Secondary Metabolites And Their Bioactivities From The Leaves Of Mineriao Stylosanthes (Stylosanthes Guianensis Cv.Mineriao)

Stylo(Stylosanthes guianensias SW)is an excellent tropical legumes(Leguminosae)forage,It takes one third of the tropical and south subtropical Artificial grass forage species in the world.It is also the most important tropical forages in the hot zone,and formed a grass industry development pattern "Alfalfa north,Stylo south".However Collectotrichum gloeosporioides makes forage and seed production declines deeply,or even total crop failure.It has become the devastating diseases of Stylo.The research of resistance to Stylo anthracnose mainly concentrated in the breeding and pesticide control.In terms of its own metabolites,Fu has explored the difference in protein between anthracnose resistance Stylo and anthracnose susceptible Stylo.The results showed that soluble protein has no inhibitory effect on Collectotrichum gloeosporioides,the role of the insoluble protein to Collectotrichum gloeosporioides could not be determined.In enzymatic activity,Under the action of Collectotrichum gloeosporioides,the activity of three enzymes:PAL,PPO and SOD in Stylosanthes guianensis cv.Mineriao increased after slightly decreased,but could not be determined in the Stylosanthes guianensis cv.CIAT2312.The secondary metabolites in Stylo have not been researched.Stylosanthes guianensis cv.Mineriao has a high resistence to Collectotrichum gloeosporioides,MSRA associated with anti-Collectotrichum gloeosporioides that cloned from Mineriao was translated into an amino acid sequence which is similar to the P450,and P450 involved in the biosynthesis of many types of compounds.Therefore,exploring the relationship between its secondary metabolites and Collectotrichum gloeosporioides inhibition is very important to MSRA,Secondary metabolites and anti-Collectotrichum gloeosporioides.The aim of this study is to have a systematic study on Secondary metabolites and biological activity in Stylosanthes guianensis cv.Mineriao.Applying the method of Ultraviolet absorption and taking Rutin as the standard curve,it was measured that the content of flavonoids in the leaves of Stylosanthes guianensis cv.Mineriao which is high anti-Collectotrichum gloeosporioides is as three times as Stylosanthes guianensis cv.Reyan No.5 which is middle anti-Collectotrichum gloeosporioides and Stylosanthes guianensis(Aubl.)Sw which is susceptible to Collectotrichum gloeosporioides.Taking advantage of various techniques and methods of chromatographic,there are 24 compounds were separated from the leaves of Stylosanthes guianensis cv.Mineriao.The structures were authenticated through the spectrum technology.The compounds are:salicylic acid(1),Medicarpin(2),aglycone of crotonionoside F(3),schensiand A(4),Pinoresinol(5),3-methoxycoumestrol(6),stigmast-5-ene-3?,7a-diol(7),glyceryl monolinoleate(8),betulinic acid(9),3-methoxy kaempferol(10),(+)-3-hydroxy-?-ionone(11),soyasapogenol B(12)?indazole(13),Styleguol(14),corchoionol C(15),?-daucosterol(16),caffeic acid methyl ester(17),Stylegu A(18),benzyl-2-O-?-D-glucopyranosyl-2,6-dihydroxybenzoate(19),(+)-isolariciresinol(20),vomifoliol(21),(+)-dehydrovomifoliol(22),5"-Isopropenyldihydrofuran-(2",3":7,8)-5,4'-dihydroxy dihydroflavonol(23),?-sitosterol(24).And the compounds 14,18 were both new compounds.They were both firstly separated from Mineriao Stylosanthes guianensis.Adopting the method of filter paper agar diffusion,the compounds 1-20,23 were screened by anti-Collectotrichum gloeosporioides(Pent.)Sacc activity tests.The result suggested that the compounds 1,2,8,12,13,14,15,17 and 18 had inhibitory activity to pathogenic bacteria of the bacillus.2 sesquiterpene compounds had good inhibitory activity.Doing experiments about F.oxysporum f.sp.vasinfectum,F.oxysporum f.sp.Cuben and F.oxysporum f.sp.niveum on the compounds 1,2,5,6,10,13,14,15 and 17.The result showed that the compound 1 had inhibitory effect on Ralstonia Solanacearum,F.oxysporum f.sp.vasinfectum,F.oxysporum f.sp.Cuben and F.oxysporum f.sp.niveum,the compound 17 had inhibitory effect on F.oxysporum f.sp.cuben.Except that the compound 2 had inhibitory effect on Verticillium dahlia Kleb and the compound 1 had inhibitory effect on Ralstonia Solanacearum and Verticillium dahlia Kleb.Using the method of MTT,the compounds 9 and 23 were screened by tumor cytotoxic activity in vitro.It was found that the compounds 9 had cytotoxic activity to Leukemia tumor cell line(K562)and Human hepatoma cell line(BEL).In accordance with the method of Ellman,the compounds 4,5,6,9,10,19,20 and 23 were tested by external neuroprotection.The result suggested that the compounds 9,10 and 23 had inhibitory activity effect to acetylcholinesterase.This research was studied Stylosanthes guianensis cv.Mineriao from the point of view of chemical composition and biological activity.The research was not only abundant the chemical composition of Stylosanthes guianensis cv.Mineriao,found that the active ingredients of Stylosanthes guianensis cv.Mineriao have the bioactivity of anti-Collectotrichum gloeosporioides(Pent.)Sacc.,anti tumor and neuroprotection,etc,but also offer a condition for establishing the way of identifying the Stylo's resistance against Collectotrichum gloeosporioides(Pent.)Sacc.by measuring the type and content differences of secondary metabolites which is anti-Collectotrichum gloeosporioides(Pent.)Sacc.At the same time,the research lays the foundation for further exploration of the relationship between MSRA and secondary metabolites.