Search Results (222)

Quinoa (Chenopodium quinoa), a stress-tolerant pseudocereal ideal for studying abiotic stress responses, was used to systematically identify optimal reference genes for qPCR normalization under gradient stresses: low temperatures (LT group: −2 °C to −10 °C), heat (HT group: 39° C to 45 °C), and drought (DR group: 7 to 13 days). Through multi-algorithm evaluation (GeNorm, NormFinder, BestKeeper, the ΔCt method, and RefFinder) of eleven candidates, condition-specific optimal genes were established as ACT16 (Actin), SAL92 (IT4 phosphatase-associated protein), SSU32 (Ssu72-like family protein), and TSB05 (Tryptophan synthase beta-subunit 2) for the LT group; ACT16 and NRP13 (Asparagine-rich protein) for the HT group; and ACT16, SKP27 (S-phase kinase), and NRP13 for the DR group, with ACT16, NRP13, WLIM96 (LIM domain-containing protein), SSU32, SKP27, SAL92, and UBC22 (ubiquitin-conjugating enzyme E2) demonstrating cross-stress stability (global group). DHDPS96 (dihydrodipicolinate synthase) and EF03 (translation elongation factor) showed minimal stability. Validation using stress-responsive markers—COR72 (LT), HSP44 (HT), COR413-PM (LT), and DREB12 (DR)—confirmed reliability; COR72 and COR413-PM exhibited oscillatory cold response patterns, HSP44 peaked at 43 °C before declining, and DREB12 showed progressive drought-induced upregulation. Crucially, normalization with unstable genes (DHDPS96 and EF03) distorted expression profiles. This work provides validated reference standards for quinoa transcriptomics under abiotic stresses. Full article

We summarize our present knowledge of the regulation of photostasis and photosynthetic performance versus photoprotection in response to vernalization and conclude that the enhanced photosynthetic performance of winter crops is due to an inherent increase in photosynthetic energy conversion efficiency induced by vernalization which translates into high seed yield in the field as well as under controlled environment conditions. This is consistent with the published data for enhanced photosynthetic performance of the only two extant terrestrial angiosperms, Colobanthus quitensis and Deschampsia antarctica, native to the frigid conditions of terrestrial Antarctica. The Cold Binding factor family of transcription factors (CBFs/DREBs) governs the enhanced photosynthetic performance of winter cereals as well as the Antarctic angiosperms. In contrast to winter crops, spring varieties survive cold environments by stimulating photoprotection at the expense of photosynthetic performance like that observed for green algae and cyanobacteria. Consequently, this minimizes the photosynthetic energy conversion efficiency of spring varieties and limits their seed yield upon cold acclimation. This review provides critical insights into the regulation of photostasis and the balance between photosynthetic performance and photoprotection in plants and how vernalization has enhanced photosynthetic energy conversion, which is essential for understanding plant adaptation to cold environments and optimizing agricultural productivity for improving crop resilience and yield in challenging climates. Full article

The type V CRISPR/Cas12f system, with its broad PAM recognition range, small size, and ease of delivery, has significantly contributed to the gene editing toolbox. In this study, enOsCas12f1 activity was detected during transient expression in rice protoplasts. The results showed that enOsCas12f1 exhibited DNA cleavage activity when it recognized TTN PAMs. Subsequently, we examined the gene editing efficiency of enOsCas12f1 in stably transformed rice plants, and the results showed that enOsCas12f1 could identify the TTT and TTC PAM sequences of the OsPDS gene, resulting in gene mutations and an albino phenotype. The editing efficiencies of TTT and TTC PAMs were 6.21% and 44.21%, respectively. Furthermore, all mutations were base deletions, ranging in size from 7 to 29 base pairs. Then, we used enOsCas12f1 to edit the promoter and 5′ UTR of the OsDREB1C gene, demonstrating that enOsCas12f1 could stably produce base deletion, mutant rice plants. Additionally, we fused the transcriptional activation domain TV with the dead enOsCas12f1 to enhance the expression of the target gene OsIPA1. Our study demonstrates that enOsCas12f1 can be utilized for rice gene modification, thereby expanding the toolbox for rice gene editing. Full article

