Yanhua Su

Shaker potassium channels play essential roles in K ⁺ uptake and distribution in plants. Studies on Shaker channels in Arabidopsis have provided a paradigmatic framework, but the rice genome encodes an additional member, OsKAT1, whose function remians poorly defined. OsKAT1-type channels are characterized by an innately short cytosolic C terminus, forming a unique clade of monocot-specific short Shakers that is prevalent in Poaceae species. In rice, OsKAT1 is predominantly expressed in the root stele. Disruption of OsKAT1 (KO mutation) leads to a significant reduction in K ⁺ secretion into the xylem sap delivered to the shoot. Patch-clamp experiments on root stele protoplasts of WT and KO plants indicate that OsKAT1 functions as an inward channel. Functional analyses in Xenopus oocytes reveal that despite activating at weakly negative voltages, OsKAT1 is intrinsically incapable of mediating substantial outward currents—a property attributed to its truncated C terminus. Together with outward K ⁺ channel activity, this feature enables stelar cells to remain significantly permeable to K ⁺ around the K ⁺ equilibrium potential. When expressed in Arabidopsis , OsKAT1 contributes to K ⁺ transport in the xylem sap, but only in wild-type plants that express SKOR, a Shaker channel specialized for this role. These results demonstrate that OsKAT1 constitutes a component of K ⁺ translocation to shoots and shed light on the energization and regulation underlying this function. Moreover, OsKAT1 is shown to improve rice adaptation to environmental conditions and enhance grain yield under field conditions through its facilitating root-to-shoot ion translocation.

Saline soils exert persistent salt stress on plants that inhibits their ability to carry out photosynthesis and leads to photosynthetic carbon (C) scarcity in plant roots and the rhizosphere. However, it remains unclear how a rhizosphere environment is shaped by photosynthetic C partitioning under saline conditions. Given that sucrose is the primary form of photosynthetic C transport, we, respectively, created sucrose transport distorted (STD) and enhanced (STE) rice lines through targeted mutation and overexpression of the sucrose transporter gene OsSUT5. This approach allowed us to investigate different scenarios of photosynthate partitioning to the rhizosphere. Compared to the non-saline soil, we found a significant decrease in soil dissolved organic carbon (DOC) in the rhizosphere, associated with a reduction in bacterial diversity when rice plants were grown under moderate saline conditions. These phenomena were sharpened with STD plants but were largely alleviated in the rhizosphere of STE plants, in which the rhizosphere DOC, and the diversity and abundances of dominant bacterial phyla were measured at comparable levels to the wildtype plants under non-saline conditions. The complexity of bacteria showed a greater level in the rhizosphere of STE plants grown under saline conditions. Several salt-tolerant genera, such as Halobacteroidaceae and Zixibacteria, were found to colonize the rhizosphere of STE plants that could contribute to improved rice growth under persistent saline stresses, due to an increase in C deposition.

K⁺ uptake to root conducting cells is a prerequisite for its release to the xylem vessels and long-distance delivery to the aerial parts, though the molecular mechanism underpinning such process remains insufficiently understood. Here we report the discovery in rice root of an essential component involved in loading of K⁺ to the stelar tissues encoded by an unusual short Shaker K⁺ channel OsK2.2. OsK2.2 represents a novel type of short Shaker channels lacking the sequences immediately downstream of the putative cyclic nucleotide binding domain that present to the C-termini of all plant Shaker channels reported to date. In silico analysis suggests that such short Shaker channels are monocot-specific and especially found in the Poaceae species. Functional assessment in Xenopus oocytes reveals that OsK2.2 uses such innate C-terminal shortness as an intrinsic strategy for maintaining large capacity of K⁺ uptake to the cells. OsK2.2 predominantly localizes to the phloem and xylem peripheral cells of rice root vasculature. Disruption of OsK2.2 function causes approximately 30% lower K⁺ in both phloem and xylem sap and reduces K⁺ accumulation to the shoots. OsK2.2 strongly contributes to the grain yield of rice under field conditions. The essential contribution of such particular type of inward Shaker channel, operating in stelar tissues, to K⁺ transport between roots and shoots and to grain yield, here evidenced in rice, may be a common trait amongst Poaceae species.

