柏大蚜自然群體內細菌的組成和多樣性分析
發(fā)布時間：2018-08-27 18:37
【摘要】：蚜蟲是重要的農林業(yè)害蟲,主要通過刺吸式口器取食植物韌皮部汁液,從而造成樹勢衰弱甚至死亡。蚜蟲體內普遍存在內共生菌為宿主昆蟲的生長發(fā)育提供必需的營養(yǎng)物質,并且在組織免疫、防御天敵等方面發(fā)揮重要的作用。為了比較同一地點柏大蚜和松大蚜體內內共生菌的組成和差異以及柏大蚜不同種群體內微生物組成是否存在差異,本論文利用16S rRNA-RFLP方法分析了同一地點柏大蚜和松大蚜體內細菌的組成以及利用T-RFLP方法分析了柏大蚜四個種群體內細菌群落的差異。主要研究結果如下:1.細菌的16S rRNA-RFLP分析的研究結果表明:柏大蚜和松大蚜體內細菌主要由其初級內共生菌Buchnera aphidicola和次級內共生菌Candidatus Serratia symbiotica組成,只是各自所占的比例存在差異;在柏大蚜中,B.aphidicola占37%,Ca.Serratia symbiotica占56%,還有少量的腸桿菌屬Enterobacter占7%;在松大蚜中,B.aphidicola占66.7%,Ca.Serratia symbiotica占33.3%。結果表明兩種蚜蟲體內的微生物組成比較簡單和相似,內共生菌種類相同但占的比例存在明顯差異。2.從三種內切酶(Afa I、Hha I和Alu I)中選出Alu I酶為柏大蚜菌群T-RFLP的理想酶切種類。因為該酶酶切產生的T-RFs數(shù)目最多而且產生的圖譜比較清晰。3.根據(jù)T-RFs條帶和峰值面積對四個種群細菌組成進行的相似性分析表明,絳帳種群(JZ)個體中極不相似比例高達39.9%,與其他三個種群表現(xiàn)為不相似,其他種群個體間細菌組成為中等相似。4.多樣性分析表明:乾陵種群多樣性指數(shù)最小(Shannon index=1.62),絳帳種群最大(Shannon index=2.50);絳帳種群優(yōu)勢度指數(shù)最大(Simpson index=0.88),其他單個種群比較接近;楊陵種群豐富度指數(shù)最大(Margalef index=1.13),其次為絳帳種群(Margalef index=1.09);馬昭種群的均勻度指數(shù)最大(Pielou index=0.58)。總之,絳帳種群與其他三個種群相比,具有較高的多樣性指數(shù)、優(yōu)勢度指數(shù)、豐富度指數(shù)以及較低的均勻度指數(shù)。5.四個種群T-RFs片段豐富度分析結果表明:從乾陵種群、楊凌種群、馬昭種群和絳帳種群中分別檢測到22種、32種、28種、60種T-RFs片段;乾陵種群、楊凌種群和馬昭種群的優(yōu)勢菌群比較明顯,但是優(yōu)勢菌群的T-RFs片段長度存在差異。絳帳種群16個個體間無明顯的優(yōu)勢菌群,個體間T-RFs片段組成存在明顯的差異;其中285bp所代表的菌群是四個地區(qū)有共有的優(yōu)勢菌群。
[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.
【學位授予單位】：西北農林科技大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S433.3
本文編號：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.
【學位授予單位】：西北農林科技大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：S433.3
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