黄孢原毛平革菌和绿色木霉平板混合培养降解稻草秸秆中不同区域差异性研究
发布时间：2018-12-20 09:14
【摘要】：21世纪以来,使用化石能源对环境造成的污染愈发严重,寻找可再生的替代能源愈发迫切。秸秆含有大量的木质纤维素,是自然界中含量最多的可再生资源,合理利用这种可再生资源成为人们关注的热点。黄孢原毛平革菌作为一种降解木质素能力最强的真菌被广为研究,含有黄孢的混合菌剂因其具有较之黄孢更为强大的降解能力成为研究的重点。绿色木霉是一种广泛存在于自然界中的木质素降解菌,因其与黄孢搭配具有较强的木质素降解能力被认为是一种理想的堆肥接种菌。本研究以稻草粉末培养基为依托,利用顶空固态微萃取等技术,对黄孢和绿色木霉的纯培养及其混合培养进行了研究,重点研究了混合培养中同一平板接触区域与非接触区域的各种差异,以期对混合培养中黄孢和绿色木霉降解木质素的机制有更深入的了解。本研究主要对黄孢和绿色木霉的纯培养及其混合培养各区域的挥发性有机物、草酸含量、木质素过氧化物酶活性和p H进行了检测,就其数据进行了比对分析。结果表明:两株菌的混合培养较其纯培养有更强的降解稻草的能力。在混合培养过程中检测到C15H24的两种同分异构体,分别属于双环倍半水芹烯和环己二烯类物质,均是倍半萜烯,证实了黄孢原毛平革菌和绿色木霉混合培养时的拮抗机制。环己二烯类物质仅在混合培养中的混合区域检测到,表明混合培养中混合区域较之非混合区域具有更强的竞争表现,分区域检测降解过程中挥发性有机物能更好的研究混合接种的降解机理。草酸的分泌可能是黄孢降解木质纤维素能力强于绿色木霉的原因之一;混合培养中各区域草酸含量变化由于受到绿色木霉的干扰总体呈下降趋势,说明真菌间的拮抗作用对真菌分泌草酸的量有较大影响。黄孢和绿色木霉纯培养状态下均能产木质素过氧化物酶,但黄孢产酶能力强于绿色木霉是黄孢降解木质纤维素能力高于绿色木霉的另一个原因;草酸是影响培养体系中p H的重要因素,其变化趋势与草酸的分泌含量呈负相关;但竞争导致混合培养中不再呈负相关。
[Abstract]:Since the 21st century, the environment pollution caused by fossil energy is becoming more and more serious, so it is more urgent to find renewable alternative energy. Straw contains a large amount of lignocellulose, which is the most abundant renewable resource in nature. The rational utilization of this renewable resource has become the focus of attention. As a kind of fungus with the strongest lignin degradation ability, P. xanthosporium has been widely studied, and the mixed fungicides containing xanthosporins have become the focus of research because of their more powerful degradation ability than that of xanthosporium. Trichoderma viride (Trichoderma viride) is a kind of lignin degrading bacteria widely existing in nature. Because of its strong lignin degradation ability with xanthosporium, Trichoderma viride is considered to be an ideal compost inoculation bacteria. The pure culture and mixed culture of Trichoderma viride and Trichoderma viridis were studied by headspace solid microextraction based on rice straw powder medium. In order to understand the mechanism of lignin degradation by Trichoderma viride and Trichoderma viride in mixed culture, the differences between the contact region of the same flat plate and the non-contact area were studied. In this study, volatile organic compounds (VOCs), oxalic acid (oxalic acid), lignin peroxidase (lignin) activity and pH were detected in pure culture and mixed culture of Trichoderma viridis and Trichoderma viridis, and the data were compared and analyzed. The results showed that the mixed culture of the two strains had stronger ability to degrade rice straw than its pure culture. Two isomers of C15H24, belonging to bicyclic hemihydrate and cyclohexadiene, were detected in mixed culture, which were sesquiterpene, which confirmed the antagonistic mechanism of P. xanthosporium and Trichoderma viridis in mixed culture. Cyclohexadienes were detected only in the mixed region of mixed culture, which indicated that the mixed region had stronger competitive performance than the non-mixed region in mixed culture. The degradation mechanism of mixed inoculation can be better studied by detecting volatile organic compounds in the process of degradation. The secretion of oxalic acid may be one of the reasons why xanthosporium can degrade lignocellulose better than Trichoderma viridis. The change of oxalic acid content in different regions of mixed culture showed a decreasing trend because of the interference of Trichoderma viridis, which indicated that the antagonism between fungi had a great effect on the amount of oxalic acid secreted by fungi. Both yellow spores and Trichoderma viridis could produce lignin peroxidase in pure culture, but the ability of xanthosporins to produce lignin peroxidase was stronger than that of Trichoderma viridis, which was another reason that the ability of degradation of lignocellulose by xanthosporium was higher than that of Trichoderma green. Oxalic acid is an important factor affecting pH in culture system, and its changing trend is negatively correlated with oxalic acid secretion, but competition leads to no negative correlation in mixed culture.
