台灣百合根圈細菌之篩選及灰黴病防治應用研究

Selection of rhizobacteria from Lilium formosanum and the application for the control of Botrytis leaf blight

劉益宏(Yi-Hung Liu)

指導教授 ： 陳昭瑩

國立臺灣大學/生物資源暨農學院/植物病理與微生物學研究所/碩士(2004年)

https://doi.org/10.6342/NTU.2004.01596

摘要

植物在受到生物或非生物因子的刺激，會使植物之生理生化發生改變，造成植物開啟多重複雜的防禦機制，使植物具有對抗非生物逆境及病原菌侵染的能力，以適應環境的變遷。目前在植物體內的防禦機制可分為先天及誘導抗病性；誘導抗病性之區分為systemic acquired resistance (SAR)、induced systemic resistance (ISR)及過敏性反應(hypersensitive response, HR)等。百合灰黴病菌(Botrytis elliptica [Berk.] Cooke)危害百合葉片及花器，為百合重要病害。為開發適用於台灣百合(Lilium formosanum Wall., Formosa lily)的生物防治菌株，自台灣百合根圈採集土壤樣品，以土壤稀釋平板法分離及純化出62株細菌菌株，並經氣相層析儀分析脂肪酸的組成及含量以鑑定菌種。將不同菌株細菌懸浮液澆灌在台灣百合根圈，篩選對百合植株沒有負面影響，甚至略有促進生長作用之菌株，在台灣百合及葵百合(Lilium ‘Star Gazer’)上進行溫室防治試驗，並在台灣百合、葵百合及阿卡波克百合(Lilium ‘Acapulco’)上進行實地田間防治試驗，結果顯示皆能有效地降低灰黴病罹病嚴重度及減少病斑的數目，顯示可以應用根圈細菌保護百合之地上部組織，避免灰黴病的危害；進一步為獲得百合抗病性表現的相關基因或分子標記，以PCR-select cDNA subtraction方法獲得百合受根圈細菌誘導表現之mRNA互補股DNAs，其中有23個cDNA選殖株在根圈細菌處理後有差異性表現。

關鍵字

生物防治 ； 台灣百合 ； 百合灰黴病菌 ； 根圈細菌 ； 誘導系統性抗病

並列摘要

Stimulations of biotic or abiotic factors alter physiological or biochemical reactions of plants to evolve multiple complex defense mechanisms for resistance to adverse stress and pathogen infection. So far, plant defense mechanisms include pre-existed and induced resistance. Induced resistance of plants includes systemic acquired resistance (SAR)、induced systemic resistance (ISR) and hypersensitive response (HR). Botrytis leaf and blossom blight caused by Botrytis elliptica (Berk.) Cooke is an important disease for the bulb and flower productions of Lilium spp. and causes severe losses of yield and value of lilies in Taiwan. For biocontrol of Lilium formosanum Wall., application of rhizobacteria was proposed for practical use. Sixty-two bacterial strains were isolated and purified from L. formosanum rhizosphere soil by dilution plate method and identified according to the fatty acid profiles by MIDI identification system. Bacterial strains promoting the growth of the seedling of Formosa lily, without negative effect, were selected. The greenhouse and field disease control assay showed that the disease severity of Botrytis leaf blight could be decreased effectively by application of bacterial suspension of Bacillus spp. and Pseudomonas spp. in the rhizosphere of lily plants. For cloning of defense-related genes or molecular markers for the induced resistance of L. formosanum, PCR-select cDNA subtraction was implemented to screen out the rhizobacterium-responsive genes. Among 193 clones, twenty-three were differentially expressed as indicated by dot blot hybridization.

並列關鍵字

biocontrol ； Botrytis elliptica ； Formosa lily ； Rhizobacteria ； induced systemic resistance (ISR)

參考文獻

Alström, S. 1991. Induction of disease resistance in common bean susceptible to halo blight bacterial pathogen after seed bacterization with rhizosphere pseudomonads. J. Gen. Appl. Microbiol. 37: 495-501.

Bagnasco, P., de la Fuente, L., Gualtieri, G., Noya, F., and Arias, A. 1998. Fluorescent Pseudomonas spp. as biocontrol agents against forage legume root pathogenic fungi. Soil. Biol. Biochem. 30: 1317-1322.

Barka, E. A., Belarbi, A., Hachet, C., Nowak, J., and Audran, J. C. 2000. Enhancement of in vitro growth and resistance to gray mould of Vitis vinifera co-cultured with plant growth-promoting rhizobacteria. FEMS Microbiol. Lett. 186: 91-95.

Benhamou, N., Kloepper, J. W., Quadt-Hallman, A., and Tuzun, S. 1996. Induction of defense-related unltrastructural modifications in pea root tissues inoculated with endophytic bacteria. Plant Physiol. 112: 919-929.

Berggren, I., van Vuurde, J. W. L., and Mårtensson, A. M. 2001. Factors influencing the effect of deleterious Pseudomonas putida rhizobacteria on initial infection of pea roots by Rhizobium leguminosarum bv. viceae. Appl. Soil. Ecol. 17: 97-105.