Molecular framework underlying stolon development in strawberry

草莓是一种非常重要的水果作物，为人类提供营养和农业产量。由于目前的草莓品种都来自传统的杂交育种努力，他们需要通过女儿克隆繁殖，以保持其独特的杂交组合。这些子克隆，被称为亚军，是由长而hortizedstem命名匍匐茎。然而，很少有人知道匍匐茎的形成是如何调节的，为什么草莓是少数几种植物赋予的分子机制进行无性繁殖通过匍匐茎。这些实验旨在揭示草莓这种特殊且非常重要的发育性状的分子机制。此外，该项目将培养下一代科学家的批判性思维，沟通以及实验设计和执行。此外，弗吉尼亚州立大学的一位教授每年夏天将带两名本科生到马里兰州大学学习DNA序列分析并进行研究。总的来说，该项目将提供知识基础和工具，以打开和关闭匍匐茎，从而提高草莓产量时，匍匐茎形成打开或增加水果产量时，匍匐茎形成关闭。该项目产生的数据，如RNA序列数据和网络分析，将通过网站和公共存放处方便获取;研究材料将与科学界共享。研究结果将有助于加深对植物发育的基本认识，并为草莓生产力的提高提供新的策略。在草莓中，通过匍匐茎进行无性繁殖的有趣模式为研究腋生分生组织细胞的命运决定提供了一个不寻常的机会，因为每个腋生分生组织要么发育成匍匐茎，要么发育成分支冠（以花终止的枝条）。先前的研究使用二倍体草莓野草莓鉴定了赤霉素的关键信号组分FveRGA 1作为匍匐茎形成的阻遏物。本实验将研究FveRGA 1如何抑制F.通过首先用CRISPR分离FveRGA 1的无效等位基因，然后表征突变体表型，来鉴定vesca。转录组和共识共表达网络将确定在腋生分生组织中表达的基因，产生匍匐茎。与腋生分生组织中的FveRGA 1直接相互作用的蛋白质将通过酵母双杂交筛选进行鉴定，并通过双分子荧光互补进行确认。将利用RNAi和转基因F.维斯卡最后，正向遗传方法将被用来定位和分离新的突变体所定义的基因，改变匍匐茎形成的能力。这些实验将为匍匐茎发育的分子基础提供新的见解，并阐明所鉴定的基因和FveRGA 1在控制匍匐茎形成中的分子相互作用机制。抑制匍匐茎产生的能力将使分支冠形成最大化，导致更多的花和更高的果实产量。从这个项目中获得的见解将提供知识和工具来打开和关闭匍匐茎，从而提高草莓的生产和产量。这个奖项反映了NSF的法定使命，并被认为是值得通过使用基金会的智力价值和更广泛的影响审查标准进行评估的支持。

Strawberry is an immensely important fruit crop contributing to human nutrition and agricultural output. As current strawberry varieties are all derived from traditional crossbreeding efforts, they need to be propagated through daughter clones to preserve their unique hybrid combination. These daughter clones, known as runners, are produced from long and hortizontal stems named stolons. However, little is known about how stolon formation is regulated and why strawberry is one of the few plants endowed with the molecular machinery to conduct asexual reproduction through stolon. The experiments are aimed at revealing the molecular mechanisms that underlie this special and highly important developmental trait of strawberry. Further, the project will train next generation scientists in critical thinking, communication, as well as experimental design and execution. Additionally, a professor at the Virginia State University will bring two undergraduate students to the University of Maryland each summer to learn DNA sequence analysis and conduct research. Overall, the project will provide knowledge base and tools to switch on and off stolon, leading to improved strawberry production when stolon formation is switched on or increased fruit yield when stolon formation is turned off. The data generated from the project such as RNA sequence data and network analyses will be easily accessible through websites and public depository; the research materials will be shared with the scientific community. The results will advance basic understanding of plant development and provide novel strategies in the improvement of strawberry productivity. In strawberry, the interesting mode of asexual reproduction through stolon provides an unusual opportunity to investigate axillary meristem cell fate determination as each axillary meristem either develops into a stolon or a branch crown (a shoot that terminates with flowers). Previous studies using a diploid strawberry Fragaria vesca identified a key signaling component of gibberellin, FveRGA1, as a repressor of stolon formation. The experiments will investigate the mechanism of how FveRGA1 represses stolon formation in F. vesca by first isolating null alleles of FveRGA1 with CRISPR and then characterizing the mutant phenotype. Transcriptome and consensus co-expression networks will identify genes expressed in the axillary meristem that gives rise to stolon. Proteins that directly interact with FveRGA1 in the axillary meristem will be identified through a yeast two-hybrid screen and confirmed by Bimolecular Fluorescence Complementation. The role of identified candidate genes in stolon development will be determined using RNAi and over-expression in transgenic F. vesca. Finally, forward genetic approach will be used to map and isolate genes defined by new mutants with altered ability in stolon formation. These experiments will provide novel insights into the molecular underpinnings of stolon development and illuminate the mechanisms of molecular interaction between the identified genes and FveRGA1 in controling stolon formation. The ablity to suppress stolon production will maximize branch crown formation, leading to more flowers and higher fruit yield. Insights derived from this project will provide knowledge and tools to switch on and off stolon, leading to improved strawberry production and yields.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.