Liquid-liquid phase separation (LLPS) has emerged as a fundamental mechanism organizing membrane-less compartments within cells, driving crucial biological processes. In plants, LLPS facilitates the spatiotemporal regulation of diverse functions, from growth and development, such as hormone signaling, photo-perception, and floral transition, to adaptive responses against abiotic and biotic stresses. This review outlines the properties and major driving forces governing LLPS, emphasizing multivalent interactions mediated by intrinsically disordered regions, repeated domains, and nucleic acids. Key influencing factors, including concentration, temperature, and ionic conditions, are discussed. We further describe a streamlined experimental workflow for studying plant LLPS, encompassing prediction, assessment, and validation. Understanding LLPS dynamics offers profound insights into plant adaptation and resilience, positioning phase separation as a pivotal regulatory paradigm in plant biology.
Argonautes (AGOs) are the effectors for the action of microRNAs (miRNAs). Plant genomes harbor large numbers of AGO genes whose functions remain to be fully understood. Here, we elucidated a function of AGO5 in the ecological model plant Nicotiana attenuata during its interactions with the specialist herbivore Manduca sexta. Plants silenced in NaAGO5 expression using inverted-repeat technology (irAGO5) were indistinguishable from the wild type (WT) in growth and development but were highly susceptible to M. sexta herbivory. M. sexta caterpillars grew faster and accumulated significantly more biomass on irAGO5 than on WT plants. Herbivory-elicited irAGO5 plants accumulated significantly lower amounts of auxin-dependent defense metabolites such as phenolamides, flavonoids, and diterpenoid glycosides, but not nicotine and trypsin protease inhibitors (TPI). Nicotine and TPI levels, which require intact jasmonate signaling, were attenuated in plants silenced in NaAGO8 expression (irAGO8). irAGO5 plants showed compromised herbivore-induced auxin levels and YUCCA gene expression but accumulated more salicylic acid; however, jasmonate accumulations were at WT levels. Exogenous auxin treatments restored resistance against M. sexta and auxin-dependent defense metabolites. Substantial temporal changes in the miRNome were observed in irAGO5 and were largely different from those in irAGO8. An AGO5-dependent miRNA-mRNA regulatory interaction network was inferred for defense-signaling components. Furthermore, double knockdowns of NaAGO5 and NaAGO8 revealed cooperative functions of the two genes during herbivory. We infer that AGO5 is a central component of the herbivore-induced smRNA pathway that modulates multiple nodes in the auxin-dependent metabolic space of the defense signaling network when N. attenuata plants interact with the specialist herbivore M. sexta.
Aphid-microbe-plant interactions are fundamental to understanding plant responses to combined biotic and abiotic stress. The grain aphid Sitobion avenae is a major pest of wheat, particularly under drought conditions. Although arbuscular mycorrhizal fungi (AMF) can enhance plant tolerance to water deficit, their effects on aphid performance across wheat cultivars differing in drought resistance remain unclear. We examined the influence of Acaulospora delicata on S. avenae performance on two wheat cultivars-Yunhan-618 (drought-resistant) and Xinong-1376 (drought-susceptible)-under well-watered and water-deficit stress conditions. Under water-deficit stress conditions, root colonization by A. delicata was higher in both Yunhan-618 and Xinong-1376 when compared to well-watered conditions. In the absence of mycorrhiza, nymphal developmental time was prolonged, especially on drought-stressed Xinong-1376 plants. AMF inoculation shortened developmental time, increased adult longevity, and enhanced fecundity of S. avenae under both water regimes. On Yunhan-618, AMF association increased intrinsic growth rate and reproductive output of this aphid. Honeydew excretion by S. avenae was greater on AMF-inoculated plants under well-watered conditions. Aphid body mass and water balance traits were generally higher on AMF-associated Yunhan-618 plants under adequate water supply. Aphids also preferentially settled on AMF-inoculated drought-susceptible wheat plants under both water regimes as compared to drought-resistant wheat plants. Overall, A. delicata enhanced plant drought resilience but simultaneously promoted aphid fitness. These findings underscore the complex and context-dependent role of AMF in shaping plant-aphid interactions, with important implications for pest dynamics under climate change.
Heterologous over expression of blueberry VcBR6OX1 triggers seedless/low-seed tomatoes via elevatingendogenous GA4, reshaping multi-hormone homeostasis and modulating hormone signal transductionpathways to regulate seed development as confirmed by transcriptome analysis. If highlight marking is needed, it is shown as follows: （1） Heterologous overexpression of the blueberry VcBR6OX1 gene can induce seedless or low-seedtomato fruits; （2） The endogenous gibberellin GA4 in transgenic plants is significantly increased, and the dynamichomeostasis of the multiple hormone network is reconfigured （3） Transcriptome analysis reveals that BR6OX1 affects seed development by regulating the hormonesignal transduction pathway. Blueberries (Vaccinium spp.) have attracted widespread attention due to their unique flavor and high antioxidant activity. However, the high seed content of some blueberry cultivars affects their texture and processing costs. The development of seedless or low-seed cultivars is essential for improving fruit quality and economic benefits. This study used bioinformatics, genetic transformation, and transcriptomic analysis to investigate the role and regulatory mechanisms of the blueberry Brassinosteroid-6-oxidase 1 (BR6OX1) gene in the formation of seedless fruits. The results revealed that the BR6OX1 gene in blueberries encodes an unstable hydrophilic protein with a conserved cytochrome P450 superfamily domain and is highly evolutionarily similar to that in closely related species. The introduction of the BR6OX1 gene into tomatoes revealed that its overexpression significantly affects the growth and development of tomato plants, resulting in seedless or low-seeded fruits. