Enhanced Plant Growth and Their Specificity of Fg-1 Isolate and Other Plant Growth-Promoting Rhizobacteria

本篇論文的目的即在鑑定分離自薑根部之綠膿桿菌Pseudomonas aeruginosa Fg-1並確定其促進嫩薑及胡瓜生長的性質，以及篩選更多分離自胡瓜及番茄之促進植物生長根棲細菌（Plant growth-promoting rhizobacteria, PGPR）供試菌株，以交互接種的方式釐清其是否有促進作物生長專一性。2001-2年間的田間試驗以Fg-1處理嫩薑造成其假莖分枝數增加，並提高產量與品質。在溫室以粉衣接種及/或澆灌法接種Fg-1于胡瓜、番茄及八種葉菜類蔬菜，結果發現僅以粉衣接種胡瓜，28天後其株高明顯優於接種523培養基之對照組。Fg-1並不能在培養皿試驗中促進胡瓜種子胚根生長；但在定溫箱試驗中可以促進2週齡胡瓜苗的根重及株高。Fg-1的細胞懸浮液在滅菌土壤中，或利用經孔徑0.22 μm Millipore mambrane過濾其培養濾液於自然土中接種仍可促進胡瓜生長，指出其促進生長的因子可能為植物生長調節劑一類，但尚未證實。若將Fg-1粉衣接種種薑和胡瓜種子則可在地下部分別纏據達60和56天，且可在胡瓜莖部內生28天，但只能纏據番茄根部14天。分別利用胡瓜及番茄從花蓮胡瓜萎凋病抑病土壤誘釣分離得到更多的促進植物生長根棲細菌（C12及C13源自胡瓜，T128及T130源自番茄），交互接種於胡瓜及番茄種子，發現C13（格藍氏陽性菌株）可促進胡瓜株高生長，但不能促進番茄，而Pseudomonas nitroreducens C12 、P. putida biotype A T128及P. aeruginosa T130則僅能促進番茄而非胡瓜生長。該結果指出本研究之供試促進植物生長根棲細菌（PGPR）菌株沒有寄主專一性，但其作物促進生長專一性是存在的。The purpose of this study was to identify Pseudomonas aeruginosa Fg-1, an bacterium isolated from ginger root, and to demonstrate its ability to promote growth of ginger and cucumber plants. More plant growth-promoting rhizobacteria (PGPR) were obtained from cucumber and tomato roots and screened by cross inoculation method to determine whether their plant growth-promoting ability had plant specificity or not. During 2001-2, field experiments resulted that Fg-1 increased pseudostem numbers of ginger plants and enhanced yields with high-quality products. Cucumber, tomato and 8 kinds of foliage vegetables were inoculated with Fg-1 by seed coating and/or drenching methods in greenhouse. The result indicated that only cucumber plant height was enhanced 28 days after seeding by seed coating approach. Fg-1 could not enhance cucumber seed emergence in Petri-dish test, but could increase its fresh root weight and plant height at 14 days old cucumber seedlings in growth chamber test. Cell suspension of Fg-1 could promote cucumber growth in autoclaved soil, and its cultural filtrate(filtered with 0.22 μm Millipore mambrane)could enhance cucumber growth in natural soil, suggested that the bacterium may release phytohormon as its growth-promoting factor, but had not proved yet. Fg-1 colonized on ginger and cucumber roots for 60 and 56 days, respectively, and was endophytic in cucumber stem for 28 days at least by seed coating approach. However, Fg-1 could not efficiently colonize on tomato root (less than 14 days) nor promote its growth. More rhizobacteria baited out from Haulian fusarium wilt suppressive soil by cucumber (C12 and C13) and tomato (T128 and T130) were submitted for cross inoculation test. The results yielded that cucumber-baited Gram-positive bacterial strain C13 could enhance cucumber growth but not tomato. In contrast, Pseudomonas nitroreducens C12, P. putida biotype A T128 and P. aeruginosa T130 could enhance tomato growth but not cucumber. These results indicated that the tested PGPR isolates in this study had no host specificity but might have growth-promoting plant specificity.中文摘要 --------------------------------------------------------------------------------- I 英文摘要 ---------------------------------------------------------------------------------- II 致謝 ---------------------------------------------------------------------------------------------- IV 目錄 --------------------------------------------------------------------------------------------- V 附表目錄 ---------------------------------------------------------------------------------- VIII 前言 ------------------------------------------------------------------------------------------- 1 前人研究 ---------------------------------------------------------------------------------- 4 材料與方法 ----------------------------------------------------------------------------- 12 一、供試病原菌及促進植物生長根棲細菌之來源與保存----- 12 1. 供試細菌之分離------------------------------------------------------------------------------------- 12 2. 供試細菌在培養基上對腐霉病菌之拮抗性及螢光性測定-------- 13 3. 促進植物生長根棲細菌之保存-------------------------------------------------------- 13 二、促進植物生長根棲細菌之篩選--------------------------------------------------------------- 14 （一）溫室試驗--------------------------------------------------------------------------------------------------------------------------------------------- 14 1. 細菌接種源之製備------------------------------------------------------------------------------- 14 2. 供試土壤之製備----------------------------------------------------------------------------------- 14 3. 應用蚯蚓排遺物促進葉菜類植物生長及其因子------------------------- 14 4. 葉菜類作物接種試驗---------------------------------------------------------------------------- 15 5. 胡瓜接種試驗---------------------------------------------------------------------------------------- 16 6. 蕃茄接種試驗 16 （二）田間試驗--------------------------------------------------------------------------------------------------- 17 1. 2001年Fg-1菌株對嫩薑生長及軟腐病之影響--------------------------- 17 a. 接種原製備-------------------------------------------------------------------------------------- 17 b. 種薑處理及種植----------------------------------------------------------------------------- 17 c. 薑軟腐病病害調查------------------------------------------------------------------------ 18 2. 2002年Fg-1菌株對嫩薑生長及軟腐病之影響---------------------------- 18 三、促進植物生長根棲細菌（PGPR）之作物促進生長專一性-------------------------------------------------------------------------------------------------------- 19 四、促進植物生長根棲細菌之鑑定---------------------------------------------- 19 五、綠膿桿菌Fg-1在作物根部之纏據---------------------------------------- 20 1. Fg-1之標定--------------------------------------------------------------------------------------------- 20 2. Fg-1在胡瓜及番茄根部及胚軸的纏據------------------------------------------- 20 3. Fg-1在薑地下部之纏據---------------------------------------------------------------------- 21 六、無菌狀態下綠膿桿菌Fg-1對胡瓜生長之影響------------------ 21 1. 過濾液試驗-------------------------------------------------------------------------------------------- 21 2. 培養皿試驗-------------------------------------------------------------------------------------------- 22 3. 溫室試驗------------------------------------------------------------------------------------------------- 22 結果 ------------------------------------------------------------------------------------------- 23 一、促進植物生長根棲細菌之分離----------------------------------------------- 23 二、促進植物生長根棲細菌之篩選---------------------------------------------------------------- 23 （一）溫室試驗------------------------------------------------------------------------------------------------------------------------------------------ 23 1. 細菌接種源之製備------------------------------------------------------------------------------- 23 2. 葉菜類作物接種試驗-------------------------------------------------------------------------------------------------------- 23 3. 胡瓜接種試驗---------------------------------------------------------------------------------------- 24 4. 番茄接種試驗---------------------------------------------------------------------------------------- 25 （二）田間試驗--------------------------------------------------------------------------------------------------- 25 1. 2001年促進嫩薑生長田間試驗--------------------------------------------------------- 25 2. 2002年促進嫩薑生長田間試驗-------------------------------------------------------- 25 三、促進植物生長根棲細菌（PGPR）之作物促進生長專一性------------------------------------------------------------------------------------------------------- 26 四、促進植物生長根棲細菌之鑑定---------------------------------------------- 26 五、綠膿桿菌Fg-1在作物根部之纏據---------------------------------------- 26 1. Fg-1之標定--------------------------------------------------------------------------------------------- 26 2. 無菌系統下Fg-1在胡瓜根部及胚軸的纏據-------------------------------- 26 3. 溫室中Fg-1在胡瓜及番茄的纏據-------------------------------------------------- 27 4. Fg-1在薑地下部之纏據---------------------------------------------------------------------- 27 六、高壓殺菌對綠膿桿菌Fg-1促進胡瓜生長之影響------------ 28 1. 過濾液試驗-------------------------------------------------------------------------------------------- 28 2. 培養皿試驗-------------------------------------------------------------------------------------------- 28 3. 滅菌土溫室試驗----------------------------------------------------------------------------------- 28 討論 -------------------------------------------------------------------------------------------- 29 引用文獻 ---------------------------------------------------------------------------------- 37 附表 ------------------------------------------------------------------------------------------- 51 附圖 --------------------------------------------------------------------------------------------- 79 附錄 -------------------------------------------------------------------------------------------- 8

Spectroscopic Characterization on Pt~(4+)Biosorption

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