Class Of Phytohormones Research Articles

GhRAP2.4 enhances drought tolerance by positively regulating the strigolactone receptor GhD14 expression in cotton (Gossypium hirsutum L.).

Drought poses significant challenges to crop productivity, necessitating a deeper understanding of plant adaptive mechanisms. Strigolactones (SLs), a class of phytohormones, have been recognized as crucial regulators in plant responses to drought, yet the specific role of SL receptor in drought tolerance in cotton (Gossypium hirsutum L.) remains underexplored. In this study, we identified and characterized DWARF14 (GhD14), a SL receptor in cotton. Silencing GhD14 in cotton compromised drought tolerance by reducing leaf relative water content and chlorophyll content, slowing stomatal closure, increasing reactive oxygen species levels, and decreasing antioxidant enzyme activities. Conversely, overexpression of GhD14 in Arabidopsis d14 mutants rescued their drought-sensitive phenotype. Further, we identified a 197-bp fragment (-697 to -894 bp) in the GhD14 promoter that plays a crucial role in the drought stress response. RELATED TO APETALA 2.4 (GhRAP2.4), a dehydration responsive element binding protein (DREB) transcription factor, binds directly to the GhD14 promoter, enhancing its transcription under drought conditions. Silencing GhRAP2.4 in cotton resulted in reduced drought tolerance. This study not only elucidates the molecular interplay between GhRAP2.4 and GhD14 in cotton's drought response but also provides a potential target for genetic modification to improve drought resilience in crops.

Multifunctional Role of Cytokinin in Horticultural Crops

Cytokinins (CKs) are a class of phytohormones identified in the early 1960s and are mainly responsible for stimulating cell division. Following the discovery, research to help understand the pluralistic roles of CKs in plant growth and stress biology increased. With their fascinating ability, CKs serve as an important element in regulating the defense–growth trade-off. Herein, we demonstrate how the CK fine-tuning the organogenesis of different parts of horticultural plants is discussed. CK’s role in tailoring reproductive biology (flowering, sex differentiation, fruit set, and fruit attributes) has been presented. An extensive explanation of the CK-mediated response of horticultural crops to abiotic (temperature, drought, and salinity) and biotic stresses (fungal, bacterial, and nematodes) is provided. Finally, we posit the unexplored roles of CKs and highlight the research gaps worth addressing.

A wheat phytohormone atlas spanning major tissues across the entire life cycle provides novel insights into cytokinin and jasmonic acid interplay.

Although numerous studies have focused on the specific organs or tissues at different development stages or under various abiotic and biotic stress, our understanding of the distribution and relative abundance of phytohormones throughout the entire life cycle of plant organs and tissues remains insufficient. Here, we present a phytohormone atlas resource covering the quantitative analysis of eight major classes of phytohormones, comprising a total of 40 hormone-related compounds, throughout the complete life cycle of wheat. In combination with transcriptome analysis, we established a Wheat Phytohormone Metabolic Regulatory Network (WPMRN). Using our WPMRN dataset and GO enrichment analysis, we swiftly characterized the function of TaLOG5-B1 in cytokinin biosynthesis. Furthermore, a detailed investigation of the WPMRN dataset uncovered transcription factor-mediated co-regulation mechanisms among different classes of phytohormones. We focused specifically on the metabolic regulatory involving cytokinin and jasmonic acid. To achieve this, we characterized genes TaLOG3-D1 and TaAOS-D1 involved in the biosynthesis of these phytohormones, along with their regulatory transcription factors TaDOF3A and TaDOF5.6B. The functions of these genes were validated in transgenic plants, revealing their ability to co-regulate radicle length. These findings serve as a case study that highlights the utility of this resource for studying phytohormone metabolic regulatory networks in cereal crops and for gaining insights into the roles of phytohormones in enhancing agronomic traits.

Dressed Up to the Nines: The Interplay of Phytohormones Signaling and Redox Metabolism During Plant Response to Drought.