Rice genome encodes ten OsDREB1 proteins that regulate tolerance to abiotic stresses such as cold and drought. OsDREB1s can bind to the C-repeat (CRT) element, dehydration response element (DRE), and GCC-box in gene promoters for transcription regulation. However, the recognition mechanism of OsDREB1s to these DNA elements remains unclear. Here, the structures of OsDREB1s were modelled using AlphaFold 3, which revealed a typical AP2 domain and a disordered KRP/RAGR motif adjacent to AP2 in all OsDREB1s. Structure modeling of OsDREB1A binding to CRT, DRE, and GCC-box showed that four Arg residues and a Glu (E66) from AP2 play important roles in binding to the major groove of DNA, while R40 in the KRP/RAGR motif was predicted to interact with the minor groove. The structure models revealed a few differences in the binding details for CRT, DRE, and GCC-box. Consistent with these predictions, OsDREB1A was evidenced to bind with the three DNA elements in slightly different affinities through EMSA experiments. Mutation analysis verified the key role of R40 and E66 in binding to CRT. Considering the highly conserved structure and sequence of the KRP/RAGR motif and AP2, we speculate that the DNA recognition mechanism found for OsDREB1A may be common for all OsDREB1s. Full article

Plant responses to heat stress include complex transcriptional networks and protein regulations in which BTB/POZ-MATH (BPM) proteins participate as a part of ubiquitin-mediated protein degradation. Arabidopsis thaliana contains six BPM genes involved in responses to environmental changes, including heat. Seedlings overexpressing BPM1 (oeBPM1), seedlings with downregulation of BPM1, 4, 5, and 6 (amiR-bpm) and wild type were exposed to 37 °C for 6 h. Treatment caused stronger decline of photosynthesis in oeBPM1 than in amiR-bpm and wild type, although all seedlings recovered after 24 h at 24 °C. The activity of the antioxidant enzymes catalase, guaiacol peroxidase, and ascorbate peroxidase remained unchanged in oeBPM1, but increased in amiR-bpm and wild type. Heat stress induced HSP70 and HSP90 in all seedlings but expression remained notably higher in amiR-bpm after recovery. DREB2A and HSFA3 expression increased in all seedlings immediately after stress, with the strongest induction in amiR-bpm. In amiR-bpm and wild type, BPM2 expression was induced immediately after exposure, while BPM1, BPM3, BPM4, and BPM6 were upregulated in wild type after recovery. In oeBPM1 seedlings, BPM4 expression decreased and BPM6 expression increased immediately after treatment at 37 °C for 6 h. The results suggest that BPM proteins modulate heat stress response by influencing photosynthesis, activation of antioxidant enzymes, accumulation of HSPs, and expression of heat-responsive genes, thus contributing to the different physiological strategies observed in A. thaliana lines with altered expression of BPM genes. 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

Drought stress limits plant survival and yield in arid regions. Uncovering the molecular mechanisms of drought tolerance is key to developing resilient crops. This study used Arabidopsis thaliana as a model to perform an in silico analysis of miRNA–mRNA interactions linked to post-transcriptional drought response. Using the MirTarget program, 274 miRNAs and 48,143 gene transcripts were analyzed to predict high-confidence miRNA–mRNA interactions based on binding free energies (−79 to −129 kJ/mole). Predicted binding sites were located in the CDS, 5′UTR, and 3′UTR regions of target mRNAs. Key regulatory interactions included ath-miR398a-c and ath-miR829-5p targeting ROS detoxification genes (CSD1, FSD1); ath-miR393a/b-5p and ath-miR167a-c-5p targeting hormonal signaling genes (TIR1, ARF6); and the miR169 family, ath-miR414, and ath-miR838 targeting drought-related transcription factors (NF-YA5, DREB1A, WRKY40). Notably, ath-miR414, ath-miR838, and the miR854 family showed broad regulatory potential, targeting thousands of genes. These findings suggest the presence of conserved regulatory modules with potential roles in abiotic stress tolerance. While no direct experimental validation was performed, the results from Arabidopsis thaliana provide a useful genomic framework for hypothesis generation and future functional studies in non-model plant species. This work provides a molecular foundation for improving drought and salt stress tolerance through bioinformatics-assisted breeding and genetic research. Full article

