GENETIC DISSECTION OF AGGRESSIVE BEHAVIOR OF HONEY BEES

蜜蜂攻击行为的基因解剖

• 批准号: 6017098
• 负责人: GREG J HUNT
• 金额: $ 10.5万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-06-01 至 2001-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6017098
• 关键词: Hymenoptera; aggression; behavioral /social science research tag; ethology; gene expression; genetic mapping; genetic markers; genetic polymorphism; genotype; insect poison; instinct; quantitative trait loci; social space; species difference; territoriality

DESCRIPTION: This is a first award application requesting funding to study the genes and gene action involved in defensive behavior in honey bees. The project will focus on the two identified QTLs recognized to affect defensive behavior in Africanized honey bee colonies. Both single gene effects as well as social interactions with those genes will be addressed by the development of behavioral assays that permit characterization of defense behavior and its components at both the whole colony and individual level. The PI lists a number of hypotheses to be tested with respect to defensive behavior. These hypotheses are: (1) that stinging behavior on individual bees is affected by the same QTLs that influence behavior at the colony level; (2) that defensive behavior per se can be partitioned into number of components through the development of specific behavioral assays and markers linked to targeted QTLs; (3) that the alleles at QTLs show genetic dominance; (4) that defensive behavioral QTLs can be precisely mapped by correlating individual level behavior with the segregation of linked markers. The PI reports a number of relevant preliminary studies and observations. Using RAPD markers, a high density map of one marker every 100 to 200 kb has been developed for the honey bee, and the PI had the major role in this development. This map has already been used to map the sex-determination gene. Preliminary studies of 172 backcross colonies derived from hybrids of Africanized and European bees have identified two QTLs (designated sting-1 and sting-2) that are associated with variation in standard behavioral assays for stinging behavior, such as the number of stings in a defined period. The proposed investigation will extend and better define this association at both the individual and colony level. This proposal first calls for the development of behavioral assays that will maximize the defensive behavioral differences that exist between Africanized and European honey bees. Emphasis will be placed on aspects of behavior that may be quantified at the individual level. Most prior studies have emphasized colony-level stinging response, and this has tended to ignore intracolony interactions in the expression of both individual and overall colony response. The defensive components to be examine are likelihood to sting first, likelihood to sting, time to respond, and total numbers of stings. One approach that will be used is the marking of individuals in synthetic colonies of mixed genetic origin. This will allow an assessment of the interaction between colony genotype composition and genotype behavior, as well as the effect of colony size on genotypic behavior. For example, if Africanized genotypes are the first to sting in defense of a colony does this increase the tendency to sting of bees of European genotype. In addition, the effects of age will be addressed as well as extrinsic features of colony environment, such as the proximity of nectar sources. In the end, those conditions and components of behavior that maximize differences between the two races of bees will be used to address questions concerning colony and individual differences, degree of dominance, and to further map putative QTLs. The proposal is very specific with respect to the origin of parental stocks, and details of all inter race crosses and backcrosses. In general, the mapping of effects at the colony and individual levels emphasizes different designs with respect to F1 and F2 backcrosses, recognizing the haplo-diploid form of inheritance in hymenoptera. Initially, the sources of the Africanized lines will be 20 colonies of bees verified to be or African origin by mitochondrial DNA analysis and morphological features. These bees will come from established and characterized feral bee swarms collected in the state of Guerrero Mexico and selected using standard behavior assays for colonies showing the strongest defensive behavior, assuming that these lines have the greatest likelihood of not segregating for defensive alleles. One European queen will be used as the source of all virgin queens for F1 crosses with recombinant drones of hybrid origin. A foundation stock of European colonies will be established from crosses between individuals from colonies with the lowest defensive scores. Basically, colony specific behaviors are examined by taking drones (males) produced from F1 hybrid females and crossing them back to identical females of either origin and thus producing individual colonies where female workers are all identical within colonies with respect to their fathers recombinant genome. Differences between colonies reflect different meiotic products derived from the hybrid females. In the study of individual differences, Within colony variation in (female) workers arises in crosses where the mother is heterozygous with respect to origin and daughters differ with respect to mei osis in the mothers germ line. The paternal genome is constant in this case. Within the colony, the association of individual components of behavior with candidate QTL markers is followed. Individual molecular marker genotypes are assessed by either sacrificing whole individual bees following behavior characterization, or in some cases by sampling stingers left after attack on a leather patch. Finally, it is proposed to map at higher resolution the two putative QTLs identified in the preliminary studies. This will be accomplished by increased saturation of the genome with four different types of markers and examining their segregation with respect to individual behaviors (the original mapping was carried out at the colony level and only involved 172 meioses). By characterizing 1000 individuals in backcrosses, it should be possible to refine the map position to within 0.3 cM with 95% confidence. It is proposed to use STS primers designed from the RAPD markers, SSCP polymorphisms and further development of another set of RAPD markers from 1000 10-mer primers, screened in bulk.

