【摘要】：近年來藥用植物內(nèi)生真菌是得到新的活性天然產(chǎn)物及研發(fā)新藥的潛在資源,具有重要的研究和開發(fā)利用價值。甘肅省藥用植物資源豐富、分布廣泛,但關(guān)于藥用植物內(nèi)生真菌的研究報道較少,因此,本論文以甘肅省貝加爾唐松草作為研究材料,從其各組織分離純化得到內(nèi)生真菌,初步研究內(nèi)生真菌次級代謝產(chǎn)物的抗氧化、抗菌活性,研究內(nèi)容和結(jié)果如下:采用組織分離法,從藥用植物貝加爾唐松草(Thalictrum baicalense Turcz.)的根,莖,葉,種子中共分離得到18株內(nèi)生真菌,根據(jù)形態(tài)進行初步鑒定,分屬于曲霉屬、頂孢霉屬、鐮刀菌屬、香柱菌屬、鏈格孢菌屬、匍柄霉屬,其中鐮刀菌屬為優(yōu)勢屬,占33.33%。制備18株內(nèi)生真菌的次級代謝產(chǎn)物,利用加熱回流法對干燥菌絲體進行提取,提取液減壓蒸干得菌絲體乙醇提取部位;用乙酸乙酯和正丁醇分別對發(fā)酵液進行萃取,各萃取3次,合并每次萃取液,減壓蒸干,得到發(fā)酵液乙酸乙酯和正丁醇萃取部位。采用DPPH自由基清除法、總還原力法、羥基自由基清除法和微量肉湯稀釋法對貝加爾唐松草內(nèi)生真菌發(fā)酵產(chǎn)物進行抗氧化及體外抑菌活性研究�？寡趸Y(jié)果表明,菌株發(fā)酵液的乙酸乙酯部位普遍具有抗氧化活性,其中菌株Y-3對DPPH自由基清除活性最強,達到91.04%,其活性僅次于Vc,菌株J-2對羥基自由基清除活性最大,達到77.86%,菌株Z-6總還原力較好,Vc的相當含量為190.2μg/mg。抑菌活性結(jié)果表明,菌株Z-1乙酸乙酯部位抑菌活性最好,對金黃色葡萄球菌和地衣芽孢桿菌的MIC值為125μg/mL,對肺炎克雷伯菌、大腸桿菌、無乳鏈球菌、銅綠假單胞菌的MIC值均為250μg/mL;菌株Z-3和Z-4正丁醇部位抑菌活性較好,對6種供試菌的MIC值都達到250μg/mL;菌株Z-7乙醇部位抑菌活性較好,對金黃色葡萄球菌、肺炎克雷伯菌、大腸桿菌和銅綠假單胞菌的MIC值均為500μg/mL,對無乳鏈球菌和地衣芽孢桿菌MIC值為1000μg/m L。利用硅膠和Sephadex LH-20凝膠柱層析,并結(jié)合薄層色譜檢識,從唐松草內(nèi)生真菌J-2發(fā)酵液的乙酸乙酯部位分離純化得到化合物,通過單體化合物的理化性質(zhì)以及核磁共振氫譜、核磁共振碳譜、質(zhì)譜等現(xiàn)代波譜學(xué)手段對其結(jié)構(gòu)進行了鑒定,初步鑒定為1-O-十六碳烷酰-2-O-(9Z-十八碳烯酰)-3-O-(9Z,12Z-十八碳二烯酰)甘油酯(1),其余化合物的的鑒定仍在繼續(xù),由于時間關(guān)系剩余部位的分離、化合物結(jié)構(gòu)鑒定和單體化合物生物活性測定將在后續(xù)工作中進行。
[Abstract]:In recent years, endophytic fungi in medicinal plants are potential resources for obtaining new active natural products and developing new drugs, which have important research, development and utilization value. Gansu Province is rich in medicinal plant resources and widely distributed, but there are few reports on endophytic fungi of medicinal plants. Therefore, in this paper, the endophytic fungi were isolated and purified from the tissues of Pinus tabulaeformis in Gansu Province. The antioxidant and antibacterial activities of secondary metabolites of endophytic fungi were studied. The contents and results were as follows: the medicinal plant (Thalictrum baicalense Turcz.) was isolated by tissue isolation method. A total of 18 endophytic fungi were isolated from roots, stems, leaves and seeds. According to their morphology, they belonged to Aspergillus, Fusarium, columella, Streptomyces and stolymyces, among which Fusarium was the dominant genus. It accounted for 33.33%. The secondary metabolites of 18 strains of endophytic fungi were prepared. The dried fungi were extracted by heating and reflux method, and the ethanol extraction parts of the fungi were obtained by vacuum evaporation of the extract. Ethyl acetate and n-butanol were used to extract the fermentation broth for three times, and the extraction solution was combined with each extraction solution and evaporated under pressure to obtain the extraction parts of ethyl acetate and n-butanol in the fermentation broth. DPPH radical scavenging method, total reduction method, hydroxyl radical scavenging method and micro broth dilution method were used to study the antioxidant and in vitro bacteriostatic activities of endophytic fungal fermentation products of Pinus tabulaeformis. The results of antioxidant activity showed that the ethyl acetate part of the fermentation broth of the strain generally had antioxidant activity, among which strain Y 鈮，

本文編號：2496290