Search Results (1,008)

Auxin (IAA), a key natural signaling molecule, plays a pivotal role in regulating plant growth, development, and stress responses. Understanding its signal transduction mechanisms is crucial for improving crop yields. In this study, we conducted a comparative transcriptome analysis of wheat leaf and root tissues treated with different concentrations of IAA (0, 1, and 50 μM). Functional enrichment analysis revealed that differentially expressed genes (DEGs) exhibited tissue-specific regulatory patterns in response to auxin. Weighted Gene Co-expression Network Analysis (WGCNA) identified receptor-like kinase genes within the MEgreen module as highly correlated with auxin response, suggesting their involvement in both root and leaf regulation. Among them, TaPBL7-2B, a receptor-like kinase gene significantly upregulated under 50 μM IAA treatment, was selected for functional validation. Ectopic overexpression of TaPBL7-2B in Arabidopsis thaliana (Col-0) enhanced auxin sensitivity and inhibited plant growth by suppressing root development and leaf expansion. In contrast, knockout of the Arabidopsis homolog AtPBL7 reduced auxin sensitivity and promoted both root and leaf growth. Transcriptome analysis of Col-0, the TaPBL7-2B overexpression line, and the pbl7 mutant indicated that TaPBL7-2B primarily functions through the MAPK signaling pathway and plant hormone signal transduction pathway. Furthermore, qRT-PCR analysis of wheat varieties with differing auxin sensitivities confirmed a positive correlation between TaPBL7-2B expression and auxin response. In conclusion, TaPBL7-2B acts as a negative regulator of plant growth, affecting root development and leaf expansion in both Arabidopsis and wheat. These findings enhance our understanding of auxin signaling and provide new insights for optimizing crop architecture and productivity. Full article

Chloroplasts, as the organelles primarily responsible for photosynthesis, require a substantial supply of iron ions. Conversely, due to Fe toxicity, the homeostasis of these ions is subject to tight regulation. Permease in chloroplast 1 (PIC1) has been identified as the primary iron importer into chloroplasts. However, previous studies suggested the existence of a distinct pathway for Fe transfer to chloroplasts, likely involving mitoferrin-like 1 (MFL1) protein. In this work, Arabidopsis MFL1 (AtMFL1) and its cucumber homolog (CsMFL1) were characterized using, among others, Arabidopsis protoplasts as well as both yeast and Arabidopsis mutants. Localization of both proteins in chloroplasts has been shown to be mediated via an N-terminal transit peptide. At the gene level, MFL1 expression profiles differed between the model plant and the crop plant under varying Fe availability. The expression of other genes involved in chloroplast Fe homeostasis, including iron acquisition, trafficking, and storage, was affected to some extent in both AtMFL1 knockout and overexpressing plants. Moreover, root growth and photosynthetic parameters changed unfavorably in the mutant lines. The obtained results imply that AtMFL1 and CsMFL1, as putative chloroplast iron transporters, play a role in both iron management and the proper functioning of the plant. Full article

Thylakoid rhodanese-like protein (TROL) serves as a thylakoid membrane hinge linking photosynthetic electron transport chain (PETC) complexes to nicotinamide adenine dinucleotide phosphate (NADPH) synthesis. TROL is the docking site for the flavoenzyme ferredoxin-NADP⁺ oxidoreductase (FNR). Our prior work indicates that the TROL-FNR complex maintains redox equilibrium in chloroplasts and systemically in plant cells. Improvement in the knowledge of redox regulation mechanisms is critical for engineering stress-tolerant plants in times of elevated global drought intensity. To further test this hypothesis and confirm our previous results, we monitored light-independent ROS propagation in the leaves of Arabidopsis wild type (WT), TROL knock-out (KO), and TROL ΔRHO (RHO-domain deletion mutant) mutant plants in situ by using confocal laser scanning microscopy with specific fluorescent probes for the three different ROS: O₂^·−, H₂O₂, and ¹O₂. Plants were grown under the conditions of normal substrate moisture and under drought stress conditions. Under the drought stress conditions, the TROL KO line showed ≈32% less O₂^·− while the TROL ΔRHO line showed ≈49% less H₂O₂ in comparison with the WT. This research confirms the role of dynamical TROL-FNR complex formation in redox equilibrium maintenance by redirecting electrons in alternative sinks under stress and also points it out as promising target for stress-tolerant plant engineering. Full article

