本文關鍵詞：棉鈴蟲和煙青蟲性信息素差異的遺傳機制　出處：《安徽大學》2017年碩士論文　論文類型：學位論文

  更多相關文章： 棉鈴蟲 煙青蟲 性信息素 遺傳家系

【摘要】：近緣種及同種不同品系間的生殖隔離現(xiàn)象一直是物種形成研究的熱點。鱗翅目許多昆蟲往往具有相同類型的性信息素化合物,并通過改變各組分的比例實現(xiàn)種間生殖隔離。近年來,陸續(xù)報道了一些涉及蛾類性信息素產(chǎn)生及其遺傳結構基礎的研究,這些工作對闡明生殖隔離遺傳變化的規(guī)律提供了重要的信息。然而,在進化過程中,由性信息素的改變導致的近緣物種形成的遺傳機制尚不清楚。棉鈴蟲(Helicoverpa armigera 和煙青蟲(Helicoverpa assulta)是鱗翅目(Lepidoptera)夜蛾科(Noctuidae)兩種重要的農(nóng)業(yè)害蟲。棉鈴蟲為高度雜食性昆蟲,危害20科近200種植物,加之其強大的繁殖力和廣泛的分布性,致使其一直成為農(nóng)業(yè)害蟲防治的難點,而煙青蟲為寡食性昆蟲,主要危害茄科植物,但其危害性和破壞力也同樣不容小覷。棉鈴蟲和煙青蟲的幼蟲和成蟲在形態(tài)結構上都非常相似,很難辨認。有研究表明棉鈴蟲雌蟲性信息素成分主要以順-11-十六碳烯醛((Z)-11-hexadecenal,Z11-16:Ald)為主,而煙青蟲的性信息素成分主要以順-9-十六碳烯醛((Z)-9-hexadecenal,Z9-16:Ald)為主,棉鈴蟲和煙青蟲中Z11-16:Ald/Z9-16:Ald比例正好相反。自然條件下,二者通過釋放不同比例的性信息素組分完成種內(nèi)化學通訊,實現(xiàn)種間生殖隔離。并且二者之間可雜交和回交,因此是研究性信息素通訊系統(tǒng)遺傳差異的絕佳模型。本研究通過兩近緣種棉鈴蟲和煙青蟲單對雜交和回交,分析親本、雜交和回交雌蛾性信息素組分,系統(tǒng)研究兩近緣種性信息素比例差異的內(nèi)在遺傳機制。首先,分別剪取棉鈴蟲和煙青蟲性腺,采用正己烷浸提,進而通過GC-MS的選擇離子方法,分別測定了棉鈴蟲和煙青蟲雌蛾性信息素主要組分Z11-16:Ald和 Z9-16:Ald,證實了棉鈴蟲性信息素以 Z11-16:Ald 為主,Z11-16:Ald/Z9-16:Ald為 96:4,而煙青蟲性信息素以 Z9-16:Ald 為主,Z11-16:Ald/Z9-16:Ald 為 4:96,為本論文的后續(xù)研究奠定了堅實的基礎。然后,開展棉鈴蟲和煙青蟲雜交和回交實驗,測定雜交代和回交代雌蛾的性信息素表型,統(tǒng)計符合棉鈴蟲和煙青蟲性信息素表型的雜交代和回交代個數(shù),推斷棉鈴蟲和煙青蟲性信息素差異的遺傳基礎。煙青蟲雌性與棉鈴蟲雄性單對雜交(SR),產(chǎn)生兩性子代,雜交代(SR)雌蛾性信息素都表現(xiàn)為棉鈴蟲表型;當雜交代(SR)雌蛾與棉鈴蟲(R)雄蛾回交時,回交代(SRR)雌性性信息素均以Z11-16:Ald為主,表現(xiàn)為棉鈴蟲表型;當雜交代(SR)雌蛾與煙青蟲(S)雄蛾回交時,回交代(SRS)雌性個體出現(xiàn)兩種性信息素表型,表型一為以Z11-16:Ald為主,表現(xiàn)為棉鈴蟲表型,表型二為以Z9-16:Ald為主,表現(xiàn)為煙青蟲表型,且二者個體數(shù)量比例為1:1。當棉鈴蟲雌性和煙青蟲雄性單對兒雜交時,雜交代(RS)全部為雄性。當雜交代(RS)雄蛾與棉鈴蟲雌蛾(R)回交時,回交代RSR)雌蛾性信息素均以Z11-16:Ald為主,表現(xiàn)為棉鈴蟲表型,而當雜交代(RS)雄蛾與煙青蟲(S)雌蛾回交時,回交代(RSS)雌蛾性信息素表型發(fā)生分離,表現(xiàn)為兩種表型:表型一為以Z11-16:Ald為主,表現(xiàn)為棉鈴蟲表型,表型二為以Z9-16:Ald為主,表現(xiàn)為煙青蟲表型,且兩表型個體數(shù)量比例接近1:1。兩個方向的雜交和回交實驗證明棉鈴蟲和煙青蟲性信息素比例差異是由單基因控制的顯隱性遺傳。進而,我們運用DNA-AFLP技術對親代、雜交代和回交代進行性信息素比例差異基因定位。通過DNA提取、酶切、連接、擴增和進一步特異性擴增,再結合以表型的數(shù)據(jù),我們共獲得了 5個分子標記。通過對5個典型標記切膠回收并測序,分別得到了其對應的核苷酸序列,為后續(xù)基因克隆和定位奠定了基礎。綜上,本研究利用兩近緣種棉鈴蟲和煙青蟲單對雜交和回交,分析親本、雜交代和回交代雌蛾性信息素組分比例,利用DNA-AFLP分子標記技術,通過棉鈴蟲和煙青蟲雜交譜系的遺傳學研究,探究了棉鈴蟲和煙青蟲性信息素比例顛倒的遺傳基礎,探討了種間隔離機制,揭示了重要害蟲(棉鈴蟲和煙青蟲)性信息素產(chǎn)生差異的遺傳機制,為后續(xù)基礎研究和發(fā)展害蟲管理新方法與新技術提供基礎。
[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.
【學位授予單位】：安徽大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S433

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