from 16-28 ℃. Its isolation rate from field specimens was high and thus, it was identified as the major pathogen of this disease. The better isolations of antagonistic bacteria were obtained from lily diseased root and their rhizosphere soil, green-power organic fertilizer and earthworm's casting amended with 1% sucrose or 1000 ppm salicylate solution. One of the antagonistic bacterium designated ADR-s-16 isolated from diseased lily plant when coated on lily scale bulblets before planting could protect the plants against Pythium spp. naturally infested in Wan-Hsin soil in greenhouse tests. However, the results in two field trails carried out in Wan-Hsin and Da-Ping-Ding experimental farms in 1998-1999 and 1999-2000, were not significant difference. One of the reasons for no significance might be due to that the scale bulblets supplied by Taiwan Sugar Company were infested with Fusarium moniliforme or still in dormancy (the incubation period in low temperature might be not long enough to break the bulb dormancy completely) and thus, the germination rate was too low in fields.en_US
dc.description.tableofcontents目錄 中文摘要-----------------------------------------------------I 英文摘要-----------------------------------------------------II 誌謝-----------------------------------------------------------III 目錄-----------------------------------------------------------IV 附表目錄-----------------------------------------------------VII 圖版目錄-----------------------------------------------------X 前言-----------------------------------------------------------1 前人研究-----------------------------------------------------5 材料與方法--------------------------------------------------10 一、病害調查-----------------------------------------------------10 二、病原菌之分離、鑑定與保存------------------------------10 1、病原菌之分離----------------------------------------------10 2、腐霉菌之鑑定----------------------------------------------10 3、病原菌之保存----------------------------------------------11 三、供試菌株、種球及栽培介質來源------------------------11 四、病原性測定--------------------------------------------------12 1、菌絲塊貼覆法----------------------------------------------12 2、種球粉衣接種法-------------------------------------------12 3、土壤混菌接種法--------------------------------------------13 a. 人工病土的製備與菌量偵測----------------------------13 b. 接種方式---------------------------------------------------13 五、溫度及種球大小對根腐病的影響------------------------13 六、拮抗細菌之篩選、測試與保存---------------------------14 1、拮抗細菌之分離--------------------------------------------14 2、拮抗性測定-------------------------------------------------14 3、生物檢定----------------------------------------------------15 4、農藥對拮抗細菌之影響-----------------------------------15 5、拮抗細菌之鑑定-------------------------------------------16 6、拮抗細菌之保存-------------------------------------------16 七、拮抗細菌之標定與存活------------------------------------16 1、拮抗細菌之標定-------------------------------------------16 2、已標定拮抗細菌抑制性之再測試-----------------------17 3、已標定拮抗細菌在百合根部群集與存活--------------17 八、溫室與田間防治試驗---------------------------------------18 1、周徑大小及儲藏時間對鱗片球發芽抽莖之影響------18 2、百合種球、土壤及介質攜帶病原菌之分析------------18 a. 組織分離---------------------------------------------------18 b. 水誘釣法---------------------------------------------------18 c. 土壤稀釋平板法-------------------------------------------18 d. 