The phosphofructokinase (PFK) gene family plays a pivotal role in glycolysis and energy metabolism in plants. This study aimed to systematically characterize the PFK gene family in Arabidopsis thaliana at the genome-wide level and to investigate the function of AtPFK2 (ATP-dependent phosphofructokinase 2) in response to salt and drought stress. Through bioinformatics analysis, 11 AtPFK genes were identified. Phylogenetic analysis revealed that these PFK genes can be classified into two subfamilies: PFK and PFP. Notably, AtPFK2 possesses a unique structure, containing only a single intron, and its promoter is enriched with stress- and hormone-responsive elements, such as ABRE and MBS. T-DNA insertion mutants (pfk2) exhibited slightly shorter roots but slightly higher fresh weight under stress conditions, whereas Arabidopsis lines AtPFK2-overexpressing (OE-PFK2) showed increased stress sensitivity, with inhibited root and leaf growth, leaf wilting, reduced malondialdehyde and chlorophyll content, and enhanced accumulation of proline and soluble sugars. Weighted gene co-expression network analysis (WGCNA) identified 14 stress-related modules, from which six core genes—LBD41, TRP3, PP2-A3, SAUR10, IAA6, and JAZ1—were selected. These genes are involved in glycine metabolism and plant hormone signaling. The results of this study indicate that AtPFK2 mediates stress responses by regulating osmoregulatory substances and hormone signaling pathways, offering new insights into the mechanisms of stress resistance in crops. Full article

Branching is a critical agronomic trait in flowering Chinese cabbage (Brassica rapa subsp. chinensis), influencing plant architecture and yield. In this study, there was a highly significant difference between CX010 (single primary rosette branches) and BCT18 (multiple primary rosette branches). Phenotypic analysis revealed significant differences in primary rosette branch numbers, with BCT18 showing up to 15 branches and CX010 displaying only one main stem branch. Genetic analysis indicated that branching was controlled by quantitative trait loci (QTL) with a normal distribution of branch numbers. Using bulked segregant analysis coupled with sequencing (BSA-seq), we identified a candidate interval of approximately 2.96 Mb on chromosome A07 linked to branching. Fine mapping narrowed this to a 172 kb region containing 29 genes, with BraA07g032600.3C (BrTCP1) as the most likely candidate. cDNA cloning of the BrTCP1 gene revealed several variations in BCT18 compared to CX010, including a 6 bp insertion, 10 SNPs, and two single-nucleotide deletions. Expression analysis indicated that BrTCP1 was highly expressed in the rosette stems of CX010 compared to BCT18, consistent with its role as a branching suppressor. The heterologous mutants in Arabidopsis confirmed the conserved role of BrTCP1 in branch inhibition. These findings reveal that BrTCP1 might be a key regulator of branching in flowering Chinese cabbage, providing insights into the molecular mechanisms underlying this trait and offering a framework for genetic improvement in Brassica crops. Full article

High nitrogen use efficiency is crucial for enhancing spinach’s tolerance to low nitrogen stress and minimizing nitrate accumulation. Here, we report that SoNRT3, a NAR2 family protein, modulates nitrate uptake and plant growth under low-nitrate conditions. SoNRT3 expression was induced by low nitrate availability in roots and prolonged nitrogen deficiency in shoots. Compared to wild-type Arabidopsis thaliana, lines overexpressing SoNRT3 exhibited higher root fresh weight, activities of nitrogen reduction/assimilation-related enzymes, tap root length, and total root diameter under low-nitrate (0.25 mM) conditions. SoNRT3 silencing reduced taproot length, lateral root number, shoot/root biomass, and ¹⁵${\mathrm{N}\mathrm{O}}_3^{-}$ uptake in spinach grown under low-nitrate conditions. SoNRT3 partially compensated for ¹⁵${\mathrm{N}\mathrm{O}}_3^{-}$ uptake in atnrt2.1 and atnrt3.1 mutants. Transcriptome analysis showed that SoNRT3 may enhance nitrate uptake and root development by promoting the expression of high-affinity nitrate transporters, nitrogen assimilation, auxin signaling, and cell differentiation. Additionally, SoNRT3 can interact with a spinach NRT2 family protein (SoNRT2a), whose transcription level was also induced by low N and N deficiency. Together, this study clarifies the key roles and regulatory network of SoNRT3 in low-nitrate tolerance, which contributes to a novel understanding of nitrate utilization in spinach. Full article

