Breeding Of Triterpenoids And Polysaccharides High-Producing Ganoderma Strains And Determination Of Biological Characteristics Of Hybrid Strains

In this study, we gathered50Ganoderma strains we gathered from China and cultured them through both bag cultivation method and liquid fermentation method. After analyzing the results of these cultivations, we selected some Ganoderma strains with outstanding biological characteristics as parent strains, and we determined the hybrid combination based on the result of phylogenetic relationship analysis of all the strains we tested. We obtained all the monokaryotic strains of our parent strains using protoplast monokaryogenesis technique, and afterward gain the hybrid strains through hybridization experiment using them. To make sure that the hybrid strains are new species, we used SRAP and ISSR DNA molecular marker techniques to identify the differences between every hybrid strain and it’s parent strains. At last, we measure the characteristics of all hybrid strains, concluding triterpenoids content and polysaccharides content, to select hybrid strains with good quality.After culture all Ganoderma strains through bag cultivation method and liquid fermentation method, we collected Ganoderma fruiting bodies and liquid-fermentative hyphae and measure their yields and triterpenoids contents and polysaccharides content to choose the top ranking strain on these indexes respectively, then combined with the result of phylogenetic relationship, the hybrid pairs of parent strains were confirmed.Monokaryotic strains of all parent strains were obtained using protoplast monokaryogenesis technique. From1206regenerated colonies, we gained107monokaryotic strains with a success rate of8.87%. From967hybrid pairs, we gained27hybrid strains with a success rate of2.79%.Through observing clamp connections and antagonistic test, hybrid strains were initially identified, then they were finally confirmed uising SRAP and ISSR DNA molecular marker techniques. All results show that the hybrid strains we obtained are different from their parent strains and the genetic characters are derived from their parent strains each.We cultivate hybrid strains through bag cultivation method and liquid fermentation method respectively. Then we measure their yields and triterpenoids content and polysaccharides content to evaluate them. As the results showed, hybrid strain0520-3is a new liquid-fermentative hyphae high-yield strain. For hybrid strain0520-5, not only intracellular polysaccharides content were significantly increased but also higher yielding through liquid fermentation compared with its parent strains.