【作者】 张龙霞；

【导师】 裴雁曦； 李惠萍；

【作者基本信息】 山西大学 ， 植物学， 2012， 硕士

【摘要】 槐树(Sophora japonica L.),适应性强,易成活,特别在吸收污染物、净化大气等方面具有独特性,成为城市绿化优选树种之一,被广泛种植用于街道绿化。但是,这种良好的绿植生态环境,也给众多有害生物提供了极好的定殖场所,其中蚧虫成为了槐树的优势危害种群,同时它极易随种苗、大树的调运,在空间上远距离传播,侵入到新的生态体系引起广泛危害,给农林生产安全造成极大威胁。因此,本实验对太原市三条主要交通路段上的槐树进行抽样分析,寻找不同的蚧虫种类,并对不同生态位上的蚧虫数量、种类进行统计学分析,从槐树与蚧虫的关系入手对蚧虫选择寄主、与共栖寄生物协同作用、生物生态学等方面进行系统的研究,以提出科学的防控措施。同时通过对蚧虫从遗传差异方面的分子生物学研究,建立快速准确的鉴定技术,以加强和保障植物检疫措施。主要内容如下：第一：实验对太原市漪汾街(幼龄槐树比较集中)、南内环(中龄槐树比较集中)、晋祠路(老龄槐树比较集中)上的槐树进行抽样分析,寻找不同的蚧虫种类,并对不同生态位上的蚧虫数量、种类进行统计学分析,之后进行种类鉴定,搞清楚我市槐树主要受哪种蚧虫的侵害,汽车尾气、灰尘等污染对蚧虫的分布是否有影响。根据我们的分析,我们已发现枣大球蚧(Eulecanium gigantea (Shinji))和皱大球坚蚧(Eulecanium kuwanai Kanda)是危害我省槐树的主要种类。第二：实验以五种危害槐树较严重的蚧虫为材料,分别用SDS法(十二烷基硫酸钠)、CTAB法(十六烷基三乙基溴化铵)、TES法(酚-氯仿)、NaC1法(氯化钠)和TNES法5种提取方法,对单只蚧虫进行全基因组DNA提取。结果表明,5种提取方法都能够获得基因组DNA,但是相比较而言,CTAB法和TES法所提取的DNA质量明显优于SDS法、NaCl法和TNES法。因此认为,CTAB法和TES法比较适合实验室提取蚧虫基因组DNA。第三：实验以五种危害槐树较严重的蚧虫为材料,采用基因扩增技术,对五种蚧虫的COI-COⅡ基因进行PCR扩增和测序,并比对其序列设计各自的特异性引物。结果表明,基于PCR扩增及直接测序的分子技术可以达到准确、快速鉴定五种蚧虫的目的。更多还原

【Abstract】 Sophora japonica L easy to survive, particularly in the absorption of pollutants, air purification and so unique, as one of city greening tree species selection, is widely grown for street greening. However, this favorable environment green plants, but also to many pest colonization provides an excellent venue which has become a locust tree scale insect populations of the advantages of harm, and it easily with the seedlings and trees transporting, Long-distance spread in space, invade a new ecological system caused widespread harm to the agricultural and forestry production caused a great threat to security.First, this experiment in Taiyuan Yi Fen Street (more concentrated in young Ash), South inner (more concentrated in the age of Ash), Jinci Road (old acacia trees are concentrated) sample survey on the acacia trees, looking for different The scale insect species, and different niche on the number of scale insect, the type used for statistical analysis, after species identification, find out what the city is mainly affected by acacia trees against scale insects, vehicle exhaust, dust and other pollution Kuwana Would affect the distribution of insects. Finally, rapid, accurate and effective quarantine methods. According to our investigation, we have found great dates the Scale Eulecanium gigantea (Shinji), and fold the big ball firmly Eulecanium kuwanai Kanda locust tree in our province is the main type of hazard.Two, Genomic DNA were extracted from five scale insect by using the sodium dodecyl Sulfate (SDS) method, the cetylt ethylammonium bromide (CTAB) method, Phenol-chloroform (TES) method, the sodium chloride (NaC1)method and TNES method respectively. The results show that the five methods were useful in extraction of genomic DNA of scale insects. But comparatively speaking, the quality of DNA samples isolated by using the CTAB method and the TES method was obviously better than that by using the other three methods. So that, our laboratory think that the CTAB and the TES method were more effective and practical method for the genomic DNA extraction of scale insect.Three, The mitochondrial COI-COII sequences were of five scale insects were were amplified by using PCR technique to analyze their sequence variation and then design their respective specific primer sequence. The results show that, based on the molecular techniques of PCR amplification and direct sequencing can achieve accurate,rapid identification of five scale insect.更多还原