| The article was aimed at some gordian techniques during G.lucidum industrial cultivation process,mainly divided into four steps.First,select the varieties of G.lucidum we need by classifying and identifying 21 Ganoderma strains.Second,screen out the G.lucidum strains which not suited for cultivae of JUNCAO and the JUNCAO formulas were not suited for cultivate G.lucidum through vitro screening test.Third,gain comprehensive analysis of the growth,commercial properties,yield and effective constituents of G.lucidum mycelium and fruiting body through industrialized cultivation test to find out the best formula and G.lucidum strains.Fourth,optimize the industrialized cultivation condition and explore how the influence regular pattern of cultivation conditions varies on different aspects of the entity to find out the best condition of cultivation,and the regular pattern of the impact of cultivation conditions on fruiting body.The main achievements are as follows:(1)Genetic relationship identification of Ganoderma strainsThe experiment by antagonistic test,ITS sequence analysis and RAPD technique to identified classification and analyzed the genetic diversity of 21 strains.The results of antagonistic test showed that,most strains had different degree of antagonism,and there was no antagonism or no extremly obvious antagonism among a few strains.The results of ITS showed that,the variation of genetic distance ranged from 0.000 to 0.072 among these strains,and the average genetic distance was about 0.039;compared with GenBank,the result showed that,No.4,No.5,No.12,No.13,No.15,No.16,No.17 and No.19 strains belonged to the spectes of G.lucidum,and the genetic relationship was relatively close.The results of RAPD showed that,the similarity coefficient was over 0.8 among the strains,it indicated that their genetic relationship were close.The similarity coefficient between strain No.1 and No.3 was reached to 0.95,so they are likely to be synonymous.In conclusion the experiment indicated that the results of three methods are consistent.It was successful to classify and identify the 21 strains,and screen out 9 G.lucidum strains.(2)Comperetive study on adaptability of G.lucidum strains in JUNCAO substrateThe tube cultivation method was used to screen and optimize the 16 JUNCAO substrates and 9 G.lucidum strains.The strains which are not suitable to cultivate using JUNCAO and the JUNCAO formulas which are not benefittial the growth of G.lucidum were excluded,by comprehensiveanalysis of the mycelial growth rate,primordium formation time,culture period,fruiting sturdy,biological conversion rate.The results showed that,G.lucidum(Ga0801,Ga0805,Ga0042,G.8)and JUNCAO formulas(No.1,No.2,No.6,No.7,No.8,No.10,No.24,No.26)were comprehensively better than the other strains and formulas.(3)Screening formulas and strains of G.lucidum on JUNCAO industrial cultivationThe experiment was carried out to cultivate G.lucidum using the preliminary screened 4 strains and 8 formulas,and the best formula and G.lucidum strain were screened out by grey relational analysis,combined the results of single cylinder output,output and input ratio,triterpene content,polysaccharide content and total sugar content.Comparaed with CK,the results showed that,the increase of effective components of 4 different G.lucidum cultivated by JUNCAO formula were relatively higher,and some evenly appear to grow exponentially.As a result,cultivating G.lucidum using is better than sawdust to some extent.No.2 JUNCAO formula was comprehensively the best,with the most comprehensive benefits.Therefore,the JUNCAO No.2 formula:Miscanthus floridulus 78%,wheat bran 20%,and gypsum 2%is the best formula.Among 4 strains of G.lucidum cultivated by No.2 JUNCAO formula,Ga0042 is the best strains with the lowest pollution rate and the best comprehensive performance.(4)The environment conditions optimization of G.lucidum industrial cultivation by JUNCAOThe experiment mainly studied two environmental factors of temperature and CO2 during G.lucidum industrial cultivation.To determin the optimum fruiting temperature and CO2 concentration by measuring the indexes of the fruiting body.The results showed that,the effect of temperature on the fruiting bodies of G.lucidum was significantly within the experimental temperature range,the pileus diameter,pileus thickness,yield,biological conversion rate of fruiting body showed a trend of rising first and then decreasing with the increase of temperature,the maximum value appeared at about 25℃.The content of triterpene of fruiting body decreased linearly with the increase of temperature,the higher the temperature was,the lower the content of the triterpene was.Conversly,the content of polysaccharide increased with the increase of temperature,the higher the temperature was,the faster the content of polysaccharide increased.As a result of comprehensive analysis,26℃ is the suitable incubation temperature,the comprehensive performance is the best under this temperature.In the CO2 test interval,the effect of CO2 on the diameter,thickness and stipe diameter of G.lucidum was significant,and they all showed a trend of increased first and then decreased with the increase of CO2 concentration.The highest value was between 0.05%and 0.06%.While the effect of CO2 on the yield of fruiting bodies was no significant.The content of triterpenoids and polysaccharide in fruiting bodies showed a trend of decreased first and then increased with the increase of CO2,the effect was significant. |
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