The immune response triggered when plant cell surface receptors recognize pathogen-associated molecular patterns (PAMPs) is known as PAMP-triggered immunity (PTI). Several studies have demonstrated that extracellular plant ferredoxin-like protein (PFLP) can enhance PTI signaling, thereby conferring resistance to bacterial diseases in various plants. The C-terminal casein kinase II (CK2) phosphorylation region of PFLP is essential for strengthening PTI. However, whether phosphorylation at this site directly enhances PTI signaling and consequently increases plant disease resistance remains unclear. To investigate this, site-directed mutagenesis was used to generate PFLPT90A, a non-phosphorylatable mutant, and PFLPT90D, a phospho-mimetic mutant, for functional analysis. Based on the experimental results, none of the recombinant proteins were able to enhance the hypersensitive response induced by the HrpN protein or increase resistance to the soft rot pathogen Pectobacterium carotovorum subsp. carotovorum ECC17. These findings suggest that phosphorylation at the T90 residue might be essential for PFLP-mediated enhancement of plant immune responses, implying that this post-translational modification is likely required for its disease resistance function in planta. To further explore the relationship between PFLP phosphorylation and endogenous CK2, the Arabidopsis insertion mutant cka2 and the complemented line CKA2R were analyzed under treatment with flg22_Pst from Pseudomonas syringae pv. tomato. The effects of PFLP on the hypersensitive response, rapid oxidative burst, callose deposition, and susceptibility to soft rot confirmed that CK2 is required for these immune responses. Furthermore, expression analysis of PTI-related genes FRK1 and WRKY22/29 in the mitogen-activated protein kinase (MAPK) signaling pathway demonstrated that CK2 is necessary for PFLP to enhance flg22_Pst-induced immune signaling. Taken together, these findings suggest that PFLP enhances A. thaliana resistance to bacterial soft rot primarily by promoting the MAPK signaling pathway triggered by PAMP recognition, with CK2-mediated phosphorylation being essential for its function. Full article

Ascorbic acid (ASC) is a furan-based lactone derived from 2-ketogluconic acid that functions as a major antioxidant and redox buffer in mature plant tissues, although its content is lower in meristematic cells. ASC is commonly considered a reactive oxygen species (ROS) scavenger; however, its role in the regulation of plant development remains unclear. Additionally, the chemical behavior of ascorbate warrants special attention during ASC supplementation in in vitro plant culture. In this study, I investigated in detail the behavior of ascorbate in plant tissue culture medium and its uptake by plants. As a secondary objective, the role of ascorbate in root growth regulation was evaluated. The effects of low ASC levels on root architecture and its interaction with auxin signaling were studied using the vtc1 and vtc2 mutants of Arabidopsis thaliana, as well as through external ascorbate supplementation. Several marker lines for auxin response/distribution were used, along with direct ascorbate measurement via HPLC. Reducing ascorbate content through mutations had no significant effect on root development or auxin signaling, whereas high-concentration ASC supplementation inhibited both auxin signaling and root development, as demonstrated using auxin response and transport markers. At the organ level, ASC supplementation significantly downregulated auxin response-mediated cell cycle activation during lateral root induction. Based on the data presented, exogenous ascorbate may regulate root development through its interaction with auxin signaling pathways. Full article

The DNA mismatch repair (MMR) system plays a crucial role in repairing DNA damage and regulating cell cycle arrest induced by cadmium (Cd) stress. To elucidate the mechanism by which miRNAs target AtMSH2 in regulating Arabidopsis’ response to Cd stress, the wild-type Arabidopsis, Atmsh2 mutant, and three miRNA-overexpressing transgenic lines were grown hydroponically in half-strength MS solution containing cadmium (Cd) at concentrations of 0, 0.5, 1, 2, and 3 mg/L for 5 days. miRNA-seq analysis, bioinformatics prediction, dual-luciferase reporter assays, and qRT-PCR results demonstrated that miR5651, miR170-3p, and miR171a-3p specifically targeted AtMSH2 and their expression levels showed a significant negative correlation. Compared to wild-type (WT) Arabidopsis, Cd stress tolerance was significantly enhanced in miRNA-overexpressing transgenic lines. Moreover, exogenous application of these three miRNAs in half-strength MS liquid medium also markedly improved Cd stress tolerance in wild-type Arabidopsis. Furthermore, the expression of these three miRNAs expression was further upregulated by Cd stress in a dose-dependent manner. Additionally, DNA damage response in miRNA-overexpressing transgenic lines was promoted based on the expression of DNA repair, DNA damage signaling, and cell cycle genes, which differed from both wild-type and Atmsh2 plants. Taken together, miR5651, miR170-3p, and miR171a-3p participated in Cd stress response and improved plant Cd tolerance by mediating the expression of AtMSH2. Our study provides novel insights into the epigenetic mechanisms of Cd tolerance in plants, which sheds light on breeding for stress resilience in phytoremediation. Full article

