问荆活性物质的提取及对番茄灰霉病菌的抑制作用

Extraction method of active compounds from Equisetum arvense and their inhibition activity against Botrytis cinerea Pers.

通过活性物质跟踪的方法,研究了问荆中的活性成分对番茄灰霉病菌的抑菌活性,结果表明:3种溶剂提取物中,氯仿提取物对番茄灰霉病菌菌丝生长和孢子萌发的抑制率最高,EC50分别为0.52和0.21mg/mL;对氯仿提取物萃取分离得到的4种不同溶剂萃取物中,乙酸乙酯组分对灰霉病菌菌丝和孢子萌发的抑制效果最佳,EC50分别为0.37和0.28mg/mL;经柱层析分离得到的12个流分中,L9流分的抑制菌丝生长作用最强,EC50为0.22mg/mL,L9流分处理灰霉病菌后发现菌丝生长稀疏,菌体细胞膜渗透性增大,电镜图片显示菌丝变形,有瘤状突起。因此,问荆提取物对灰霉病菌具有很强的抑制效果,为新型植物源农药的开发提供了一定基础。

The active substances with fungicidal activity were extracted from Equisetum arvense with three different solvents,petrolium ether,chloroform and ethanol.Crude extract was separated and purified by column chromatography.The active fractions were tracked by fungicidal activity against Botrytis cinerea Pers.The results indicated that the strongest inhibition on the spore germination(SG)and mycelial growth(MG)of Botrytis cinerea were observed in chloroform extract,and the EC50of SG and MG were 0.52and 0.21mg/mL,respectively.Subsequent purification with different polarity solvents produced four fractions by solid-liquid extraction.Amongst those fractions,the one extracted with ethyl acetate showed the highest SG and MG inhibition activity,and the EC50 were 0.37and 0.28 mg/mL,respectively.Silca gel column was used to further separate the ethyl acetate fraction and total of 12sub-fractions were collected.Sub-fraction L9could effectively inhibit MG,and the EC50 was 0.22mg/mL.When treated with L9,fewer amounts of mycelia were produced and the permeability of the cell membrane of mycelia was increased.The mycelia transformation and tumors on the hyphae were determined with electron microscope image.The present research results indicate that Equisetum arvense possess high fungicidal activity and can be development as a natural pesticide.