AKT2 potassium (K⁺) channels are members of the plant Shaker family which mediate dual-directional K⁺ transport with weak voltage-dependency. Here we show that OsAKT2 of rice (Oryza sativa) functions mainly as an inward rectifier with strong voltage-dependency and acutely suppressed outward activity. This is attributed to the presence of a unique K191 residue in the S4 domain. The typical bi-directional leak-like property was restored by a single K191R mutation, indicating that this functional distinction is an intrinsic characteristic of OsAKT2. Furthermore, the opposite R195K mutation of AtAKT2 changed the channel to an inward-rectifier similar to OsAKT2. OsAKT2 was modulated by OsCBL1/OsCIPK23, evoking the outward activity and diminishing the inward current. The physiological relevance in relation to the rectification diversity of OsAKT2 was addressed by functional assembly in the Arabidopsis (Arabidopsis thaliana) akt2 mutant. Overexpression (OE) of OsAKT2 complemented the K⁺ deficiency in the phloem sap and leaves of the mutant plants but did not significantly contribute to the transport of sugars. However, the expression of OsAKT2-K191R overcame both the shortage of phloem K⁺ and sucrose of the akt2 mutant, which was comparable to the effects of the OE of AtAKT2, while the expression of the inward mutation AtAKT2-R195K resembled the effects of OsAKT2. Additionally, OE of OsAKT2 ameliorated the salt tolerance of Arabidopsis.

Maize grows in nitrate-dominated dryland soils, but shortly upon localized dressing of nitrogen fertilizers, ammonium is retained as a noticeable form of nitrogen source available to roots. Thus in addition to nitrate, the absorption of ammonium can be an important strategy that promotes rapid plant growth at strong nitrogen demanding stages. The present study reports the functional characterization of three root-expressed ammonium transporters (AMTs), aiming at finding out functional and regulatory properties that correlate with efficient nitrogen acquisition of maize. Using a stable electrophysiological recording method we previously established in Xenopus laevis oocytes that integrates the capture of currents in response to voltage ramps with onsite stability controls, we demonstrate that all three ZmAMT1s engage NH₄⁺ uniporting as ammonium uptake mechanisms. The K_m value for ZmAMT1.1a, 1.1b, or ZmAMT1.3 is, respectively, 9.9, 15.6, or 18.6 μM, indicating a typical high-affinity transport of NH₄⁺ ions. Importantly, the uptake currents of these ZmAMT1s are markedly amplified upon extracellular acidification. A pH drop from 7.4 to 5.4 results in a 140.8%, 64.1% or a 120.7% increase of ammonium uptake activity through ZmAMT1.1a, 1.1b, or ZmAMT1.3. Since ammonium uptake by plant roots accompanies a spontaneous acidification to the root medium, the functional promotion of ZmAMT1.1a, 1.1b, and ZmAMT1.3 by low pH is in line with the facilitated ammonium uptake activity in maize roots. Furthermore, the expression of the three ZmAMT1 genes is induced under ammonium-dominated conditions. Thus we describe a facilitated ammonium uptake strategy in maize roots by functional and expression regulations of ZmAMT1 transporters that may coordinate with efficient acquisition of this form of nitrogen source when available.

AtAMT1;3 is a major contributor to high-affinity ammonium uptake in Arabidopsis roots. Using a stable electrophysiological recording strategy, we demonstrate in Xenopus laevis oocytes that AtAMT1;3 functions as a typical high-affinity NH₄⁺ uniporter independent of protons and Ca²⁺. The findings that AtAMT1;3 transports methylammonium (MeA⁺, a chemical analog of NH₄⁺) with extremely low affinity (K_m in the range of 2.9–6.1 mM) led to investigate the mechanisms underlying substrate binding. Homologous modeling and substrate docking analyses predicted that the deduced substrate binding motif of AtAMT1;3 facilitates the binding of NH₄⁺ ions but loosely accommodates the binding of MeA⁺ to a more superficial location of the permeation pathway. Amongst point mutations tested based on this analysis, P181A resulted in both significantly increased current amplitudes and substrate binding affinity, whereas F178I led to opposite effects. Thus these 2 residues, which flank W179, a major structural component of the binding site, are also important determinants of AtAMT1;3 transport capacity by being involved in substrate binding. The Q365K mutation neighboring the histidine residue H378, which confines the substrate permeation tunnel, affected only the current amplitudes but not the binding affinities, providing evidence that Q365 mainly controls the substrate diffusion rate within the permeation pathway.