Plants must effectively respond to various environmental stimuli to achieve optimal growth. This is especially relevant in the context of climate change, where drought emerges as a major factor globally impacting crops and limiting overall yield potential. Throughout evolution, plants have developed adaptative strategies for environmental stimuli, with plant hormones and reactive oxygen species (ROS) playing essential roles in their development. Hormonal signaling and the maintenance of ROS homeostasis are interconnected, playing indispensable roles in growth, development, and stress responses and orchestrating diverse molecular responses during environmental adversities. Nine principal classes of phytohormones have been categorized: auxins, brassinosteroids, cytokinins, and gibberellins primarily oversee developmental growth regulation, while abscisic acid, ethylene, jasmonic acid, salicylic acid, and strigolactones are the main orchestrators of environmental stress responses. Coordination between phytohormones and transcriptional regulation is crucial for effective plant responses, especially in drought stress. Understanding the interplay of ROS and phytohormones is pivotal for elucidating the molecular mechanisms involved in plant stress responses. This review provides an overview of the intricate relationship between ROS, redox metabolism, and the nine different phytohormones signaling in plants, shedding light on potential strategies for enhancing drought tolerance for sustainable crop production.

Nitrogen deficiency influences strigolactone levels in basal parts of shoots and shoot branching phenotype in Arabidopsis thaliana.

Strigolactones (SLs) are allelochemicals attracting both symbiotic arbuscular mycorrhizal (AM) fungi and root parasitic plants and a novel class of phytohormones inhibiting lateral shoot branching. In general, nutrient deficiency significantly promotes SL production in roots and exudation into the rhizosphere, and also induces suppressed shoot branching phenotype in host plants of AM fungi including rice, tomato, maize, etc. Nutrient deficiencies also inhibit the shoot branching in Arabidopsis thaliana (hereafter Arabidopsis), a non-host of AM fungi, while the level of carlactone, a non-canonical SL and the SL precursor for the other SLs, was reported to be unaffected. Because Arabidopsis SLs including CL and methyl carlactonoate,, are highly unstable, relationships between SL levels and shoot branching in Arabidopsis remain elusive. Herein, we demonstrate that nitrogen deficiency increases SL levels in the basal part of shoots in Arabidopsis and lateral shoot branching appears to be strongly regulated by these SLs.

Molecular cloning and characterization of a brassinosteriod biosynthesis-related gene PtoCYP90D1 from Populus tomentosa

Brassinosteroids (BRs), one of the major classes of phytohormones are essential for various processes of plant growth, development, and adaptations to biotic and abiotic stresses. In Arabidopsis, AtCYP90D1 acts as a bifunctional cytochrome P450 monooxygenase, catalyzing C-23 hydroxylation in the brassinolide biosynthetic pathway. The present study reports the functional characterizations of PtoCYP90D1, one of the AtCYP90D1 homologous genes from Populus tomentosa. The qRT-PCR analysis showed that PtoCYP90D1 was highly expressed in roots and old leaves. Overexpression of PtoCYP90D1 (PtoCYP90D1-OE) in poplar promoted growth and biomass yield, as well as increased xylem area and cell layers. Transgenic plants exhibited a significant increase in plant height and stem diameter as compared to the wild type. In contrast, the CRISPR/Cas9-generated mutation of PtoCYP90D1 (PtoCYP90D1-KO) resulted in significantly decreased biomass production in transgenic plants. Further studies revealed that cell wall components increased significantly in PtoCYP90D1-OE lines but not in PtoCYP90D1-KO lines, as compared to wild-type plants. Overall, the findings indicate a positive role of PtoCYP90D1 in improving growth rate and elevating biomass production in poplar, which will have positive implications for its versatile industrial or agricultural applications.

Changes in the Endogenous Phytohormone Status of Drought-Stressed Impatiens walleriana Hook. f. Plants Treated with Methyl Jasmonate

The response of plants to changing environmental conditions is mediated by endogenous phytohormones of which concentration and activity can be altered by the exogenous application of plant growth regulators (PGRs). The present study was conducted to investigate the effects of foliar application of distilled water (ddH2O) and methyl jasmonate (MeJA—5 and 50 µM) under drought stress (15 and 5% of soil water content—SWC) on the endogenous phytohormone status of Impatiens walleriana. The following classes of phytohormones were analyzed by HPLC–MS: abscisic acid (ABA) and its metabolic intermediates, jasmonates, salicylic acid (SA), cytokinins (CKs), and auxin indole-3-acetic acid (IAA). The results showed that plant leaves sprayed with 50 μM MeJA during severe drought (5% SWC) had the highest concentrations of ABA and jasmonic acid (JA). In addition, the plant leaves treated with 50 μM MeJA at 15% SWC showed a significantly increased JA content. In plant groups treated with ddH2O and 50 µM MeJA, the content of another jasmonate, jasmonic acid-isoleucine (JA-Ile), increased at 5% SWC. Interestingly, foliar application of MeJA did not induce changes in endogenous MeJA content in I. walleriana leaves. Similarly, no changes in SA or CK content were observed in any of the treated plant groups. On the other hand, an increase in the auxin IAA was observed in plants sprayed with ddH2O and 50 µM MeJA at 5% SWC. The results from this study indicate that MeJA foliar application could significantly affect the endogenous phytohormonal status of I. walleriana, which changes are fundamental in overall plant drought experience. According to the obtained results, the concentration of ABA and jasmonates changed the most significantly, indicating a possible overlap of their signaling pathways, as well as interaction with other phytohormones.

