【作者】 陈晶；

【导师】 陈国宏；

【作者基本信息】 扬州大学 ， 动物遗传育种与繁殖， 2008， 硕士

【摘要】 本研究以来自于南昌、黄山、桐庐、费县、宜兴、武夷山等6个华东地区的中华蜜蜂(中蜂)群体以及平湖浆蜂、苏王1号等两个意大利蜜蜂(意蜂)群体共523只(形态测定500只)工蜂为研究素材,利用23对从西方蜜蜂中筛选出的微卫星引物,分析其基因组DNA在23个微卫星位点上的遗传变异,同时测量10个主要形态性状,以期用微卫星DNA多态性与形态学相结合的方法来分析华东境内8个蜜蜂群体的遗传多样性。主要研究结果如下:1、对于测定的10个形态特征,群体间存在着极显著的差异(P <0.01)。两个意大利蜜蜂群体平湖浆蜂和苏王1号在前翅长、前翅宽、前翅面积、b值、第3背板长上均与其它6种中蜂差异显著(P <0.05),且第4背板及第3+4背板总长与除桐庐中蜂外的其他中蜂群体均差异显著(P <0.05)。蜜蜂翅长、翅宽、翅面积、吻长及其背板长等性状间均有显著相关。单独进行形态特征聚类与进行主成分分析后再聚类的结果差异不大,与环境因素结合后聚类表明,环境因子对蜜蜂形态特征的影响非常明显,因此应将生态因子作为蜜蜂群体区分的一个重要方面。2、利用所选的23个微卫星位点对6个中蜂群体以及平湖浆蜂、苏王1号进行多态性检测,均得到了特异性扩增结果,共检测到430个等位基因,平均每个位点的等位基因数为18.696个,单个位点的等位基因数从6 (BI366)到34 (AP043)不等。所有群体的期望杂合度为0.702至0.939,PIC值为0.653至0.936。平均期望杂合度为0.844±0.069,PIC值为0.828±0.080。23个微卫星位点均具有较高的多态性。单个位点偏离Hardy-Weinberg平衡的群体数从5-10不等。群体间分化系数为33.6% (P <0.001),所有位点都极显著地贡献于这一结果(P <0.001)。杂合子缺失水平很高为0.572 (P <0.001)。平湖浆蜂、苏王1号与6个中蜂群体间也存在着极显著的遗传分化,群体间的Reynolds’遗传距离从0.215(平湖浆蜂-苏王1号)到0.683(费县中蜂-武夷山中蜂)不等,而Nm值变异范围为从0.255(费县中蜂-武夷山中蜂)到1.041(平湖浆蜂-苏王1号)之间。中华蜜蜂与意大利蜜蜂均表现出较高的群体杂合度和丰富的遗传多样性。3、利用23对微卫星标记对8个群体的亲缘关系进行分析,NJ系统发生树显示,8个蜜蜂群体可以分为两大类,即中华蜜蜂、意大利蜜蜂各一大类,宜兴中蜂聚在所有中蜂的最外部。黄山中蜂与桐庐中蜂的基因流动值为0.821,在6个中蜂中基因流动值最大。平湖浆蜂和苏王1号与其它中华蜜蜂群体的总体遗传关系都比较远。皖南山区的黄山中蜂与江淮地区的宜兴中蜂遗传差异性较大。武夷山中蜂同其它5个群体的基因流动值很小,是一个相对隔绝的群体。4、利用微卫星DNA分析6个中华蜜蜂群体遗传距离与地理距离的关联性,6个中蜂群体的遗传距离和地理距离回归公式:FST/(1-FST) =–0.2804 + 0.8722ln (d),以及Mantel’s检验的结果(P = 0.104)并不能为遗传距离与地理距离之间的显著联系提供足够的证据。说明华东6个中蜂群体的地理分布可能不是影响其群体遗传结构的决定因素。5、苏王1号群体杂合度和遗传多样性高于平湖浆蜂,两者遗传距离较近,而且彼此之间有较大的基因流动。更多还原

