人参根际土壤微生物多样性研究

发布时间：2018-03-04 02:26

本文选题：人参　切入点：根际微生物　出处：《延边大学》2015年硕士论文　论文类型：学位论文

【摘要】：人参Panax ginseng C.A.Meyer),是我国名贵药用植物,也是吉林省主要特产之一。长白山区以其独特的地理位置和气候条件,成为吉林省人参主产区。人参属于忌地性极强的植物,存在连作障碍,长期采用伐林栽参,影响可持续发展。因此,人参产业的发展受到了阻碍,解决人参忌连作问题迫在眉睫。大多数学者认为连作障碍与土壤理化性状变劣、微生物体系的变化存在密切关系。但从土壤改良方面只能从一定程度上缓解连作的症状,病害发生、产量低的问题仍然存在。本实验从根际微生物的角度进行分析,利用微生物纯培养、PCR-DGGE技术、基因克隆方法开展人参忌连作问题研究,重点研究真菌对植物病害的影响,以期为人参病害预防、病理研究以及开发防治人参病害杀菌剂提供基础。研究的主要结论如下：1、确立了人参根际土壤真菌ITS区PCR反应体系：25μL反应体系DNA浓度52.5 ng、dNTPs浓度0.20 mM、Mg2+浓度1.50 mM、引物浓度4.80μmol搭配效果最佳。2、筛选出适合真菌DGGE指纹图谱分析的引物ITS1/ITS4、ITS1-GC/ITS2,并确定真菌DGGE凝胶单体浓度为12%-6%,变性剂浓度为60%-40%。细菌分析采用引物314F-GC/518R,DGGE凝胶单体浓度为8%,变性剂浓度为60%-40%。3、研究结果表明根际真菌与非根际真菌种类存在明显差异,并且真菌种类随着种植年限的增加而变少。随着种植年限的增加同种真菌在数量上也发生了变化,表现出变少的趋势,而且有少量新的真菌滋生。人参根际细菌与非根际细菌多样性指数变化不明显,但有参地细菌量明显多于无参地,后期滋生的细菌有可能存在致病菌。4、通过基因克隆测序得到68条测序结果,将测序结果系统分析,BLAST对比得出人参根际土壤中分别有菌根真菌(Mycorrhizal fungus)、担子菌属(Basidiomycota sp.)、真子囊菌门(Euascomycete sp.)、枝孢属(Euascomycete sp.)、被孢霉属(Mortierella alpina)、子囊菌属(Ascomycete sp.)地丝霉属(Geomyces sp.)、类球囊霉属(Paraglomus laccatum)、隐球菌(Cryptococcus podzolicus) 、子囊菌(Ascomycota sp.) 。
[Abstract]:Ginseng Panax ginseng C.A. Meyerian is one of the most valuable medicinal plants in China and one of the main specialties of Jilin Province. Changbai Mountain area, with its unique geographical location and climatic conditions, has become the main producing area of Panax ginseng in Jilin Province. Therefore, the development of ginseng industry has been hindered, so it is urgent to solve the problem of ginseng non-continuous cropping. Most scholars think that continuous cropping obstacle and soil physical and chemical properties become worse. The changes of microbial system are closely related. But in the aspect of soil improvement, the symptoms of continuous cropping can only be alleviated to a certain extent, the problems of disease occurrence and low yield still exist. This experiment is analyzed from the point of view of rhizosphere microorganism. In order to prevent ginseng disease, PCR-DGGE technique and gene cloning method were used to study the effects of fungi on plant diseases. The main conclusions of the study are as follows: 1. The PCR reaction system of ginseng rhizosphere fungi in ITS region was established. The reaction system of PCR was established. The concentration of DNA was 52.5 渭 L and the concentration of DNA was 0.20 mg / mg _ 2 and the concentration of primer was 1.50 mm. The best concentration of 4.80 渭 mol was 4.80 渭 mol. The primers ITS1 / ITS4 / ITS1-GC / ITS2 were selected, and the concentration of fungal DGGE gel monomer was 12-6, denaturant concentration was 60-40.The bacterial analysis was conducted with primer 314F-GCr518RDGGE gel monomer concentration 8 and denaturant concentration 8. The results showed that there were significant differences between the species of rhizosphere fungi and non-rhizosphere fungi. With the increase of planting years, the number of fungi of the same species also changed, showing a trend of decreasing. The diversity index of ginseng rhizosphere bacteria and non-rhizosphere bacteria was not obvious, but the number of ginseng rhizosphere bacteria was more than that of non-ginseng soil. It is possible that the bacteria bred in the later period may have pathogenic bacteria. 68 sequences were obtained by gene cloning and sequencing. The systematic analysis of sequencing results showed that Mycorrhizal fungus, Basidiomycota sp., Euascomycete sp., Euascomycete sp., Mortierella alpina., ascomycete sp., Mycorrhizal Fungi, Ascomycete sp., Euascomycecete sp., Mortierella alpinaanus, Ascomycete sp., respectively, were found in the rhizosphere soil of Panax ginseng, and that the genus Geomyces sp., Basidiomycota sp., Euascomycete sp., Euascomycete sp., Mortierella alpinaanus, Ascomycete sp.) were found in the rhizosphere soil of Panax ginseng. Of the genus Paraglomus laccatumus, Cryptococcus podzolicus, Ascomycota sp.)
【学位授予单位】：延边大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S567.51;S154.3

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