Plant Reproduction

In angiosperms, pollen tubes deliver sperm cells to the ovule and communicate with the external environment as they elongate through the pistils. Although pollination alters Ca2+ conditions within the pistil, the effects of extracellular Ca2+ fluctuations on pollen tube growth and guidance remain largely unknown. In this study, we visualized intracellular Ca2+ dynamics using a semi-in vivo assay with the Ca2+-sensitive fluorescent protein GCaMP6s to investigate how pollen tubes respond to changes in extracellular Ca2+ levels. We found that the Ca2+ levels in the apical region of the pollen tubes reflected the extracellular Ca2+ concentrations. The pollen tube growth rate increased depending on the Ca2+ concentration in the growth medium. However, excessive Ca2+ affected the polar growth of pollen tubes. At elevated Ca2+ concentrations of 10 mM, the pollen tube exhibited coiling behavior and failed to maintain directional growth toward the ovule. Moreover, we provided the first evidence that Ca2+ oscillations are not restricted to the apical region but propagate as a wave, reaching 30-50 m from the apex toward the basal regions. As the pollen tube approached the ovule, it coincided with a substantial elevation in Ca2+ levels, which appeared to drive the accelerated nuclear migration toward the tube apex. Our findings demonstrate that the extracellular Ca2+ environment directly regulates intracellular Ca2+ levels in pollen tubes, thereby influencing their growth and guidance.

3-PHOSPHOINOSITIDE-DEPENDENT PROTEIN KINASE1 (PDK1), a conserved master regulator of AGC kinases, is encoded by two redundant genes in Arabidopsis thaliana, PDK1 and PDK2. pdk1 pdk2 mutants exhibit a broad range of defects, including apolar or arrested pollen tube growth, a phenotype also observed in agc1.5 agc1.7 mutants. Pollen-specific expression of constitutively active AGC1.5 in pdk1 pdk2 restores polar pollen tube growth, indicating that PDK1 functions upstream of redundant AGC1.5/AGC1.7 signaling in this process. In contrast, the PDK1 splice variant PDK1S0, lacking the phospholipid-binding PH domain, cannot restore polar pollen tube growth. Our results indicate a key role for the phospholipid PI(4,5)P2 in recruiting PDK1 through its PH domain to establish polar pollen tube growth, as PI(4,5)P2 marks the pollen germination initiation site together with PDK1, it forms a dome at the plasma-membrane of the pollen tube tip beneath which PDK1 remains largely cytosolic and exhibits reciprocal feedback regulation with the PDK1-AGC1.5/1.7 kinases. Defects in endocytosis and actin organization further support that phospholipid-dependent PDK1-AGC signaling maintains pollen tube growth polarity.

Pollen tubes are dynamic tip-growing cells that deliver sperm nuclei to female gametes in flowering plants, allowing for sexual reproduction and seed formation. Actin and microtubule cytoskeletons both play important roles in directional pollen tube growth and guidance. While actin dynamics are well-studied in pollen tubes, the role of microtubules and the interactions between these two cytoskeletal filaments are less well understood. To address this knowledge gap, we imaged growing Arabidopsis thaliana pollen tubes co-expressing fluorescently-labeled tubulin and actin markers and observed partial co-localization of actin and microtubule filaments. We found that treatment with microtubule disrupting drugs did not affect the actin cytoskeleton. In contrast, when actin filaments were depolymerized, microtubules in the medial region of pollen tubes were disrupted, while microtubules at the cell cortex remained intact. Thus, the microtubule cytoskeleton in A. thaliana pollen tubes relies on the actin cytoskeleton in a spatially dependent manner. Furthermore, we utilized native expression of the microtubule plus-end binding protein EB1b to track microtubule orientation in growing pollen tubes. We found the microtubule array to be largely parallel, with plus ends growing away from the tube apex. Together, these findings offer new insights into the dynamics and organization of microtubules in growing pollen tubes and the interactions between actin filaments and microtubules.

