應用兩種鏈黴菌綜合防治南方根瘤線蟲病害與茄科細菌性斑點病之潛力探討

黃棨揚

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應用兩種鏈黴菌綜合防治南方根瘤線蟲病害與茄科細菌性斑點病之潛力探討

Studies on the potency of applying two species of Streptomyces with chemicals to control Meloidogyne incognita and Xanthomonas vesicatoria

黃棨揚 , Masters　　Advisor：蔡東纂　　Co-advisor ：陳珮臻

繁體中文

鏈黴菌；綜合防治；南方根瘤線蟲；茄科細菌性斑點病； Streptomyces ； Integrated pest managemant ； Meloidogyne incognita ； Xanthomonas vesicatoria

Abstract │ Reference (117) │ Altmetrics

Abstract 〈TOP〉

摘要
本研究測試兩種鏈黴菌Streptomyces plicatus B4-7 (以下簡稱B4-7)、S. saraceticus SS31 (以下簡稱SS31)對南方根瘤線蟲、茄科細菌性斑點病的防治機制，並測試兩個鏈黴菌結合化學藥劑後防治病害之潛力。結果發現鏈黴菌B4-7對測試真菌Pytihum aphanidermatum、Rhizoctonia solani AG4 HGII、Sclerotium rolfsii、Fusarium solani Vn4-2與Alternaria brassicicola有較高生長抑制率。而SS31與B4-7皆 可抑制五種細菌病害。將鏈黴菌B4-7、SS31與4種殺線蟲劑、1種殺蟲劑、4種殺菌劑於PSA平版上測試藥劑相容性，結果顯示殺蟲劑與殺線蟲劑無論是同時與鏈黴菌施用或是延後24小時施用，皆不會影響鏈黴菌生長。但四種殺菌劑在同時施用下會對兩種鏈黴菌不等的抑制效果，其中嘉賜銅、氫氧化銅對B4-7有強烈抑制效果，而SS31則會被鏈黴素抑制，但是延後24小時施用藥劑皆可以提升鏈黴菌與測試藥劑之相容性。以1%燕麥、2%米糠、2%黃豆砂糖三種培養液進行生物防治菌增量，發現B4-7燕麥培養液處理根瘤線蟲卵後得到最低卵孵化率47.8%，且最大有效稀釋濃度為30倍。B4-7三種培養液浸泡南方根瘤線蟲二齡幼蟲的處理組比起SS31處理組皆有較多的線蟲失去活性。將二齡線蟲先以稀釋30倍的B4-7培養液浸泡兩天後，再將無活性蟲體移至無菌水中一天，結果顯示二齡線蟲經B4-7黃豆砂糖培養液處理有19.1%死亡率，另有14.5%的J2可在清水中恢復泳動力，表示二齡線蟲經B4-7培養液處理後，蟲體不活化可能為麻痺而非死亡。為了測試由麻痺回復之蟲體是否仍具有侵染能力，將B4-7燕麥培養液處理後之二齡線蟲接種至菜豆根系，五天後進行根系染色，結果顯示二齡線蟲入侵數分別為13.2與15.4隻，與清水處理J2對照組之42.3與37.6隻相比，數目上有顯著差異，在空心菜以B4-7燕麥培養液處理後，可降低土壤內二齡線蟲數與根系上結瘤率。鏈黴菌SS31之黃豆砂糖培養液在雙層NA平板對茄科細菌性斑點病拮抗效果最佳，有最大抑制圈20.10 mm。將帶菌種子以SS31三種基質的培養液分別浸泡30分鐘後，結果顯示鏈黴菌SS31之2%黃豆砂糖與1%燕麥培養液可使種子帶菌率降為0%。於溫室條件下測試化學防治與鏈黴菌之間共同施用的防治效果，發現SS31黃豆砂糖培養液單獨施用處理組得到最低的致病率23.70%與發病嚴重度7.00%，如鏈黴菌與鏈黴素或三元硫酸銅同時施用則會降低防治茄科細菌性斑點病能力。錯開生物防治菌與化學藥劑施用時間，先施用SS31再施用化學藥劑，比起同時施用組其防治效果在統計上顯著較佳。本研究結果顯示鏈黴菌Streptomyces plicatus B4-7、Streptomyces saraceticus SS31具有生物防治潛力，且可與化學藥劑結合使用，未來可導入茄科病害的綜合防治管理策略中。

