柏大蚜自然群体内细菌的组成和多样性分析
发布时间：2018-08-27 18:37
【摘要】：蚜虫是重要的农林业害虫,主要通过刺吸式口器取食植物韧皮部汁液,从而造成树势衰弱甚至死亡。蚜虫体内普遍存在内共生菌为宿主昆虫的生长发育提供必需的营养物质,并且在组织免疫、防御天敌等方面发挥重要的作用。为了比较同一地点柏大蚜和松大蚜体内内共生菌的组成和差异以及柏大蚜不同种群体内微生物组成是否存在差异,本论文利用16S rRNA-RFLP方法分析了同一地点柏大蚜和松大蚜体内细菌的组成以及利用T-RFLP方法分析了柏大蚜四个种群体内细菌群落的差异。主要研究结果如下:1.细菌的16S rRNA-RFLP分析的研究结果表明:柏大蚜和松大蚜体内细菌主要由其初级内共生菌Buchnera aphidicola和次级内共生菌Candidatus Serratia symbiotica组成,只是各自所占的比例存在差异;在柏大蚜中,B.aphidicola占37%,Ca.Serratia symbiotica占56%,还有少量的肠杆菌属Enterobacter占7%;在松大蚜中,B.aphidicola占66.7%,Ca.Serratia symbiotica占33.3%。结果表明两种蚜虫体内的微生物组成比较简单和相似,内共生菌种类相同但占的比例存在明显差异。2.从三种内切酶(Afa I、Hha I和Alu I)中选出Alu I酶为柏大蚜菌群T-RFLP的理想酶切种类。因为该酶酶切产生的T-RFs数目最多而且产生的图谱比较清晰。3.根据T-RFs条带和峰值面积对四个种群细菌组成进行的相似性分析表明,绛帐种群(JZ)个体中极不相似比例高达39.9%,与其他三个种群表现为不相似,其他种群个体间细菌组成为中等相似。4.多样性分析表明:乾陵种群多样性指数最小(Shannon index=1.62),绛帐种群最大(Shannon index=2.50);绛帐种群优势度指数最大(Simpson index=0.88),其他单个种群比较接近;杨陵种群丰富度指数最大(Margalef index=1.13),其次为绛帐种群(Margalef index=1.09);马昭种群的均匀度指数最大(Pielou index=0.58)。总之,绛帐种群与其他三个种群相比,具有较高的多样性指数、优势度指数、丰富度指数以及较低的均匀度指数。5.四个种群T-RFs片段丰富度分析结果表明:从乾陵种群、杨凌种群、马昭种群和绛帐种群中分别检测到22种、32种、28种、60种T-RFs片段;乾陵种群、杨凌种群和马昭种群的优势菌群比较明显,但是优势菌群的T-RFs片段长度存在差异。绛帐种群16个个体间无明显的优势菌群,个体间T-RFs片段组成存在明显的差异;其中285bp所代表的菌群是四个地区有共有的优势菌群。
[Abstract]:Aphids are important pests in agriculture and forestry. They mainly feed on plant phloem sap through sucking mouthparts, which results in debilitating and even death of trees. Endosymbionts are common in aphids which provide essential nutrients for the growth and development of host insects and play an important role in tissue immunity defense of natural enemies and so on. In order to compare the composition and difference of symbiotic bacteria in the different populations of cypress aphid and pine aphid at the same location, and whether there were differences in microbial composition among different populations of cypress aphid, In this paper, 16s rRNA-RFLP method was used to analyze the bacterial composition in the same site and the differences of bacterial communities among the four populations were analyzed by T-RFLP method. The main results are as follows: 1. The results of 16s rRNA-RFLP analysis showed that the bacteria were mainly composed of their primary endosymbiosis bacteria (Buchnera aphidicola) and secondary endosymbionts (Candidatus Serratia symbiotica), but their proportions were different. B. aphidicola accounted for 37% of Ca. Serratia symbiotica, and a small amount of Enterobacter Enterobacter accounted for 7%, and B. aphidicola accounted for 66.7% Ca. Serratia symbiotica for 33. 3%. The results showed that the microorganism composition of the two aphids was simple and similar, and the species of endosymbionts were the same but the proportion of them was obviously different. Alu I enzyme was selected from three endonuclease species (Afa IHha I and Alu I) as the ideal species of T-RFLP. Because the enzyme produced the largest number of T-RFs and produced a clearer map. 3. 3. According to the similarity analysis of the bacterial composition of the four populations based on the T-RFs bands and the peak area, it was found that the extremely dissimilar proportion of the (JZ) individuals in the crimson population was as high as 39.9%, which was not similar to the other three populations. The composition of bacteria among individuals of other populations was moderately similar. 4. The diversity analysis showed that the diversity index of Qianling population was the smallest (Shannon index=1.62), the maximum of Jiangdan population (Shannon index=2.50), the most dominant index (Simpson index=0.88) of Jiangdan population, and the same of other individual populations. The population richness index (Margalef index=1.13) of Yangling population was the largest, followed by Margalef index=1.09 population, and the largest evenness index (Pielou index=0.58) of Ma Zhao population. In a word, compared with the other three populations, the population has higher diversity index, dominance index, richness index and lower evenness index. The results of T-RFs fragment richness analysis of four populations showed that 22 species 32 species, 28 species and 60 species of T-RFs fragments were detected from Qianling population, Yang Ling population, Ma Zhao population and Jiangdan population, respectively, Qianling population, Qianling population, The dominant microflora of Yang Ling population and Ma Zhao population were obvious, but the T-RFs fragment length of dominant microflora was different. There was no obvious dominant flora among 16 individuals in Jiangdan population, and there were obvious differences in the composition of T-RFs fragments among individuals, among which 285bp represented a common dominant flora in four regions.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S433.3
