It has been almost 25 years since the first report of the gene encoding a high-affinity phosphate transporter （PT）, PH084, in yeast. Since then, an increasing number of yeast PH084 homologs as well ...as other genes encoding proteins with phosphate （Pi） transport activities have been identified and functionally characterized in diverse plant species. Great progress has been made also in deciphering the molecular mechanism underlying the regulation of the abundance and/or activity of these genes and their products. The regulatory genes affect plant Pi homeostasis commonly through direct or indirect regulation of the abundance of PTs at different levels. However, little has been achieved in the use of PTs for developing genetically modified crops with high phosphorus use efficiency （PUE）. This might be a consequence of overemphasizing Pi uptake from the rhizosphere and lack of knowledge about the roles of PTs in Pi transport and recycling within the plant that are required to optimize PUE. Here, we mainly focused on the genes encoding proteins with Pi transport activities and the emerging understanding of their regulation at the transcriptional, posttranscriptional, translational, and post-translational levels. In addition, we propose potential strategies for effective use of PTs in improving plant growth and development.
Summary
In agricultural soils, amino acids can represent vital nitrogen (N) sources for crop growth and yield. However, the molecular mechanisms underlying amino acid uptake and allocation are poorly ...understood in crop plants. This study shows that rice (Oryza sativa L.) roots can acquire aspartate at soil concentration, and that japonica subspecies take up this acidic amino acid 1.5‐fold more efficiently than indica subspecies. Genetic association analyses with 68 representative japonica or indica germplasms identified rice Lysine‐Histidine‐type Transporter 1 (OsLHT1) as a candidate gene associated with the aspartate uptake trait. When expressed in yeast, OsLHT1 supported cell growth on a broad spectrum of amino acids, and effectively transported aspartate, asparagine and glutamate. OsLHT1 is localized throughout the rice root, including root hairs, epidermis, cortex and stele, and to the leaf vasculature. Knockout of OsLHT1 in japonica resulted in reduced root uptake of amino acids. Furthermore, in 15N‐amino acid‐fed mutants versus wild‐type, a higher percentage of 15N remained in roots instead of being allocated to the shoot. 15N‐ammonium uptake and subsequently the delivery of root‐synthesized amino acids to Oslht1 shoots were also significantly decreased, which was accompanied by reduced shoot growth. These results together provide evidence that OsLHT1 functions in both root uptake and root to shoot allocation of a broad spectrum of amino acids in rice.
Significance Statement
Amino acids can serve as important nitrogen sources in agricultural fields, but little is known about the underlying mechanisms of amino acid uptake and allocation in crops. This study conducted genetic association analyses of 15N‐aspartate uptake among rice accessions, combined with genetic and biochemical analyses, and revealed that Oryza sativa Lysine‐Histidine‐type Transporter 1 (OsLHT1) functions in both root uptake and root to shoot allocation of a broad spectrum of amino acids.
Crop productivity relies heavily on nitrogen (N) fertilization. Production and application of N fertilizers consume huge amounts of energy, and excess is detrimental to the environment; therefore, ...increasing plant N use efficiency (NUE) is essential for the development of sustainable agriculture. Plant NUE is inherently complex, as each step-including N uptake, translocation, assimilation, and remobilization-is governed by multiple interacting genetic and environmental factors. The limiting factors in plant metabolism for maximizing NUE are different at high and low N supplies, indicating great potential for improving the NUE of current cultivars, which were bred in well-fertilized soil. Decreasing environmental losses and increasing the productivity of crop-acquired N requires the coordination of carbohydrate and N metabolism to give high yields. Increasing both the grain and N harvest index to drive N acquisition and utilization are important approaches for breeding future high-NUE cultivars.
•Pi starvation signal network with OsPHR2 as the central regulator.•PHO2-mediated ubiquitination are important bypass for Pi signaling.•SPX domain-containing proteins are essential in Pi ...homeostasis.•Diverse roles and regulation of Pi transporters.•Improvement of tolerance to low Pi stress in rice.