BackgroundRAB Guanine Nucleotide Dissociation Inhibitors (RAB GDIs) are important vesicle transport regulators in eukaryotes, participating in the functional cycle of RAB GTPases by stabilizing their non-active GDP-conformation. AimsWe address the importance of the three Arabidopsis thaliana RAB GDI paralogs by genetic and developmental analyses and put these results into the seed plants evolution context. MethodsWe use methods of genetics, microscopy and phylogenetics. ResultsOur genetic analyses of Arabidopsis T-DNA insertional mutants confirm recent CRISPR alleles data indicating lethality of double gdi1 gdi2 mutants, and our microscopic data point to embryo development arrest in double mutant seeds. We also confirm the involvement of GDI2 and GDI3 in pollen tube growth. Moreover, our data show that GDI1 also contributes to proper pollen function. Our phylogenetic analysis reveals independent diversification of RAB GDIs in Gymnosperms and Angiosperms, with early specialization of an Angiosperm reproduction-and gametophyte-related clade. ConclusionsIn Arabidopsis, RAB GDI1 and 2 are important for the vegetative growth while RAB GDI2 and 3 are vital for reproduction. Evolution of the RAB GDI family reflects the evolution of seed plants. HighlightsRAB GDIs are vital for plant growth and reproduction and act redundantly. Even the low-transcribed RAB GDI1 isoform contributes to the proper pollen function. Two RAB GDI clades evolved in early Angiosperms.

CoverCress is a new winter annual oilseed crop developed from field pennycress within the past 20 years. Field pennycress is commonly considered to be self-pollinated but little basic research has been published and there is some misalignment of conclusions. Our experience working with pennycress plant growth in greenhouse and field conditions over the past 13 years suggests that outcrossing is uncommon. We conducted lab, greenhouse, and field experiments to strengthen the body of work. Pollen viability kinetics analysis showed that longevity of pollen viability is negatively impacted by increasing temperatures and by direct exposure to light. Samples treated at 4C declined to 50% viability in 12 hours while it took just 2.5 hrs at 37C, and 1.6 hrs in full sunlight on a cool early April day. Cross-pollination was absent among greenhouse-grown plants flowering inside an agitated plastic pollen-containment covering. Across greenhouse tests, high rates of cross-pollination occurred only in an emasculation treatment that rendered flowers male sterile and opened the pistil to cross-fertilization. Field trials designed to measure pollen flow distance using a trackable fae1 knockout reporter gene failed to show detectable movement of pollen under field conditions in two locations. This data strongly suggests that domesticated field pennycress may be considered a self-pollinated crop and managed as such.

The nucleus is the characteristic organelle for eukaryotic organisms. Unlike the classic textbook view of static two-dimensional nuclei, nuclear shape is dynamic inside the live cell. The alteration or deformed nuclear shape is the hallmark of cancer in animal cells and environmental stress in plants. The nuclear envelope proteins interact with chromatin to regulate gene expression. Unfortunately, we have limited knowledge about the impact of abiotic stress on nuclear shape, movement, and chromatin dynamics. To circumvent this issue, we are utilizing a dual fluorescently tagged marker lines - nuclear envelope protein and chromatin - to perform live cell imaging in the model plant Arabidopsis thaliana root. The live cell imaging was performed in control and salt-stressed conditions. We utilized these captured movies to analyze through open-source image processing software Fiji/ImageJ with the help of the TrackMate plugin. Using this method, we have demonstrated that chromatin velocity is decreased in salt-treated conditions. This method will be widely applied to quantitative live cell imaging of nuclear shape and chromatin dynamics during plant development and environmental stress. SummaryThis process aims to simultaneously record nucleus and chromatin dynamics in Arabidopsis thaliana roots and investigate changes in these dynamics in response to developmental and environmental cues.

BackgroundMelastomes are well known for their striking diversity in stamen morphologies mostly adapted to buzz pollination by bees. The variously modified connective appendages and heteranthery in the family have fascinated botanists for more than two centuries and a variety of functions associated with pollination have been discovered for these staminal traits over the years. The repeated evolutionary shifts in these traits have been linked to pollinator shifts, likely contributing to diversification in the family. The evolutionary lability of staminal traits, especially the connective morphology, led us to hypothesize that these traits might be controlled by relatively simple genetic mechanisms and we here take the first steps to test this hypothesis by using a comparative transcriptomics approach with Arthrostemma ciliatum as our model. We also tested the functional significance of heteranthery and whether the classical division of labour hypothesis holds true for this species by comparing the number, size and viability of pollen in the two stamen types. ResultsStaminal development of this species was studied and suitable stages for transcriptome comparisons were identified. Differential expression analyses between the morphologically distinct stamen whorls at four developmental stages showed the differential expression of several transcripts involved in stamen development/elongation. Pollen comparisons between the two whorls showed that the antepetalous/inner whorl stamens have a significantly higher number of pollen and higher germination rates while the antesepalous/outer whorl stamens have significantly larger pollen. ConclusionsWe identified Jasmonate and Gibberellin signalling pathway genes (JAZ, GID1, DELLA and ARF homologs), EPF/EPFL family genes, autophagy related genes (VPE homologs) and S Locus ELF homologs as putative candidates involved in causing staminal dimorphism in A. ciliatum. Our results indicate that, for the heterantherous morph of this species, the shorter stamens (antepetalous/inner whorl) have both pollinating and feeding functions contradicting the division of labour theory. We also report the possible existence of heterostyly in A. ciliatum as an outbreeding mechanism.

