Characterizing the molecular and developmental basis of environmental versus genetic trait variation in aphids

描述蚜虫环境与遗传性状变异的分子和发育基础

基本信息

批准号：
9145717
负责人：
JENNIFER A BRISSON
金额：
$ 30.33万
依托单位：
UNIVERSITY OF ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-09-17 至 2020-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9145717
关键词：
Address Affect Aphids Biological Models Bypass CRISPR/Cas technology Complex Cues Data Development Disease susceptibility Ecdysone Embryo Employee Strikes Endocrine Environment Environmental Risk Factor Exhibits Experimental Models Female Gene Expression Gene Expression Profiling Genes Genetic Genetic Polymorphism Genetic Recombination Genetic Variation Genomics Goals Health Height Hormonal Human Human Characteristics In Situ Hybridization Injection of therapeutic agent Investigation Knowledge Lead Learning Life Cycle Stages Lighting Link Maps Measures Methods Mission Modeling Molecular Morphology Mothers Mutate Nature Pathway interactions Phenotype Pisum Pisum sativum Positioning Attribute Prevalence Process Production Publications Regulator Genes Research Role Signal Transduction Staging System Testing Time Translating Variant Weight Wing Work X Chromosome asexual base developmental genetics dimorphism experience fascinate genome sequencing human disease innovation knock-down male mutant novel offspring programs steroid hormone trait

项目摘要

DESCRIPTION (provided by applicant) Phenotypic plasticity affects an array of human characteristics, from height and weight to disease susceptibility. Yet the mechanisms by which environmental inputs are detected and then alter developmental programs are not well understood, and experimental models for studying this link are limited. This project addresses the fascinating problem of how both the environment and genetic variation influence the development of alternative phenotypes using an innovative model system, the pea aphid. This species offers an unparalleled opportunity to examine the role of nature and nurture in phenotype determination: it exhibits dramatically different winged and wingless morphs that are induced by environmental conditions in asexual females and by a single unidentified genetic locus in males. Thus, strikingly, two dimorphisms, each under different control mechanisms, exist within this single species. Here we propose a comprehensive investigation of the developmental and genetic basis of both dimorphisms, the first study of its kind, with a research plan that builds upon extensive preliminary data. In Aim 1 we will identify and characterize the wing polymorphism locus to illuminate how this single locus can cause dramatic differences in morphology in males. Our methods include association mapping using genome sequencing, gene expression level profiling across development using qRT-PCR, gene expression spatial profiling using in situ hybridization at targeted developmental stages, and mutant analysis using the CRISPR/Cas9 system. In Aim 2, we will study the hormonal basis of wing determination to ascertain how environmental cues are translated into phenotypic differences in females. We will test the role of a steroid hormone, ecdysone, in this process. We will measure ecdysone titer levels in induced mothers (winged offspring producers) versus uninduced mothers (wingless offspring producers), and experimentally manipulate their offspring phenotypes via injection of ecdysone. Further, we will measure expression levels of ecdysone-responsive genes via qRT-PCR in the embryos of mothers receiving or not receiving wing-inducing signals. In Aim 3, we will examine the extent to which there are shared mechanisms underlying the environmentally induced and genetically controlled dimorphisms, to establish how the two dimorphisms are related at the mechanistic level. We will approach this problem by looking for an effect of mutating the male polymorphism gene on the female alternative morphs and by measuring gene expression of ecdysone-responsive genes in winged and wingless male aphids via qRT-PCR. Overall, this research will result in a model of how both environmental and genetic factors influence similar alternative phenotypes. We anticipate that a major finding of this work will be the illumination of how environmental signals are bypassed when there is an evolutionary transition to genetic control of phenotypic variation.

描述（由申请人提供）表型可塑性会影响从身高和体重到疾病敏感性的一系列人类特征。然而，检测到环境输入并更改发展计划的机制尚不清楚，并且研究此链接的实验模型受到限制。该项目解决了使用创新的模型系统PEA蚜虫影响环境和遗传变异如何影响替代表型的发展的令人着迷的问题。该物种提供了一个无与伦比的机会来检查自然和护士在表型确定中的作用：它表现出明显不同的有翼和无翅的形态，这些变态是由无性雌性中环境条件以及男性单一的独立遗传基因座的环境条件引起的。该物种提供了一个无与伦比的机会来检查自然和护士在表型确定中的作用：它表现出明显不同的有翼和无翅的形态，这些变态是由无性雌性中环境条件以及男性单一的独立遗传基因座的环境条件引起的。令人惊讶的是，在这个单一物种中存在两个二态性，每个二态性都存在于不同的控制机制下。在这里，我们提出了对两种二态性的发展和遗传学的全面研究，这是对同类的第一个研究的基础，该研究的研究计划以广泛的初步数据为基础。在AIM 1中，我们将确定并表征机翼多态性基因座，以阐明该单个基因座如何导致男性形态的巨大差异。我们的方法包括使用基因组测序，使用QRT-PCR跨发育的基因表达水平分析，基因表达在靶向发展阶段进行原位杂交的基因表达空间分析以及使用CRISPR/CAS9系统的突变体分析。在AIM 2中，我们将研究机翼决心的马基础，以确定如何将环境线索转化为女性的表型差异。我们将在此过程中测试类固醇骑马ecdysone的作用。我们将测量诱发母亲（有翼的后代生产者）与未诱发的母亲（无翅的后代生产者）中的ecdysone滴度水平，并通过注射ecdysone来实验操纵其后代表型。此外，我们将通过QRT-PCR测量ecdysone响应基因的表达水平，这些母亲的胚胎接受或不接受诱导翼信号。在AIM 3中，我们将研究具有环境诱导和遗传控制的二态性的共同机制的程度，以确定两种二态性如何在机械级别相关。我们将通过寻找突变雄性多态性基因对女性替代形态的影响以及通过QRT-PCR中ecdysone反应基因的基因表达来解决这个问题。总体而言，这项研究将导致一个模型，即环境因素和遗传因素如何影响相似的替代表型。我们预计，这项工作的一个重大发现将是对环境信号的绕过对表型变异的遗传控制的进化过渡时的绕过的照明。