【作者】 吴庆禹；

【导师】 迟德富； 宋瑞清；

【作者基本信息】 东北林业大学 ， 森林保护学， 2007， 博士

【摘要】 异色瓢虫Harmonia axyridis(Pallas)是一种重要的捕食性天敌昆虫，也是一个典型的多型性物种，它是蚜虫、松干蚧、粉蚧等害虫的主要天敌。现在正被人们广泛释放用来对蚜虫和蚧虫进行生物防治。本文主要从形态学水平、生态水平、分子水平对异色瓢虫不同色斑类型的遗传多样性进行了研究，揭示出异色瓢虫不同色斑类型间在遗传多样性方面的差异，为深入了解异色瓢虫的生存状况、分类地位、进化历史和亲缘关系，预测其未来的发展趋势，更有效地进行生物综合防治奠定了基础。(1)对2178只异色瓢虫的斑型进行统计的结果，共发现126种色斑类型，其中有15种色斑型在以前的报道中没有记录。多样性分析的结果，Monk指数、Simpson多样性指数以及Shannon-Wiener指数分别为0.0579，0.901和1.56。(2)对黄底型异色瓢虫的斑点的概率进行了统计，其中，斑点3的概率最高，而斑点9和斑点1／2的概率则较小。不同的色斑型异色瓢虫间，雌雄比例差别很大；尤其是在黄底型类群中，随着斑点数的增多，类型中雄性比例减小。对异色瓢虫主要的类群之间的体长、体宽进行了比较研究，结果显示在体长和体宽方面差异显著，在长宽比方面差异极显著；说明不同色斑类型间，在虫体的大小和形状上存在一定的差异。(3)通过扫描电镜的观察，描述了异色瓢虫不同色斑类型触角形态结构，以及触角上感器的种类和数量；在不同色斑型间进行了对比，结果显示异色瓢虫不同色斑类型之间存在一些差异。(4)异色瓢虫不同色斑类型越冬成虫的生态分布与其活动的林型、栖息植物、发生时间、气候变化等有直接关系。在同一时间同一地域数量和分布寄主上存在一些差异。(5)异色瓢虫不同色斑类型越冬成虫的空间分布型为聚集分布，并符合负二项分布。(6)本试验用改良CTAB法、饱和NaCl法以及CTAB法与SDS法相结合3种方法，进行综合设计，探索出一套操作简便、快速、准确的高质量异色瓢虫DNA提取和纯化技术。(7)从40条RAPD随机引物中筛选出12条具有多态性的引物，对9种异色瓢虫不同色斑类型进行RAPD分析，共检测到多态位点169个，位点范围在100bp～2000bp之间，多态位点百分率在34.32％～65.68％之间；从40个引物中筛选出13条条带清晰且多态性丰富的ISSR引物，对13种异色瓢虫不同色斑类型进行ISSR分析，共扩增出127条ISSR标记带，其中多态性条带120条，多态性百分率94.49％。(8)RAPD和ISSR分析的一致性和可信度：由RAPD和ISSR分析算得的两组相似系数之间相关性系数r=0.928(n=127)，达到极显著水平。表明应用这两种技术对异色瓢虫遗传多样性与亲缘关系的分析具有较高的一致性和可信度。但是这两种方法的聚类结果在个别色斑型分类上仍存在一定的差异。(9)依据RAPD分析结果，对供试的9种异色瓢虫不同色斑类型进行聚类分析，结果表明：它们之间的遗传相似系数在0.46～0.58之间；依据ISSR分析结果，对供试的13种异色瓢虫不同色斑类型进行聚类分析，它们之间的遗传相似系数在0.36～0.60之间。综上所述，通过从形态学水平、生态水平、分子水平对异色瓢虫不同色斑类型的遗传多样性进行比较，异色瓢虫不同色斑类型间在遗传多样性方面存在一定的差异，为人类更好地保护、开发和利用天敌昆虫资源提供了科学依据。更多还原

【Abstract】 Harmonia axyridis (Pallas) is an important predator insect, as well as a typical polymorphic species, it is also the main natural enemy insects of aphids, matsumura pine scale, mealybugs and other pests. Now being widely released for biological control of aphids and scale insects.This paper mainly study on genetic diversity of different splash stain of H. axyridis from the morphological, ecological, molecular level, and reveals their differences in genetic diversity, then can deeply understand the living conditions, Classification, evolutionary history and relationships of H. axyridis, forecast its future development trends, and establish the foundation for more effective biological integrated prevention and treatment.(1) Through survey on the spots of 2178 units of H. axvridis, 126 splash types were found, among them 15 splash types--15 spot types were not recorded in the past report. The results of diversity analysis showed that Monk index, Simpson diversity index and Shannon-Wiener index was 0.0579.0.901 and 1.56 differently.(2) The statistical results of probability of type showed that a spot 3 was the highest, while spots 9 and 1/2 were smaller. Different splash types of H, axyridis had a great difference in the proportion of male and female; particularly in the yellow-bottom groups, with the number of spots increase, the proportion of male types reduced. Through correlation study between length and width of the main groups of H. axyridis, the results showed significant differences in body length and width, and the aspect ratio varied significantly; the different splash types had a certain body size and shape variations.(3) Observed by scanning electron microscopy, antenna structure and the types and quantities of sensors on antennae of different splash types of H. axyridis were described; and a comparison between different splash types was made, the results showed that there were some differences between units of H. axyridis.(4) The ecological distribution, forest types, habitat plants, the time of its occurrence and climate change of overwintering adults of different splash types of H. axyridis were directly related. Its number and distribution hosts also had some differences at the same time and same terrain.(5) The spatial distribution of overwintering adults of different splash types of H. axyridis was mass distribution, and also accord with the negative binomial distribution.(6) In this experiment, the modified CTAB method, saturated NaC1 method and SDS and CTAB combining method were used for cornprehensive design, a set of simple, rapid, accurate, high-quality DNA extraction and purification technology was explored. (7) 12 polymorphic primers were selected from 40 random RAPD primers, through RAPD analysis on 9 different splash types of H. axyridis, 169 polymorphic sites were detected, the sites lie between 100 bp and 2000 bp, and its percentage lie between 34.32 and 65.68; 13 ISSR primers whose bands were clear and rich in polymorphism were selected from 40 primers. Through ISSR analysis on 13 different splash types of H. axyridis, 127 ISSR bands were amplified, including 120 polymorphic bands, and the polymorphic percentage is 94.49.(8) The consistency and credibility of ISSR and RAPD analysis: There was a high correlation between RAPD and ISSR (r=0.928, n=127), indicating that the two methods were highly consistant and suitable for detecting genetic diversity and genetic relationships in H. axyridis. However, both kinds of clustering analysis have some difference in individual splash types.(9) According to the clustering analysis of RAPD results of 9 different splash types of H. axyridis, the results showed that the genetic similarity index was between 0.46 and 0.58. The genetic similarity index is between 0.36 and 0.60 based on the ISSR analysis of the results.In short, through comparison on genetic diversity of different splash types ofH. axyridis from morphological, ecological and molecular level, different types have some differences in genetic diversity. It provided scientific basis for protection, development and use of natural enemy insects for human.更多还原