Plant Pathology and Microbio..
Non Technical Summary
Global climate change can bring about environmental disasters such as drought which causesevere negative impacts oncrop heath and productivity. Thisreducedcrop outputresults in economic loss for producersand challenges the food security ofour growing population. The ability of a crop plant to efficiently uptake water and essential nutrients from the soil using its root system often dictates its ability to withstand abiotic stressors and aid in maximizingyield potential. Root growth and development are regulated in partby the plant growth hormone auxin, althought there islittle known about the genetic behind this phenomena.We have identified the auxin response factor, ZmARF27 to be a possible key regulator of root structure inmaize.Through the use of molecular-based technologies I aim tocharacterize this gene to uncover auxin's role in root development. I propose to identify biological pathways and downstream (both direct and indirect) targets of this genetic regulator as well as identify interacting proteins crucial for its functionality. The use of sequencing-based technology will allow me to not only quantifiably measure fluctuations in known genetic pathways but also identify and discover new elements connecting auxin signaling and root formation.The ultimate goal of this project is to establish a connection between the maize root and auxin and lay the ground work for future studies. Understanding their connection and how to harness their capabilities willallow scientists and plant breeders the opportunity to generate more stress tolerant and higher yielding varieties.
Animal Health Component
Research Effort Categories
Goals / Objectives
The overall goal of this project is to characterizethe molecular function of the maize transcription factorAUXIN RESPONSE FACTOR 27 (ZmARF27) and understandits role inregulating auxin signaling based control of root architecture.Objectives:1) Identify auxin responsive target genes that are bound and regulated by ZmARF272) Quantify gene expression changes caused by ZmARF27 in response to auxin3) Elucidate the protein interactors of ZmARF27 that fine tune its transcriptional activity
This project will be conducted using discovery based research methods to test the hypothesis that ZmARF27 positively drives root growth by mediating auxin signaling in maize.Efforts will include the use of molecular techniques to identify genes impacted by ZmARF27 as well as proteins critical to its functionality. More specifically, the efforts will include Chromatin Immuno Precipitation Sequencing (ChIP-Seq), QuantSeq 3' mRNA Sequencing, and protein:protein binding via TurboID analyzed by Liquid Chromatography coupled Mass Spectrometry. ChIP-Seq will be performed on maize root protoplast transformed with ZmARF27 and then treated with or without the auxin indole-3-acetic acid (IAA). Chromatin will be isolated, ZmARF27 pulled down and bound DNA targets will be sequenced and direct targets of ZmARF27 will be identified. QuantSeq 3' mRNA Sequencing will be performed on zmarf27 mutant root and wild type root tissue treated with or without IAA. Obtained reads will be mapped and the obtained counts will be used in differential expression analysis to determine significantly up and down regulated genes.TurboID experiments will be performed on maize root protoplast transformed with a ZmARF27-TurboID plasmid and treated with IAA as described in the ChIP-Seq method. Following treatment the protoplasts will be lysed and the proteins interacting with ZmARF27 will be pulled down and digested into peptides. The peptides will then be ran on a mass spectrometer and obtained spectra will be searched and statistically analyzed to determine enriched protein interactors of ZmARF27.Efforts will include publication of the data in a peer reviewed journal and presentation of research findings at lab meetings and international research conferences.