Isolation And Production Of Triptolide-producing Endophytic Fungi From Tripterygium Wilfordii Hook.f.

Tripterygium wilfordii(Tripterygium wilfordii Hook.F.)Is Celastraceae vine,is a precious traditional medicinal plants,the use of over 700 years of history,a wide range of pharmacological effects in the country,mainly distributed in China 's Fujian,Zhejiang,Anhui,Jiangxi and Taiwan provinces.Endophytic fungi plants commonly found in a variety of healthy plant tissue or organ,in the long-term life of the host plant mutualism body influence each other to form.Micro-ecological systems in the plant endophyte long life in plants,an important impact on the growth of the host plant physiology,relevant studies have shown that plants have beneficial endophytic fungi growth,enhance host plant resistance,enhanced host plant metabolism a variety of biological effects,and some can also be synthesized endophyte same or similar host plant secondary metabolites.Currently,the main source of Tripterygium to digging wild resource-based,and the limited nature of resources tripterygium number,which led to dwindling wildlife resources tripterygium serious impact on local biodiversity and ecological balance.In this paper,the separation of the production of triptolide endophytic fungi,the fungi tripterygium interaction between students,macroporous endogenous fungal effects of synthetic resin triptolide,students interact with endogenous fungal bacteria in triptolide,etc.contents were studied in order to deepen their understanding of Tripterygium endophyte seek an effective way to produce triptolide protect tripterygium wildlife resources.The main contents are as follows:(1)Tripterygium endophytic fungi isolated and identified.In this study,the tripterygium endophytic fungi were isolated,purified,liquid culture broth analysis and detection,and endophytic fungi isolated were screened to obtain a strain producing activity of triptolide strain NS-1.The fungal morphology combined with molecular biology,the strain was initially identified as Fusarium sp.(2)Study of Interaction of Endophytic Fungi to Induce the Synthesis of the Triptolide from Tripterygium wilfordii Hook.f.In this study,the interaction through the establishment of endophyte co-culture system,the synthesis of triptolide screened strongest combination of advantages NS-1+NS-8,which is a synthesis of triptolide cultured alone NS-1 of 1.56 times.After different NS-8 prepared as a crude extract(fermented solution P,the crude mycelium extract Q),added to the end of the exponential growth of the NS-1,in order to study the effects of NS-8 addition on NS-1.It was found that the fermentation broth P greatly promoted the NS-1 synthesis of triptolide,the accumulation of extracellular triptolide in the first seven days maximum 0.2039?g/ml,4.2 times for the control group;mycelium Q in the form of crude extracts did not induce experimental NS-1 to promote the growth,but inhibit growth to a certain extent its normal biological experimental group NS-1 smaller than the CK,CK glucose consumption rate is also lower than the extracellular a prime production outside also significantly lower than CK,the amount of intracellular a prime with CK similar.(3)Macroporous tripterygium endophyte effect on the synthesis of triptolide resin.This study showed that the use of macroporous resin fermentation of NS-1 system in the TPL adsorption,can significantly promote the ability of NS-1 synthesis TPL.The results show that in the NS-1 liquid culture first four days,adding 2.0g NKA-9 resin adsorption harvested after 30h adsorption effect of TPL 's best.(4)By the endophyte tripterygium NS-1 with the isolated 23 endophytic bacteria were cultured tripterygium found endophytic bacteria LY-3 ? NS-1 A synthetic hormone has a good role in promoting co-culture of both the synthesis of triptolide is 1.41 times cultured alone synthesized NS-1 a prime volume.The LY-3 preparation into different elicitors(active elicitor H,inactivated elicitor S),added to the end of the exponential growth of NS-1,in order to study the effects of LY-3 addition on NS-1.The results showed that the active elicitors H joined to the accumulation of triptolide in the culture medium 24h after joining TPL accumulation reaches a maximum 0.0069ug/ml,1.41 times for the control group;join inactivated elicitor S in the experimental group,the accumulation of triptolide promoting effect is not significant,also joined in 24h after reaching the maximum,but its maximum 0.0051 ug/ml was only 1.04 times in CK.