五种螟蛾线粒体基因组序列的测定及螟蛾总科系统发育研究

发布时间：2018-06-27 13:29

本文选题：鳞翅目 + 螟蛾总科　；参考：《陕西师范大学》2015年硕士论文

【摘要】：螟蛾总科是鳞翅目的第二大类群,其大多数种类都是农林及仓储害虫,特别是钻蛀类螟蛾。大面积的虫害给农林业造成严重的经济损失,因此螟蛾类昆虫的防治一直是相关研究的重点。随着分子生物学研究技术的发展,线粒体基因组的研究在螟蛾总科中逐渐开展,但主要集中在单个的线粒体基因组的特点分析,很少有对螟蛾总科昆虫的线粒体基因组特点及系统发育关系进行全面深入的分析。本研究通过LA-PCR技术结合引物步移法,对褐巢螟Hypsopygia regina、黄纹银草螟Pseudargyria interruptella、双斑薄翅螟Evergestis junctalis、橙黑纹野螟Tyspanodes striata和豆荚野螟Maruca vitrata的线粒体基因组序列进行测定,并对上述5种新序列进行注释分析。联合GenBank中已释放的20种螟蛾总科昆虫的线粒体基因组数据,对螟蛾总科昆虫线粒体基因组进行比较分析,并以蛋白质编码基因(PCGs)为数据集,以雅库巴果蝇Drosophila yakuba为外群,利用最大似然法和贝叶斯推理法对螟蛾总科的系统发育关系进行重建并探讨分析。所得主要结论为：1.褐巢螟、黄纹银草螟、双斑薄翅螟、橙黑纹野螟和豆荚野螟线粒体基因组全序列长度分别为15212 bp、15231 bp、15438 bp、15255 bp和1 5200 bp,其中豆荚野螟为目前已测螟蛾总科昆虫完全序列中最短的物种。线粒体基因组的组成和结构与已报道的螟蛾总科昆虫一致,为闭合环状双链DNA分子,包括13个蛋白质编码基因、22个tRNA基因、2个rRNA基因和一个A+T富集区。与鳞翅目蝙蝠蛾等原始类群昆虫相比,tRNAMet基因的排列顺序有差别,即由tRNAIle-tRNAGln-tRNAMet变为tRNAMet-tRNAIle-tRNAGln。2.双斑薄翅螟全线粒体基因组,除A+T富集区非编码区外,在基因tRNAAla和tRNAArg、ND3和tRNAAla之间分别存在114 bp和62 bp的基因间隔,其均为插入的串联重复序列；在tRNAPro和ND6之间存在长达88 bp的基因重叠序列。3.5种螟蛾线粒体基因组的碱基组成均具有明显的AT偏向性,其中褐巢螟的AT含量较低,为78.65%；橙黑纹野螟的AT含量最高,为81.38%；豆荚野螟、黄纹银草螟和双斑薄翅螟的AT含量分别为80.86%、79.40%和81.08%。4.蛋白质编码基因的起始密码子的使用情况：5种螟蛾的COI基因均以CGA起始,其余基因则以ATN起始；终止密码子的使用：5种螟蛾的COII基因都以单独的T终止,黄纹银草螟的ND5基因以不完全终止密码子TA终止,其他的PCGs则以完全终止密码子TAN终止。5.5种螟蛾的tRNA二级结构,除基因tRNASer(AGN)的二级结构缺DHU臂外,其余21个tRNA均能形成典型的三叶草结构。且tRNA二级结构存在一定数量的碱基错配,包括U-U和G-U两种类型,主要为G-U错配。6.5种螟蛾的A+T富集区具有极高的AT含量,褐巢螟、黄纹银草螟、双斑薄翅螟、橙黑纹野螟和豆荚野螟分别为94.86%、90.99%、94.30%、94.51%和93.62%。并且5种昆虫的A+T富集区中均存在相关的保守元件ATTTA、ATAG、poly-T及长短不一的微卫星结构。7.已知25种螟蛾的PCGs的密码子UUA(L)、AUU(L)、UUU(F)、AUA(M)的使用频次较高,平均为465.4、415.5、344.5、256.5。PCGs的氨基酸使用情况与密码子使用情况一致,使用量从高到低依次为亮氨酸、异亮氨酸、苯丙氨酸、蛋氨酸。8.ML法和BI法构建的系统发育树的拓扑结构大体相同,与传统分类一致,两棵系统发育树都很好的支持了草螟科和螟蛾科的单系性,同时还有力地支持了建树所包括的11个亚科(螟蛾科4亚科,草螟科7亚科)的单系性,在ML系统树中,各个亚科的相对位置表现为：((((斑螟亚科+螟蛾亚科)+丛螟亚科)+蜡螟亚科)+((((草螟亚科+苔螟亚科)+禾螟亚科)+(水螟亚科+薄翅螟亚科))+(斑野螟亚科+野螟亚科)))。
[Abstract]:The family of Lepidoptera is the second largest group of Lepidoptera, most of which are agroforestry and storage pests, especially borer moth. The large area of insect pests cause serious economic loss to agroforestry. Therefore, the prevention and control of borer moth has been the focus of related research. With the development of molecular biology research, mitochondrial genome The study was carried out gradually in the family of borer mothmoidea, but mainly focused on the characteristics of single mitochondrial genome. Few of the mitochondrial genome characteristics and phylogenetic relationship of borer mothmoidea were thoroughly analyzed. The LA-PCR technique combined with primer stepping method to the Hypsopygia Regina of the brown nest borer and the P of the Yellow Rice stem borer. Seudargyria interruptella, the mitochondrial genome sequence of the Evergestis junctalis, the Tyspanodes striata of the orange moth borer and the Maruca vitrata of the pod borer, and the annotated analysis of the 5 new sequences. The mitochondrial genome data of the 20 species of borer, which have been released in GenBank, are used for the total of the moth moth. The mitochondrial genome of the family insect was compared and analyzed with the protein encoding gene (PCGs) as the data set and the Drosophila yakuba of the YAC Drosophila melanogaster as the outer group. The phylogenetic relationship of the family of borer moth was reconstructed by the maximum likelihood method and Bias reasoning method. The main conclusions were as follows: 1. Brown nest borer, yellow rice stem