【作者】 王铮；

【导师】 张日俊；

【作者基本信息】 中国农业大学 ， 动物营养与饲料科学， 2004， 硕士

【摘要】 本研究以药用真菌灵芝为主要研究对象，对灵芝的液体深层发酵工艺进行优化，运用原生质体技术进行了高产糖肽复合物新菌株的选育，以期提高深层发酵胞外PSPc的产量，研究了PSPc对肉仔鸡的生产性能和对免疫功能的影响。 试验一研究了灵芝液体深层发酵培养基中营养因子和非营养因子对PSPc分泌的影响，试验证明较高的碳氮比有利于PSPc的分泌，而且纤维素、半纤维素、Mg、K、P等营养因子对于灵芝的发酵过程中PSPc的分泌都有着一定的影响，并且确定了比较适合的添加量为KH₂PO₄1.5g／L，MgSO₄·7H₂O 0.75g／L，最佳的C／N为50：1，添加纤维素和半纤维素、核酸可以促进PSPc的产量。以此为根据确定了以豆粕、玉米和麦麸为主要成分的培养基，以利于深层发酵PSPc的生产。在初步的发酵试验中PSPc的含量最高可以达到2.5mg／ml。 试验二运用原生质体技术，进行高产PSPc新菌株的选育，（1）通过对灵芝和香菇原生质体的制备和再生的条件进行了探索，可以使原生质体的数量达到10⁷／ml，为利用原生质体进行紫外线诱变和原生质体融合试验打下了较好的基础并提供了丰富的试验材料。并利用荧光标记的方法，对灵芝和香菇的原生质体的融合条件和现象进行了观察和研究。（2）利用紫外线诱变选育出了两株在固体培养基上菌丝体生长速度和PSPc产量较高的新菌株（具体生产性能还有待进一步研究）。 试验三研究了液体深层发酵生产的PSPc对肉仔鸡的生产性能和免疫功能的影响。测定了不同生长阶段肉仔鸡的日增重、饲料转化率、牛血清白蛋白抗体、淋巴细胞转化率、死亡率、胸肌率、腿肌率和腹脂率。添加0.3％灵芝糖肽复合物饲料添加剂对肉仔鸡前期（28天）的生长性能最好较对照组的日增重提高5％，存在极显著差异（p＜0.01），但是后期（47天）的生长性能与对照组差异不显著。添加0.3％和0.5％灵芝糖肽复合物对于肉仔鸡的免疫功能的增强效果是显著差异的。更多还原

【Abstract】 This paper optimizes the conditions for the submerged fermentation process of Ganoerma lucidum and at the same time studies the application of the protoplast fusion technique on the breeding of variety which can produce high yield Polysaccharide Peptide Complex (PSPc) under submerged fermentation conditions. In addition, the influences the PSPc on the performance and immune function in broiler chickens have been studied.Effects of the medium nutritional and non-nutritional ingredients on the excretion of PSPc have been investigated. The results showed that relatively higher C/N ratio was favorable to PSPc formation. And cellulose, hemicellulose, Mg, K. and P have a great influence on the growth of mycelia and the production of PSPc during fermentation. The optimal median with soybean powder, maize powder and wheat bran as the main gradients (C/N=50:l), and the optimal addition amount of KH2PO4(1.5g/L), MgSCV7H2O(0.75g/L) are confirmed. The fermentation experiment shows that the maximum yield of PSPc is 2.5mg/ml.Protoplast fusion technique is used to breed the Ganoerma lucidum varieties which possess the steady property of high productivity. (l)The conditions of separation and regeneration of Ganoderma lucidum protoplast and Lentinula edodes protoplast are studied in detail. Results show that the highest production amounts to 107/ml. This study has laid the foundation for the following protoplast fusion and UV induced mutagenesis researchs. The fusion conditions between Ganoderma lucidum and Lentinus edodes have been studied by fluorescence labeled method. (2)By carrying out the UV induced mutagenesis of Ganoderma lucidum’s protoplast, we obtain 2 mutants with higher mycelium growth rate and higher yield of PSPc in solid culture.The influences of the PSPc obtained from submerged fermentation on the immune function and performance of broiler chickens have been studied in the third experiment. The average weight gain, feed conversion rate, bovine serum albumin(BSA) antibody value, T-lymphocyte proliferation, breast meat rate and leg meat rate have been tested. The animal test shows that the broiler chickens fed with the 0.3% fermentation broth for 28 days exhibit the best performance : 5% higher than the control group(p<0.01). The promoting effect of feeding 0.3% and 0.5% PSPc on the immune function of the broiler chickens is different.更多还原