Peptide Signals Regulate Nitrogen Deficiency Adaptation of Dicotyledonous Model Plants.

Nitrogen is a crucial macroelement essential for plant growth and development. In Arabidopsis Thaliana, classical phytohormones such as auxin and cytokinin orchestrate local and systemic signalling networks coordinate plant growth and development in response to nitrogen deficiency. Nowadays, emerging signalling pathways involving small peptides like CLAVATA3/EMBRYO SURROUNDINGR REGION (CLE) and C-TERMINALLY ENCODED PEPTIDE (CEP) and their corresponding kinase receptors, also regulate Arabidopsis' adaptation to nitrogen scarcity. Unlike Arabidopsis, which adapts to nitrogen deficiency by changing root development, legumes have the unique ability to form nitrogen-fixing root nodules through symbiotic interactions with soil rhizobia. During the symbiotic nodulation in Medicago, CLE and CEP peptides and their receptors consist of an autoregulatory network governing the number of nodules in accordance with the soil nitrogen level. Additionally, other plant peptides, such as phytosulfokine (PSK) and root meristem growth factors (RGF), have been identified as new regulators of leguminous root nodule development under nitrogen-limited condition. However, the precise mechanism by which these peptides coordinate nitrogen deficiency response and the development of nitrogen-fixing organs remains to be fully elucidated. This review summarises the adaptive strategies of dicotyledons to nitrogen deficiency, with a particular focus on the regulation of Medicago nitrogen-fixing nodule development by the peptides.

Five unaddressed questions about cytokinin biosynthesis.

Cytokinins, a class of phytohormones, play crucial roles in regulating plant growth and stress responses through finely tuned feedback loops involving metabolic and signaling cascades. Cytokinin metabolism modulates the abundance of these biologically active molecules. Over the past 25 years, studies have identified key genes involved in cytokinin biosynthesis and inactivation pathways. Nevertheless, several gaps remain in our understanding, particularly regarding the movement of intermediate metabolites between subcellular compartments and the discrepancy between the product of adenosine phosphate-isopentenyltransferase (IPT) and the substrate preferences of subsequent reactions. In addition, recent gene discoveries related to lonely guy (LOG)-independent pathways suggest a spatial extension of cytokinin biosynthesis into the apoplast. Other intriguing issues remain to be addressed, i.e., elucidating the synthetic pathway for cis-zeatin and unraveling the molecular mechanisms governing selective substrate use by the cytokinin biosynthetic enzyme tumor morphology root (Tmr) derived from the phytopathogen Agrobacterium tumefaciens during crown gall formation. Further studies are needed to reveal a fully comprehensive picture of cytokinin metabolism.

Near-Infrared Fluorescent Carbon Nanotube Sensors for the Plant Hormone Family Gibberellins

Gibberellins (GAs) are a class of phytohormones, important for plant growth, and very difficult to distinguish because of their similarity in chemical structures. Herein, we develop the first nanosensors for GAs by designing and engineering polymer-wrapped single-walled carbon nanotubes (SWNTs) with unique corona phases that selectively bind to bioactive GAs, GA3 and GA4, triggering near-infrared (NIR) fluorescence intensity changes. Using a new coupled Raman/NIR fluorimeter that enables self-referencing of nanosensor NIR fluorescence with its Raman G-band, we demonstrated detection of cellular GA in Arabidopsis, lettuce, and basil roots. The nanosensors reported increased endogenous GA levels in transgenic Arabidopsis mutants that overexpress GA and in emerging lateral roots. Our approach allows rapid spatiotemporal detection of GA across species. The reversible sensor captured the decreasing GA levels in salt-treated lettuce roots, which correlated remarkably with fresh weight changes. This work demonstrates the potential for nanosensors to solve longstanding problems in plant biotechnology.