Background/Objectives: Drought stress is a significant environmental challenge that affects plant growth and productivity. Methods: In this study, an underutilized and better drought stress tolerance genotype of Cucumis melo var. melo, i.e., AHK-200, was investigated for drought tolerance potential, with special emphasis on various morphological, physiological, biochemical, and molecular parameters. Results: Our findings show that AHK-200 demonstrates superior drought tolerance with an enhanced root length, better water retention capacity, and stable cell membrane integrity under water deficit conditions. Physiologically, AHK-200 exhibited minimal reduction in relative water content (RWC) and photosynthetic efficiency (PN), along with increased stomatal conductance (gs) and chlorophyll content and reduced photoinhibition under drought stress. Biochemically, AHK-200 showed higher antioxidant enzyme activity (APX, CAT, SOD, GR, POD) and osmolyte accumulation (proline), which are critical for mitigating oxidative stress. At the molecular level, drought-related genes such as DREB2C, DREB2D, and RD22 were upregulated, supporting AHK-200 resilience to drought stress. Additionally, AHK-200 displayed elevated mineral concentrations, including Na, K, Ca, and Fe, which are essential for cellular homeostasis and stress adaptation. Conclusions: Overall, our study provides a comprehensive understanding of the drought tolerance mechanisms in AHK-200, highlighting its potential for use in breeding drought-tolerant genotypes in cucurbits and related crops. This research could guide future efforts in gene manipulation and transgenic development aimed at enhancing drought resistance and yield potential in crop plants. Furthermore, DREB2C, DREB2D, and RD22 transcription factors regulate many pathways related to stress; the overexpression of these genes may open a new avenue in melon improvement against drought stress. Full article

Drought stress is a devastating natural stress that threatens crop productivity and quality. Mitigating the adverse effects of drought stress on wheat is a key object in agriculture. C-repeat binding transcription factor/DROUGHT RESPONSE ELEMENT BINDING FACTOR 1 (CBF/DREB1) transcription factors are well known for their role in cold acclimation. However, the involvement of CBF genes in drought stress and the mechanisms underlying their function remain poorly understood. In this study, 81 CBFs were identified in wheat, which were further clustered into four distinct lineages based on phylogenetic analysis. Chromosomal localization indicated that most CBF genes were dispersed across chromosome 5. We identified three homoeologous genes (TaCBF14A, TaCBF14B, and TaCBF14D) that were simultaneously upregulated under drought stress based on RNA-seq analysis. According to the high expression after drought stress, TaCBF14B was selected for further functional analysis. Subcellular localization and transcriptional activation activity analysis indicated that TaCBF14B likely functions as a transcription factor involved in drought stress tolerance. Overexpression of TaCBF14B in Arabidopsis enhanced the primary root growth by 13.49% (OE1), 12.56% (OE2), and 19.53% (OE3) under 200 mM mannitol treatment, and 21.65% (OE1), 16.63% (OE2), and 28.13% (OE3) under 250 mM mannitol treatment compared to WT. Meanwhile, the water loss rate of transgenic lines was 56% in WT leaves, but only 44%, 50%, and 40% in OE1, OE2, and OE3 lines, respectively. Compared to the wild type, POD activities of OE1, OE2, and OE3 were significantly increased by 42.94%, 29.41%, and 62.52%, respectively. And the Pro activities in OE1, OE2, and OE3 were significantly increased by 16.33%, 5.18%, and 29.09%, respectively, compared to the wild type. Additionally, the MDA content in OE1, OE2, and OE3 was significantly reduced by 40.53%, 15.81%, and 54.36%, respectively. Further analysis showed that the transgenic lines were hypersensitive to abscisic acid (ABA), and exhibited increased expression of AtABI3. We speculate that TaCBF14B plays an important role in enhancing drought tolerance. In summary, our findings provide new insights into the functional roles of CBF genes in drought stress tolerance. Full article

