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A preliminary study of the skullcap endophytic fungi

Author: ZhangLiGuang
Tutor: WeiXiYing
School: Shaanxi Normal University
Course: Microbiology
Keywords: Scutellaria Endophytic fungi Baicalin Orthogonal optimization
CLC: S567.239
Type: Master's thesis
Year: 2011
Downloads: 101
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Abstract


Endophyte all of those life history or a life in healthy plant tissues and its fungus does not cause significant illness. Recent studies show that part of the endophytic fungus capable of producing the host plant and the same or similar biological activity of the substance, thus becoming one of the sources of new drugs. As a novel microbial resources, the endophytic fungal secondary metabolites and gradually become a hot topic in the bio-pharmaceutical, biological control of pests and diseases and industrial fermentation. Skullcap is one of the commonly used herbs have a calming, blood pressure, anti-bacterial, anti-viral, anti-tumor effect, attending febrile disease, upper respiratory tract infection, Hyperactivity cough, damp, jaundice, pneumonia, dysentery embolism. Medicinal plant Scutellaria object, its endophytic fungi were studied, and the results are as follows: (1) using conventional microbiological isolation from medicinal plants skullcap health disease-free tissue isolated 17 endophytic fungi, which the root of 3, 7 stems, leaves a flower 6. (2) cup and saucer method for the determination of the skullcap endophytic fungi fermentation broth of E. coli (Escherichia coli) A70, Bacillus subtilis (Bacillus subtilis) A99, Staphylococcus aureus (Staphylococcus aureus) 26003 and other 10 kinds of indicator bacteria inhibition activity. Experimental results show that the majority of endophytic fungi fermentation broth have different degrees of inhibitory activity on at least one indicator bacteria (including 4 strains endophytic fungi (J1 J4 J5 G2) on a variety of directions bacterial shows a wide antibacterial spectrum). (3) by thin layer chromatography (TLC) and high performance liquid chromatography (HPLC) analysis method of detecting endogenous fungal metabolites. TLC detection results show that, there are two students fungi (J1 and H3 cytoplasmic) metabolic product with standard product yellow baicalensis glycosides have the same Rf value, indicates that further HPLC detection results and J1 and H3 metabolism product with baicalin standard products have the same retention time. Determine the initial strain J1 and H3 can produce the same active ingredient to the host plant Scutellaria baicalin. (4) According to the J1 and H3 strain colony morphology characteristics and microscopic observation, combined with the ITS nucleic acid sequence analysis, the preliminary identification of the strain J1 and H3 are deuteromycetes door, Penicillium species. (5) to select higher production baicalin content J1 strain optimization of media and culture conditions. Through the the J1 strain commonly used liquid medium mycelial growth and baicalin generation, found bean sprouts juice body medium not only conducive to the growth of mycelium, but also conducive to the accumulation of baicalin. Univariate analysis to optimize laboratory fermentation medium J1 strain to determine the best C source, N source is sucrose and ammonium nitrate. The by the the strain J1 fermentation conditions orthogonal to optimize the results, selected three factors, the concentration of C source impact on biomass, N source followed by concentration, pH value with minimal impact. Baicalin yield the most influential factor is the concentration of C source N source followed by concentration, pH, minimal impact. The orthogonal optimization experiments the J1 strain optimization of fermentation conditions are as follows: for maximum biomass concentration of sucrose 4.0% ammonium nitrate concentration of 0.15%, pH 7.0; baicalin in order to obtain maximum yield, sucrose concentration of 4.0%, the ammonium nitrate concentration of 0.25%, pH 6.0. Because the the orthogonal results show that for baicalin cumulative conditions and the mycelium growing conditions vary, the final optimization of fermentation: cultivation of mycelial growth medium until the strain into the stable, you can appropriate to increase the concentration of N source, the lower the pH to improve the yield of baicalin.

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