Fine-Scale Genetic Mapping and Sequence Analysis of Genes Involved in Honey Bee Stinging Behavior

蜜蜂叮咬行为相关基因的精细定位和遗传分析。行为性状的遗传结构(基因数量及其效应)是行为生态学领域的一个重要考虑因素，但对此知之甚少。蜜蜂在这方面可能是有用的，因为它正被公认为行为和行为遗传学研究的模式生物。作为一种行为遗传模型，蜜蜂具有易于培养、社会行为、单倍体雄性以及迄今报道的后生动物最高的重组率等优势。较高的减数分裂重组率有利于构建详细的遗传图谱，并根据基因图谱上的位置克隆基因，因为遗传距离与交叉率直接相关。这项拟议的项目旨在研究特定基因和信息素交流在蜜蜂防御行为中的作用。以前的研究确定并绘制了数量性状基因座(QTL)，这些QTL标记了影响群体水平行为性状的基因的位置，即蜜蜂在行为测试中沉积的叮咬数量。此外，还确定了影响报警信息素产生的QTL。随后发现其中两个“刺痛”QTL影响个体蜜蜂的守卫和刺痛行为，从而证实了QTL对行为的影响。已确认的两个QTL中的一个也可能对报警信息素的产生产生影响。一个QTL影响一种与蜂群叮咬反应相关的未知报警信息素成分的水平，该QTL与这个防御行为QTL相近。这项拟议的研究将进一步表征QTL对个体和群体行为表型的影响，并试图确定受每个基因影响的行为的特定组成部分。此外，与单倍体雄性杂交将允许进一步的遗传解剖，以更准确地定位基因的位置，并最终获得这些基因的序列。这些杂交将产生一系列蜜蜂，每一种蜜蜂都是不同单倍体雄性蜜蜂的后代。在QTL附近的关键区域，单倍体父本会有所不同。拟议研究的下一步是识别跨越包含QTL的区域的蜜蜂DNA的大型细菌人工染色体(BAC)克隆。例如，如果影响行为特征的基因可以精确地定位到遗传距离的5厘米器官，这相当于大约250千碱基(KB)的蜜蜂DNA。由于目前蜜蜂BAC文库的平均克隆大小为113KB，因此只有几个BAC克隆就可以跨越这250kb的区域。然后，将使用测序和生物信息学分析来确定这些QTL的候选基因。

Fine-scale mapping and genetic analysis of genes involved in honey bee stinging behavior.ABSTRACT The genetic architecture (numbers of genes and their effects) of behavioral traits is an important consideration for the field of behavioral ecology, yet little is known on this topic. The honey bee may be useful in this regard because it is becoming recognized as a model organism for behavioral and behavioral genetic studies. As a behavioral genetic model, the bee has the advantages of ease of culture, social behavior, haploid males, and the highest recombination rate ever reported for a metazoan. The high meiotic recombination rate facilitates the construction of detailed genetic maps and cloning genes based on their positions on genetic maps because genetic distance is directly related to crossover rates. The proposed project seeks to study the role of specific genes and pheromonal communication in honey bee defensive behavior. Previous studies identified and mapped quantitative trait loci (QTL) that mark the positions of genes that influence a colony-level behavioral trait, the numbers of stings that bees deposited in a behavioral assay. QTL that influence alarm pheromone production were also identified. Two of the "stinging" QTL were subsequently found to influence the guarding and stinging behavior of individual bees, thus confirming the QTL effects on behavior. One of the two confirmed QTL also may have an influence on alarm pheromone production. A QTL that influences levels of an unknown alarm pheromone component that correlated with colony stinging response mapped close to this defensive-behavior QTL. The proposed research would further characterize QTL effects on individual and colony behavioral phenotypes and seek to identify specific components of behavior that are influenced by each gene. In addition, crosses with haploid males would allow for further genetic dissection to pinpoint the location of genes more precisely and ultimately obtain the sequence of these genes. These crosses would generate lines of bees, each of which is descended from a different haploid male. The haploid fathers will differ from each other in key regions near the QTL. The next step in the proposed research is to identify large bacterial artificial chromosome (BAC) clones of honey bee DNA that span the region containing the QTL. For example, if the gene influencing the behavioral trait can be pinpointed to 5 centimorgans of genetic distance, this represents about 250 kilobases (Kb) of honey bee DNA. This 250 Kb region can be spanned by just a few BAC clones because the current honey bee BAC library has average clone sizes of 113 Kb. Sequencing and bio-informatic analyses would then be used to identify candidate genes for these QTL.