Potassium transporter TRH1/KUP4 contributes to distinct auxin-mediated root system architecture responses

In plants, auxin transport and development are tightly coupled, just as hormone and growth responses are intimately linked in multicellular systems. Here we provide insights into uncoupling this tight control by specifically targeting the expression of TINY ROOT HAIR 1 (TRH1), a member of plant HAK/KUP/KT transporters that facilitate potassium uptake by co-transporting protons, in Arabidopsis root cell files. Use of this system pinpointed specific root developmental responses to acropetal versus basipetal auxin transport. Loss of TRH1 function shows tiny root hairs and defective root gravitropism, associated with auxin imbalance in the root apex. Cell file-specific expression of TRH1 in the central cylinder rescued trh1 root agravitropism, whereas positional TRH1 expression in peripheral cell layers, including epidermis and cortex, restored trh1 root hair defects. Applying a systems-level approach, the role of RAP2.11 and RSL5 transcription factors in root hair development was verified. Furthermore, ERF53 and WRKY51 transcription factors were overrepresented upon restoration of root gravitropism supporting involvement in gravitropic control. Auxin has a central role in shaping root system architecture by regulating multiple developmental processes. We reveal that TRH1 jointly modulates intracellular ionic gradients and cell-to-cell polar auxin transport to drive root epidermal cell differentiation and gravitropic response. Our results indicate the developmental importance of HAK/KUP/KT proton-coupled K+ transporters.

EARLY STARVATION 1 Is a Functionally Conserved Protein Promoting Gravitropic Responses in Plants by Forming Starch Granules

Starch granules in the endodermis of plant hypocotyls act as statoliths that promote hypocotyl negative gravitropism—the directional growth of hypocotyls against gravity—in the dark. To identify the molecular components that regulate hypocotyl negative gravitropism, we performed a mutagenesis screen and isolated reduced gravitropic 1 (rgv1) mutants that lack starch granules in their hypocotyl endodermis and show reduced hypocotyl negative gravitropism in the dark. Using whole genome sequencing, we identified three different rgv1 mutants that are allelic to the previously reported early starvation 1 mutant, which is rapidly depleted of starch just before the dawn. ESV1 orthologs are present in starch-producing green organisms, suggesting ESV1 is a functionally conserved protein necessary for the formation of starch granules. Consistent with this, we found that liverwort and rice ESV1 can complement the Arabidopsis ESV1 mutant phenotype for both starch granules and hypocotyl negative gravitropism. To further investigate the function of ESV1 in other plants, we isolated rice ESV1 mutants and found that they show reduced levels of starch in their leaves and loosely packed starch granules in their grains. Both Arabidopsis and rice ESV1 mutants also lack starch granules in root columella and show reduced root gravitropism. Together, these results indicate ESV1 is a functionally conserved protein that promotes gravitropic responses in plants via its role in starch granule formation.

ACORBA: Automated workflow to measure Arabidopsis thaliana root tip angle dynamic

Plants respond to the surrounding environment in countless ways. One of these responses is their ability to sense and orient their root growth toward the gravity vector. Root gravitropism is studied in many laboratories as a hallmark of auxin-related phenotypes. However, manual analysis of images and microscopy data is known to be subjected to human bias. This is particularly the case for manual measurements of root bending as the selection lines to calculate the angle are set subjectively. Therefore, it is essential to develop and use automated or semi-automated image analysis to produce reproducible and unbiased data. Moreover, the increasing usage of vertical-stage microscopy in plant root biology yields gravitropic experiments with an unprecedented spatiotemporal resolution. To this day, there is no available solution to measure root bending angle over time for vertical-stage microscopy. To address these problems, we developed ACORBA (Automatic Calculation Of Root Bending Angles), a fully automated software to measure root bending angle over time from vertical-stage microscope and flatbed scanner images. Moreover, the software can be used semi-automated for camera, mobile phone or stereomicroscope images. ACORBA represents a flexible approach based on both traditional image processing and deep machine learning segmentation to measure root angle progression over time. By its automated nature, the workflow is limiting human interactions and has high reproducibility. ACORBA will support the plant biologist community by reducing time and labor and by producing quality results from various kinds of inputs.