描述：这是要求资助学习的第一个奖项申请 蜜蜂防御行为涉及的基因和基因作用。 这 项目将集中于确定的QTL，以影响防御性 非洲蜜蜂殖民地的行为。 两个单基因效应 以及与这些基因的社交互动将由 制定允许国防表征的行为测定 行为及其在整个群体和个人层面上的组成部分。 PI列出了许多有关防御性的假设 行为。 这些假设是：（1）单个蜜蜂上的刺痛行为是 受影响在菌落水平上影响行为的相同QTL的影响； （2） 这种防御行为本身可以分为组件数量 通过开发特定的行为分析和标记 目标QTL； （3）QTL的等位基因显示遗传优势； （4）那 防御行为QTL可以通过相关的个人来精确映射 链接标记的分离的水平行为。 PI报告了许多相关的初步研究和观察。 使用RAPD标记，每100至200 kb的高密度图具有一个标记 是为蜂蜜蜜蜂开发的，PI在此中具有主要作用 发展。 该地图已被用来绘制性别确定性 基因。 对源自杂种的172个反向交叉菌落的初步研究 非洲和欧洲蜜蜂已经确定了两个QTL（指定的Sting-1 与标准行为变化有关的sting-2） 刺痛行为的测定法，例如定义的刺激数量 时期。 拟议的调查将扩展并更好地定义 个人和殖民地层面的关联。 该建议首先要求开发行为测定法 最大化非洲人之间存在的防御行为差异 和欧洲蜜蜂。 重点将放在行为方面 可以在个人级别进行量化。 大多数先前的研究都有 强调殖民地的刺痛反应，这往往忽略 个体和整体表达中的圆内相互作用 殖民地反应。 要检查的防御组件是可能性的 首先刺痛，刺痛的可能性，回应时间和总数 刺。 一种将使用的方法是个人的标记 混合遗传起源的合成菌落。 这将允许评估 菌落基因型组成与基因型之间的相互作用 行为以及菌落大小对基因型行为的影响。 为了 例如，如果非洲化的基因型是第一个捍卫一个 殖民地这样做会增加欧洲蜜蜂刺痛的趋势 基因型。 此外，将解决年龄的影响以及 殖民地环境的外在特征，例如花蜜的接近度 来源。 最后，那些行为的条件和组成部分 最大化蜜蜂种族之间的差异将用于解决 有关殖民地和个体差异，优势程度的问题， 并进一步映射推定的QTL。 该提案在父母股票的起源方面非常具体， 以及所有种族交叉和反向交叉的细节。 通常， 在殖民地和个体层面的效果映射强调不同的 关于F1和F2反向交叉的设计，识别单倍体二倍体 膜翅目遗传的形式。 最初， 非洲线将是被证实为或非洲的20个殖民地 线粒体DNA分析和形态特征的起源。 这些蜜蜂 将来自收集在 格雷罗墨西哥的状态，并使用标准行为分析选择 假设这些线条表现出最强的防御行为 有最大的可能性不隔离防御等位基因。 一 欧洲女王将用作F1的所有处女皇后的来源 与混合起源的重组无人机交叉。 基础股票 欧洲殖民地将从个人之间的十字架建立 防守得分最低的殖民地。 基本上，通过服用无人机（男性）来检查菌落特定的行为 由F1杂种女性产生，并将其交叉回相同的女性 源头，因此产生了女工的单个殖民地 与父亲重组有关的殖民地中都相同 基因组。 殖民地之间的差异反映了不同的减数分裂产品 源自杂种女性。 在研究个体差异时， 在（女）工人的殖民地变异中发生在十字架中 母亲在起源方面是杂合的，女儿的不同 尊重母亲种系中的元素症。 父亲基因组是 在这种情况下，恒定。 在殖民地中，个人的关联 遵循候选QTL标记的行为组成部分。 个人 通过牺牲整体评估分子标记基因型 遵循行为表征的个体蜜蜂，或者在某些情况下 在攻击皮革补丁后留下的抽样刺痛器。 最后，提议以较高分辨率映射两个推定的QTL 在初步研究中确定。 这将通过 具有四种不同类型的标记的基因组饱和度增加，并且 研究他们对个人行为的种族隔离（ 原始映射是在殖民地层进行的，仅涉及172 Meioses）。 通过表征1000个人在反向交叉中，应该是 可以以95％的置信度将地图位置完善到0.3厘米以内。 建议使用由RAPD标记设计的SSCP设计的STS引物 多态性和进一步发展的另一组RAPD标记 1000 10-Mer底漆，批量筛选。