Molecular mechanism of brassinosteroids involved in root gravity response based on transcriptome analysis

BackgroundBrassinosteroids (BRs) are a class of phytohormones that regulate a wide range of developmental processes in plants. BR-associated mutants display impaired growth and response to developmental and environmental stimuli.ResultsHere, we found that a BR-deficient mutant det2-1 displayed abnormal root gravitropic growth in Arabidopsis, which was not present in other BR mutants. To further elucidate the role of DET2 in gravity, we performed transcriptome sequencing and analysis of det2-1 and bri1-116, bri1 null mutant allele. Expression levels of auxin, gibberellin, cytokinin, and other related genes in the two mutants of det2-1 and bri1-116 were basically the same. However, we only found that a large number of JAZ (JASMONATE ZIM-domain) genes and jasmonate synthesis-related genes were upregulated in det2-1 mutant, suggesting increased levels of endogenous JA.ConclusionsOur results also suggested that DET2 not only plays a role in BR synthesis but may also be involved in JA regulation. Our study provides a new insight into the molecular mechanism of BRs on the root gravitropism.

Chemistry of Strigolactones, Key Players in Plant Communication.

Today, the use of artificial pesticides is questionable and the adaptation to global warming is a necessity. The promotion of favorable natural interactions in the rhizosphere offers interesting perspectives for changing the type of agriculture. Strigolactones (SLs), the latest class of phytohormones to be discovered, are also chemical mediators in the rhizosphere. We present in this review the diversity of natural SLs, their analogs, mimics, and probes essential for the biological studies of this class of compounds. Their biosynthesis and access by organic synthesis are highlighted especially concerning noncanonical SLs, the more recently discovered natural SLs. Organic synthesis of analogs, stable isotope-labeled standards, mimics, and probes are also reviewed here. In the last part, the knowledge about the SL perception is described as well as the different inhibitors of SL receptors that have been developed.

Strigolactones promote flowering by inducing the miR319-LA-SFT module in tomato

Strigolactones are a class of phytohormones with various functions in plant development, stress responses, and in the interaction with (micro)organisms in the rhizosphere. While their effects on vegetative development are well studied, little is known about their role in reproduction. We investigated the effects of genetic and chemical modification of strigolactone levels on the timing and intensity of flowering in tomato (Solanum lycopersicum L.) and the molecular mechanisms underlying such effects. Results showed that strigolactone levels in the shoot, whether endogenous or exogenous, correlate inversely with the time of anthesis and directly with the number of flowers and the transcript levels of the florigen-encoding gene SINGLE FLOWER TRUSS (SFT) in the leaves. Transcript quantifications coupled with metabolite analyses demonstrated that strigolactones promote flowering in tomato by inducing the activation of the microRNA319-LANCEOLATE module in leaves. This, in turn, decreases gibberellin content and increases the transcription of SFT. Several other floral markers and morpho-anatomical features of developmental progression are induced in the apical meristems upon treatment with strigolactones, affecting floral transition and, more markedly, flower development. Thus, strigolactones promote meristem maturation and flower development via the induction of SFT both before and after floral transition, and their effects are blocked in plants expressing a miR319-resistant version of LANCEOLATE. Our study positions strigolactones in the context of the flowering regulation network in a model crop species.

The protein phosphatase qGL3/OsPPKL1 self-regulates its degradation to orchestrate brassinosteroid signaling in rice

Brassinosteroids (BRs) are a class of phytohormones that regulate plant growth and development. In previous studies, we cloned and identified PROTEIN PHOSPHATASE WITH KELCH-LIKE1 (OsPPKL1) as the causal gene for the quantitative trait locus GRAIN LENGTH3 (qGL3) in rice (Oryza sativa). We also showed that qGL3/OsPPKL1 is mainly located in the cytoplasm and nucleus and negatively regulates BR signaling and grain length. Because qGL3 is a negative regulator of BR signaling, its turnover is critical for rapid response to changes in BRs. Here, we demonstrate that qGL3 interacts with the WD40-domain-containing protein WD40-REPEAT PROTEIN48 (OsWDR48), which contains a nuclear export signal (NES). The NES signal is crucial for the cytosolic localization of OsWDR48 and also functions in the self-turnover of qGL3. We show that OsWDR48 physically interacts with and genetically acts through qGL3 to modulate BR signaling. Moreover, qGL3 may indirectly promote the phosphorylation of OsWDR48 at the Ser-379 and Ser-386 sites. Substitutions of both phosphorylation sites in OsWDR48 to non-phosphorylatable alanine enhanced the strength of the OsWDR48–qGL3 interaction. Furthermore, we found that brassinolide can promote the accumulation of non-phosphorylated OsWDR48, leading to strong interaction intensity between qGL3 and OsWDR48. Taken together, our results show that OsWDR48 facilitates qGL3 retention and induces degradation of qGL3 in the cytoplasm. These findings suggest that qGL3 self-modulates its turnover by binding to OsWDR48 to regulate its cytoplasmic localization and stability, leading to efficient orchestration of BR signal transduction in rice.

