一年蓬提取物的抗菌活性
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摘要
为了研制安全、高效、经济的植物源杀菌剂,以入侵植物一年蓬(Erigeron annuus)为原料,2种植物病原真菌——黄瓜枯萎病菌(Fusarium oxysporium)和番茄早疫病菌(Alternaria solani)的抑制率为指标,通过单因素及响应面实验,优化得到一年蓬抑菌活性成分的最佳提取工艺:提取时间6 min、料液质量浓度1∶60 g/mL、提取温度60℃;所得提取物依次经石油醚、氯仿、乙酸乙酯、正丁醇萃取得各萃取相,其中乙酸乙酯相对2种病原菌抑制率均最高;乙酸乙酯对黄瓜枯萎病菌及番茄早疫病菌的最小抑菌浓度(MIC)分别为0.75,1.50 mg/mL,对2种病原菌的最小杀菌浓度(MBC)皆为36 mg/mL;经离体叶片实验,乙酸乙酯对黄瓜枯萎病和番茄早疫病均具有一定的预防和治疗效果.表明一年蓬提取物的乙酸乙酯相具有开发成植物源杀菌剂的潜力.
The chemical fungicides were regarded as efficient and economical fungicides, however, long-term usage of chemical fungicides would cause refractory degradation, pesticide residues, spread, environmental pollution, food safety issues and so on. In order to develop a kind of plant-derived fungicide with safe, efficient and economic features, Erigeron annuus of invasive plant were used as raw material. With inhibitory activity on two fungi Fusarium oxysporium and Alternaria solani as indicator, the optimal extraction conditions for E. annuus were determined as microwave power of 500 W, time of 6 min, solid-iquid ratio of 1 ∶60 g/mL, extraction temperature of 60 ℃. Under the optimal conditions, the water-soluble extracts of E. annuus were sequentially extracted with petroleum ether, chloroform, ethyl acetate and n-butyl alcohol. The ethyl acetate fractions showed the highest inhibitory activity on these two fungi, the minimal inhibition concentration(MIC) against F. oxysporium and A. solani were 0.75 mg/mL and 1.50 mg/mL, respectively. And minimal bactericidal concentration(MBC) against F. oxysporium and A. solani were both 36 mg/mL. Furthermore, its vivo assay showed good preventive and curative effects against cucumber wilt and tomato early blight. It was concluded that ethyl acetate fractions of the extract from E. annuus had the potential to develop into plant-derived fungicides.
The chemical fungicides were regarded as efficient and economical fungicides, however, long-term usage of chemical fungicides would cause refractory degradation, pesticide residues, spread, environmental pollution, food safety issues and so on. In order to develop a kind of plant-derived fungicide with safe, efficient and economic features, Erigeron annuus of invasive plant were used as raw material. With inhibitory activity on two fungi Fusarium oxysporium and Alternaria solani as indicator, the optimal extraction conditions for E. annuus were determined as microwave power of 500 W, time of 6 min, solid-iquid ratio of 1 ∶60 g/mL, extraction temperature of 60 ℃. Under the optimal conditions, the water-soluble extracts of E. annuus were sequentially extracted with petroleum ether, chloroform, ethyl acetate and n-butyl alcohol. The ethyl acetate fractions showed the highest inhibitory activity on these two fungi, the minimal inhibition concentration(MIC) against F. oxysporium and A. solani were 0.75 mg/mL and 1.50 mg/mL, respectively. And minimal bactericidal concentration(MBC) against F. oxysporium and A. solani were both 36 mg/mL. Furthermore, its vivo assay showed good preventive and curative effects against cucumber wilt and tomato early blight. It was concluded that ethyl acetate fractions of the extract from E. annuus had the potential to develop into plant-derived fungicides.
引文
[1]王彦杰.番茄早疫病病原菌鉴定及抗病种质资源筛选[D].哈尔滨:东北农业大学,2004.
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[3]李明桃.番茄早疫病的发生规律及其防治技术[J].中国瓜菜,2014,27(4):73-74.
[4]闫霜,吴洪生,周晓冬,等.黄瓜枯萎病生物防治研究进展[J].山东农业科学,2011,(1):86-92.
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[8]黄华,郭水良.外来植物加拿大一枝黄花生理指标的季节动态及其适应意义[J].浙江师范大学学报(自然科学版),2005,28(2):201-205.
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[10]张应烙,龙婉婉,尹彩萍.21种植物花提取物抑菌活性的筛选研究[J].河南农业科学,2005,34(8):60-62.
