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.

瘿是由另一种生物（如细菌、真菌、线虫或昆虫）诱导的植物上的肿瘤样生长。昆虫诱导的植物虫瘿代表了自然界中最惊人和最复杂的种间相互作用之一，几个世纪以来吸引了博物学家、生态学家和进化生物学家的兴趣。越来越多的胆囊形成昆虫也成为重要的经济农业害虫，减少世界范围内的作物产量。令人惊讶的是，昆虫瘿诱导和生长的一般机制尚不清楚。本研究的目的是通过利用木蜂在活橡树上的螫螫系统来揭示进化上保守的瘿形成的分子机制，在这个系统中，许多种木蜂成功地对同一株橡树进行了螫螫。该系统允许进行强大的自然实验，以测试刺痛生物如何操纵植物基因组的普遍性，并为研究植物控制策略以对抗刺痛食草动物和其他操纵宿主植物防御和免疫的害虫产生新的假设。此外，胆囊研究对于基因工程的进步具有巨大的潜力，因为胆囊的形成涉及同时操纵植物中的数千个基因-这是目前技术（例如RNAi， CRISPR）所不可能的。在这项研究过程中，pi将与德克萨斯南方大学（一个历史悠久的黑人大学）的合作者共同主办一系列研讨会，并开发一系列实验室；在新墨西哥大学招收暑期本科生研究人员，这是一所联邦认可的少数族裔服务机构；并积极参与休斯顿和阿尔伯克基多个机构的科学研究的公众参与。以前对昆虫诱导的植物虫瘿的分子研究一次只关注一个物种，这就造成了系统发育上不同的宿主和昆虫的拼凑，这使得确定虫瘿背后的共同潜在机制变得具有挑战性。我们开发的学习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表型独特的瘿。该系统使我们能够减少来自不同胆囊表型（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7种乔木蜂中鉴定潜在的效应分子。通过转录组学和蛋白质组学，黄蜂诱导胆汁诱导所需的基因将在毒液腺（负责胆汁诱导）和幼虫唾液腺（负责胆汁生长）中被鉴定出来。在目标2中，我们将确定核心植物对木质素galler的反应。通过将多种胆囊形态的基因表达与发育过程中与其匹配的对照非胆囊组织进行比较，我们可以分离出胆囊间的表型变异，以确定启动和维持胆囊生长所需的核心基因和途径的改变。在Aim 3中，我们的研究结果将通过对20多个先前发表的包括大豆、小麦、水稻、葡萄和蓝莓在内的几种农业害虫的胆囊转录组进行大型荟萃分析，来评估在其他胆囊系统中的保护作用。这将确保我们的结果将通知整个研究社区。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值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.