The development, evolution and function of the Bullseye petal pattern in the Hibiscus trionum complex

主题：农业与粮食安全芙蓉复合体中“牛眼”花瓣图案的发展、进化和功能：确定导致不同花型的调控机制总体目标：了解造成木槿花及其近亲独特的牛眼图案的色素在近端和远端花瓣区域局部积累的过程。了解不同的牛眼几何形状在调节植物-传粉者相互作用中的作用。该项目进一步分为三个目标。目的1：确定调控花瓣发育过程中局部色素产生的调控和信号机制，以形成牛眼模式。系统发育；克隆；定量聚合酶链式反应表达；转基因过表达和基因敲除系；酵母杂交。目的2：鉴定和比较不同牛眼模式的品种，以了解导致牛眼进化的发育和分子变化。模式变异的形态特征：色素提取；花瓣图案和特征的图像分析；紫外线测量；潜在的模式进化的分子变化：H.t.richardsonii转录组；H.t.richardsonii转录组；H.T.商业和H.t.richardsonii品种的qPCR；基因分离和这些品系的转基因。目的3：探讨牛眼模式在调节花与传粉者相互作用中的作用。具体测试牛眼大小变化对传粉者行为的影响，并评估三种模式元素(色素、细胞形状和角质层装饰)中每一种对牛眼功能的相对贡献。问题1：测试牛眼模式大小的变化是否影响熊蜂(Bombus Terrestris)的相互作用通过差异条件实验确定熊蜂是否能够区分不同大小的着色区域的牛眼模式，并确定熊蜂所能感知的最小牛眼大小差异。使用偏好测试来确定熊蜂是否偏爱具有给定大小范围内的牛眼图案的花。问题2：测试色素和结构元素对牛眼功能的贡献通过测试授粉者是否偏爱由结构图案元素或仅由色素图案元素组成的图案来确定或元素的组合，因此在与花的交互过程中，它们是否使用结构或着色，或两种特征类型的组合作为提示。

Theme: Agriculture and Food SecurityThe development, evolution and function of the "Bullseye" petal pattern in the Hibiscus trionum complex: Identifying the regulatory mechanisms leading to diverse flower patterns Overall aim: Understand the processes accounting for localised accumulation of pigments in proximal and distal petal regions of H.trionum and its close relatives which create their distinctive bullseye pattern. Understanding the role of different bullseye geometries in mediating plant-pollinator interactions. The project is further split into three aims. Aim 1: Identify the regulatory and signalling mechanisms regulating local pigment production in developing petals to generate a bullseye pattern.Computational and Molecular techniques are used to investigate MYB and bHLH candidate genes. Phylogenies; cloning; qPCR for expression; transgenic over-expressor and knock-out lines; Yeast Hybrids. Aim 2: Characterise and compare varieties that differ in their bullseye patterns to understand the developmental and molecular changes accounting for bullseye evolution.Morphological characterisation of pattern variants: pigment extractions; image analysis of petal patterns and features; UV measurements; Molecular changes underlying pattern evolution: H.t.richardsonii transcriptome; qPCR in H.t.commercial and H.t.richardsonii varieties; gene isolation and transgenics in these lines. Aim 3: Explore the role of bullseye pattern in mediating flower-pollinator interaction. Specifically testing the impact of bullseye size variation on pollinator behaviour and assess the relative contribution of each of the three pattern elements (pigmentation, cell shape and cuticular ornamentation) to bullseye function.Question1: Test if variation in bullseye pattern size influences bumblebee (Bombus terrestris) interactions Determine with differential conditioning experiments whether bumblebees can distinguish bullseye patterns exhibiting pigmented regions of different sizes and determine the smallest bullseye size difference bumblebees can perceive.Determine with preference tests if bumblebees favour flowers with a bullseye patterns from a given size range.Question 2: Test the contribution of pigmentation and structural elements to bullseye functionDetermine with preference tests whether pollinators favour patterns composed of structural or pigmentation pattern elements alone, or elements in combination, and therefore whether they use structural or pigmentation, or a combination of both feature types, as cues during an interaction with a flower.