Parallel Abstract 〈TOP〉

Abstract
	Streptomyces plicatus B4-7 and S. saraceticus SS31 were tested for their antagonism ability to several important plant pathogens and assessed the feasibility of combining pesticides applications in the IPM program. The biocontrol mechanism of the tested Streptomyces strains on Southern root-knot nematode (Meloidogyne incognita) and bacterial leaf spot disease (Xanthomonas axonopodis pv vesicatoria) were also studied. Streptomyces plicatus B4-7 had better inhibition on the growth of Pytihum aphanidermatum, Rhizoctonia solani AG4 HGII, Sclerotium rolfsii, Fusarium solani Vn4-2 and Alternaria brassicicola than SS31. Both SS31 and B4-7 had inhibition on the  five bacterial pathogens such as Xanthomonas axonopodis pv vesicatoria, X. vesicatoria, X. axonopodis subsp. citri, X. campestris pv. campestris, Acidovorax avenae subsp. citrulli, Pectobacterium carotovorum subsp. carotovorum. Both B4-7 and SS31 strain could not inhibit Acidovorax avenae subsp. citrulli.The two Streptomyces strain were tested with 4 bactericides, 1 insecticide, 4 nematicides for their compatibility. The results showed that insecticide Avermectin and nematicides were compatible with two Streptomyces strains in the in-vitro test. But the four bactericides all inhibited the growth of the 2 biocontrol Streptomyces strains. Copper hydroxide and Kasugamycin-copper oxychloide strongly inhibited B4-7 growth, and Streptomycin inhibited SS31. Postponing the bactericide application 24 hrs could lower the inhibition of four bactercides on Streptomyces B4-7 and SS31. Three media, 1% oatmeal, 2% rice bran, 2% soybean sucrose were used to culture B4-7 and SS31. The egg hatching rate of Southern root-knot nematodes that treated with 30 times dilution B4-7 Oatmeal culture broths had lowest egg hatching rate 47.8%. The J2 of Southern root-knot nematode treated with B4-7 culture broths had higher immobile rates than J2 treated with SS31 broths. When the immobile J2 were moved to sterile water to recover for one day, the results showed that 14.5% immobile J2 could recover in water. To test whether the paralyzed J2 when recovered would have the ability to invade the host, J2 treated with 30X B4-7 oatmeal culture broths were inoculated onto common bean root, and evaluated five days after inoculation. The penetration number of J2 when treated with B4-7 Oatmeal culture broth was 13.2 and 15.4 in 2 experiment, which were significantly different from the blank treatment. S. plicatus B4-7 oatmeal culture broth can control M. incognita on water spinach. Streptomyces SS31 soybean sucrose culture broth had largest inhibit zone 20.1 mm which had the best inhibition on Xanthomonas axonopodis pv.vesicatoria.on double layer NA agar. The seeds coated with Xanthomonas axonopodis pv.vesicatoria. were treated with three culture broths of Streptomyces SS31 for 30 mins, the results indicated that SS31 soybean sucrose and oatmeal culture broths can completely inhibit Xanthomonas axonopodis pv vesicatoria on the contaminated seed. In a greenhouse trail, the single treatment of SS31 soybean sucrose culture broth to control Xanthomonas axonopodis pv vesicatoria on tomato resulted the lowest disease incidence 23.7% and disease severity 7.0%. Appling SS31 culture broth with Streptomycin or Tribasic copper sulfate at the same time resulted higher disease incidence, but postponing pesticides application could improve disease control efficacy. Our studies showed that Streptomyces plicatus B4-7 and Streptomyces saraceticus SS31 were suitable bioagents that might be combined with pesticides application in the integrated pest management of solanaceous crops diseases.

Reference ( 117 ) 〈TOP〉

Asaka, O., and Shoda, M. 1996. Biocontrol of Rhizoctonia solani damping-off of tomato with Bacillus subtilis RB14. Applied and Environmental Microbiology 62(11): 4081-4085.
Akhtar, M., and Malik, A. L. 2000. Roles of organic soil amendments and soil organisms in the biological control of plant-parasitic nematodes: a review. Bioresource Technology 74(1): 35 - 47.
Aghighi, S., Shahidi Bonjar, G. H., and Saadoun, I. 2004. First report of antifungal properties of a new strain of Streptomyces plicatus (strain101) against four Iranian phytopathogenic isolates of Verticillium dahliae, a new horizon in biocontrol agents. Biotechnology (Faisalabad) 3(1): 90-97.
Agrios, G. N. 2005. Plant pathology. 5th ed. Academic Press CA, USA, 922 pp.

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