三种野生食药用真菌的驯化和培养特性研究

发布时间：2018-03-19 01:36

本文选题：茶呰子叶状层菌　切入点：粉托鬼笔　出处：《鲁东大学》2017年硕士论文　论文类型：学位论文

【摘要】：茶呰子叶状层菌、粉托鬼笔、短裙竹荪均是野生食药用真菌。据研究,茶呰子叶状层菌与金银花的功效有相似之处,可用于消炎、解毒、止痛等,目前已被制成保健品制剂,对各类炎症及感冒有极好的疗效。粉托鬼笔首先由本实验室进行研究,在本实验室中已栽培出子实体,但尚未进行大规模的栽培。短裙竹荪是一种味道极佳的食药用真菌,具有降血脂,调节脂肪酸预防高血压的作用。目前,这三种野生食药用真菌采摘量有限,尚未进行过系统的菌丝培养研究,也限制了它们的药理研究。本论文进行了茶呰子叶状层菌、粉托鬼笔的菌种分离及鉴定、固体培养特性和液体发酵培养条件及菌丝化学成分含量研究。短裙竹荪的固体培养和发酵罐发酵培养条件及菌丝化学成分含量研究。为以后进行大规模的工业发酵生产做好准备工作。首先对采集的野生茶呰子叶状层菌子实体进行菌种分离及菌种鉴定,确定组织分离所得菌丝体即为茶呰子叶状层菌菌丝体。采用固体皿培法研究固体培养特性:发现茶呰子叶状层菌菌丝体在麦麸、麦芽汁、胡萝卜、PDA、GPC、GPY固体培养基上均能生长,在麦芽汁固体培养基上生长速度最快,在第50天时的麦麸固体培养基上生物量最大,在第40天时GPY培养基上的单位质量的菌丝体中三萜含量为0.39%,考虑获得三萜的总量最高,应选择60天的麦麸培养基;茶呰子叶状层菌对碳源和有机氮源的适应性较强,在麦芽糖、果糖、木糖、乳糖上生长较好,除了在尿素中不能生长,在其他氮源中均能生长。采用摇瓶发酵法筛选发酵培养基:茶呰子叶状层菌在只有碳源+氮源+生长因子的培养液中也能生长,但在麦芽汁培养液中生长最好,麦芽汁培养液适合作为茶呰子叶状层菌的种子培养液和发酵培养液;在摇瓶发酵中,三萜含量不低于1%,多糖含量在15%以上,第7天的生物量达到最大,同时采用HPLC测得腺苷获得量最多。随着液体发酵时间的延长,茶呰子叶状层菌菌球颜色逐渐由乳白色变为棕褐色,且发酵液颜色也逐渐变为棕褐色,并有一股清新的果香味。本研究采用组织分离法对粉托鬼笔菌种进行菌种分离并进行分子生物学鉴定得出分离所得菌丝体为粉托鬼笔。粉托鬼笔在不同固体培养基上均能生长,在麦麸、麦芽汁、胡萝卜、PDA固体培养基上生长较好,粉托鬼笔对碳源和氮源的适应性较好,在不同碳源、氮源条件下,其菌丝的生长速度和气生菌丝的形态有较大差异。通过不同培养液及不同碳源及氮源对液体发酵培养基的筛选发现:PDA培养液适合作为粉托鬼笔的种子培养液,在10L的发酵罐中投料7L,蔗糖140g,大豆蛋白胨42g,酵母浸粉70g,接种量5%,温度26℃,转速150rpm,通气量1:0.3,培养至58h适合放罐。菌丝体中多糖含量达18.02%。适合作为判断放罐标准的指标是菌丝生物量及发酵液总糖含量。短裙竹荪菌丝体在所试不同培养上均能生长,在GPY、PDA、麦芽汁、麦麸、胡萝卜固体培养基上生长较好,在GPY固体培养基上生长速度最快,生物量最大,为145.50mg。短裙竹荪对碳源和有机氮源的适应性很强。短裙竹荪液体种子培养基为PDA培养液,在10L的发酵罐中投料7L,蔗糖140g,大豆蛋白胨42g,酵母浸粉70g,接种量5%,温度26℃,转速150rpm,通气量1:0.2,培养至90h适合放罐。菌丝体中多糖含量达17.92%。
[Abstract]:Tea Zi Zi leaflike layer fungus, phallus hadriani, Dictyophora duplicata is wild edible and medicinal fungi. According to the study, tea Zi sub leaflike layer fungus and honeysuckle efficacy is similar, can be used for detoxification, anti-inflammatory, analgesic, has now been made health care preparation, has an excellent efficacy of inflammation and cold. Phallus hadriani first by this laboratory study in our laboratory has cultivated a fruiting body, but not for large-scale cultivation. Dictyophora duplicata edible fungus is an excellent taste, with lowering blood pressure, regulating fatty acid to prevent hypertension. At present, these three kinds of wild edible and medicinal fungi picking limited that has not yet been studied culture system of mycelium, also limits the pharmacological research on them. This paper tea Zi sub leaflike layer fungus, bacteria isolation and identification of phallus hadriani, solid culture and liquid fermentation conditions of mycelium and chemical Research contents. Solid culture and culture of Dictyophora duplicata conditions and chemical composition content of mycelium fermentation. For industrial fermentation production of large-scale preparations. Firstly, wild tea Zi sub leaflike layers sporophore collected bacteria isolation and identification, determine the tissue isolated from mycelium is tea Zi Zi the leaflike layer fungus mycelium. The solid plate culture method on solid culture characteristics: Tea Zi sub leaflike layer fungus mycelium in wheat bran, malt juice, carrot, PDA, GPC, GPY solid medium growth can in wort solid medium, the fastest growth rate in