Ammonium transporter (AMT)-mediated acquisition of ammonium nitrogen from soils is essential for the nitrogen demand of plants, especially for those plants growing in flooded or acidic soils where ammonium is dominant. Recent advances show that AMTs additionally participate in many other physiological processes such as transporting ammonium from symbiotic fungi to plants, transporting ammonium from roots to shoots, transferring ammonium in leaves and reproductive organs, or facilitating resistance to plant diseases via ammonium transport. Besides being a transporter, several AMTs are required for the root development upon ammonium exposure. To avoid the adverse effects of inadequate or excessive intake of ammonium nitrogen on plant growth and development, activities of AMTs are fine-tuned not only at the transcriptional level by the participation of at least four transcription factors, but also at protein level by phosphorylation, pH, endocytosis, and heterotrimerization. Despite these progresses, it is worth noting that stronger growth inhibition, not facilitation, unfortunately occurs when AMT overexpression lines are exposed to optimal or slightly excessive ammonium. This implies that a long road remains towards overcoming potential limiting factors and achieving AMT-facilitated yield increase to accomplish the goal of persistent yield increase under the present high nitrogen input mode in agriculture.

Background: Overuse of nitrogen fertilizers is often a major practice to ensure sufficient nitrogen demand of high–yielding rice, leading to persistent NH₄⁺ excess in the plant. However, this excessive portion of nitrogen nutrient does not correspond to further increase in grain yields. For finding out the main constraints related to this phenomenon, the performance of NH₄⁺ excess in rice plant needs to be clearly addressed beyond the well-defined root growth adjustment. The present work isolates an acute NH₄⁺ excess condition in rice plant from causing any measurable growth change and analyses the initial performance of such internal NH₄⁺ excess. Results: We demonstrate that the acute internal NH₄⁺ excess in rice plant accompanies readily with a burst of reactive oxygen species (ROS) and initiates the downstream reactions. At the headstream of carbon production, photon caption genes and the activity of primary CO₂ fixation enzymes (Rubisco) are evidently suppressed, indicating a reduction in photosynthetic carbon income. Next, the vigorous induction of glutathione transferase (GST) genes and enzyme activities along with the rise of glutathione (GSH) production suggest the activation of GSH cycling for ROS cleavage. Third, as indicated by strong induction of glycolysis / glycogen breakdown related genes in shoots, carbohydrate metabolisms are redirected to enhance the production of energy and carbon skeletons for the cost of ROS scavenging. As the result of the development of these defensive reactions, a carbon scarcity would accumulatively occur and lead to a growth inhibition. Finally, a sucrose feeding cancels the ROS burst, restores the activity of Rubisco and alleviates the demand for the activation of GSH cycling. Conclusion: Our results demonstrate that acute NH₄⁺ excess accompanies with a spontaneous ROS burst and causes carbon scarcity in rice plant. Therefore, under overuse of N fertilizers carbon scarcity is probably a major constraint in rice plant that limits the performance of nitrogen.

Background: Overuse of nitrogen fertilizers is often a major practice to ensure sufficient nitrogen demand of high–yielding rice, leading to persistent NH4+ excess in the plant. However, this excessive portion of nitrogen nutrient does not correspond to further increase in grain yields. For finding out the main constraints related to this phenomenon, the performance of NH4+ excess in rice plant needs to be clearly addressed beyond the well-defined root growth adjustment. The present work isolates an acute NH4+ excess condition in rice plant from causing any measurable growth change and analyses the initial performance of such internal NH4+ excess. Results: We demonstrate that the acute internal NH4+ excess in rice plant accompanies readily with a burst of reactive oxygen species (ROS) and initiates the downstream reactions. At the headstream of carbon production, photon caption genes and the activity of primary CO2 fixation enzymes (Rubisco) are evidently suppressed, indicating a reduction in photosynthetic carbon income. Next, the vigorous induction of glutathione transferase (GST) genes and enzyme activities along with the rise of glutathione (GSH) production suggest the activation of GSH cycling for ROS cleavage. Third, as indicated by strong induction of glycolysis / glycogen breakdown related genes in shoots, carbohydrate metabolisms are redirected to enhance the production of energy and carbon skeletons for the cost of ROS scavenging. As the result of the development of these defensive reactions, a carbon scarcity would accumulatively occur and lead to a growth inhibition. Finally, a sucrose feeding cancels the ROS burst, restores the activity of Rubisco and alleviates the demand for the activation of GSH cycling. Conclusion: Our results demonstrate that acute NH4+ excess accompanies with a spontaneous ROS burst and causes carbon scarcity in rice plant. Therefore, under overuse of N fertilizers carbon scarcity is probably a major constraint in rice plant that limits the performance of nitrogen.