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Multiple function Bacillus spp. on control of Pak-choi and strawberry anthracnose
|關鍵字:||Bacillus;多功能;拮抗作用;誘導抗病;Bacillus;multiple function;antagonism;induced systemic resistance||引用:||Adam, M., Heuer, H., and Hallmann, J. 2014. Bacterial antagonists of fungal pathogens also control root-knot nematodes by induced systemic resistance of tomato plants. PLOS ONE 9: e90402. doi: 10.1371/journal.pone.0090402. Aktar, M. W., Sengupta, D., and Chowdhury, A. 2009. Impact of pesticides use in agriculture: their benefits and hazards. Interdisciplinary Toxicology 2: 1-12. Alabouvette, C., Olivain, C., and Steinberg, C. 2006. Biological control of plant diseases: the european situation. European Journal of Plant Pathology 114: 329-341. Andrade Pinto, J. M., Souza, E. A., and Oliveira, D. F. 2010. Use of plant extracts in the control of common bean anthracnose. Crop Protection 29: 838-842. Ann, P. J., Lu, L. S., Chuang, T. Y., and Koa, C. W. 1998. Effect of fruit bagging and mulching on control of mango fruit anthracnose disease. Plant Pathology Bulletin 7: 19-26. 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大部分的Bacillus屬細菌被認為對環境與植物是安全且有益的，於防治作物病害，因具拮抗能力、促進植物生長、競爭及誘導抗病的能力，被視為極具發展潛力的生物防治菌；而含兩個以上防治機制的Bacillus屬菌株，陸續被證實具較佳防治炭疽病效果。對峙試驗結果顯示，自香蕉植體內所得108株Bacillus屬菌株中，有12個菌株具有較佳抑制4種真菌病原菌絲生長的效果。以定量方式測試此12株菌株抑制炭疽病菌菌絲生長的能力，結果顯示，抑制白菜炭疽病菌效果均較草莓炭疽病菌佳。觀察具拮抗性Bacillus屬菌株對炭疽病菌的菌絲形態影響，指出接觸到拮抗菌株之白菜炭疽病菌菌絲變粗且尖端呈現膨大之情形。此外，12株拮抗性菌株中，8株菌株可被檢測到產生伊枯草菌素 (iturinA) 的ituD與lpa-14基因，另有6株可被檢測到產生表面素 (surfactin) 的sfp基因。於生物膜形成測試，12株具拮抗能力菌株皆可形成濃度高的生物膜，唯具形成生物膜濃度與拮抗能力並無明顯正相關。溫室試驗中，連續3週施用108 cfu/ml之12株菌株懸浮液後，有4株菌株可顯著提高白菜地上部鮮重達26%至100%，地下部鮮重則無顯著差異。於誘導抗病試驗結果指出，接種白菜炭疽病菌5天前，分別澆灌108 cfu/ml之Bacillus菌株發酵液於土壤中，其中有3株菌株可顯著降低罹病度34.9%至60.0%；於接種草莓炭疽病菌3天前，分別噴施108 cfu/ml之Bacillus菌株發酵液於第4葉上，其中有5株菌株病斑直徑可從15.5 mm顯著降低為3.6至6.9 mm。評估多功能菌株防治白菜炭疽病菌之效果，顯示經具拮抗與誘導抗病特性R8-25菌株處理白菜植株後，白菜炭疽病罹病度較處理只具拮抗能力之PS6-5菌株低。評估多功能菌株防治草莓炭疽病效果，顯示處理具拮抗與誘導抗病能力菌株之防治效果與具單一拮抗能力菌株無差異，且防治效果皆不穩定。以16S rDNA與rpoB gene對12株具拮抗性Bacillus屬菌株鑑定，9株屬於B. amyloliquefaciens，另3株則屬於B. subtilis。本研究證實，多功能Bacillus屬菌株於白菜炭疽病的防治效果確實較單一功能菌株佳，雖在草莓防治上不明顯，未來可進一步探討最適接種平台，並分析多功能菌株誘導植物抗病性產生之相關反應。
Most of Bacillus species are often considered to be ecofriendly and beneficial to the environment and plants. They are thought to be the most potential candidates as biological control agents due to their capability to reduce crop diseases, including production of antimicroorganism compounds, plant growth promoting activity, resistance-induced activity and competition. Moreover, multiple function Bacillus strains have been reported and showed the potential to control anthracnose diseases. The dual culture test showed that 12 of 108 Bacillus strains from banana plants have ability against 4 plant fungal pathogens. For evaluation of antagonistic Bacillus strains on inhibition of mycelial growth, 12 antagonistic Bacillus strains showed better inhibition ability to C. higginsianum PA-01 than C. gloeosporioides CSG8-2 and E3. Scanning electronic microscopy observation of C. higginsianum PA-01 treated by antagonistic Bacillus strain revealed significant morphological alternation in hyphae, such as bulging and tip swelling. Moreover, 8 of 12 antagonistic Bacillus strains contained iturin A producing gene (ituD and lpa-14 gene), and 6 of them contained surfactin producing gene (sfp gene) by PCR detections. For biofilm formation, all 12 antagonistic strains exhibited strong activity to produce biofilm, while concentration of biofilm seemed not associated with antagonistic ability. Under greenhouse condition, among these 12 antagonistic Bacillus strains, 4 strains significantly increased the shoot fresh weight of pak-choi after soil drench treatment continued 3 weeks, but no difference in root fresh weight. The resistance-induced test showed 3 strains have ability to induce pak-choi against anthracnose by drenching 108 cfu/ml of fermentation broth 5 days before inoculation of C. higginsianum PA-01, which could significantly reduce disease severity 34.9 to 60.0%; and 5 strains have ability of inducing strawberry against anthracnose by spraying 108 cfu/ml of fermentation broth on the forth leaf 3 days before inoculation of C. gloeosporioides E3, which could significantly reduce diameter of lesion from 15.5 mm to 3.6 ~ 6.9 mm. For evaluation efficacy of multiple function Bacillus strain on control of Pak-choi anthracnose, Bacillus strain with antifungal and resistance inducing activity showed better control efficacy than Bacillus strain with antifungal activity only on Pak-choi. Efficacy of control on strawberry treated with Bacillus strain with antifungal and resistance inducing or antifungal only was not significant and not stable. Furthermore, 12 antagonistic strains were identified as B. amyloliquefaciens (9 strains), and the others were B. subtilis (3 strains) based on 16S rDNA and rpoB gene. In this study, multiple function Bacillus strains exactly showed better efficacy on control of Pak-choi anthracnose, though it was not stable on strawberry. Further, develop a stable inoculation platform and defense responses activated by multiple function Bacillus strain on Pak-choi will be carried out in the future.
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