Auxin and Root Gravitropism: Addressing Basic Cellular Processes by Exploiting a Defined Growth Response

Root architecture and growth are decisive for crop performance and yield, and thus a highly topical research field in plant sciences. The root system of the model plant Arabidopsis thaliana is the ideal system to obtain insights into fundamental key parameters and molecular players involved in underlying regulatory circuits of root growth, particularly in responses to environmental stimuli. Root gravitropism, directional growth along the gravity, in particular represents a highly sensitive readout, suitable to study adjustments in polar auxin transport and to identify molecular determinants involved. This review strives to summarize and give an overview into the function of PIN-FORMED auxin transport proteins, emphasizing on their sorting and polarity control. As there already is an abundance of information, the focus lies in integrating this wealth of information on mechanisms and pathways. This overview of a highly dynamic and complex field highlights recent developments in understanding the role of auxin in higher plants. Specifically, it exemplifies, how analysis of a single, defined growth response contributes to our understanding of basic cellular processes in general.

Allelopathic root inhibition and its mechanisms

Allelopathy represents a valuable biochemical strategy in plant-plant interactions among different plants, e.g. among crops and weeds. It is an important strategy in the colonization of many invasive alien plants. Allelopathic plants affect the growth of other plants in the vicinity through the release of secondary metabolites (allelochemicals) into the soil. In particular, many allelochemicals suppress the root growth of target plants, but little is known about the mechanism involved in root growth inhibition. In this review, we will highlight the mechanism of root suppression involving: (i) Alterations in auxin homeostasis affecting polar auxin transport and root gravitropism, (ii) Biochemical and physiological processes in inhibited roots associated with oxidative stress due to direct production and accumulation of reactive oxygen species or suppression of antioxidative response and (iii) The ultrastructural modifications in root tip exposed to allelochemicals that drastically suppress the cell division and eventually lead to shorter roots of target plants.

GENERATION OF POTATO PLANTS WITH AN EDITED GENOME AND ALTERED STARCH METABOLISM

Potato plants with the edited genome were obtained. Edited lines are characterized by changes in root gravitropism, aboveground part development and starch metabolism.

An R2R3-MYB Transcription Factor PgFLP Directly Activates PgPIN10 to Regulate the GSA of Adventitious Root in Pomegranate

Background: The self-rooted seedling is widely used in pomegranate planting industry currently; However, the root system of self-rooted seedling is shallow and poor cold resistance. Therefore, the study of the molecular mechanisms of pomegranate adventitious root gravitropism is very important for developing deep-rooted pomegranate cultivars.Results: We report the pomegranate FOUR LIPS (PgFLP) that play an key role in regulating the gravitropic set-point angle of pomegranate adventitious root in response to gravity signal. In our study, PgFLP directly regulates the transcriptional expression of PgPIN10 by binding to its promoter, thus regulating the GSA of adventitious root in pomegranate. Additionally, the 35S::PgFLP show stronger gravitational response than wild-type, leading to a smaller GSA in Arabidopsis lateral roots, indicating that PgFLP participates in regulating the GSA of adventitious root via PgPIN10 in pomegranate. Conclusion: Our results confirm that the transcriptional regulation of PgPIN10 by R2R3-MYB transcription factor PgFLP in setting the gravitropic set-point angle of pomegranate adventitious root in response to gravity signal.

TheArabidopsisNRT1/PTR FAMILY protein NPF7.3/NRT1.5 is an indole-3-butyric acid transporter involved in root gravitropism

Active membrane transport of plant hormones and their related compounds is an essential process that determines the distribution of the compounds within plant tissues and, hence, regulates various physiological events. Here, we report that theArabidopsisNITRATE TRANSPORTER 1/PEPTIDE TRANSPORTER FAMILY 7.3 (NPF7.3) protein functions as a transporter of indole-3-butyric acid (IBA), a precursor of the major endogenous auxin indole-3-acetic acid (IAA). When expressed in yeast, NPF7.3 mediated cellular IBA uptake. Loss-of-functionnpf7.3mutants showed defective root gravitropism with reduced IBA levels and auxin responses. Nevertheless, the phenotype was restored by exogenous application of IAA but not by IBA treatment.NPF7.3was expressed in pericycle cells and the root tip region including root cap cells of primary roots where the IBA-to-IAA conversion occurs. Our findings indicate that NPF7.3-mediated IBA uptake into specific cells is required for the generation of appropriate auxin gradients within root tissues.