CAJ | 학술논문

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-oxabi-cyclo[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。
본연구지재분리、감정황화초목서수침제액중적주요화감물질。채용색보분리、박층분석화생물활성검측상결합적방법분리출료활성교강적화감물질조분，병통과기질련용（GC-MS）대잠재적화감물질진행료정성분석。결과표명，황화초목서수침제액적을산을지상중함유다충화감물질，상대함량최고적위향두소，상대함량교고적유6-（3-Hydroxy-but-1-enyl）-1，5，5-trimethyl-7-oxabicyclo[4，1，0]heptan-2-ol、이경분병필남동、2，6-이숙정기대갑분、이추삼갑기배병완、린산삼갑지등물질；장을산을지상진일보분리득도3층혼합물，상층혼합물중함량상대최고적위향두소；중층혼합물중함량상대교고적유이추삼갑기배병완、6-（3-Hydroxy-but-1-enyl）-1，5，5-trimethy-7-oxabi-cyclo[4，1，0]heptan-2-ol、2，6-이숙정기대갑분、갑기배무순등물질；하층혼합물중함량상대교고적유이추삼갑기배병완、갑기배무순、배기기추화련등물질。을산을지상급기분리후적각층혼합물처리대다화흑맥초충자맹발화유묘생장균구유일정적화감작용，기중，을산을지상처리대다화흑맥초화감억제작용최강，기처리대다화흑맥초충자맹발화유묘생장적억제작용현저강우기분층후적각층혼합물처리（P＜0．05）；기차시상층혼합물처리，기처리대다화흑맥초충자맹발화유묘생장적억제작용현저강우기타2층（P＜0．05）；하층혼합물처리대다화흑맥초적충자맹발화유묘생장칙구유명현적촉진작용（P＞0．05）。이상결과제시，황화초목서수침제액중화감물질충류비상다，향두소적함량최고，차향두소함량고적조분처리대흑맥초적억제작용취강，추측향두소응위황화초목서수침제액중적주요화감물질지일。
This study aimed to isolate and identify the main allelochemicals of aqueous extract of yellow sweet clover (Melilotusofficinalis).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,tri-methyl 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-Hydrox-y-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-tri-methyl cyclopropane,1-Methylcyclopentanol and cyclohexyl-phosphonous dibromide had higher relative con-tents .The ethyl acetate phase and the fractions separated from it all showed allelopathic effect on the seed ger-mination and seedling growth of Italian ryegrass,in which the ethyl acetate phase treatment showed the stron-gest 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.