Study On The Separation And Identification Of Allelochemicals Of The First And Second Chromatography Fractions Of Alcohol Extracts From Parsley Rotten Root Material And Their Induced Resistance On Cucumber Wilt

In this study,parsley rotten root and rhizosphere soil（collectively referred to as parsley rotten root material）were used as test materials.40%alcohol was used as extractant to extract,and obtain two kinds of extract.Through column chromatography,10 fractions were collected respectively.Each fraction was mixed with PDA in the molten state.Then Fusarium oxysporium f sp.cucumerinum was inoculated it after solidification,so as to screen the best fraction for allelopathy effect.The eight best fractions collected from the first and second chromatography were detected by GC-MS to identify the main components and contents.Finally,the eight best fractions were used to study the induced resistance by the root-drenching method in the field.The research results of this study are as follows:1.Through two successive column chromatography,four fractions were collected respectively,eight of which were the best fractions.The fungal colony diameter of each fraction was significantly different from that of alcohol control（40%alcohol）and total control（sterile water）.The allelopathic effect I of RRE9 and RRE10 were 35.73%and 36.47%,and the allelopathic effect Ⅱ were 32.16%and 32.68%;the allelopathic effect I of RRE94,RRE910,RRE101 and RRE102 were 31.92%,34.37%,36.06%and 37.54%,and the allelopathic effect Ⅱ were 26.93%,29.58%,31.37%and 32.98%.The allelopathic effect I of RRSE6 and RRSE10 were 34.62%and 35.19%,and the allelopathic effect Ⅱ were 30.35%and 31.67%,the allelopathic effect of RRSE65,RRSE68,RRSE108 and RRSE109 were 45.15%,44.59%,50.32%and 50.15%,and the allelopathic effect Ⅱ were 41.51%,40.91%,47.03%and 46.85%.2.The first and second alcohol extracts of parsley rotten root materials had a significant effect on the growth and metabolism of Fusarium oxysporum.The activity of cell wall degrading enzyme（including cellulase,β-glucosidase and pectinase）was significantly different from that of control;the content of fusaric acid was significantly different from that of control;（3）the content of MDA and soluble protein was significantly different from that of control;（5）the activity of protective enzyme system（including SOD,cat,pod and PPO）was significantly different from that of control.3.GC-MS analysis was carried out on the eight best fractions of the second alcohol extracts of parsley rotten root materials,and the following allelochemials were detected respectively:Alkanes,phenols,esters and ethers are the main allelochemials in the seco-nd alcohol extracts of parsley rotten root,including 3,3-dimethylhexane（24.60%）,2-bromo nony lhydride（8.22%）,2,4-dimethylheptane（33.5%）,3,5-di-tert-buty lphenol（61.2%）,2,4-di-tert-butylbenzoic acid-5-hydroxy-n-amyl ester（17.4%）,3-meth yl-3-et-hyl-1,2-cyclopropane diamine（1.22%）,ursolic acid（1.78%）,N-4-ethoxyphe nyl-ace-tamide)（28.6%）,2-hydroxy-2-methyl-succinic acid（20.2%）,2,4,7-trishydro xy-1-napht-halenone（18.6%）,methoxyphenamine（13.5%）,Benzoyl metronidazole（9.55%）,M-flu-orophenylalanine（8.2%）,ethaneperoxoic acid,1-cyano-1-[2-（2-phe nyl-1,3-dioxolan-2-yl）ethyl]pentyl ester（2.75%）,diheptyl phthalate（1.58%）,（2E）-3-（4-methylphenyl）-1-p-henyl-2-propene-1-one（1.02%）,2-ethylhexyl vinyl ether（5.79%）,1,2-dipentyldithiane-1,1,2-trioxides（2.8%）,Allyl octyl ether（1.39%）,Glutaric acid（2,4-di-tert-butylph-enyl）monoester（8.05%）,N-[4-（diethylamino）phenyl]ac etamide（1.48%）,Alpha asp-artate phenylalanine（40,7%）,Decanedioicacid,1-methyl ester（11.5%）.The best fractions of the second alcohol extracts of parsley rotten root rhi-zosphere soil are mainly acids,esters and ketones,among which 3,5-di-tert-but y-lphenol（27.8%）and 2,4-di-tert-butylphenol（23.5%）also exist.It also include-s:methyl benzoate（22.33%）,7-chloro-2,3-dihydro-3-hydroxy-5-phenyl-lh-1,4-be nzo-diazepine-2-one（0.19%）,3,4,5-trihydroxy（propyl benzoate 0.83%）,2,4-dime thox-ybenzoic acid（5.63%）,2,Ethyl 4-dimethylbenzoate（23.38%）,allylnonyl o xalat-e（2.96%）,4-methyl-1-undene（0.73%）,decylamine（0.64%）,p-tert-butylphe nylgl-ycidyl ether（0.48%）,5-methyl-3,3-diphenyl-3h-pyrazole（0.23%）,9-ethylant hrac-ene（0.06%）,2,2-trifluoroacetamide（0.4%）,2,5-dimethoxycinnamic acid（21.81%）,n-π（TERT butoxycarbonyl）-L-glutamic acid（1.81%）,3-[2-[4-（6-fluoro-1,2-benzoxazol-3,yl）-1-piperidinyl]ethyl]ethyl]-6,7,8,9-tetrahydro-2-methyl₄ h-pyridin[1,2-α]pyrimidine-4-ketone（1.03%）,5-o-coffee acyl quinic acid（0.36%）,6-fluoro-3-trifluo-trifluomethyl benzoic acid-2-acyl-4,6-dichlorophenyl ester（0.05%）,4-dihydroxybenzoic acid（35.58%）,spirulinic acid（7.94%）,trimethylph osphoryl acetate（4.38%）,1’-（4-chlorophenylsulfonyl）-[1,4’]pipe-ridinyl-4 ’-f ormamide（1.08%）and diethyl malonate（26.63%）.The allelochemicals detected from parsley rotten root materials are mainly esters and acids.4.RRE94,RRE910,RRE101,RRE102,RRSE65,RRSE68,RRSE108 and RRSE1098 of second alcohol extracts of parsley rotten root materials were used to induce cucumber seedlings.The disease index and the effect of induced resistance were as follows:on the 9th day,the disease index of RRE94,RRE910,RRE101,RRE102,RRSE65,RRSE68,RRSE108 and RRSE1098 were 32.65%,32.25%,31.49%,30.89%,30.46%,29.62%,30.16%and 28.76%respectively;the induced resistance effect I were 55.09%,55.56%,56.64%,57.62%,58.11%,59.42%,58.48 and 60.42%;the induced resistance effect Ⅱ were 45.18%,45.77%,47.08%,48.21%,48.42%48.86%,50.39%,49.3%and 51.69%.