borer, double. The complete sequence length of the mitochondrial genome of the thin wing borer, the orange striped wild borer and the pod borer were 15212 BP, 15231 BP, 15438 BP, 15255 BP and 15200 BP, in which the pod borer was the shortest species in the complete sequence of the survey of the borer moth. The composition and structure of the mitochondrial genome were the same as that of the reported borer moth. The cyclic double chain DNA molecule, including 13 protein coding genes, 22 tRNA genes, 2 rRNA genes and one A+T enrichment area, has the difference in the sequence of tRNAMet gene in the order of the tRNAMet gene, which is transformed from tRNAIle-tRNAGln-tRNAMet to the whole mitochondrial genome of the tRNAMet-tRNAIle-tRNAGln.2. bis thin wing borer, except for A. In the non coding region of the +T enriched region, there were 114 BP and 62 BP intervals between gene tRNAAla and tRNAArg, ND3 and tRNAAla respectively, and all of them were inserted tandem repeats. There was an overlap sequence of 88 BP between tRNAPro and ND6, and the base composition of the mitochondrial genome of the.3.5 species of borborx borborx had obvious AT bias. The AT content of the nesting borer was 78.65%, and the AT content of the orange moth borer was the highest, 81.38%. The AT content of the pod borer, the yellow rice stem borer and the pod borer was 80.86%, 79.40% and 81.08%.4., respectively: the COI gene of the 5 species began with CGA, and the other genes began with ATN. The use of terminating codons: the COII gene of the 5 kinds of borer is terminated by a separate T. The ND5 gene of the yellowworm moth is terminated by the incomplete terminating codon TA, and the other PCGs terminates the tRNA two structure of the.5.5 species by completely terminating the codon TAN. Except for the two stage structure of the gene tRNASer (AGN), the other 21 tRNA can form a code. There are a number of base mismatches in the tRNA two structure, including two types of base mismatch, including two types of G-U and G-U. The A+T enrichment area of the G-U mismatch.6.5 species has a high AT content, the brown nest borer, the yellow rice stem borer, the double spotted borer, the orange striped wild borer and the pod borer, respectively, 94.86%, 90.99%, 94.30%, 94.51% and 93.62%. and 5, respectively. There are related conserved elements ATTTA, ATAG, poly-T, and different microsatellite structures in the A+T enrichment areas of the species..7. is known as the PCGs codon UUA (L) of the 25 species of borer moth, AUU (L), UUU (F), and the average use of amino acids is in accordance with the use of codons. The topological structure of the phylogenetic tree constructed by leucine, isoleucine, phenylalanine, methionine.8.ML and BI was the same as that of the traditional taxonomy. The two phylogeny trees supported the monophyletic of the family albonididophysi and borer, and also supported the 11 subfamilies (4 subfamilies of the family borbididididididus). In the ML system tree, the relative position of each subfamily in the ML system tree is as follows: ((((((((((moth, moth, moth, moth) + subfamily with parafferia borer) + (((((((((of subfamily borer + moss Ming Yake) + grasses subfamily) + (subfamily of water borer + thin wing borer) + (subfamily borbel and wild borer subfamily)).
【学位授予单位】：陕西师范大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S433.4

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