【作者】 张斌；

【导师】 贺一原；

【作者基本信息】 中南林业科技大学 ， 林学， 2021， 硕士

【摘要】 目前对扩散多型现象的研究主要集中在翅多型方面,飞行肌和飞行行为多型的研究极少。长颚斗蟋Velarifictorus aspersus具翅二型现象,前期研究中发现,长翅型雌雄成虫存在可遗传的飞行行为多型现象,因此,本研究检测了不同飞行能力的长翅雌成虫在飞行肌及体内主要营养物质的差异,分析其飞行行为多型的代谢基础;对不同飞行能力雌成虫进行转录组测序,为进一步研究其飞行行为多型的分子机制奠定基础。主要获得以下结果:(1)羽化后第五天不同飞行能力的长翅雌成虫飞行后的主要飞行能源物质(甘油三酯、蛋白质、糖原、总脂含量)存在显著差异。(2)通过对飞行后长颚斗蟋V.aspersus长短翅雌成虫进行转录组测序,共获得95.64Gb Clean Data,各样品均达到6.02 Gb以上。获得46404个转录本,最高长度为44099bp,平均长度为1405.13bp,CG含量为40.57%;对转录本进行聚类和组装分析后共得到36496个Unigenes,最高长度为44099bp,平均长度为1411.02bp,CG含量为41.1%。(3)与NCBI蛋白质数据库、Swiss-Prot蛋白质数据库、KEGG数据库和COG数据库进行比对,能够用已知的生物学功能注释的总共有16584个Unigenes,注释率为45.33%,注释到GO数据库中的51个功能亚类。经Nr数据库比对,进行同源性分析,注释物种匹配到序列最多的是隐穴白蚁Cryptotermes secundus,匹配率为 25.59%。(4)对不同飞行能力的雌成虫转录组分析表明,飞行能力弱的长翅雌成虫与短翅雌成虫间筛选出5650个差异表达基因,其中上调基因数量为3178条,下调基因数量为2472条;飞行能力弱与强的长翅雌成虫间筛选出873个差异表达基因,其中上调基因数量为492条,下调基因数量为381条。差异表达基因进行GO富集分析,飞行能力弱的长翅雌成虫与短翅雌成虫间的差异表达基因主要富集在细胞的组分这一大类功能上,飞行能力弱与强的长翅雌成虫间差异表达基因主要富集在分子功能这一大类功能上。差异表达基因进行KEGGPATHWAY富集分析,飞行能力弱的长翅雌成虫与短翅雌成虫间的差异基因主要富集在代谢这个大的通路上。飞行能力弱与强的长翅雌成虫间差异表达基因同样也是主要富集在在代谢这个大的通路上。本研究首次对长颚斗蟋V.aspersus雌成虫进行转录组测序分析,为蟋蟀飞行行为多型性的研究提供了分子基础。更多还原

【Abstract】 At present,the research on diffusion polymorphism mainly focuses on wing polymorphism,and there are very few studies on flight muscles and flight behavior polymorphism.The Velarifictorus aspersus has winged dimorphism.In previous studies,it was found that long-winged female and male adult worms have heritable polymorphism of flight behavior.Therefore,this study tested the long-winged female adults with different flight abilities in flight muscles and The differences in the main nutrients in the body are analyzed to analyze the metabolic basis of their flight behavior polytypes;transcriptome sequencing of female adults with different flight abilities will lay the foundation for further research on the molecular mechanisms of their flight behavior polytypes.The main results are as follows:(1)The main flight energy materials(triglycerides,protein,glycogen,total lipid content)of long-winged female adults with different flight abilities on the fifth day after emergence are significantly different.(2)The transcriptome sequencing of the female adults of the V.aspersus long-winged wing after flight,a total of 95.64Gb Clean Data was obtained,and each sample reached 6.02 Gb or more.Obtained 46404 transcripts,the highest length was 44099 bp,the average length was 1405.13 bp,and the CG content was 40.57%.After clustering and assembly analysis of the transcripts,a total of 36496 Unigenes were obtained,the highest length was 44099 bp,and the average length was 1411.02 bp.The CG content is 41.1%.(3)Comparing with NCBI protein database,Swiss-Prot protein database,KEGG database and COG database,a total of 16,584 Unigenes can be annotated with known biological functions,with an annotation rate of 45.33%,and annotated into the GO database 51 functional subcategories.After comparing with the Nr database and conducting homology analysis,the annotated species with the most matching sequence is the crypt termite Cryptotermes secundus,with a matching rate of 25.59%.(4)The transcriptome analysis of female adults with different flight abilities showed that 5650 differentially expressed genes were screened between long-winged female adults with weak flight ability and short-winged female adults,of which the number of up-regulated genes was 3178 and the number of down-regulated genes was 2472.873 differentially expressed genes were screened among long-winged female adults with weak and strong flying ability.Among them,the number of up-regulated genes was 492 and the number of down-regulated genes was 381.Differentially expressed genes were analyzed by GO enrichment.The differentially expressed genes between long-winged female adults with weak flight ability and short-winged female adults were mainly enriched in the function of cell components,and long-winged females with weak flight ability and strong long wings Differentially expressed genes among female adults are mainly enriched in molecular functions.The differentially expressed genes were analyzed by KEGG PATHWAY enrichment analysis.The differential genes between long-winged female adults and short-winged female adults with weak flight ability were mainly enriched in the large pathway of metabolism.Differentially expressed genes between long-winged female adults with weak and strong flying ability are also mainly enriched in the large pathway of metabolism.In this study,the transcriptome sequencing analysis of female adults of V aspersus was the first to provide a molecular basis for the study of cricket flight behavior polymorphism.更多还原