Rice is one of the most important cereal crops feeding a large segment of the world's population. Inefficient utilization of phosphate (Pi) fertilizer by the plant in rice production increases cost and pollution. Developing cultivars with improved Pi use efficiency is essential for the sustainability of agriculture. Pi uptake, translocation and remobilization are regulated by complex molecular mechanisms through the functions of Pi transporters (PTs) and other downstream Pi Starvation Induced (PSI) genes. Expressions of these PSI genes are regulated by the Pi Starvation Response Regulator (OsPHR2)-mediated transcriptional control and/or PHO2-mediated ubiquitination. SPX-domain containing proteins and the type I H+-PPase AVP1 involved in the maintenance and utilization of the internal phosphate. The potential application of posttranscriptional regulation of PT1 through OsPHF1 for Pi efficiency is proposed.
Cellular pH homeostasis is regulated through the activities of N transporters and proton pumps affecting proton production or consumption during root acquisition, short and long-distance transport, ...and assimilation of N.
Abstract
The enzymatic controlled metabolic processes in cells occur at their optimized pH ranges, therefore cellular pH homeostasis is fundamental for life. In plants, the nitrogen (N) source for uptake and assimilation, mainly in the forms of nitrate (NO3–) and ammonium (NH4+) quantitatively dominates the anion and cation equilibrium and the pH balance in cells. Here we review ionic and pH homeostasis in plant cells and regulation by N source from the rhizosphere to extra- and intracellular pH regulation for short- and long-distance N distribution and during N assimilation. In the process of N transport across membranes for uptake and compartmentation, both proton pumps and proton-coupled N transporters are essential, and their proton-binding sites may sense changes of apoplastic or intracellular pH. In addition, during N assimilation, carbon skeletons are required to synthesize amino acids, thus the combination of NO3– or NH4+ transport and assimilation results in different net charge and numbers of protons in plant cells. Efficient maintenance of N-controlled cellular pH homeostasis may improve N uptake and use efficiency, as well as enhance the resistance to abiotic stresses.
Plant nitrate transporters were first identified and functionally characterized more than 20 years ago. They are encoded at least by four gene families, NRT1 (NPF), NRT2, CLC, and SLAC1/SLAH. In this ...review, we overview the functions of the nitrate transporters in relation to their potential use as targets for improving crop nitrogen use efficiency. These functions include their roles in root architecture and nutrient acquisition; vacuole nitrate and protein storage; nutrient allocation from source to sink; sensing both abiotic and biotic stresses; the ionic balance of nitrate with potassium, chloride and cellular pH; and the circadian clock-regulated carbon and nitrogen balance. We provide and discuss some examples of the use of nitrate transporter genes and their regulators in improving plant growth and development, nitrogen use efficiency, and resistance to some abiotic stresses. We propose several strategies for effectively using nitrate transporters to achieve higher crop yields and nitrogen use efficiency by using gene transformation or genome editing or molecular marker-assisted breeding.
OsNRT1.1a is a low-affinity nitrate（NO_3~-） transporter gene. In this study, another mRNA splicing product, OsNRT1.1b,putatively encoding a protein with six transmembrane domains, was identified ...based on the rice genomic database and bioinformatics analysis. OsNRT1.1a/OsNRT1.1b expression in Xenopus oocytes showed OsNRT1.1a-expressing oocytes accumulated ~（15）N levels to about half as compared to OsNRT1.1bexpressing oocytes. The electrophysiological recording of OsNRT1.1b-expressing oocytes treated with 0.25 mM NO_3~- confirmed ~（15）N accumulation data. More functional assays were performed to examine the function of OsNRT1.1b in rice. The expression of both OsNRT1.1a and OsNRT1.1b was abundant in roots and downregulated by nitrogen（N） deficiency. The shoot biomass of transgenic rice plants with OsNRT1.1a or OsNRT1.1b overexpression increased under various N supplies under hydroponic conditions compared to wild-type（WT）. The OsNRT1.1a overexpression lines showed increased plant N accumulation compared to the WT in 1.25 mM NH_4NO_3 and 2.5 mM NO_3~- or NH_4~＋ treatments, but not in 0.125 mM NH_4NO_3.However, OsNRT1.1b overexpression lines increased total N accumulation in all N treatments, including 0.125 m M NH_4NO_3,suggesting that under low N condition, OsNRT1.1b would accumulate more N in plants and improve rice growth, but also that OsNRT1.1a had no such function in rice plants.