Fruit development is a typical angiosperm feature that greatly facilitates seed dispersal. Despite extensive studies on the gene regulatory network underlying pod shattering in the dry Arabidopsis fruit and the ripening process in the fleshy tomato fruit, it is yet unclear if a conserved regulatory network acts in early fruit development. Here, we investigated the roles of Petunia x hybrida (petunia) FRUITFULL (FUL), SHATTERPROOF (SHP) and APETALA 2 (AP2) homologs, three types of transcription factors repeatedly associated with fruit development and/or ripening. Petunia is closely related to tomato but produces dry dehiscent fruits like Arabidopsis. Our functional analysis revealed that the three petunia FUL-like genes, PETUNIA FLOWERING GENE (PFG), FLORAL BINDING PROTEIN 26 (FBP26) and FBP29, redundantly regulate endocarp development. They promote the formation of regularly shaped inner endocarp cells, probably via auxin/brassinosteroid signalling and cell wall modification. Furthermore, we discovered that the SHP-like gene FLORAL BINDING PROTEIN 6 (FBP6) has an opposite role, promoting more mesocarp-shaped endocarp cells, indicating that the FUL-like and SHP-like genes act antagonistically in early pericarp development. Finally, we show that the AP2-like genes REPRESSOR OF B-FUNCTION 1 (ROB1), ROB2 and ROB3 are crucial factors in petunia fruit development. rob1 rob2 rob3 mutants completely fail to dehisce and show major defects in pericarp patterning. The ROB transcription factors repress the activity of the FUL-like genes, and have, together with FBP6, an opposite effect on auxin and brassinosteroid signalling genes. Our study suggests that a module consisting of antagonistically acting TFs, including co-orthologs of AP2, FUL and SHP, regulates early pericarp patterning, at least partially via auxin and brassinosteroids.

In flowering plants, pollen tubes communicate with ovular cells to achieve precise one-to-one pollen tube reception. The final step of this communication between the pollen tube and synergid cells has been extensively investigated and visualized by calcium imaging. Synergid cells exhibit characteristic cytoplasmic calcium concentration oscillations, which are thought to play a critical role in pollen tube reception. However, their significance and relationship with calcium dynamics in the entire ovule remain unclear. Here, we show, using the calcium sensor GCaMP6s, that proteins involved in asparagine-linked glycosylation (N-linked glycosylation) are required for normal calcium oscillations in synergid cells but are not essential for pollen tube reception. Using a semi-in vivo assay in Arabidopsis thaliana, we found that the amplitude of these oscillations prior to rapid pollen tube growth across the filiform apparatus was reduced in mutants lacking the oligosaccharyltransferase (OST) 3/6 subunit or alpha1,2-glucosyltransferase (ALG) 10, both of which are involved in N-linked glycosylation. Notably, these mutants did not exhibit reduced fertility attributable to defects in the female gametophyte but instead showed a polytubey phenotype due to a sporophytic defect. These findings suggest that N-linked glycans mediate communication between synergid cells and the pollen tube and indicate that the typical pattern of calcium oscillations in synergid cells is not essential for triggering pollen tube rupture. Furthermore, we show that sporophytic tissues of the ovule exhibit calcium waves that propagate toward the funiculus in correlation with pollen tube contact and rupture, implying that ovular tissues can potentially transmit these signals distantly beyond the ovule. Together, these findings reveal previously unrecognized intercellular calcium signaling and its significance in pollen tube reception by the ovule.

The objective of this study is to determine the quality and define the different classes of honeys produced in the Ivorian forest region according to their pollen content. This involves the analysis of five honey samples from the sub-prefecture of Cechi. Four of the honey samples were wild-harvested, and one was from experimental beekeeping in the Cechi reserve. A total of 54 pollen taxa were identified. The most represented botanical families are: Fabaceae (9 species, or 16.67%), Apocynaceae, and Combretaceae, each with 5 species, or 9.27%. The pollen taxon richness of the honeys varies from 18 to 34 taxa. Most are polyfloral honeys, with the exception of the honey from the reserve, which contains 66.13% Bridelia micrantha pollen (Euphorbiaceae), a monofloral honey. These samples contain highly variable pollen content and fall into three categories of honey: honeys rich in pollen, honeys very rich in pollen, and honeys extremely rich in pollen, attesting to their high quality and natural origin.