the fiftieth day wheat bran solid medium maximum biomass, on the fortieth day GPY three mycelium cultured on terpene content of unit mass of the body is 0.39%, the highest total consideration for three terpene, should choose 60 days of wheat bran medium; tea Zi Zi The leaflike layer fungus with carbon and organic nitrogen in adaptability, maltose, fructose, xylose, galactose on growth is better, in addition to not grow in urea, can grow in other nitrogen sources were screened. The fermentation medium by shaking flask fermentation method: Tea Zi sub leaflike layer fungus can grow in culture only the liquid carbon source nitrogen source + + growth factor, but the best growth in liquid culture wort, wort culture as tea Zi sub leaflike layer fungus seed culture and liquid fermentation liquid for liquid; in shaking flask, three terpene content of not less than 1%, the content of polysaccharide in the above 15%. The biomass of seventh days to reach the maximum, while the HPLC measured the amount received most. With the extension of adenosine fermentation time, tea Zi sub leaflike layer fungus fungus ball color gradually from white to brown, and the fermentation liquid color gradually turned into brown, and have a fresh fruit flavor in this research. Using tissue isolation method of phallus hadriani bacteria strains isolated and molecular biological identification shows that isolated from mycelium as phallus hadriani. Phallus hadriani in different solid medium were used in the growth of wheat bran, malt juice, carrot, PDA solid medium growth is good, supporting adaptability for carbon powder stinkhorn and the nitrogen source is better in different carbon source, nitrogen source conditions, the mycelium growth rate and mycelium morphology are quite different. The screening medium of liquid fermentation by different medium and different carbon source and nitrogen source showed that PDA culture medium is suitable as a phallus hadriani seed in medium. Fermentation tank feed 7L, 10L sucrose 140g, soybean peptone 42g, yeast extract 70g, inoculum 5%, 26 degrees Celsius temperature, ventilation speed 150rpm, 1:0.3, 58h culture to fit tank. 18.02%. suitable for judging draw-off biaoda polysaccharide content in mycelium Standard index is the total sugar content of mycelia biomass and the fermentation liquid. The skirt of dictyophoraindusiata different culture could grow, in GPY, PDA, malt, wheat bran, carrots grown on solid medium is better in GPY solid medium, the fastest growth rate, the highest biomass and adaptability of 145.50mg. skirt Dictyophora of carbon source and organic nitrogen source is very strong. Dictyophora duplicata liquid seed culture medium for PDA medium in the fermentation tank feed 7L 10L, sucrose 140g, soybean peptone 42g, yeast extract 70g, inoculum 5%, 26 degrees Celsius temperature, ventilation speed 150rpm, 1:0.2, to 90h for training put the tank. 17.92%. content of polysaccharide in mycelium

【学位授予单位】：鲁东大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S646

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