【学位授予单位】：湖南大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：TQ914.3;O636.2
[Abstract]:Since the 21st century, the environment pollution caused by fossil energy is becoming more and more serious, so it is more urgent to find renewable alternative energy. Straw contains a large amount of lignocellulose, which is the most abundant renewable resource in nature. The rational utilization of this renewable resource has become the focus of attention. As a kind of fungus with the strongest lignin degradation ability, P. xanthosporium has been widely studied, and the mixed fungicides containing xanthosporins have become the focus of research because of their more powerful degradation ability than that of xanthosporium. Trichoderma viride (Trichoderma viride) is a kind of lignin degrading bacteria widely existing in nature. Because of its strong lignin degradation ability with xanthosporium, Trichoderma viride is considered to be an ideal compost inoculation bacteria. The pure culture and mixed culture of Trichoderma viride and Trichoderma viridis were studied by headspace solid microextraction based on rice straw powder medium. In order to understand the mechanism of lignin degradation by Trichoderma viride and Trichoderma viride in mixed culture, the differences between the contact region of the same flat plate and the non-contact area were studied. In this study, volatile organic compounds (VOCs), oxalic acid (oxalic acid), lignin peroxidase (lignin) activity and pH were detected in pure culture and mixed culture of Trichoderma viridis and Trichoderma viridis, and the data were compared and analyzed. The results showed that the mixed culture of the two strains had stronger ability to degrade rice straw than its pure culture. Two isomers of C15H24, belonging to bicyclic hemihydrate and cyclohexadiene, were detected in mixed culture, which were sesquiterpene, which confirmed the antagonistic mechanism of P. xanthosporium and Trichoderma viridis in mixed culture. Cyclohexadienes were detected only in the mixed region of mixed culture, which indicated that the mixed region had stronger competitive performance than the non-mixed region in mixed culture. The degradation mechanism of mixed inoculation can be better studied by detecting volatile organic compounds in the process of degradation. The secretion of oxalic acid may be one of the reasons why xanthosporium can degrade lignocellulose better than Trichoderma viridis. The change of oxalic acid content in different regions of mixed culture showed a decreasing trend because of the interference of Trichoderma viridis, which indicated that the antagonism between fungi had a great effect on the amount of oxalic acid secreted by fungi. Both yellow spores and Trichoderma viridis could produce lignin peroxidase in pure culture, but the ability of xanthosporins to produce lignin peroxidase was stronger than that of Trichoderma viridis, which was another reason that the ability of degradation of lignocellulose by xanthosporium was higher than that of Trichoderma green. Oxalic acid is an important factor affecting pH in culture system, and its changing trend is negatively correlated with oxalic acid secretion, but competition leads to no negative correlation in mixed culture.
【学位授予单位】：湖南大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：TQ914.3;O636.2
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