土球法------------------------------------------------------19 3、接種源製備與種球處理-----------------------------------19 4、溫室防治試驗-----------------------------------------------19 5、1998 ~ 1999 年萬興農場田間試驗----------------------20 6. 1999 ~ 2000年大坪頂農場田間與溫室試驗------------21 結果----------------------------------------------------------23 一、病害調查與病原菌分離------------------------------------23 二、腐霉菌之鑑定------------------------------------------------23 三、病原性測定--------------------------------------------------25 四、溫度及種球大小對根腐病的影響------------------------25 五、拮抗細菌之篩選與測試------------------------------------26 六、拮抗細菌之標定與存活------------------------------------27 七、溫室與田間防治試驗---------------------------------------27 1、周徑大小及儲藏時間對鱗片球發芽抽莖之影響-------27 2、百合種球、土壤及介質攜帶病原菌之分析-------------28 3、接種源製備與種球處理----------------------------------------28 4、溫室試驗----------------------------------------------------28 5、1998 ~ 1999 年萬興農場田間試驗----------------------29 6、1999 ~ 2000年大坪頂農場田間與溫室試驗-----------30 討論----------------------------------------------------------31 參考文獻----------------------------------------------------37 附錄----------------------------------------------------------79 附表目錄 表一、Table 1.表一、從不同地區百合罹病株分離得到之可疑病原菌(1997-1998年)Table 1. Pathogens isolated from lily diseased plants in different areas during 1997-1998-----------50 表二、Table 2.表二、從百合鱗片球罹病株分離得到之可疑病原菌(1998-2000年)Table 2. Pathogens isolated from diseased plants of lily scale bulblets during 1998-2000--------51 表三、Table 3.表三、腐霉菌對百合鱗片球之病原性測定Table 3. Pathogenicity of Pythium spp. to lily scale bulblets------------------------------------------52 表四、Table 4.表四、從不同來源分離得到對百合根部病原菌具拮抗性之細菌菌株比例Table 4. The ratio of bacteria antagonistic to soil-borne pathogens of lily isolated from different sources---------------------------------53 表五、Table 5.表五、拮抗細菌對百合鱗片球形成之影響Table 5. Effect of antagonistic bacteria on the formation of bulblets from lily scales in greenhouse------------------------------------------55 表六、Table 6.表六、農藥對拮抗細菌之影響Table 6. Effect of pesticides on the growth of antagonistic bacteria------------------------56 表七、Table 7.表七、利用抗生素標定拮抗細菌菌株Table 7. Using antibiotics to label antagonistic bacteria on nutrient agar------------------------57 表八、Table 8.表八、經抗生素標定之拮抗菌株對百合根部病原菌之抑之抑制性Table 8. The inhibition zone between the labeled antagonistic bacteria and pathogens on CV8 agar------------------------------------------------58 表九Table 9表九、周徑大小及儲藏時間對鱗片球發芽率之影響Table 9. Effect of circumference and incubation period of lily scale bulblets on their germination in Da-Ping-Ding soil-------------59 表十、Table 10.表十、種植前百合種球之可疑病原菌分離Table 10. Pathogens isolated from lily bulbs before planting----------------------------------------60 表十一、Table 11.表十一、萬興和大坪頂土壤及百合栽培介質之可疑病原菌分離Table 11. The isolation of pathogens from Wan-Hsin, Da-Ping-Ding soils and lily cultivating medium------------------------------61 表十二、Table 12.表十二、拮抗細菌處理百合鱗片球並種於含有 Pythium splendens 之人工病土中之發病率Table 12. Effect of antagonistic bacteria on root rot of lily scale bulblets planted in soils infested with Pythium splendens------------ 62 表十三、Table 13.表十三、在溫室中拮抗細菌對百合鱗片球生長之影響Table 13. Effect of antagonistic bacteria on the growth of scale bulblets in greenhouse---63 表十四、Table 14.表十四、在萬興農場拮抗細菌對百合鱗片球植株生長之影響Table 14. Effect of antagonistic bacteria on the growth of lily scale bulblets at Wan-Hsin experimental farm----------------------------64 表十五、Table 15.