本文選題：豆類絲核菌 + 培養(yǎng)液��； 參考：《西北農(nóng)林科技大學》2006年碩士論文

【摘要】： 目的:旨在探索豆類絲核菌菌絲體及其次級代謝產(chǎn)物的成分,分析其各成分的用途,為開拓豆類絲核菌的新用途奠定理論基礎。 方法:1.定期用馬鈴薯葡萄糖瓊脂培養(yǎng)基(PDA)移接豆類絲核菌,使之保持旺盛的生長力。2.將特定的豆類絲核菌菌株用改良的Czapek’s培養(yǎng)基進行培養(yǎng),獲得豆類絲核菌培養(yǎng)液和豆類絲核菌菌絲體。3.用不同的方法對菌絲體和次級代謝產(chǎn)物分別進行處理。①對豆類絲核菌菌絲體的處理:將自然干燥的豆類絲核菌菌絲體用0.5%的鹽酸進行浸泡24h后應用40KHz超聲波清洗器在70℃處理30min,收集提取液,如此重復三次,合并提取液。將提取液在60℃條件下減壓濃縮,備用。用2mol/L鹽酸調(diào)提取液為pH 2,加入新鮮配制的雷氏鹽飽和水溶液,有沉淀生成,濾集生成的沉淀,并將沉淀溶于丙酮,濾除不溶物,向丙酮溶液中加入硫酸銀飽和水溶液,使沉淀分解。過濾后向濾液中加入一定量氯化鋇溶液,過濾,濾液用2mol/L氨水將之調(diào)pH 10,揮干得到褐色粘稠狀帶有特異性香味的物質(zhì)。將其在90℃下減壓升華得到白色晶體。②對豆類絲核菌培養(yǎng)液的處理:方法一,取1000ml預處理完備的豆類絲核菌培養(yǎng)液,在60℃減壓濃縮,濃縮液調(diào)pH 2,用正丁醇萃取10次,得到有機相和酸水液。將酸水液調(diào)至pH 10,再用正丁醇萃取10次得到水相和有機相。有機相用0.2mol?L的稀硫酸萃取,得到水相和有機相。將水相用濃氨水堿化后濃縮至干,所得浸膏減壓升華得到白色粉末狀物質(zhì)。方法二,取1000ml預處理完備的豆類絲核菌培養(yǎng)液,調(diào)pH 10,用H2O:CH2Cl2 (3∶1, v/v )萃取,棄去CH2Cl2層,將水層調(diào)pH 10后,再用H2O∶CH2Cl2(3∶1, v/v )萃取,棄去CH2Cl2層,將水層調(diào)pH 7,濃縮后上732型強酸性苯乙烯系陽離子交換樹脂,先用去離子水沖洗,后用1mol/LNH4OH洗脫,收集氨洗液,調(diào)pH 7,濃縮后凍干,得到淺黃色粉末,將之減壓升華得到白色粉末狀物質(zhì)。方法三,取1000ml預處理完備的豆類絲核菌培養(yǎng)液,調(diào)pH7,上大孔吸附樹脂,反復富集5次,靜置12h,用滴水沖洗,洗2個柱體積,收集水洗脫液,再用乙醇洗脫,收集約2個柱體積的醇水混合洗脫液,最后收集2個柱體積的醇洗脫液,分別減壓濃縮各洗脫段的液體,經(jīng)TLC檢測,合并醇洗液,減壓回收乙醇,余液凍干,得到褐色粉末。TLC則可檢出,與標準品對照,均呈現(xiàn)淡紫色,但由于純度較差難以升華。 結(jié)果:經(jīng)過熔點測定儀和GC-MS檢測,初步鑒定豆類絲核菌菌絲體中可能含有蒲公英甾醇、α-香樹脂醇、大狼毒素以及兩種未知的化合物。這幾種物質(zhì)均是首次從豆
[Abstract]:Objective: to explore the composition of the mycelium and its secondary metabolites of Rhizoctonia legume, and to analyze the uses of each component, so as to lay a theoretical foundation for the development of new uses of Rhizoctonia legume. Method 1: 1. The strain of Rhizoctonia legume was transfered on potato glucose Agar medium (PDAs) regularly to maintain its vigorous growth ability. 2. The specific strains of Rhizoctonia legume were cultured on the modified Czapeks medium to obtain the culture medium of Rhizoctonia legume and the mycelia of Rhizoctonia legume. Treatment of mycelium and secondary metabolites by different methods. 1 treatment of mycelium of Rhizoctonia legume: soaking naturally dried Rhizoctonia mycelium with 0.5% hydrochloric acid for 24 hours and then using 40kHz ultrasonic cleaning The scrubber was treated at 70 鈩，

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