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Jez, Joseph M. ; Topp, Christopher N. (Ed.)A plants’ water and nutrients are primarily absorbed through roots, which in a natural setting is highly dependent on the 3-dimensional configuration of the root system, collectively known as root system architecture (RSA). RSA is difficult to study due to a variety of factors, accordingly, an arsenal of methods have been developed to address the challenges of both growing root systems for imaging, and the imaging methods themselves, although there is no ‘best’ method as each has its own spectrum of trade-offs. Here, we describe several methods for plant growth or imaging. Then, we introduce the adaptation and integration of three complementary methods, root mesocosms, photogrammetry, and electrical resistance tomography (ERT). Mesocosms can allow for unconstrained root growth, excavation and preservation of 3-dimensional RSA, and modularity that facilitates the use of a variety of sensors. The recovered root system can be digitally reconstructed through photogrammetry, which is an inexpensive method requiring only an appropriate studio space and a digital camera. Lastly, we demonstrate how 3-dimensional water availability can be measured using ERT inside of root mesocosms.
Jez, Joseph M. ; Topp, Christopher N. (Ed.)Structural biologists rely on X-ray crystallography as the main technique for determining the three-dimensional structures of macromolecules; however, in recent years, new methods that go beyond X-ray-based technologies are broadening the selection of tools to understand molecular structure and function. Simultaneously, national facilities are developing programming tools and maintaining personnel to aid novice structural biologists in de novo structure determination. The combination of X-ray free electron lasers (XFELs) and serial femtosecond crystallography (SFX) now enable time-resolved structure determination that allows for capture of dynamic processes, such as reaction mechanism and conformational flexibility. XFEL and SFX, along with microcrystal electron diffraction (MicroED), help side-step the need for large crystals for structural studies. Moreover, advances in cryogenic electron microscopy (cryo-EM) as a tool for structure determination is revolutionizing how difficult to crystallize macromolecules and/or complexes can be visualized at the atomic scale. This review aims to provide a broad overview of these new methods and to guide readers to more in-depth literature of these methods.
Jez, Joseph M. ; Topp, Christopher N. (Ed.)Single-cell RNA-seq is a tool that generates a high resolution of transcriptional data that can be used to understand regulatory networks in biological systems. In plants, several methods have been established for transcriptional analysis in tissue sections, cell types, and/or single cells. These methods typically require cell sorting, transgenic plants, protoplasting, or other damaging or laborious processes. Additionally, the majority of these technologies lose most or all spatial resolution during implementation. Those that offer a high spatial resolution for RNA lack breadth in the number of transcripts characterized. Here, we briefly review the evolution of spatial transcriptomics methods and we highlight recent advances and current challenges in sequencing, imaging, and computational aspects toward achieving 3D spatial transcriptomics of plant tissues with a resolution approaching single cells. We also provide a perspective on the potential opportunities to advance this novel methodology in plants.