Previously, the chitin receptor-interacting protein kinase LIK1 (LysM receptor kinase 1/CERK1-interacting kinase) was shown to play an important role in regulating chitin signaling and plant defense. A limited proteolysis proteomics study revealed several LIK1-derived peptides that showed differential abundance between ATP-treated and mock-treated Arabidopsis samples, suggesting a possible involvement of LIK1 in extracellular ATP (eATP) signaling. To explore this possibility, LIK1 mutants were obtained and examined for their response to ATP. The results showed that mutations in LIK1 significantly reduced the expression of eATP-responsive genes. In addition, LIK1 was found to interact with the eATP receptor P2K1 and to be phosphorylated by it. The LIK1 protein was localized to the plasma membrane and its gene expression appeared to be ubiquitous. Collectively, these findings indicate that LIK1 not only contributes to chitin signaling but also participates in eATP signaling, highlighting its potential role as a shared component in multiple signaling pathways to regulate plant responses to diverse internal and external cues.

Non-structural carbohydrates (NSC) stored in the stem play a crucial role in supporting yield formation in rice. However, internode morphological determinants of NSC accumulation are unclear. This study aimed to clarify the relationship between internode morphology and NSC accumulation and to identify a robust morphological indicator for evaluating NSC accumulation capacity. Two years of field experiments were conducted using multiple cultivars. The NSC content was quantified for individual internodes and at the whole-plant culm level, and its relationships with internode morphological traits were analyzed. Since the upper internodes (UIN; first and second internodes) and lower internodes (LIN; third and subsequent internodes) exhibited contrasting roles in NSC accumulation, a novel index was introduced, the volume composition ratio (VCR) of UIN/LIN, which represents their relative volumetric contributions within a culm. The VCR of UIN/LIN showed the strongest correlation with culm NSC and high reproducibility across years, outperforming simple morphological traits. Manipulation of internode development using plant growth regulators demonstrated that altering VCR effectively modified culm NSC accumulation. Accordingly, the VCR of UIN/LIN serves as a robust morphological indicator of culm NSC accumulation capacity, providing a practical framework for improving NSC accumulation to achieve high and stable yield performance in rice. HighlightThis novel internode structural index robustly predicts the culm non-structural carbohydrate accumulation capacity, providing a practical morphological indicator for improving yield stability in rice.

Atmospheric nitrogen (N) deposition is increasingly affecting global ecosystems, with nitrate contributing a growing proportion alongside ammonium. However, the interaction between N forms and leaf developmental stage in shaping physiological and metabolic strategies in Chinese fir remains poorly understood. In this study, a field experiment was conducted to explore the physiological and metabolic responses of young and old leaves to ammonium and nitrate N addition. Our findings showed that N addition enhanced photosynthetic performance in young leaves, with a stronger effect from nitrate. In contrast, old leaves exhibited limited photosynthetic response but accumulated higher non-structural carbohydrates and showed elevated N assimilation enzyme activities, particularly under nitrate addition. Phytohormone profiles varied between leaf ages, with young leaves having higher auxin levels while old leaves exhibiting increased abscisic and salicylic acid contents under N addition. Additionally, N addition induced differential reprogramming of amino acid metabolism, with age-dependent accumulation patterns. Metabolomic analysis identified key amino acids involved in coordinating carbon-nitrogen metabolism. These results highlighted the complementary metabolic strategies by young and old leaves of Chinese fir under contrasting N forms addition and emphasized the importance of considering both N form and leaf age in optimizing N management for sustainable plantation practices. HighlightsO_LINitrate enhanced photosynthesis in young Chinese fir leaves more effectively than ammonium. C_LIO_LIOld leaves prioritized C storage and N assimilation under N addition, especially nitrate. C_LIO_LIComplementary metabolic strategies between leaf ages optimized resource use under different N forms addition. C_LI

