导　　师： 陈盛禄;钟伯雄

学科专业： 090504

授予学位： 博士

作　　者： ;

机构地区： 浙江大学

摘　　要： 该研究采用微卫星dna分析技术、基于混合线性模型的条件和非条件方差分析方法,对浙农大1号浆蜂(ea)、本地意大利蜜蜂(eb)、原种意大利蜜蜂(ee)进行了分子遗传和数量遗传的研究.主要研究内容包括:微卫星dna的等位基因频率、多态信息含量(pic)、群体杂合度(h)、遗传距离、蜂王浆高产dna标记分析;应用条件和非条件方差分析方法对蜂群产浆量、台浆量、台基接收率、咽下腺及蜂群繁殖力进行发育遗传研究,得出如下结论:1.通过对10对荧光引物进行pcr扩增、图谱扫描和基因分型,在10个微卫星位点在ea、ee、eb中扩增出2~15个等位基因不等,共扩增到96个等位基因,平均每个位点扩增9.6个,其中有48个(占总数的50﹪)等位基因在ee、eb和ea中是不相同的,在ea、eb、ee中分别发现6个、6个、12个特有等位基因;10个微卫星位点在3蜂种中共扩增到的等位基因数分别为75、61、66,平均每个位点的等位基因为7.5、6.1、6.6.表明微卫星dna在3蜂种中存在较高的多态性(基因多样性),同时也说明ea经过长期的地理隔离和人工选择,其遗传基础已与ee和eb发生了不同程度的改变.2.该研究的10个微卫星位点在ea、ee、eb中的平均pic分别为0.57、0.53、0.57,均大于0.5,差异不显著.pic是衡量片段多态性的指标,按照pic的评判标准,10个微卫星位点均多态性良好,说明ea与ee和eb一样具有高度的群体内遗传多样性.3.群体基因杂合度的分析表明,ea、ee、eb的杂合度分别为0.61、0.57、0.60,差异不显著.群体杂合度表示被检测的位点上群体中的杂合子频率.4.品种之间的内氏遗传距离可较准确地反映群体间的亲缘关系,结果表明ea与ee间的距离为0.25,ea与eb间的距离为0.16,ee与eb间的距离为0.14.由此可见,ea和ee间的亲缘关系较远,ee和eb以及eb和ea的亲缘关系较近.5.通过对10个微卫星位点等位基因频率的分析发现,在6个微卫星位点上7个等位基因频率在ee、eb、ea中依次增加,3个微卫星位点的3个等位基因的频率在ee、eb、ea依次降低,1个微卫星位点上的1个等位基因在ee和eb中频率较高,但ea没有.6.采用条件及非条件遗传效应方差的统计分析方法,分析蜂群产浆量、台浆量、台基接受率3性状的遗传特征及成对性状之间的关系.成对性状之间的相关研究表明,蜂群产浆量和台浆量之间在各个时期存在显著或极显著的遗传相关,说明2性状基因效应之间的协同作用是一致的;而蜂群产浆量和台基接受率之间在大多时期存在显著的遗传相关,但在一些时期没有相关,2性状之间的基因效应协同作用较差.7.应用条件和非条件分析的方法对3品种西方蜜蜂的蜂群繁殖力和工蜂咽下腺重量进行了发育遗传研究. When human beings were aware of the natural medical functions of royal jelly in the 1950s, its composition and pharmacological effects were investigated comprehensively worldwide. The production technology underwent continuous studies since the 1960s, whereas, China is always leading to this field. China has now been produced around 2000 tons of royal jelly each year, accounting for the 90 per cent of the world's total output. This is attribute to Chinese style of royal jelly production technology led by high royal jelly producing bee, thus making the royal jelly production as much as 10 times higher than last decade in China. Not only makes it a big contribution to humankind's health, but also brings a production revolution to beekeeping. The genetic characteristics of the high royal jelly bee specific to China, however, have not been investigated so far. So, the researches focusing on this area will be actively promoted honeybee breeding, and reasonable resources exploitation and protection. The approaches of microsatellite DNA and conditional and unconditional genotypic variances based on mixed liner model were employed to assay the molecular and quantitative genetic features of the 3 species of western honeybees/(//4pis mellifera L./), namely, pure Italy bee/(Ee/), Chinese Italy bee/(Eb/) and the high royal jelly producing bee/(Ea/). The main research contents of this project were involved in allele frequencies, genotypic frequencies, polymorphysm information content/(PIC/), population heterozygosity, genetic distance and the microsatellite DNA genetic markers of the high royal jelly producing bee by microsatellite DNA; meanwhile the conditional and unconditional genotypic variances were conducted to analyze the developmental genetics characteristics of the royal jelly production, royal jelly in each cell, acceptance of queen cell cups, the weight of hypopheryngeal gland of worker bees and the colony population size. The principal results of this studies are as followings: 1. Through PCR amplification, gene scanning and genotyping at 10 microsatellite loci, 2-15 alleles can be detected at different loci among Ee, Eb and Ea. There were total 96 alleles amplified among 10 loci in Ee, Eb and Ea with an average of 9.6 alleles were found in each locus. Among the 96 alleles, 48 of it were different in Ee, Eb and Ea. Moreover, there were 6, 6 and 12 alleles specific to Ea, Eb and Ee, respectively. There were 75, 61,66 alleles amplified at 10 loci with an average allele 7.5, 6.1 and 6.6 at each loci in Ee, Eb and Ea respectively. These results showed that there were higher polymorphysim or genetic diversity among Ee, Eb and Ea, indicating that the genetic features of Ea were different from that of Ee and Eb because of geographical isolation and human selections. 2. The average polymorphysim information content/(PIC/) of Ee, Eb and Ea at 10 loci were 0.57,0.53, 0.57 respectively with no differences. The PIC is index of judging the polymorphysim of the fragments. By the standard, the 10 loci had a high polymorphysim amplified in Ee, Eb and Ea. It suggested that Ea had a similar high genetic diversity to Ee and Eb. 3. Heterozygosity analysis indicated that Ee, Eb and Ea were 0.61, 0.57, 0.60 respectively with no differences. Heterozygosity is a heterozygote frequency in each locus. The lower heterozygosity, the higher genetic purity in the population indicated a lower genetic variances and diversity in the population. Thus it can be seen that there were similar high heterozygosity among Ea, Ee and Eb. The results also suggested that the excellent royal jelly production of the Ea maybe better performing the hybrid vogor. 4. The Nei's genetic distance can correctly reveal the genetic relationship among the populations. The results showed that the distance between Ea and Ee, Ea and Eb, Ee and Eb were 0.25, 0.16, 0.14 respectively. It suggested that the affiliation Ea and Ee was far away, while Ea and Eb, Ee and Eb were close. This was coherent to the evolutionary history of the Ea, that was Ea selected from Eb, while Eb

关 键 词： 蜜蜂 蜂王浆 微卫星 遗传效应方差 蜂群繁殖力 基因型方差

分 类 号： [Q953]

领　　域： [生物学]