本文编号：2208102
[Abstract]:Aphids are important pests in agriculture and forestry. They mainly feed on plant phloem sap through sucking mouthparts, which results in debilitating and even death of trees. Endosymbionts are common in aphids which provide essential nutrients for the growth and development of host insects and play an important role in tissue immunity defense of natural enemies and so on. In order to compare the composition and difference of symbiotic bacteria in the different populations of cypress aphid and pine aphid at the same location, and whether there were differences in microbial composition among different populations of cypress aphid, In this paper, 16s rRNA-RFLP method was used to analyze the bacterial composition in the same site and the differences of bacterial communities among the four populations were analyzed by T-RFLP method. The main results are as follows: 1. The results of 16s rRNA-RFLP analysis showed that the bacteria were mainly composed of their primary endosymbiosis bacteria (Buchnera aphidicola) and secondary endosymbionts (Candidatus Serratia symbiotica), but their proportions were different. B. aphidicola accounted for 37% of Ca. Serratia symbiotica, and a small amount of Enterobacter Enterobacter accounted for 7%, and B. aphidicola accounted for 66.7% Ca. Serratia symbiotica for 33. 3%. The results showed that the microorganism composition of the two aphids was simple and similar, and the species of endosymbionts were the same but the proportion of them was obviously different. Alu I enzyme was selected from three endonuclease species (Afa IHha I and Alu I) as the ideal species of T-RFLP. Because the enzyme produced the largest number of T-RFs and produced a clearer map. 3. 3. According to the similarity analysis of the bacterial composition of the four populations based on the T-RFs bands and the peak area, it was found that the extremely dissimilar proportion of the (JZ) individuals in the crimson population was as high as 39.9%, which was not similar to the other three populations. The composition of bacteria among individuals of other populations was moderately similar. 4. The diversity analysis showed that the diversity index of Qianling population was the smallest (Shannon index=1.62), the maximum of Jiangdan population (Shannon index=2.50), the most dominant index (Simpson index=0.88) of Jiangdan population, and the same of other individual populations. The population richness index (Margalef index=1.13) of Yangling population was the largest, followed by Margalef index=1.09 population, and the largest evenness index (Pielou index=0.58) of Ma Zhao population. In a word, compared with the other three populations, the population has higher diversity index, dominance index, richness index and lower evenness index. The results of T-RFs fragment richness analysis of four populations showed that 22 species 32 species, 28 species and 60 species of T-RFs fragments were detected from Qianling population, Yang Ling population, Ma Zhao population and Jiangdan population, respectively, Qianling population, Qianling population, The dominant microflora of Yang Ling population and Ma Zhao population were obvious, but the T-RFs fragment length of dominant microflora was different. There was no obvious dominant flora among 16 individuals in Jiangdan population, and there were obvious differences in the composition of T-RFs fragments among individuals, among which 285bp represented a common dominant flora in four regions.
【学位授予单位】：西北农林科技大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S433.3
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相关期刊论文 前2条
1 吕增芳;;花椒树蚜虫危害严重的原因及防治建议[J];山西果树;2010年01期
2 葛朝红;孟建朝;田海燕;;棉田蚜虫的发生与防治[J];种子科技;2011年01期
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