| A wild Ganoderma strain was screened and purified in this study, and was named Rongbao Lingzhi No.1. Using RAPD and ITS sequence technique, we studied genetic characteristics between Rongbao Lingzhi No.1and main cultivate Ganoderma strains; And then, the taxonomic position and agronomic traits of Rongbao Lingzhi No.l was determined. At the same time, the main medicinal component content in different parts of the fruit body of Ganoderma lucidum species was detected. The main results are as follows:1. Five Wild Ganoderma lucidum fruit bodies were collected from Guangyuan, Yaan, Emei and Panzhihua, these strains were isolated and purified. Based on the growth status, a good candidate Ganoderma strain with potential application was screened and named as Rongbao Lingzhi No.l, and the genetic aspects was determined and compared with other main cultivated cultivars. The results showed that the genetic difference existed between Rongbao Lingzhi No.l and other cultivars, and the full ITS sequence was655bp, and the phylogenic tree was constructed, the results revealed that Rongbao Lingzhi No.1belong to Ganoderma lingzhi.2. The agronomic traits of Rongbao Linzhi No.1were determined. The results showed that its mycelium can in the range of15-35℃, and the optimum growth temperature was about25-301℃; and the growth pH ranged pH4to pH9, and optimum growth pH was pH5-7. The optimal mother culture was enriched PDA medium; and the proper stock culture and three-grade culture consisted of sawdust73%, wheat bran20%, corn powder5%, superphosphate1%, CaSO41%.3. The fruit bodies of Rongbao lingzhi No.l cultivar, South Korea Ganoderma cultivar, American Ganoderma cultivar, and Japanese Ganoderma cultivar were separated into context, tube and stipe, and polysaccharide content of each part was detected by anthrone-sulfuric acid method. The results showed that the total polysaccharide amount was as the follows, South Korea Ganoderma was0.7994%, American Ganoderma cultivar was0.7737%, Rongbao lingzhi No.l cultivar was0.7560%, and Japanese Ganoderma cultivar was0.6970%, there was not significant difference between the Ganoderma cultivars; the polysaccharide content of four strains mainly distributed in tube of fruit body, and the polysaccharide content in context and stipe is lower.4. The factors of HPLC method for determine the ganoderic acid A was explored, and the ganoderic acid A content in different parts of Ganoderma fruit bodies was detected. The optimal chromatographic conditions was:UltimateTM XB-C18used as chromatographic column, and the column temperature was25℃, mobile phase was acetonitrile (B)-acetic acid solution (A), the gradient elution was:0min,20%B,5min36%B,40min,37%B,50min,80%B,60min,90%B, flow rate:0.800ml/min, detection wavelength:254nm. The results showed that the ganoderic acid A content in context of Rongbao lingzhi No.1was4.50mg/g, which was much higher than the other cultivars. The ganoderic acid A content in different parts of the fruit body varied as the following order, context> stipe> tube.5. To establish a HPLC method for the determination of Ganoderma species in different parts of the four nucleosides (uridine, inosine, guanosine, adenosine) content. Chromatographic conditions:UltimateTM XB-C18column, the column temperature was25℃, mobile phase of methanol (B)-0.1%acetic acid in water (A), gradient elution:0min1%B,60min,5%B, flow rate:1.000ml/min, detection wavelength:254nm. The content of each species of the four nucleosides ingredients are:the tubes> context> Stipe, inosine was measured only in Rongbao Linzhi No.l,about0.100mg/g, the total amount of the the four nucleosides content order:South Korea Ganoderma(1.728mg/g)> American Ganoderma (1.592mg/g)> Japanese Ganoderma(1.569mg/g)> Rongbao Linzhi No.1(0.523mg/g). |
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