[11]余柯达,路梅,叶美娟,等.茶花和佛手胶孢炭疽菌对不同类型杀菌剂敏感性研究[J].浙江师范大学学报(自然科学版),2014,37(3):334-338.
[12]MARAN J P,SWATHI K,JEEVITHA P,et al.Microwave-assisted extraction of pectic polysaccharide from waste mango peel[J].Carbohyd Polym,2015,123:67-71.
[13]SHAHWAR D,UR-REHMAN S,AHMAD N,et al.Antioxidant activities of the selected plants from the family Euphorbiaceae,Lauraceae,Malvaceae and Balsaminaceae[J].Afr J Biotechnol,2010,9(7):1086-1096.
[14]SAMAGA P V,RAI V R,RAI K M L.Bionectria ochroleuca NOTL33-an endophytic fungus from Nothapodytes foetida producing antimicrobial and free radical scavenging metabolites[J].Ann Microbiol,2014,64(1):275-285.
[15]CHEN Z F,HE B,ZHOU J A,et al.Chemical compositions and antibacterial activities of essential oils extracted from Alpinia guilinensis against selected foodborne pathogens[J].Ind Crop Prod,2016,83:607-613.
[16]吴翠霞.白藜芦醇及其衍生物的抑菌活性及对番茄早疫病菌的作用机理研究[D].泰安:山东农业大学,2012.
[17]吴双.伏立康唑对植物病原真菌的作用及机理研究[D].哈尔滨:东北农业大学,2013.
[18]方建新.野生一年蓬的开发利用[J].资源开发与市场,2006,22(5):474-475.
[2]赵帅,杜春梅,田长彦.黄瓜枯萎病综合防治研究进展[J].中国农学通报,2014,30(7):254-259.
[3]李明桃.番茄早疫病的发生规律及其防治技术[J].中国瓜菜,2014,27(4):73-74.
[4]闫霜,吴洪生,周晓冬,等.黄瓜枯萎病生物防治研究进展[J].山东农业科学,2011,(1):86-92.
[5]卜元卿,孔源,智勇,等.化学农药对环境的污染及其防控对策建议[J].中国农业科技导报,2014,16(2):19-25.
[6]YOON M Y,CHA B,KIM J C.Recent trends in studies on botanical fungicides in agriculture[J].Plant Pathology Journal,2013,29(1):1-9.
[7]XIE G W,CHEN W G,LIN M Z,et al.Ecological risk assessment with MCDM of some invasive alien plants in China[J].Communications in Computer and Information Science,2009,35:580-587.
[8]黄华,郭水良.外来植物加拿大一枝黄花生理指标的季节动态及其适应意义[J].浙江师范大学学报(自然科学版),2005,28(2):201-205.
[9]VELMURUGAN P,MIN C,LEE S M,et al.Antimicrobial fabrication of cotton fabric and leather using green-synthesized nanosilver[J].Carbohyd Polym,2014,106(12):319-325.
[10]张应烙,龙婉婉,尹彩萍.21种植物花提取物抑菌活性的筛选研究[J].河南农业科学,2005,34(8):60-62.
[11]余柯达,路梅,叶美娟,等.茶花和佛手胶孢炭疽菌对不同类型杀菌剂敏感性研究[J].浙江师范大学学报(自然科学版),2014,37(3):334-338.
[12]MARAN J P,SWATHI K,JEEVITHA P,et al.Microwave-assisted extraction of pectic polysaccharide from waste mango peel[J].Carbohyd Polym,2015,123:67-71.
[13]SHAHWAR D,UR-REHMAN S,AHMAD N,et al.Antioxidant activities of the selected plants from the family Euphorbiaceae,Lauraceae,Malvaceae and Balsaminaceae[J].Afr J Biotechnol,2010,9(7):1086-1096.
[14]SAMAGA P V,RAI V R,RAI K M L.Bionectria ochroleuca NOTL33-an endophytic fungus from Nothapodytes foetida producing antimicrobial and free radical scavenging metabolites[J].Ann Microbiol,2014,64(1):275-285.
[15]CHEN Z F,HE B,ZHOU J A,et al.Chemical compositions and antibacterial activities of essential oils extracted from Alpinia guilinensis against selected foodborne pathogens[J].Ind Crop Prod,2016,83:607-613.
[16]吴翠霞.白藜芦醇及其衍生物的抑菌活性及对番茄早疫病菌的作用机理研究[D].泰安:山东农业大学,2012.
[17]吴双.伏立康唑对植物病原真菌的作用及机理研究[D].哈尔滨:东北农业大学,2013.
[18]方建新.野生一年蓬的开发利用[J].资源开发与市场,2006,22(5):474-475.
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