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MeJA Strains of Hericium 5L0433 Ergosterol Synthesis and Analysis of Subtracted cDNA Library of ESTs

Author: LiYingXue
Tutor: ZhanYaGuang
School: Northeast Forestry University
Course: Forest Biological Engineering
Keywords: Hericium (Hericium erinaceus Pers.) Mycelium Ergosterol Methyl jasmonate Subtractive library
CLC: S646.9
Type: Master's thesis
Year: 2011
Downloads: 36
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Abstract


In this study, the mycelia of Hericium (Hericium erinaceus Pers.) As experimental material, the The hedgehog MYCELIA shake flask cultures production ergosterol regulation method. And by suppression subtractive hybridization (Suppression subtractive hybridization, SSH) to build a hedgehog mycelium forward subtractive library. Specific genes from the subtractive library screening and preliminary bioinformatics analysis. The main findings are as follows: 1, the establishment of the MeJA induction hedgehog 5L0433 mycelium production of ergosterol technology Hericium liquid fermentation production of ergosterol can use the following training techniques to improve the production of ergosterol: medium: maltose 20g, phosphate the potassium dihydrogen 3G, magnesium sulfate 1.5 g, yeast extract 10g, water 1000ml pH NATURAL. After inoculation, 25 ° C stationary culture 2d, 25 ° C, 150r/min shaking Eight days after adding sterile 50 micromol · L-1MeJA, induced by the 11th day of harvest mycelium the the ergosterol production capacity reaches its maximum value 1.882mg/100ml. The MeJA Hericium 5L0433 ergosterol biosynthesis MeJA (methyl jasmonate) elicitor promote the accumulation of ergosterol reveals: the MeJA induction child add the hedgehog liquid fermentation mycelia growth, ergosterol content , growth reflects a certain role in promoting. Mycelium yield and ergosterol identified, 50gmol L-1 optimal the MeJA concentration of the hedgehog mycelium induced. MeJA induced the first 8d the mycelium highest 0.96g/100ml. The MeJA 10μmol · L-1, 50μmol · L-1, 100μmol · L-1, 200μmol · L-1 concentrations 50gmol · L-1 and 100gmol · L-1 mycelium body yield to 0.31 g / the 100ml and 0.33g/100ml significantly higher than the control (0.27g/100ml). The MeJA concentration for In 50gmol · L-1, the dry mycelial ergosterol content up to 1.603mg · g-1 is the control (1.260mg · g-1) 1.27 times. 3, subtractive library of specific genes in the initial bioinformatics analysis was successfully constructed MeJA induced hedgehog forward subtracted library. Using BLAST software abatement to library screening to specific genes ESTs fragment sequences in GenBank sequence homology comparison, while the use of COG functional classification and GO functional classification of sequence preliminary bioinformatics analysis. In Forward library MhmrA, GO functional classification results show that the cellular components function in the library with these three cells, organelles, protein complexes. Molecular function library contains antioxidant activity, catalytic activity, signal transduction activity, translational regulation of activity, biological pathways including cell library program, these four categories of the physiological process, regulation of biochemical processes, response to stimuli. Forward library MhmrA of COG functional classification results show that the view from the fragment the proportion of the various functional genes, GO functional classification results show that methyl jasmonate-induced gene expression in the ergosterol process involved in post-translational modification protein turnover, carbohydrate transport and metabolism, energy production and conversion, amino acid transport and metabolism, inorganic ion transport and metabolism, coenzyme transport and metabolism 15.

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