表十五、在萬興農場拮抗細菌對百合公斤球植株生長之影響Table 15. Effect of antagonistic bacteria on the growth of lily yearling bulbs at Wan-Hsin experimental farm----------------------------65 表十六、Table 16.表十六、萬興農場田間試驗收穫種球之可疑病原菌分離Table 16. Pathogens isolated from lily bulbs harvested from Wan-Hsin experimental farm---66 表十七、Table 17.表十七、百合鱗片球在大坪頂農場種植後之發芽率Table 17. Germination of lily scale bulblets grown in soil and lily cultivating medium at Da-Ping-Ding experimental farm-----------------67 表十八、Table 18.表十八、在大坪頂農場塑膠布室籃裝介質區拮抗細菌對百合鱗片球植株生長之影響Table 18. Effect of antagonistic bacteria on the growth of lily scale bulblets grown in lily cultivating medium in greenhouse at Da-Ping-Ding experimental farm-----------68 表十九、Table 19.表十九、在大坪頂農場露地紅土區拮抗細菌對百合鱗片球植株生長之影響Table 19. Effect of antagonistic bacteria on the growth of lily scale bulblets grown in soil at Da-Ping-Ding experimental farm--------69 表二十、Table 20.表二十、在中興大學溫室籃裝介質區拮抗細菌對百合鱗片球植株生長之影響Table 20. Effect of antagonistic bacteria on the growth of lily scale bulblets grown in lily cultivating medium in National Chung Hsing University greenhouse----------------70 表二十一Table 21.表二十一、在中興大學溫室籃裝紅土區拮抗細菌對百合鱗片球植株生長之影響Table 21. Effect of antagonistic bacteria on the growth of lily scale bulblets grown in Da-Ping-Ding soil in National Chung Hsing University greenhouse----------------71 表二十二Table 22.表二十二、大坪頂農場田間試驗收穫種球之可疑病原菌分離Table 22. Pathogens isolated from lily bulbs harvested from Da-Ping-Ding experimental farm------72 圖版目錄 圖一、Fig. 1.圖一、不同月份從大坪頂農場土壤中 Pythium spp.之誘釣率(1999-2000年)。Fig 1. The baiting rate of Pythium spp. from Da-Ping-Ding field soil in different months during 1999-2000.-------------------------------73 圖二、Fig. 2.圖二、腐霉菌引起百合根腐病之病徵:(A)於鱗片球上造成莖腐及猝(B)於僅長一葉片植株上造成根腐、葉片黃化及萎凋。Fig 2. Symptoms of root rot on lily scale bulblets caused by Pythium spp.. (A) Stem rot and damping-off; (B) Root rot, leaf yellowing and wilt.------------------------74 圖三、Fig. 3.圖三、溫度和株齡對百合鱗片球根腐病發病率之影響。Fig 3. Effect of temperature and seedling age of lily scale bulblet on disease incidence of root rot caused by Pythium spp..-------------75 圖四、Fig. 4.圖四、拮抗細菌在百合根部之存活。Fig 4. Survival of antagonistic bacteria on lily roots (cv. Casa Blanca) planted in Wan-Hsin and Da-Ping-Ding soils. ----------76 圖五、Fig. 5.圖五、拮抗細菌於523培養基不同時間之O.D.值和菌液濃度。Fig 5. Concentration and O.D. value of antagonistic bacteria cultured in 523 liquid medium for hours.------------------------------77 圖六、Fig. 6.圖六、拮抗細菌處理鱗片球種於溫室萬興土壤中。於 6 個月後收穫之種球,左=處理無菌水;中=處理523 液體培養基;右=處理拮抗細菌 ADR-s-16。Fig 6. Lily bulbs harvested from scale bulblets treated with distilled water (left); 523 liquid medium (middle) and antagonistic bacterium ADR-s-16 (right) grown in Wan-Hsin soil in greenhouse for 6 months.--------------------------78zh_TW
dc.language.isoen_USzh_TW
dc.publisher植物病理學系zh_TW
dc.subjectLily scale bulbleten_US
dc.subject百合鱗片球zh_TW
dc.subjectPythium spp.en_US
dc.subjectBiological controlen_US
dc.subjectSubclinical pathogenen_US
dc.subjectFusarium moniliformeen_US
dc.subject腐霉菌zh_TW
dc.subject生物防治zh_TW
dc.subject潛隱性病害zh_TW
dc.subject苗枯病菌zh_TW
dc.title腐霉菌引起百合鱗片球根腐病之鑑定及生物防治zh_TW
dc.titleIdentification and Biocontrol of Lily Scale Bulblet Root Rot Caused by Pythium spp.en_US
dc.typeThesis and Dissertationzh_TW
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