The current study assessed the synergistic effects of PGPR and Si in alleviating drought stress in rice. Bacteria were isolated from Phragmites australis inhabiting an urban riverbank. Among the isolated strains, Pseudomonas qingdaonensis NZ 1 showed promising results under in vitro drought stress induced by PEG-6000 (−0.28 MPa). To further investigate the synergistic effect of Pseudomonas qingdaonensis NZ 1 and silicon, a plant growth experiment was conducted comprising the control (dH₂O) and plants treated with NZ 1, Si (1 mM), and NZ 1+Si under normal and drought stress conditions. The results revealed that NZ 1+Si-treated plants showed improved plant growth parameters, chlorophyll contents, relative water contents, antioxidant potential, and nutrient uptake under normal and drought conditions. Moreover, endogenous abscisic acid and jasmonic acid levels were substantially reduced, while the salicylic acid level was increased in NZ 1+Si-treated plants. Moreover, the relative expression of the ABA metabolic genes OsNCED3 and OsCYP707A6 and transcription factors OsbZIP23 and OsDREB1B were significantly altered. Furthermore, the leaf Si, calcium, potassium, and phosphorus contents were increased in NZ 1+Si-treated drought-stressed plants, along with the upregulation of OsLSi1. The combined application of NZ 1 and Si offers a sustainable agricultural strategy to effectively mitigate the detrimental effects of drought. Full article

Alfalfa (Medicago sativa) is a globally distributed economic legume crop used for forage and ecological restoration. We aimed to explore the mechanisms underlying the cold acclimation observed in this species. Our results for fall plant growth showed that non-dormant alfalfa (SD) maintained a vigorous growth rate compared to that of fall-dormant alfalfa (ZD); however, the winter survival rate of ZD was higher than that of SD. Among the ZD samples, the starch content first accumulated and then decreased; the sucrose content was consumed first along with simultaneous raffinose accumulation, which was followed by sucrose content accumulation, with consistent changes in the corresponding related synthase and hydrolase activity. SD exhibited the opposite trend. The transcriptome data showed that most of the differentially expressed genes were involved in carbon metabolism (ko01200), amino acid biosynthesis (ko01230), and starch and sucrose metabolism (ko00500). Our data clearly show that alfalfa’s cold acclimation mechanism is a complex process, with the establishment of stable carbon homeostasis; sucrose is first converted into starch and raffinose, and then, starch is converted into sucrose, which enables alfalfa’s cold resistance. The process is accompanied by CBF/DREB1A TF regulation. This study provides important insights into the cold acclimation mechanisms of alfalfa. Full article

Low temperature significantly contributes to the medicinal quality of Conyza blinii. The CBF/DREB1-dependent cold-responsive signaling pathway is a major contributor to plant cold stress resistance. However, whether the CBF/DREB1 signaling pathway affects the terpenoid metabolism of C. blinii under cold stress remains to be explored. Here, we systematically identified and analyzed the impact of CbCBFs on the terpenoid metabolism of C. blinii. The results showed that three CbCBFs and CbICE1 were identified based on the transcriptome. The functions significantly correlated with CbCBFs encompass plant hormones and stress responses. Co-expression analysis revealed that key genes in BR and JA signaling pathways were correlated with CbCBFs. Among them, CbCBF2 is the predominant factor under low-temperature conditions and is significantly positively correlated with oleanolic acid. Overexpression of CbCBF2 significantly upregulated the catalase gene CbβAS and increased oleanolic acid content in the leaves. These results indicate that CbCBF2 can act as a major regulatory factor to promote the synthesis of oleanolic acid by integrating JA and BR signaling under low temperature conditions. Full article

Loropetalum chinense is a significant small tree and ornamental shrub known for its colorful foliage and is widely used in landscaping in tropical and subtropical regions. This study aimed to establish an efficient, tissue culture-independent genetic transformation system for L. chinense. Cuttings from two varieties, ‘Xiangnong Xiangyun’ and ‘Hei Zhenzhu’, were infected with different strains of Agrobacterium rhizogenes. The results showed that the K599 strain significantly induced hairy roots in both varieties, with ‘Xiangnong Xiangyun’ demonstrating a higher survival rate (60%), rooting rate (51.66%), and hairy root induction efficiency (45%) compared to ‘Hei Zhenzhu’. Based on these findings, ‘Xiangnong Xiangyun’ and the K599 strain were selected for further optimization through an orthogonal L9 (3³) experiment, which focused on optimizing the infection solution composition, bacterial concentration, and infection duration, Finally, the genetic transformation system established at the beginning of the experiment was validated on ‘Xiangnong Xiangyun’ plants using the pre-screening LcDREB-43 gene of our group. Among these factors, infection duration was identified as the most influential for improving transformation efficiency. The optimal conditions were determined as an infection solution containing MES solution, a bacterial concentration of OD₆₀₀ = 0.8, and a 15 min infection duration. Under these optimized conditions, the survival rate, rooting rate, induction efficiency, and transformation efficiency reached 86.67%, 70%, 61.67%, and 43.33%, respectively. Furthermore, the transgenic plants with LcDREB-43 overexpression and pCAMBIA1305-GFP were obtained through the established transformation system, the authenticity of the system was proved, and the production application was carried out through phenotypic observation, molecular identification, and auxiliary verification of physiological indicators. Full article

