Collaborative Research: How to manipulate a plant? Testing for conserved effectors and plant responses in gall induction and growth using a multi-species comparative approach.

虫胆是由另一种生物(即细菌、真菌、线虫或昆虫)诱导的植物上的肿瘤状生长。昆虫诱导的植物虫害是自然界中最令人惊叹和最复杂的种间相互作用之一，几个世纪以来一直吸引着自然学家、生态学家和进化生物学家的兴趣。越来越多地，形成虫害的昆虫也成为重要的经济农业害虫，降低了世界范围内的作物产量。令人惊讶的是，昆虫虫胆诱导和生长背后的一般机制尚不清楚。这一建议旨在通过利用鲤虫黄蜂在活橡树上的叮咬系统来揭示进化上保守的胆汁形成的分子机制，在这种系统中，许多种类的鲤虫成功地将同一植物取胆。这个系统允许强大的自然实验来测试恼人的生物如何操纵植物基因组的普遍性，并产生新的假设，以研究针对恼人的食草动物和其他操纵寄主植物防御和免疫的害虫的植物控制策略。此外，恼人的研究对于基因工程的发展具有巨大的潜力，因为胆汁形成涉及植物中数千个基因的同时操作--这是目前的技术所不可能的(例如，RNAi、CRISPR)。在这项研究过程中，PI将与德克萨斯南方大学(HBCU)的合作者共同主办一系列研讨会，并与合作者开发一系列实验室；在联邦承认的少数族裔服务机构新墨西哥大学招募夏季本科生研究人员；以及积极参与休斯顿和阿尔伯克基多个机构的公共科学研究。以前对昆虫诱导的植物虫害的分子研究一次集中在一个物种上，这造成了系统发育不同的宿主和恼人的昆虫的拼凑，这使得识别恼人背后的共同潜在机制变得具有挑战性。我们开发的研究系统代表了这个问题的解决方案；其中七个不同的黄蜂进化谱系都在同一寄主植物物种上形成了表型独特的虫胆。该系统使我们能够降低来自不同胆汁表型(例如，颜色、毛发、大小、起始组织)的噪音，以确定这种相互作用的核心分子机制。在目标1中，我们将在七种鲤科黄蜂中识别潜在的效应分子。通过转录组和蛋白质组，将在毒腺(负责胆汁诱导)和幼虫唾液腺(负责胆汁生长)中鉴定诱导虫胆所需的黄蜂基因。在目标2中，我们将确定核心植物对鲤鱼属植物的反应。通过比较多种虫胆形态与其相匹配的对照未受刺激的组织在整个发育过程中的基因表达，我们可以分离出虫胆之间的表型变异，以确定启动和维持虫胆生长所需的核心基因和途径的变化。在目标3中，我们在橡木上的结果将通过对之前发表的20多个五倍子转录本进行大型荟萃分析来评估在其他伤害系统中的保护作用，这些转录本包括大豆、小麦、水稻、葡萄和蓝莓上的几种农业害虫。这将确保我们的结果将告知整个令人难堪的研究社区。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

A gall is a tumor-like growth on a plant that is induced by another organism (i.e., bacteria, fungi, nematode, or insects). Insect-induced plant galls represent one of the most amazing and complex interspecific interactions in nature, capturing the interest of naturalists, ecologists and evolutionary biologists for centuries. Increasingly, gall-forming insects also act as economically important agricultural pests, reducing crop yields worldwide. Surprisingly, the general mechanisms behind insect gall induction and growth are unknown. This proposal aims to uncover the evolutionarily conserved molecular mechanisms of gall formation by harnessing the galling system of cynipid wasps on live oaks, in which many species of cynipids successfully gall the same plant. This system allows for powerful natural experiments to test the generality of how galling organisms manipulate plant genomes and to generate new hypotheses for investigating plant control strategies against galling herbivores and other pests that manipulate host plant defense and immunity. Additionally, galling research has great potential for the advancement of genetic engineering because gall-formation involves the simultaneous manipulation of thousands of genes in the plant – not possible with current technologies (e.g., RNAi, CRISPR). During the course of this research the PIs will co-host a seminar series and develop a series of labs with collaborators at Texas Southern University, a historically black college (HBCU); recruit summer undergraduate researchers at University of New Mexico, a federally recognized minority-serving institution; as well as actively participate in public engagement of scientific research in multiple institutions in Houston and Albuquerque.Previous molecular studies on insect-induced plant galls have focused on one species at a time, which has created a patchwork of phylogenetically divergent hosts and galling insects that has made it challenging to identify common underlying mechanisms behind galling. The study system we have developed represents a solution to this problem; wherein seven distinct evolutionary lineages of wasp all form phenotypically unique galls on the same host plant species. This system gives us the power to reduce the noise from different gall phenotypes (e.g., color, hairs, size, starting tissue) to determine the core molecular mechanisms of this interaction. In Aim 1, we will identify potential effector molecules in seven species of cynipid wasps. Through transcriptomes and proteomes, the wasp genes needed to induce gall induction will be identified in the venom gland (responsible for gall induction) and larval salivary glands (responsible for gall growth). In Aim 2, we will determine the core plant response to cynipid gallers. By comparing the gene expression of the multiple gall morphologies to their matched control ungalled tissues throughout development, we can separate out phenotypic variation across galls to identify alterations in core genes and pathways required to initiate and maintain gall growth. In Aim 3, our results in oak will be assessed for conservation in other galling systems by conducting a large meta-analysis with 20+ previous published gall transcriptomes including several agricultural pests on soybean, wheat, rice, grape, and blueberry. This will ensure that our results will inform the entire galling research community.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.