Title

鏈黴菌Streptomyces sp. S1菌株生物製劑於強化殺菌劑防治荔枝露疫病Peronophythora litchii之應用性

Translated Titles

Application of Streptomyces sp. strain S1 as a bioagent for strengthening the effectiveness of fungicides for controlling Peronophythora litchii on Litchi

Authors

曾曉慈

Key Words

荔枝露疫病 ; 鏈黴菌 ; 生物防治 ; 殺菌劑 ; Peronophythora litchii ; Streptomyces sp. ; biocontrol ; fungicide

PublicationName

中興大學植物病理學系所學位論文

Volume or Term/Year and Month of Publication

2016年

Academic Degree Category

碩士

Advisor

曾德賜

Content Language

繁體中文

Chinese Abstract

荔枝露疫病由病原Peronophythora litchii所引起,此病害為影響台灣荔枝產量及品質最主要的限制因子。目前慣性管理荔枝的商業化生產主要仰賴施用已知多是傾向廣譜性作用而非專一於卵菌綱特殊標靶之化學殺菌劑。近期監控報導指出在台灣一些廣譜性作用殺菌劑對於P. litchii的控制效果降低。現在迫切需要發展有效控制病害,同時提升環境友善及減低對農藥需求的防治措施,以提高荔枝產業的國際競爭力。本篇研究主要目的為探討整合具拮抗性之Streptomyces sp. S1菌株作為添加劑來強化化學殺菌劑的防治藉此降低對其的需求之可能性。Streptomyces sp. S1菌株已知展現優良潛力作為控制植物病害尤其以土棲真菌或類真菌之微生物殺菌劑。其對於病害防治的效果主要作用部分為超寄生作用,已證實與幾丁質分解酵素及葡聚醣分解酵素的活性有關。廣效篩選化學抗性,在濃度高達1000 ppm下,S1的生長不受14種普遍應用於卵菌綱供試殺菌劑的影響。在14種供試殺菌劑中,S1混合滅達樂80 ppm、氟比來40ppm、曼普胺40ppm、達滅芬1 ppm應用於P. litchii的菌絲生長均具加成抑制的效果。在抑制孢囊發芽的情況,S1分別混合應用滅達樂10 ppm、三氟敏 1 ppm、克收欣 1 ppm及安美速1 ppm表現出顯著加成效果。還有荔枝離葉於接種前24小時處理S1分別混合滅達樂、氟比來、賽座滅及達滅芬,降低感染的發生趨近於零。對抗P. litchii侵染荔枝果實也表現具加成之效果,稀釋10倍濃度的氟比來、賽座滅及達滅芬各別混合S1均展現顯著強化效果。再來於離葉系統測試,噴灑處理100倍S1發酵稀釋液,有效阻止病勢發展長達12小時。掃描式電子顯微鏡檢視由S1發酵液處理後,對於目標病原造成菌絲及孢囊外觀產生型變。從S1菌絲侵入露疫孢囊及造成菌絲扭轉證實產生超寄生作用。然而,以AlamarBlue® 染劑作為分子探針,檢測S1培養液結合賽座滅添加處理對P.lithii能量代謝影響,其不具對還原力降低之加成效果。綜合上述,累積證據指出S1可以作為綜合管理荔枝露疫病的一大助力。如此結合生物製劑與殺菌劑,可有效強化病害防治,並降低殺菌劑的使用量,減少對環境的破壞。

English Abstract

Downy blight disease caused by Peronophythora litchii has long been a major limiting factor severely impacting the yield and quality of litchi in Taiwan. The control measures currently practiced in litchi commercial production rely mainly on application of chemical pesticide mostly known with broad-spectrum activity rather than that specifically targeting only oomycetes. In recent monitoring study theses broad-spectrum fungicide have been reported lower effect on controlling of P. litchii. Development of effective disease control measure with improved environmental friendliness and reduced need of pesticide application is urgently needed for strengthening the international competitiveness of the litchi industry. The main objective of this study was to explore the feasibility of integration of an antagonistic Streptomyces sp. strain S1 as an additive to strengthen the effectiveness of chemical control thus to reduce the need of its application. Additional benefit includes reduction of chemical residues, environmental impact and risk of resistance. The Streptomyces sp. strain S1 was shown of great potential as a microbial bio-fungicide for the control of plant diseases-especially those caused by soil borne fungal or fungal-like pathogens. The effectiveness of disease control was known due greatly in part to the mycoparasitic effect rendered by the superior chitinase and glucanase activities of the bacteria. Screening of chemical resistance indicated that the growth of S1 was not affected by a total of 14 tested fungicides widely adapted for oomycetes at the concentration up to 1000 ppm. Among these 14 tested fungicides, the mixed application of metalaxyl at 80ppm, fluopicolide at 40 ppm, mandipropamide at 40ppm or dimethomorph at 1ppm, each respectively together with S1 showed additive inhibitory effect of the bacteria on the mycelial growth of P. litchii. As for inhibitory effect on sporangium germination, the mixed application of metalaxyl at 10 ppm, trifloxystrobin at 1 ppm, kresoxim-methyl at 1ppm or amisulbrom at 1ppm, each respectively together with S1 also showed remarkable additive effectiveness. And a pretreatment of metalaxyl, fluopicolide, cyazofamid, and dimethomorph each respectively together with S1 on detached leaves 24 hours before inoculation reduced the infection to nearly nil. An additive control effectiveness against P. litchii infection on litchi fruits was also demonstrated where that fluopicolide, cyazofamid and dimethomorph each respectively at 10X-dilution dose were sprayed with the addition of S1 broth culture. Futhermore, on a detached leaf system, spray treatment of S1 at 100X-dilution was shown to deter the disease development for 12hr. Scanning electron microscopy showed morphogenetic impact with S1 broth treatment caused substantial damages to the oomycete mycelium and sporangium. The mycoparastic effect was manifested by S1 penetrated in sporangium and caused twist mycelium. However, with the addition of AlamarBlue® as indicative molecular probe, the culture filtrate from S1 combined with cyazofamid treatment on the energy metabolism of P. litchii was demonstrated no strengthen effect. Taken together, the accumulated evidence indicated clearly the value of S1 as an aid for the integrated management of the downy mildew disease on litchi. Such combined use of biocontrol agent with fungicides can strengthen effect on disease control and reduce the dosage of fungicides, thereby reducing environmental risks.

Topic Category 農業暨自然資源學院 > 植物病理學系所
生物農學 > 植物學
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