Phytohormones in a universe of regulatory metabolites: lessons from jasmonate.

Small-molecule phytohormones exert control over plant growth, development, and stress responses by coordinating the patterns of gene expression within and between cells. Increasing evidence indicates that currently recognized plant hormones are part of a larger group of regulatory metabolites that have acquired signaling properties during the evolution of land plants. This rich assortment of chemical signals reflects the tremendous diversity of plant secondary metabolism, which offers evolutionary solutions to the daunting challenges of sessility and other unique aspects of plant biology. A major gap in our current understanding of plant regulatory metabolites is the lack of insight into the direct targets of these compounds. Here, we illustrate the blurred distinction between classical phytohormones and other bioactive metabolites by highlighting the major scientific advances that transformed the view of jasmonate from an interesting floral scent to a potent transcriptional regulator. Lessons from jasmonate research generally apply to other phytohormones and thus may help provide a broad understanding of regulatory metabolite-protein interactions. In providing a framework that links small-molecule diversity to transcriptional plasticity, we hope to stimulate future research to explore the evolution, functions, and mechanisms of perception of a broad range of plant regulatory metabolites.

The N-terminal α2 helix element is critical for the activity of the rice transcription factor MYC2.

Jasmonates (JAs) are a class of phytohormones that play a crucial role in plant growth, development, and environmental stress responses. Central to JA signaling are the MYC2-type transcription factors, as they activate the expression of JA-responsive genes. We previously used CRISPR-Cas9-based genome editing to engineer rice OsMYC2 and yielded a mutant (myc2-5) with a single amino acid (aa) deletion (75I) outside the known functional domains of the protein. This myc2-5 mutant also showed some JA-deficient phenotypes, promoting us to investigate how 75I deletion affects JA responses. The mutation is found in the α2 helix element at the N-terminal of OsMYC2. The deletion of 75I in OsMYC2 rendered plants deficient in most of the JA responses, including root growth, leaf senescence, spikelet development, and resistance to pathogens and herbivores. Biochemical assays revealed that the 75I deletion markedly reduced OsMYC2 protein accumulation, subsequently diminishing its transcriptional activity. However, the deletion did not influence the protein's subcellular localization, DNA-binding capability, or its interactions with JAZ transcriptional repressors and the Mediator complex subunit MED25. Additionally, the screening of seven other deletions in the α2 helix further reinforces the importance of this protein element. Our results highlight the significance of the α2 helix in the N-terminus for OsMYC2's functionality, primarily through modulating its protein levels. This insight expands our knowledge of JA signaling and opens new avenues for research into the yet-to-be-explored domains of the MYC2 protein, with the potential to tailor JA responses in rice and other plant species.

A Model of the Full-Length Cytokinin Receptor: New Insights and Prospects.

Cytokinins (CK) are one of the most important classes of phytohormones that regulate a wide range of processes in plants. A CK receptor, a sensor hybrid histidine kinase, was discovered more than 20 years ago, but the structural basis for its signaling is still a challenge for plant biologists. To date, only two fragments of the CK receptor structure, the sensory module and the receiver domain, were experimentally resolved. Some other regions were built up by molecular modeling based on structures of proteins homologous to CK receptors. However, in the long term, these data have proven insufficient for solving the structure of the full-sized CK receptor. The functional unit of CK receptor is the receptor dimer. In this article, a molecular structure of the dimeric form of the full-length CK receptor based on AlphaFold Multimer and ColabFold modeling is presented for the first time. Structural changes of the receptor upon interacting with phosphotransfer protein are visualized. According to mathematical simulation and available data, both types of dimeric receptor complexes with hormones, either half- or fully liganded, appear to be active in triggering signals. In addition, the prospects of using this and similar models to address remaining fundamental problems of CK signaling were outlined.

