西芹醇层物对黄瓜枯萎病菌的化感作用及化感物质鉴定

Allelopathic Effects of Fresh Parsley Root Ethanol Extracts on Fusarium oxysporum f.sp. cucumberinum and Allelochemicals Identification

枯萎病是黄瓜栽培过程中较为严重的真菌病害,目前对该病防治非常困难。生产中发现西芹与黄瓜轮作能够降低黄瓜枯萎病的发病率。为探明西芹鲜根中对黄瓜枯萎病菌F.oxysporum f.sp.cucumerinum有抑制作用的化感物质成分,利用连续柱层析法对西芹鲜根乙醇浸提液进行了4次柱层析,每次层析后获得的流分与黄瓜枯萎病菌共培养,测定菌落直径并计算其化感效果,以化感抑制效果筛选最佳流分;然后将最佳流分与黄瓜枯萎病菌在液体培养基中共培养,测定其菌体分泌的镰刀菌酸的含量;最后对第4次层析最佳流分进行GC-MS检测。结果表明,第4次层析最佳流分作用于黄瓜枯萎病菌后,对其菌落生长及分泌镰刀菌酸的能力均产生抑制作用;化感效果分别达到了54.59%、52.35%、51.55%和50.97%,镰刀菌酸含量下降到38.09%、59.54%、36.78%和36.65%;第4次层析最佳流分中,共鉴定出酸类、酯类、酚类、醇类及含氮化合物等5类16种化感物质,分别为十六烷基胺、16-甲基十七烷酸甲酯、十六酸、2,4-二甲基-苯并[b]喹啉、D-异薄荷醇、2,6-二叔丁基对甲酚、α-阿拉伯呋喃糖苷、硬脂醇乙酸酯、9,12-十八碳二烯酸、2-羟基丙酸、顺-9-十六(碳烯酸)、月桂酸乙酯、芴-2-偶氮-2,4-二羟基苯、甘露糖醇月桂酸酯、薄荷呋喃、4-甲氧基水杨酸。研究获得的结果可为西芹提取物防控黄瓜枯萎病提供理论依据。

In field production of cucumber Cucumis sativus, it is relatively difficult to control wilt caused by Fusarium oxysporum f. sp. cucumberinum. However, the extent of infection was reduced by crop rotation with parsley Apium graveloens. To identify inhibitory allelochemicals released into the soil by parsley crop, ethanol extracts from fresh parsley roots were examined after a repeated series of purification （four cycles） in column chromatography using a column with silicone coating. We used a bioassay that incorporated the various column fractions into PDA medium and co-cultured the plates with F. oxysporum. By measuring colony diameter and spores germination rate, we screened the best fractions in terms of allelopathic inhibition effect and identified further allelochemicals of the fourth chromatography using GC-MS. Optimal fractions （RE21101, RE2955, RE6235 and RE6865） obtained from the fourth cycle of chromatography showed considerable allelopathic effect. The allelopathic inhibition effects of the four optimal fractions on hypha growth compared with the fourth cycle tomographic control （CK4）, were 54.59%, 52.35%, 51.55% and 50.97%, respectively. In comparison to CK4, the contents of fusaric acid respectively were decreased to 38.09%, 59.54%, 36.78% and 36.65%. Total 16 types allelochemicals, including N,N-dimethyl-1-hexadecanmine, heptadecanoic acid, 1,6-methy-, methyl ester, hexadecanoic acid, 2,4-dimethyl-benzo[b]quinoline, （＋）-isomenthol, butylated, hydroxyl, alpha-arabinofuranoside, acetic acid octadecyl ester, 9,12-octadecadienoic acid, 2-hydroxy acid, cis-9-hexadecyl （carbon） acid, fluoren-2-azo-2, 4-dithdroxybenzene, mannitol laurate, menthofuran, 4-menthoxysalicylaldehyde, can be classified into 5 forms matters including organic acids, phenols, alcohols, esters, and nitrogenous compounds, were identified by the GC-MS analysis. The results provided the needed theoretical basis for the prevention and control Fusarium wilt of cucumber by using extracts of parsley.