High temperatures pose significant challenges to rice plants’ growth and their associated endophytic bacteria. Understanding how these bacteria respond to heat stress is vital. We assessed the potential of five endophytic bacterial strains derived from Oryza sativa—Bacillus tequilensis LB3, B. coagulans LB6, B. paralicheniformis AS9, B. pumilus LB16, and B. paranthracis i40C—to mitigate heat stress effects on rice plants. These strains demonstrated robust abilities in producing indole-3-acetic acid (IAA) and siderophores, nitrogen fixation, and solubilization of phosphate and potassium. Under high-temperature conditions, they significantly enhanced rice plant growth, with increases in plant length of up to 78% at 40 °C. Notably, LB6 showed the highest biomass increase (195%). The strains also improved chlorophyll SPAD values, an indicator of reduced heat stress effects and improved plant health. Phytohormone profiling and biochemical analyses revealed significant increases in abscisic acid (ABA) levels, reduced lipid peroxidation (MDA), and elevated osmoprotectant proline accumulation under heat stress. Inoculated plants exhibited up to 539 ng g⁻¹ of ABA (vs. 62 ng g⁻¹ in uninoculated controls), a 68% reduction in MDA (indicating less oxidative damage), and enhanced proline synthesis, collectively suggesting improved stress adaptation. These changes were linked to bacterial IAA production and nutrient modulation, which alleviated heat-induced physiological decline. These findings underscore the potential of these endophytes as biofertilizers to improve rice resilience under heat stress. Among the strains, LB6 exhibited superior performance, offering the greatest promise for heat-stress mitigation in rice production. This study advances our understanding of phytohormonal, heat stress signaling, and chemical processes underlying bacterial-mediated thermotolerance, providing a foundation for sustainable agricultural strategies. Future research can explore morphological and biochemical analyses, stress-responsive gene expression (e.g., HSPs, DREBs, and APX) linked to thermotolerance, and the combined effects of selected strains with fertilizers in high-temperature rice cultivation. Full article

Dehydration-responsive element-binding (DREB) transcription factors are ubiquitous in plants and regulate plant growth, development, signal transduction, and responses to stress, particularly drought stress. However, DREB genes in flax have not previously been studied. This study conducted a comprehensive and systematic analysis of the DREB gene family in flax (Linum usitatissimum L.). A total of 59 LuDREB genes were identified in Longya-10 (a breeding variety), with an uneven distribution across all 15 chromosomes. Further analysis revealed significant variations among LuDREB members, with predictions indicating that these proteins are hydrophilic and localized in the nucleus and cytoplasm. A phylogenetic analysis classified the LuDREB genes into six subgroups, a classification further supported by gene structure and motif composition. Members within the same subgroup exhibited structural conservation, suggesting functional redundancy. The duplication analysis identified 30 pairs of segmentally duplicated LuDREB genes and one pair of tandemly duplicated genes, indicating that segmental duplication was the primary driver of LuDREB gene expansion. A comparative collinearity analysis revealed that most LuDREB genes had orthologs in other plant species, suggesting that this gene family has remained relatively conserved throughout evolution. Cis-acting element analysis identified numerous hormone- and stress-responsive elements in LuDREB promoters, and the quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) results confirmed the role of all LuDREB genes in drought stress response. In addition, transcriptome analysis revealed that LuDREB49 and LuDREB56 exhibited high expression levels in the capsules, whereas LuDREB3 and LuDREB36 showed significantly higher expression levels in the stems, suggesting that these LuDREB genes may have specialized functions in capsule or stem development. Collectively, this study provides a comprehensive overview of LuDREB genes, offering valuable insights into their roles in flax growth, development, and stress responses. Full article