Weeds present a pervasive challenge in agricultural fields. The integration of herbicide-resistant crops with chemical weed management offers an effective solution for sustainable weed control while reducing labor inputs, particularly in large-scale intensive farming systems. Consequently, the development of herbicide-resistant cultivars has emerged as an urgent priority. In this study, we found that the G311E mutant gene of Arabidopsis MATE (multidrug and toxic compound extrusion) family transporter DTX6, designated DTX6m, confers robust resistance to bipyridyl herbicides paraquat and diquat in rice. DTX6m-overexpression lines exhibited marked resistance to these two herbicides, tolerating diquat concentrations up to 5 g/L, which is five-fold higher than the recommended field application dosage. Agronomic assessments demonstrated that grain yields of DTX6m-overexpressing plants were statistically equivalent to those of wild-type plants. Moreover, the plants displayed beneficial phenotypic changes, such as accelerated flowering and a slight reduction in height. Seed morphometric analysis indicated that in comparison with the wild-type control, DTX6m-transgenic lines exhibited altered grain dimensions while maintaining consistent 1000-grain weight. Nutritional assays further demonstrated that DTX6m increased the levels of free amino acids in seeds, while normal protein and starch contents were retained. Collectively, these results establish that DTX6m effectively boosts rice resistance to paraquat and diquat, validating DTX6m as a candidate gene for engineering plant herbicide resistance and also implying a potential role for DTX6m in amino acid homeostasis in plants. Full article

The APETALA2/ethylene response factor (AP2/ERF) is a class of plant-specific transcription factors, among which the dehydration-responsive element-binding protein (DREB) subfamily has been widely reported to enhance plant resistance to abiotic stresses. A high-temperature-related gene, Apium graveolens DREBA6b (AgDREBA6b; accession number: OR727346), was previously cloned from a heat-tolerant celery variety. In this study, we transformed this gene into Arabidopsis thaliana using an Agrobacterium rhizogenes-mediated method to explore its function. The results showed that overexpressing AgDREBA6b in Arabidopsis thaliana significantly improved plant growth under high-temperature stress (38 °C) compared to the dreb mutant and wild-type (WT) plants. The anatomical structure of the leaves revealed that the number and degree of stomatal openings in the overexpressed plants were significantly higher than those in the WT and dreb plants, suggesting that AgDREBA6b enhances stomatal opening. Additionally, the chlorophyll content, chlorophyll fluorescence properties, proline (Pro), malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) activities were higher in the transgenic plants, indicating better stress tolerance. qPCR analysis showed that four heat tolerance-related genes (AtHSP98.7, AtHSP70-1, AtAPX1, and AtGOLS1) were upregulated in the transgenic plants, with higher expression levels than in WT and mutant plants. This study provides valuable genetic resources for understanding the molecular mechanisms of celery’s heat tolerance and offers insights for breeding heat-tolerant celery varieties. Full article

The AP2/ERF transcription factor ABSCISIC ACID INSENSITIVE 4 (ABI4) plays diverse roles in plant development and responses to abiotic stress. However, its potential involvement in regulating anthocyanin biosynthesis is not fully understood. In this study, three different loss-of-function abi4 alleles (abi4-1, abi4-2, and abi4-101) were employed to investigate the role of ABI4 in the regulation of anthocyanin accumulation in Arabidopsis seedlings. These abi4 mutants exhibited significantly increased anthocyanin accumulation, which was associated with elevated expression of genes involved in anthocyanin biosynthesis. HY5 (LONG HYPOCOTYL 5), a central component of photomorphogenesis, acts as a key light-regulated molecular switch. Further analysis revealed that ABI4 requires HY5 to function as a negative regulator of anthocyanin biosynthesis. Additionally, loss of ABI4 resulted in heightened light sensitivity, leading to increased light-induced chlorophyll accumulation and chloroplast development, along with upregulation of photosynthesis-related genes. Interestingly, the light-hypersensitive phenotype of abi4 mutants was partially rescued by the loss of HY5 function. Taken together, these findings demonstrate that ABI4 negatively regulates anthocyanin accumulation in Arabidopsis seedlings through a HY5-dependent light signaling pathway. Full article

