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草业学报 ›› 2014, Vol. 23 ›› Issue (5): 184-192.DOI: 10.11686/cyxb20140521

• 论文 • 上一篇    下一篇

黄花草木樨水浸提液中潜在化感物质的分离、鉴定

邬彩霞*,刘苏娇,赵国琦   

  1. 扬州大学动物科学与技术学院,江苏 扬州 225009
  • 收稿日期:2014-04-15 出版日期:2014-10-20 发布日期:2014-10-20
  • 作者简介:邬彩霞(1978-),女,内蒙古临河人,博士,讲师。E-mail:cxwu@yzu.edu.cn
  • 基金资助:

    国家自然科学基金(31101764)和江苏省高校自然科学基金(11KJB230004)资助

Isolation and identification of the potential allelochemicals in the aqueous extract of yellow sweet clover (Melilotus officinalis)

WU Cai-xia,LIU Su-jiao,ZHAO Guo-qi   

  1. College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China
  • Received:2014-04-15 Online:2014-10-20 Published:2014-10-20

摘要:

本研究旨在分离、鉴定黄花草木樨水浸提液中的主要化感物质。采用色谱分离、薄层分析和生物活性检测相结合的方法分离出了活性较强的化感物质组分,并通过气质联用(GC-MS)对潜在的化感物质进行了定性分析。结果表明,黄花草木樨水浸提液的乙酸乙酯相中含有多种化感物质,相对含量最高的为香豆素,相对含量较高的有6-(3-Hydroxy-but-1-enyl)-1,5,5-trimethyl-7-oxabicyclo[4,1,0]heptan-2-ol、二氢苯并吡喃酮、2,6-二叔丁基对甲酚、二溴三甲基环丙烷、磷酸三甲酯等物质;将乙酸乙酯相进一步分离得到3层混合物,上层混合物中含量相对最高的为香豆素;中层混合物中含量相对较高的有二溴三甲基环丙烷、6-(3-Hydroxy-but-1-enyl)-1,5,5-trimethy-7-oxabicyclo[4,1,0]heptan-2-ol、2,6-二叔丁基对甲酚、甲基环戊醇等物质;下层混合物中含量相对较高的有二溴三甲基环丙烷、甲基环戊醇、环己基溴化膦等物质。乙酸乙酯相及其分离后的各层混合物处理对多花黑麦草种子萌发和幼苗生长均具有一定的化感作用,其中,乙酸乙酯相处理对多花黑麦草化感抑制作用最强,其处理对多花黑麦草种子萌发和幼苗生长的抑制作用显著强于其分层后的各层混合物处理(P<0.05);其次是上层混合物处理,其处理对多花黑麦草种子萌发和幼苗生长的抑制作用显著强于其他2层(P<0.05);下层混合物处理对多花黑麦草的种子萌发和幼苗生长则具有明显的促进作用(P>0.05)。以上结果提示,黄花草木樨水浸提液中化感物质种类非常多,香豆素的含量最高,且香豆素含量高的组分处理对黑麦草的抑制作用就强,推测香豆素应为黄花草木樨水浸提液中的主要化感物质之一。

Abstract:

This study aimed to isolate and identify the main allelochemicals of aqueous extract of yellow sweet clover (Melilotus officinalis). Chromatographic separation, TLC analysis and bioassay test were conducted to isolate the potential allelochemicals of the aqueous extract of yellow sweet clover. GC-MS were used to identify the potential allelochemicals of the aqueous extracts of yellow sweet clover. The results showed that the ethyl acetate phase of yellow sweet clover aqueous extracts containing a variety of allelochemicals, in which coumarin had the highest relative content, then followed by 6-(3-Hydroxy-but-1-enyl)-1,5,5-trimethyl-7-oxabicyclo-[4,1,0]heptan-2-ol, chromanone,2,6-Di-tert-butyl-4-methylphenol, 2-Bromo-1,1,3-trimethyl cyclopropane, trimethyl phosphate, and other substances. The ethyl acetate phase was further separated into three fractions, which were the upper layer, middle layer and lower layer. The chemical which had the highest relative content of the upper layer is also coumarin. In the middle layer, 2-Bromo-1,1,3-trimethyl cyclopropane, 6-(3-Hydroxy-but-1-enyl)-1,5,5-trimethy-7-oxabicyclo[4,1,0]heptan-2-ol, 1-Methylcyclopentanol, trimethyl phosphate and 2-Bromo-1,1,3-trimethyl cyclopropane had higher relative contents. In the lower layer, 2-Bromo-1,1,3-trimethyl cyclopropane, 1-Methylcyclopentanol and cyclohexyl-phosphonous dibromide had higher relative contents. The ethyl acetate phase and the fractions separated from it all showed allelopathic effect on the seed germination and seedling growth of Italian ryegrass, in which the ethyl acetate phase treatment showed the strongest inhibition on seed germination and seedling growth of Italian ryegrass, and significantly stronger than all the treatments of the fractions separated from it (P<0.05). The upper layer treatment showed the second strongest inhibition and significantly stronger than the other two layers (P<0.05). While the lower layer treatment showed significant promoting effects on seed germination and seedling growth of Italian ryegrass (P<0.05). These results suggested that many allelochemicals existing in the aqueous extract of yellow sweet clover, in which coumarin had the highest relative content, and the treatments with higher content of coumarin concentration showed stronger inhibition effect on Italian ryegrass, which also suggested that coumarin should be one of the main allelochemicals in the aqueous extract of yellow sweet clover.

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