Photosynthesis accounts for most of the final grain yield in rice, making improvements in radiation use efficiency (RUE) a key strategy for enhancing productivity. Agronomically, RUE is defined as the biomass produced per unit of total solar radiation or photosynthetically active radiation intercepted by the canopy. However, the interaction between carbon and nitrogen metabolism plays a critical role in determining plant growth and grain yield. Assimilated nitrogen is required for the synthesis of photosynthetic pigments and enzymes, while the reduction of nitrate (NOLL) and nitrite (NOLL), as well as the assimilation of ammonium (NHLL), depend on the reducing power and carbon skeletons generated by photosynthesis. In this study, two high-yielding rice (Oryza sativa) cultivars--an indica-type (El Paso 144) and a japonica-type (INIA Parao) were subjected to two nitrogen treatments (3 mM and 9 mM NOLL/NHLL) and two light intensities (850 and 1500 mol mL{superscript 2} sL{superscript 1}). A strong interaction between light intensity and nitrogen metabolism was observed, with contrasting responses between subspecies. These differences reflect a coordinated regulation of carbon assimilation and primary nitrogen metabolism. The results provide new insights into the metabolic strategies underlying nitrogen compound accumulation under variable irradiance. Such knowledge is essential for improving nitrogen fertilizer use efficiency and yield performance in elite rice genotypes cultivated under commercial field conditions.

AO_SCPLOWBSTRACTC_SCPLOWNitrogen fertilizers are essential for crop productivity but cause environmental harm, necessitating the development of cultivars that thrive under limited nitrogen. This study investigates the transcriptomic response to nitrate in Arabidopsis thaliana (a model dicot), Brachypodium distachyon (a model Pooideae), and Hordeum vulgare (barley, a domesticated Pooideae) to identify conserved and species-specific molecular mechanisms. Using RNA-seq after 1.5 and 3 hours of nitrate treatment, we found that core nitrate-responsive biological processes - such as nitrate transport, assimilation, carbon metabolism, and hormone signaling - are largely conserved across species. However, comparative analysis at gene level based on orthology revealed specificities between the species. For instance, rRNA processing was uniquely stimulated in Arabidopsis, while cysteine biosynthesis from serine and gibberellin biosynthesis were specifically regulated in Brachypodium and barley. Orthologs of key nitrate-responsive genes (e.g., NRT, NLP, TCP20) exhibited variable regulation, reflecting potential adaptations linked to domestication or nutrient acquisition strategies. These findings highlight the importance of integrating model and crop species to uncover targets for improving nitrogen use efficiency in cereals. The study provides a pipeline integrating gene ontology and orthology analyses to compare transcriptomic responses between species.

Fruit softening is primarily determined by modifications in the cell wall architecture, which are mediated by the coordinated activity of cell wall-degrading enzymes. In strawberry (Fragaria x ananassa Duch.), transgenic suppression of the genes encoding the pectinases polygalacturonase (PG), {beta}-galactosidase ({beta}Gal), and rhamnogalacturonan lyase (RGLyase), reduced fruit softening. In this study, we evaluated the fruit firmness phenotype of selected transgenic lines across several harvest years, from 3 to 9 years depending on the line, and analyzed the cell wall composition of ripe fruits using a carbohydrate microarray. Multiple linear regression analysis revealed that fruit firmness was significantly affected by both genotype and harvest year, while fruit size and soluble solids content showed no significant contribution. All pectinase-silenced lines exhibited increased firmness relative to the wild type, with those lines with PG down-regulated showing the most significant effects, followed by B-Gal and RGLyase fruits. The firmer phenotype was maintained stably in all the transgenic lines during the different years analyzed. Carbohydrate microarray analyses of sequentially extracted cell wall fractions demonstrated that transgenic ripe fruits retained higher levels of low methyl-esterified homogalacturonan (HG) and rhamnogalacturonan I (RG-I) epitopes compared to wild-type ripe fruits, resembling the composition of white-stage control fruits. Principal component analysis of microarray data revealed a clear separation between wild-type ripe fruits and transgenic lines, with the latter clustering near the earlier developmental stages of wild-type fruits. Correlation analysis further revealed positive associations between increased firmness and the abundance of high-methylated HG pectic epitopes in the water fraction, recognized with JIM7, and low-methylated HG abundance in the rest of the fractions (JIM5, LM18, and LM19). Overall, these results suggest that suppressing pectinase genes alters pectin remodeling during ripening, resulting in the retention of structurally intact pectin domains and increased fruit firmness. These genes are therefore excellent candidates for the improvement of this key quality trait in strawberry.