LTR retrotransposons are widespread genomic elements that significantly impact genome structure and function. In Arabidopsis thaliana, the EVD LTR retrotransposon encodes a GAG protein essential for retrotransposon particle assembly. Here, we present a comprehensive analysis of the structural features, intracellular localization, and transcriptomic effects of the EVD GAG (evdGAG) protein. Using AlphaFold3, we identified canonical capsid (CA-NTD and CA-CTD) and nucleocapsid (NC) domains, with predicted disordered regions likely facilitating oligomerization. Transient expression of GFP-tagged evdGAG in protoplasts of A. thaliana and distant plant species (Nicotiana benthamiana and Helianthus annuus) revealed the formation of multiple large cytoplasmic aggregates resembling retrosomes, often localized near the nucleus. Stable overexpression of evdGAG in wild-type and ddm1 mutant backgrounds induced significant transcriptomic changes, including up-regulation of stress response and defense-related genes and downregulation of photosynthesis and chloroplast-associated pathways. Importantly, genes linked to stress granule formation were also up-regulated, suggesting a role for evdGAG in modulating cellular stress responses. Our findings provide novel insights into the cellular and molecular properties of plant retrotransposon GAG proteins and their influence on host gene expression. Full article

BOL/DRNL/ESR2, an AP2/ERF transcription factor, regulates early organ development in Arabidopsis (Arabidopsis thaliana). Its loss of function causes flower organ defects, while its overexpression induces green callus formation in roots without the addition of hormones. Jasmonates, plant hormones known as major players in stress responses, also regulate some aspects of organ development (e.g., stamen development and plant and organ growth). Here, we studied the interaction between BOL and the JA pathway. We found that exogenous application of methyl jasmonate (MeJA) partially rescued the stamen phenotypes in bol-cr mutants, linking BOL and JA-mediated stamen development. Moreover, MeJA treatments in wild-type plants partially mimicked some bol-D mutant phenotypes like reduced rosette and root size, while JA inhibition restored wild-type leaf curvature, suggesting an alteration in JA homeostasis in the gain-of-function mutant. BOL overexpression caused increased JA levels, whereas bol loss-of-function plants had reduced levels. Furthermore, inducible BOL activity led to downregulation of a JA-responsive marker. Finally, JA biosynthesis inhibition affected BOL-induced root callus formation and led to an expansion of the BOL expression domain in roots. Our findings indicate that BOL modulates parts of the JA pathway and that feedback from the JA pathway appears to affect expression of the transcription factor. Full article

Peanut (Arachis hypogaea L.), a vital oilseed and cash crop, faces yield limitations due to abiotic stresses. The 9-cis-epoxycarotenoid dioxygenase (NCED) enzyme, a key enzyme in abscisic acid (ABA) biosynthesis regulating plant development and stress responses, remains mechanistically uncharacterized in peanut abiotic stress tolerance. In this study, we isolated a novel gene, AhNCED4, from the salt-tolerant mutant M24. The expression of AhNCED4 was strongly induced by NaCl, PEG6000, and ABA in peanut huayu20. Overexpression of AhNCED4 enhanced salt and drought tolerance in Arabidopsis. Transgenic overexpression of AhNCED4 improved salt and stress resistance through upregulated ROS-scavenging genes superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) with elevated enzymatic activities while reducing malondialdehyde (MDA), superoxide anion (O²⁻), and hydrogen peroxide (H₂O₂) accumulation compared to wild-type plants. Further research showed that the chlorophyll fluorescence parameters of transgenic lines were significantly increased, while light damage was significantly reduced. These findings establish AhNCED4 as a critical regulator of stress adaptation and an excellent candidate gene for resistance breeding in peanut. Full article