【作者】 唐森森；

【导师】 李大全；

【作者基本信息】 石河子大学 ， 动物遗传育种与繁殖， 2006， 硕士

【摘要】 远缘杂交在研究物种的形成、进化、系统发育和遗传等生物学问题上具有重要意义。鸡与鹌鹑杂交是属间杂交,其杂交种胚胎早期出现大量的死亡,并且杂交种出壳以前雌性胚胎全部死亡,这使鸡与鹌鹑杂交成为研究异种生殖细胞不亲和性和杂交种不育的典型。随着对鸡与鹌鹑远缘杂交研究的深入,迫切需要从分子生物学角度来解释引起杂交种早期胚胎死亡的原因。为了探讨鸡与鹌鹑杂交种胚胎mRNA在发育早期的表达情况,以及mRNA特异性表达基因对于早期死亡所起的作用,展开本试验。本试验以鸡(♂)和鹌鹑(♀)正交组合杂交种的早期胚胎作为研究对象,首先从鹌鹑W染色体上设计引物,对杂交种胚胎性别进行鉴定。结果雌性胚胎获得特异性目的条带,雄性胚胎没有得到条带,通过性别鉴定,发现存活胚胎中大部分为雄性,那么死亡的胚胎则雌性占多数。观察胚胎发育情况,发现杂交种胚胎在孵化早期67-91h,胚胎生长呈平缓增长的趋势,72-79h的杂交种胚胎生长速度相对较快,杂交种胚胎在孵化到第72h胚胎死亡率明显高于其他时段,而74h胚胎死亡率低于其他时段。推测杂交种胚胎死亡的时间集中在69-72h,这一时间段胚胎的死亡,是导致整个孵化过程胚胎死亡率升高的主要原因。其次,对入孵67—91h的鸡与鹌鹑杂交种进行mRNA差异显示,筛选出杂交种胚胎发育早期的差异表达基因,结果表明,获得的胚胎不同发育时期六个基因片段与鸡的mRNA具有高度同源性,说明这些差异表达的基因为已知基因。其中,孵化69h胚胎经比对,与鸡雌激素受体结合位点相关抗原mRNA具有99%同源性,且表达时间比已知表达时间提前了12小时,很可能是造成杂交种胚胎早期大量死亡的原因之一。更多还原

【Abstract】 Cross breeding is the main breeding method and the important approach of incubate the new species. Chicken and quail intergeneric hybridization is a classic distant hybridization in fowl. In fowl science, chicken and quail belong to Aves, Galliformes, Phasiamidae, Gallus and Coturnix, and this hybridization combination has become the example of researching the incompatibility of different germ cell and sterile of distant hybridization. In the point of molecular biology the mRNA has different display of certain organism in different development stages and different cellular types. So analysis the function of gene expression is the key point in early embryonic development.The experiment investigated chicken(♂) and quail’s(♀) early hybridization embryos. First, we observed the embryo and proved that when the fertilized egg hatched for 67-91h, the embryo’s growth had the gentle increase trend . Most embryo dead concentrated in hatching for 69-72h, and the death in this phase was the main reason of the whole hatching progress.Second, In order to sexing the chicken and quail’s hybridization, I designed a pair of primers referencing the Wpkci gene in quail’s W chromosome, and the gene is special in sexing the quail. The primer amplified the cDNA of chicken and quail hybridization’s embryo and obtained two special bands in female one, while male one received nothing. The experiment has built a method to sexing the hybridization of chicken and quail in early stage of embryo development.Third, we applied mRNA Differential Display to screening differential genes during early embryonic development of chicken and quail hybridization, cloned and analysised these sequences’relationship with the embryonic development of hybridization at the early stage, and presumed the main infection to the death of the early embryo. The result shows that six genes have high homoeology with several chicken mRNA, Gallus gallus AT rich interactive domain 3C mRNA, hypothetical protein mRNA, estrogen receptor binding site associated antigen mRNA , similar to peptidyl prolyl isomerase E mRNA. Especially we found the estrogen receptor binding site associated antigen mRNA may be have main infection with the early embryonic development.更多还原