Fusion of gametes possessing meiotically reduced (haploid) chromosome complements is the main pathway of propagation among eukaryotes. However, duckweeds, the smallest angiosperms, propagate mainly vegetatively, and meiosis has not yet been documented in detail for this plant family. The more surprising was the recent evidence of rather frequent interspecific hybrids and triploid clonal accessions which became obvious by genome size measurements, genomic in situ hybridization (GISH) and combined plastid and nuclear DNA markers. These observations indicated sexual propagation involving reduced as well as unreduced male and female gametes in Lemna minor and L. turionifera leading to allodiploid and allotriploid hybrids (MT, MMT, MTT) and autotriploid L. minor (MMM) accessions. Here, we i) documented the meiotic stages of Lemna species for the first time; ii) provided evidence of unreduced male gametes through fluorescent in situ hybridization (FISH) with single locus probes; iii) determined their abundance in different individuals and iv) hypothesized about the reasons of unreduced male gamete formation. These findings open new insights into the modes of sexual reproduction and evolution of duckweeds which may be useful for future breeding efforts in this emerging crop.

Plant genome sequences provide access to the gene repertoire of a species. This facilitates basic research, biotechnological processes, or horticultural applications. Here, we present the genome sequence of Begonia manicata and unravel the genes underlying the pigmentation of red structures emerging from its leaves and stems. Structural genes of the anthocyanin biosynthesis and corresponding regulatory genes were discovered to be upregulated in these red structures suggesting that the pigmentation is caused by the accumulation of anthocyanins. Our work provides a resource for future studies on pigmentation of Begoniaceae.

Increasing attention is being focused on the glycemic index (GI) of daily food for humans, and the resistant starch content (RSC) is an important indicator of GI for starch-rich staple foods. In recent years, some studies revealed that the loss function of single or multiple key enzymes in the primary pathway of starch synthesis substantially increases RSC in rice, such as starch branching enzyme IIb (BEIIb) and soluble starch synthase IIIa (OsSSIIIa). However, a noteworthy negative characteristic of these high RSC mutants is the substantially increased amylose content (AC). AC as a major determinator of rice eating quality, must not be higher than an acceptable limit for most consumers. To solve this problem, in this study, we adopted two promoter editing (PE) editing strategies to develop rice germplasms with a better balance of RSC and AC: one is to edit the promoter of BEIIb in a low AC rice variety, another is to edit the promoter of Waxy (Wx) gene in a BEIIb loss of function mutant. Using AC[≤]20%, which is the range of premium quality rice in China as a criteria, we finally obtained 2 homozygous lines with significantly increased RSC ([≥]5%) in the NG46 background by promoter editing of BEIIb and 1 homozygous line in the YouTang2 (YT2, a BEIIb mutant) background by promoter editing of Wx gene. Further analysis revealed that AC and the amount of long-chain branches of amylopectin are positively correlated with RSC in the population of BEIIb PE lines. However, unexpectedly, the Wx PE-line with substantially decreased AC (17.7%) also showed significantly increased RSC (16.9%). Our study not only produces useful germplasms for the high RSC rice breeding in the future but also provides an insight into understanding the relationship between AC and RSC in defective BEIIb rice.

C4 photosynthesis is a CO2-concentration mechanism that separates CO2 fixation between two cell types, thereby reducing photorespiration and making C4 plants more efficient than their C3 counterparts. While the C4 cycle has evolved multiple times across different genera, this study evaluates very closely related C3 and C4 species within the genus Flaveria. Apart from their carbon metabolism, C4 plants also possess adaptations in their mineral nutrition. One key nutrient which is also directly involved in photosynthesis is phosphorus. It is absorbed by the plant in the form of inorganic phosphate and is an essential component of DNA, ATP, lipids, and carbohydrates. In the Flaveria C4 species, but not in the C3 species, phosphate limitation was shown to affect the dark reactions of photosynthesis. This study investigates how phosphate deficiency impacts the light reactions in C3 and C4 Flaveria plants. We observed a differential response in the functionality of photosynthetic energy conversion between the two species. When exposed to a limited phosphate supply, the C3 species reduced its linear electron transport rate while dissipating excess energy through high-energy quenching, which was regulated by a higher pH gradient across the thylakoid membrane. In contrast, the C4 species did not regulate its photosynthetic light reaction under phosphate limitation. Instead, it exhibited increased stress levels, evidenced by a stronger biomass reduction and the induction of stress markers in the leaves. Additionally, this study uncovered an acceleration in NPQ relaxation during phosphate limitation, regardless of the photosynthesis type. HighlightPhosphate deficiency reduced linear electron transport rates and induced dissipation of excess energy through non-photochemical quenching in the C3 Flaveria species, while in the C4 species, despite elevated stress levels, the photosynthetic light reactions were unaffected.