Arsenic (As) contamination in paddy soil can cause phytotoxicity and elevated As accumulation in rice grain. Rice varieties vary in As uptake and tolerance, but the underlying mechanisms remain ...unclear. In this study, the aus variety Kasalath was found to be more tolerant to arsenate As(V) than the japonica variety Nipponbare, but the two varieties showed similar arsenite As(III) tolerance. Nipponbare took up more phosphate (Pi) and As(V) than Kasalath. The expression of genes for Pi transporters or Pi homeostasis regulation was quantified. Nipponbare showed 2- to 3-fold higher expression of the Pi transporter genes OsPT2 and OsPT8 than Kasalath. Two ospt8 mutants were isolated from the Kasalath background and compared with an ospt8 mutant in the Nipponbare background. Mutation in OsPT8 in both backgrounds decreased As(V) uptake by 33–57%, increased As(V) tolerance assayed by root elongation by >100-fold, and abolished the varietal differences in As(V) uptake and tolerance. The results show that OsPT8 plays a key role in As(V) uptake and that As(V) uptake mediated by OsPT8 exerts a profound toxic effect on root elongation. The results also suggest that differential OsPT8 expression explains the varietal differences in As(V) uptake and tolerance between Kasalath and Nipponbare.
Low availability of nitrogen (N) is often a major limiting factor to crop yield in most nutrient-poor soils. Arbuscular mycorrhizal (AM) fungi are beneficial symbionts of most land plants that ...enhance plant nutrient uptake, particularly of phosphate. A growing number of reports point to the substantially increased N accumulation in many mycorrhizal plants; however, the contribution of AM symbiosis to plant N nutrition and the mechanisms underlying the AM-mediated N acquisition are still in the early stages of being understood. Here, we report that inoculation with AM fungus Rhizophagus irregularis remarkably promoted rice (Oryza sativa) growth and N acquisition, and about 42% of the overall N acquired by rice roots could be delivered via the symbiotic route under N-NO₃⁻ supply condition. Mycorrhizal colonization strongly induced expression of the putative nitrate transporter gene OsNPF4.5 in rice roots, and its orthologs ZmNPF4.5 in Zea mays and SbNPF4.5 in Sorghum bicolor. OsNPF4.5 is exclusively expressed in the cells containing arbuscules and displayed a low-affinity NO₃⁻ transport activity when expressed in Xenopus laevis oocytes. Moreover, knockout of OsNPF4.5 resulted in a 45% decrease in symbiotic N uptake and a significant reduction in arbuscule incidence when NO₃⁻ was supplied as an N source. Based on our results, we propose that the NPF4.5 plays a key role in mycorrhizal NO₃⁻ acquisition, a symbiotic N uptake route that might be highly conserved in gramineous species.
The HAK/KUP/KT transporters have been widely associated with potassium (K) transport across membranes in both microbes and plants. Here, we report the physiological function of OsHAK16, a member ...belonging to the HAK/KUP/KT family in rice (Oryza sativa L.). Transcriptional expression of OsHAK16 was up-regulated by K deficiency or salt stress. OsHAK16 is localized at the plasma membrane. OsHAK16 knockout (KO) dramatically reduced root K net uptake rate and growth at both 0.1 mM and 1 mM K supplies, while OsHAK16 overexpression (OX) increased total K uptake and growth only at 0.1 mM K level. OsHAK16-KO decreased the rate of rubidium (Rb) uptake and translocation compared to WT at both 0.2 mM and 1 mM Rb levels. OsHAK16 disruption decreased while its overexpression increased K concentration in root slightly but in shoot remarkably. The relative distribution of total K between shoot and root decreased by 30% in OsHAK16-KO lines and increased by 30% in its OX lines compared to WT. OsHAK16-KO diminished K uptake and K/Na ratio, while OsHAK16-OX improved K uptake and translocation from root to shoot, resulting in increased sensitivity and tolerance to salt stress, respectively. Expression of OsHAK16 enhanced the growth of high salt-sensitive yeast mutant by increasing its K but no Na content. Taking all these together, we conclude that OsHAK16 plays crucial roles in maintaining K homeostasis and salt tolerance in rice shoot.
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