Resolving binding pathways and solvation thermodynamics of plant hormone receptors

Plant hormones are small molecules that regulate plant growth, development, and responses to biotic and abiotic stresses. They are specifically recognized by the binding site of their receptors. In this work, we resolved the binding pathways for eight classes of phytohormones (auxin, jasmonate, gibberellin, strigolactone, brassinosteroid, cytokinin, salicylic acid, and abscisic acid) to their canonical receptors using extensive molecular dynamics simulations. Furthermore, we investigated the role of water displacement and reorganization at the binding site of the plant receptors through inhomogeneous solvation theory. Our findings predict that displacement of water molecules by phytohormones contributes to free energy of binding via entropy gain and is associated with significant free energy barriers for most systems analyzed. Also, our results indicate that displacement of unfavorable water molecules in the binding site can be exploited in rational agrochemical design. Overall, this study uncovers the mechanism of ligand binding and the role of water molecules in plant hormone perception, which creates new avenues for agrochemical design to target plant growth and development.

Highly sensitive analysis of photoregulation and dynamic distribution of phytohormones based on nanoconfined liquid phase nanoextraction

BackgroundAs a vital energy source, light is one of the most significant environmental signals for plants' growth and development. The crosstalk amongst phytohormones regulated by light exhibits quantitative dynamic changes, but methodologies to analyze their distribution during plant growth are still limited. Rapid, highly sensitive, low-invasive detection and simultaneous assessment of the levels of multiple classes of phytohormones have important phytology applications, however the existing sample pretreatment strategies remain intricate, laborious, and far from being developed for in vivo high-sensitivity testing. (81) ResultsWe applied a nanoconfined liquid phase nanoextraction (NLPNE) technique based on acidified carbon nanofibers (ACNFs) in combination with LC-ESI-MS/MS for highly sensitive analysis of acidic phytohormones’ photoregulation and dynamic distribution. In this system, the mass transfer ability of analytes entering the nanoconfined space is significantly improved given the nanoconfined effect. In particular, the accelerated and strong adsorption of alkaline compounds to the ACNFs surface provide minimum interference for acidic compounds (photosensitive phytohormones), which facilitates their simple, fast, and selective quantification with improved sensitivity. The ACNFs-NLPNE strategy achieved quantitative enrichment of multi-class phytohormones in less than 5 min, and detection limits down to 0.49 fg mL−1. Moreover, we monitored the phytohormone changes under red and blue monochromatic light with relative standard deviations <13.4 %. The results further indicated that short-time red light regulation promoted Lepidium sativum L. growth while blue light inhibited it. (141) SignificanceA nanoconfinement effect-based sample pretreatment platform was developed for monitoring photoregulation phytohormones dynamic distribution with higher sensitivity and stability. Our findings highlighted the importance of the NLPNE approach in providing an accurate plant crosstalk information at the molecular level, which opens a promising avenue for investigating internal hormonal responses to external stimuli. (52).

Low phosphorus promotes NSP1–NSP2 heterodimerization to enhance strigolactone biosynthesis and regulate shoot and root architecture in rice

Phosphorus is an essential macronutrient for plant development and metabolism, and plants have evolved ingenious mechanisms to overcome phosphate (Pi) starvation. However, the molecular mechanisms underlying the regulation of shoot and root architecture by low phosphorus conditions and the coordinated utilization of Pi and nitrogen remain largely unclear. Here, we show that Nodulation Signaling Pathway 1 (NSP1) and NSP2 regulate rice tiller number by promoting the biosynthesis of strigolactones (SLs), a class of phytohormones with fundamental effects on plant architecture and environmental responses. We found that NSP1 and NSP2 are induced by Oryza sativa PHOSPHATE STARVATION RESPONSE2 (OsPHR2) in response to low-Pi stress and form a complex to directly bind the promoters of SL biosynthesis genes, thus markedly increasing SL biosynthesis in rice. Interestingly, the NSP1/2–SL signaling module represses the expression of CROWN ROOTLESS 1 (CRL1), a newly identified early SL-responsive gene in roots, to restrain lateral root density under Pi deficiency. We also demonstrated that GR244DO treatment under normal conditions inhibits the expression of OsNRTs and OsAMTs to suppress nitrogen absorption but enhances the expression of OsPTs to promote Pi absorption, thus facilitating the balance between nitrogen and phosphorus uptake in rice. Importantly, we found that NSP1p:NSP1 and NSP2p:NSP2 transgenic plants show improved agronomic traits and grain yield under low- and medium-phosphorus conditions. Taken together, these results revealed a novel regulatory mechanism of SL biosynthesis and signaling in response to Pi starvation, providing genetic resources for improving plant architecture and nutrient-use efficiency in low-Pi environments.