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紫色链霉菌MT7及其胞外代谢产物对产毒柑橘和木瓜腐烂真菌的生物防治

Biological control of toxigenic citrus and papaya-rotting fungi by Streptomyces violascens MT7 and its extracellular metabolites.

作者信息

Choudhary Bharti, Nagpure Anand, Gupta Rajinder K

机构信息

University School of Biotechnology, Guru Gobind Singh Indraprastha University, Dwarka, New Delhi, India.

出版信息

J Basic Microbiol. 2015 Dec;55(12):1343-56. doi: 10.1002/jobm.201500323. Epub 2015 Jul 27.

Abstract

An Indian indigenous, Loktak Lake soil isolate Streptomyces violascens MT7 was assessed for its biocontrol potential both in vitro and in vivo against toxigenic fruit-rotting fungi. Strain MT7 exhibited broad-spectrum antifungal activity against various pathogenic postharvest fungi of citrus and papaya. In shake-flask fermentation, antagonist S. violascens MT7 highly produced extracellular antifungal metabolites in early stationary growth phase in glucose-yeast extract-malt extract (M93) broth. Both extracellular culture fluid (ECF) and its n-butanol extract showed significant broad-spectrum fungal mycelial inhibition of several tested fruit-rotting fungi. Antifungal metabolite was found to be heat stable, nonpeptidic, and polyene type antibiotic. The lowest minimum inhibitory concentration (MIC) of n-butanol extract against Colletotrichum gloeosporioides MTCC 9664 and Aspergillus niger MTCC 281 was 0.0312 and 0.0625 mg/ml, respectively. Purification of n-butanol extract through silica gel chromatography resulted in partial purification of bioactive metabolite and the TLC autobiography revealed the presence of single antifungal metabolite with Rf value of 0.755. In vivo bioassays demonstrated the biocontrol potential of tested biocontrol agents on fruit-rotting fungi. Use of cell suspension of S. violascens MT7, extracellular metabolite(s), and n-butanol extract significantly (p < 0.05) reduced sour-rot development on Citrus reticulata Blanco (oranges) and soft-rot development on papaya fruits. Therefore, these results strongly suggest a high potential for application of S. violascens MT7 and its extracellular metabolites as an effective eco-friendly alternative to synthetic fungicides for controlling toxigenic citrus and papaya-rotting fungi.

摘要

对一株从印度本地洛克塔克湖土壤中分离得到的链霉菌——紫色链霉菌MT7,进行了体外和体内抗产毒果实腐烂真菌的生防潜力评估。菌株MT7对柑橘和木瓜的多种采后致病真菌表现出广谱抗真菌活性。在摇瓶发酵中,拮抗菌紫色链霉菌MT7在葡萄糖 - 酵母提取物 - 麦芽提取物(M93)肉汤的稳定生长期早期大量产生细胞外抗真菌代谢产物。细胞外培养液(ECF)及其正丁醇提取物对几种受试果实腐烂真菌均表现出显著的广谱真菌菌丝抑制作用。发现抗真菌代谢产物具有热稳定性、非肽类且为多烯型抗生素。正丁醇提取物对胶孢炭疽菌MTCC 9664和黑曲霉MTCC 281的最低最小抑菌浓度(MIC)分别为0.0312和0.0625 mg/ml。通过硅胶柱色谱法对正丁醇提取物进行纯化,得到了生物活性代谢产物的部分纯化产物,薄层色谱自传显示存在单一抗真菌代谢产物,其Rf值为0.755。体内生物测定证明了受试生防菌对果实腐烂真菌的生防潜力。使用紫色链霉菌MT7的细胞悬液、细胞外代谢产物和正丁醇提取物可显著(p < 0.05)减少布兰科柑橘(橙子)上酸腐病的发生以及木瓜果实上软腐病的发生。因此,这些结果强烈表明紫色链霉菌MT7及其细胞外代谢产物具有很高的应用潜力,可作为一种有效的生态友好型替代品,用于替代合成杀菌剂来控制产毒的柑橘和木瓜腐烂真菌。

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