棉铃虫和烟青虫性信息素差异的遗传机制

发布时间：2017-12-26 17:12

本文关键词：棉铃虫和烟青虫性信息素差异的遗传机制　出处：《安徽大学》2017年硕士论文　论文类型：学位论文

更多相关文章： 棉铃虫 烟青虫 性信息素 遗传家系

【摘要】：近缘种及同种不同品系间的生殖隔离现象一直是物种形成研究的热点。鳞翅目许多昆虫往往具有相同类型的性信息素化合物,并通过改变各组分的比例实现种间生殖隔离。近年来,陆续报道了一些涉及蛾类性信息素产生及其遗传结构基础的研究,这些工作对阐明生殖隔离遗传变化的规律提供了重要的信息。然而,在进化过程中,由性信息素的改变导致的近缘物种形成的遗传机制尚不清楚。棉铃虫(Helicoverpa armigera 和烟青虫(Helicoverpa assulta)是鳞翅目(Lepidoptera)夜蛾科(Noctuidae)两种重要的农业害虫。棉铃虫为高度杂食性昆虫,危害20科近200种植物,加之其强大的繁殖力和广泛的分布性,致使其一直成为农业害虫防治的难点,而烟青虫为寡食性昆虫,主要危害茄科植物,但其危害性和破坏力也同样不容小觑。棉铃虫和烟青虫的幼虫和成虫在形态结构上都非常相似,很难辨认。有研究表明棉铃虫雌虫性信息素成分主要以顺-11-十六碳烯醛((Z)-11-hexadecenal,Z11-16:Ald)为主,而烟青虫的性信息素成分主要以顺-9-十六碳烯醛((Z)-9-hexadecenal,Z9-16:Ald)为主,棉铃虫和烟青虫中Z11-16:Ald/Z9-16:Ald比例正好相反。自然条件下,二者通过释放不同比例的性信息素组分完成种内化学通讯,实现种间生殖隔离。并且二者之间可杂交和回交,因此是研究性信息素通讯系统遗传差异的绝佳模型。本研究通过两近缘种棉铃虫和烟青虫单对杂交和回交,分析亲本、杂交和回交雌蛾性信息素组分,系统研究两近缘种性信息素比例差异的内在遗传机制。首先,分别剪取棉铃虫和烟青虫性腺,采用正己烷浸提,进而通过GC-MS的选择离子方法,分别测定了棉铃虫和烟青虫雌蛾性信息素主要组分Z11-16:Ald和 Z9-16:Ald,证实了棉铃虫性信息素以 Z11-16:Ald 为主,Z11-16:Ald/Z9-16:Ald为 96:4,而烟青虫性信息素以 Z9-16:Ald 为主,Z11-16:Ald/Z9-16:Ald 为 4:96,为本论文的后续研究奠定了坚实的基础。然后,开展棉铃虫和烟青虫杂交和回交实验,测定杂交代和回交代雌蛾的性信息素表型,统计符合棉铃虫和烟青虫性信息素表型的杂交代和回交代个数,推断棉铃虫和烟青虫性信息素差异的遗传基础。烟青虫雌性与棉铃虫雄性单对杂交(SR),产生两性子代,杂交代(SR)雌蛾性信息素都表现为棉铃虫表型;当杂交代(SR)雌蛾与棉铃虫(R)雄蛾回交时,回交代(SRR)雌性性信息素均以Z11-16:Ald为主,表现为棉铃虫表型;当杂交代(SR)雌蛾与烟青虫(S)雄蛾回交时,回交代(SRS)雌性个体出现两种性信息素表型,表型一为以Z11-16:Ald为主,表现为棉铃虫表型,表型二为以Z9-16:Ald为主,表现为烟青虫表型,且二者个体数量比例为1:1。当棉铃虫雌性和烟青虫雄性单对儿杂交时,杂交代(RS)全部为雄性。当杂交代(RS)雄蛾与棉铃虫雌蛾(R)回交时,回交代RSR)雌蛾性信息素均以Z11-16:Ald为主,表现为棉铃虫表型,而当杂交代(RS)雄蛾与烟青虫(S)雌蛾回交时,回交代(RSS)雌蛾性信息素表型发生分离,表现为两种表型:表型一为以Z11-16:Ald为主,表现为棉铃虫表型,表型二为以Z9-16:Ald为主,表现为烟青虫表型,且两表型个体数量比例接近1:1。两个方向的杂交和回交实验证明棉铃虫和烟青虫性信息素比例差异是由单基因控制的显隐性遗传。进而,我们运用DNA-AFLP技术对亲代、杂交代和回交代进行性信息素比例差异基因定位。通过DNA提取、酶切、连接、扩增和进一步特异性扩增,再结合以表型的数据,我们共获得了 5个分子标记。通过对5个典型标记切胶回收并测序,分别得到了其对应的核苷酸序列,为后续基因克隆和定位奠定了基础。综上,本研究利用两近缘种棉铃虫和烟青虫单对杂交和回交,分析亲本、杂交代和回交代雌蛾性信息素组分比例,利用DNA-AFLP分子标记技术,通过棉铃虫和烟青虫杂交谱系的遗传学研究,探究了棉铃虫和烟青虫性信息素比例颠倒的遗传基础,探讨了种间隔离机制,揭示了重要害虫(棉铃虫和烟青虫)性信息素产生差异的遗传机制,为后续基础研究和发展害虫管理新方法与新技术提供基础。
[Abstract]:The reproductive isolation between the marginal species and the same species has been a hot spot in the study of species formation. Many insects of Lepidoptera often have the same type of sex pheromone compound and achieve interspecific reproductive isolation by changing the proportion of each component. In recent years, a number of studies have been reported on the generation and genetic structure of sex pheromones, which provide important information for elucidating the laws of genetic variation in reproductive isolation. However, in the evolutionary process, sex pheromone changed the genetic mechanism of species leads to the formation of is not clear. The cotton bollworm (Helicoverpa armigera and Helicoverpa assulta) are two important agricultural pests of the Lepidoptera family of nocturum (Noctuidae). The cotton bollworm is highly polyphagous insects, harm of the approximately 200 species of plants in 20 families, with its strong fecundity and wide distribution, which has become a difficult agricultural pest control, and tobacco budworm is oligophagous insect, the main harm Solanaceae, but its harmfulness and damaging the same can not be underestimated. The larvae and adults of the Helicoverpa armigera and the adults are very similar in shape and structure, and are difficult to identify. Studies have shown that the cotton bollworm female sex pheromone components mainly along the -11- sixteen carbon aldehyde ((Z) -11-hexadecenal, Z11-16:Ald), and the tobacco budworm sex pheromone components mainly in cis -9- sixteen carbon aldehyde ((Z) -9-hexadecenal, Z9-16:Ald), cotton bollworm and tobacco budworm in proportion to Z11-16:Ald/Z9-16:Ald on the contrary. Under natural conditions, the two groups accomplish intraspecific chemical communication by releasing different proportions of sex pheromone components to achieve interspecific reproductive isolation. And the two can cross and backcross, so it is a perfect model for the study of genetic differences in the sex pheromone communication system. In this study, the sex pheromone components