【Abstract】 23 microsatellite markers from Apis mellifera ligustica were used to analyzed the genetic diversity of Apis cerana cerana populations and Apis mellifera ligustica, the six populations of Apis cerana cerana were from Nanchan, Huangshan, Tonglu, Feixian, Yixing, Wuyi, respectively. The two populations of Apis mellifera ligustica were Pinghu royal jelly bee and Suwang No.1 bee. At the same time, the ten main indexes of external morphology such as the length, width and area of the anterior wing, the cubital vein indexes, the length of proboscis and the length of terga 3+4 were detected. Genetic variability within populations and genetic differentiation among poputions were estimated. The main results were summarized as follows.1. There existed significant differences(P <0.05) among 10 characteristics of 8 populations in East-China. It had significant differences(P <0.05) betwwen Pinghu royal jelly bee and Suwang Suwang No.1 bee in length of bee right forewing, width of bee right forewing, area of the anterior wing, length of bee right forewing cubital vein b, bee length of tergum 3. The correlation between morphology features was significant (P <0.05) in length of bee right forewing, width of bee right forewing, area of the anterior wing, length of proboscis, length of tergum. Cluster by morphological characteristics and principal component analysis did not differ greatly. Clustered by morphological characteristics and ecological factors, which showed that ecological factors on the impact of bees morphological characteristics was very obvious. It suggested that the ecological factor was also an important side on the population classificiation.2. Twenty-three pairs of microsatellite primers were tested in six Apis cerana cerana populations and two Apis mellifera ligustica populations. 23 pairs of microsatellite primers can be successfully amplified. The results showed that 430 alleles were found in 23 microstaellite loci, the number of alleles per locus ranged from 6 (BI366) to 34 (AP043), mean number of effective alleles was 18.696. The gene heterozygosity (He) of all populations ranged from 0.702 to 0.939, and the average value was 0.844±0.069, respectively. The polymorphism information content (PIC) ranged from 0.653 to 0.936, and the average value was 0.828±0.080. All 23 microsatellite loci in this study showed high levels of polymorphism. The number of populations deviated from Hardy-Weinberg equilibrium per locus ranged from 5 to 10. In the all populatons, the genetic differentiation among populations, measured as FST value was 33.6% (P <0.001), and all loci were contributed to this differentiation significantly (P <0.001). Significant genetic differentiation was also observed between two subspecies of Apis cerana cerana and Apis mellifera ligustica. And the deficit of heterzygote was observed very high (0.572) (P <0.001). Reynolds’distance values varied between 0.215 ( Pinghu royal jelly bee and Suwang No.1 bee pair) and 0.683 ( Apis cerana cerana from Feixian and Wuyi pair), The Nm value was ranged from 0.255 ( Apis cerana cerana from Feixian and Wuyi pair) to1.041 ( Pinghu royal jelly bee and Suwang No.1 bee pair).3. The phylogenetic relationship among 8 bee populations were analyzed, and an un-rooted consensus tree was constructed using the Neighbour-Joining method. The tree topology revealed two main clusters. 6 Apis cerana cerana populaitions formed one cluster, and Apis cerana cerana from Yixing was found in the outside. Pinghu royal jelly bee and Suwang No.1 were formed the second cluster. The Nm value of 0.821 between Apis cerana cerana Huangshan and Tonglu was largerest in 6 Apis cerana cerana populaitions. The evolutional dendrogram of Pinghu royal jelly bee and Suwang No.1 was further than any Apis cerana cerana populaitions. It appeared rather farer phylogenetic relationship with Apis cerana cerana from Huangshan and Yixing. The Nm value of Apis cerana cerana from Wuyi and the other Apis cerana cerana populaitions was small, the Apis cerana cerana from Wuyi may be segregated.4. The geographical elements may own to the relationship for particular population pairs, however, the equation FST/(1-FST) =–0.2804 + 0.8722ln (d) and the results from Mantel’s test (P=0.104) did not provide enough support for a significant correlation between the genetic and the geographical pair wise distances. The results concluded that the geographical distributing may not be the determinant influence on the genetic structure of Apis cerana cerana populations.5.The genetic diversity of Suwang No.1 bee was higher than Pinghu royal jelly bee. The results indicated that the populations of Pinghu royal jelly bee and Suwang No.1 bee had a tendency towards intermixing.更多还原