具生理活性平榛内生菌株的筛选与诱变育种研究

【作者】 李敏华；

【导师】 金涛；

【作者基本信息】 黑龙江大学 ， 遗传学， 2018， 硕士

【摘要】 内生菌可产生与宿主植物相同或类似的活性成分,且种类繁多、繁殖速度快、代谢可塑性强。为筛选出具有生理活性的生产候选菌株,本试验以平榛树皮为材料,分离纯化得到41株内生真菌。采用高效液相色谱分析及质谱鉴定,从中筛选到1株产辅酶Q10的内生真菌(JH-1a),经分子生物学鉴定,并结合形态学特征分析,确定JH-1a菌株为钩状木霉菌(Trichoderma hamatum),并初步进行紫外诱变育种,筛选到一株性状优良的突变菌株。经过细胞病变抑制实验和CCK-8细胞毒性测定实验证实1株产黄酮内生菌(Jl-5)具有抗PRRSV活性,并从3个方面对其抗PRRSV机制进行初步研究,证实内生菌Jl-5对PRRSV具有直接灭活作用、阻断吸附作用以及抑制复制作用。以上研究,为开发保健品和抗病毒药物奠定了基础。更多还原

【Abstract】 Endophytes produce the same or similar active ingredients as host plants,and have a wide variety of species with the feature of fast reproduction speed and high metabolic plasticity.In order to select the candidate of production strains with physiological activity,41 strains of endophytic fungus were isolated and purified from the bark of Corylus heterophylla Fisch.One endophytic fungi yielding coenzyme Q10(JH-1a)was screened out by high performance liquid chromatography(HPLC)combined with mass spectrum(MS).Through molecular biological identification and research on the morphological characteristics,JH-1a strain was identified as Trichoderma hamatum.Moreover,a mutant strain with superior traits was obtained by preliminary ultraviolet breeding.The anti-PRRSV activity of Jl-5 strain,the endophytic fungi yielding flavonoids,was confirmed through the Cytopathic-effect inhibition assay and the CCK-8 method.The anti-PRRSV mechanism was preliminarily studied from three aspects,and it was confirmed that the endophytic fungi Jl-5 can directly inactivate PRRSV,block PRRSV adsorption and inhibit PRRSV replication.The research result provides a theoretical basis for developing health products and antiviral drugs.更多还原