of parents, crosses and backcross females were analyzed by single crosses and backcrosses of two closely related species of Helicoverpa armigera and tobacco bollworm, and the intrinsic genetic mechanism of two closely related sex pheromone ratios was systematically studied. First, we cut H.armigera and h.assulta gonad, using n-hexane extraction, and then through the selection method of GC-MS ion, Helicoverpa armigera and h.assulta female sex pheromone components Z11-16:Ald and Z9-16:Ald were determined respectively, confirmed the cotton bollworm sexual pheromone Z11-16:Ald is the main Z11-16:Ald/Z9-16:Ald, 96:4, and smoke green insect pheromones Z9-16:Ald, Z11-16:Ald/Z9-16:Ald 4:96, has laid a solid foundation for the follow-up study of this paper. Then, carry out H.armigera and h.assulta hybridization and backcross experiments, determination of hybrid generation and backcross female moth sex pheromone phenotype, with statistics of H.armigera and h.assulta sex pheromone phenotype of hybrid and backcross generation number, the genetic basis of inferring H.armigera and h.assulta sex pheromone difference. The tobacco budworm Helicoverpa armigera single male and female hybrid (SR), produce offspring of both sexes, hybrid generation (SR) of female sex pheromone showed bollworm phenotype; when the hybrid generation (SR) and female moths of Helicoverpa armigera (R) male moth backcross, backcross (SRR) female information in all with Z11-16: Ald, showed bollworm phenotype; when the hybrid generation (SR) female moth and tobacco budworm (S) male moth backcross, backcross (SRS) two female sex pheromone phenotype, a phenotype for Z11-16:Ald based, performance for the cotton bollworm to the phenotype phenotype. Z9-16:Ald is the main performance for the tobacco budworm, phenotype, and the two individual number ratio of 1:1. When the cotton bollworm and tobacco budworm female male single pair of hybrid, hybrid generation (RS) all male. When the hybrid generation (RS) male moth and H. armigera females (R) backcross, backcross generation RSR) female sex pheromone were dominated by Z11-16:Ald, showed bollworm phenotype, while hybrids (RS) male moth and tobacco budworm (S) female moth backcross, backcross (RSS) female sex pheromone table type separation performance for two phenotypes: a phenotype to Z11-16:Ald dominant, showed bollworm phenotype, the phenotype of Z9-16:Ald based, performance for the tobacco budworm phenotype, and two of the phenotypic individual number ratio close to 1:1. The cross and backcross experiments in two directions showed that the difference in the sex pheromone ratio of cotton bollworm and tobacco was a recessive inheritance controlled by single gene. Furthermore, we used DNA-AFLP technology to locate the sex pheromone ratio differentially gene in parental, hybrids and metasomatism. 5 molecular markers were obtained by DNA extraction, enzyme digestion, connection, amplification and further specific amplification, and then combined with phenotypic data. The corresponding nucleotide sequences of 5 typical markers were recovered and sequenced, which laid the foundation for the subsequent gene cloning and location. In summary, two species of Helicoverpa armigera and h.assulta of hybridization and backcrossing using the research, analysis, and hybrid backcross parent female sex pheromone component ratio, using DNA-AFLP molecular markers by genetic studies of H.armigera and h.assulta hybrid lineages, exploring the genetic basis of cotton bollworm and tobacco budworm sex pheromone proportion upside down, to explore the interspecific isolation mechanism, reveals the important pests (Helicoverpa armigera and h.assulta) sex pheromone production mechanism of genetic differences, provide the foundation for the follow-up of new technology and methods based on research and development of pest management.
【学位授予单位】：安徽大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S433

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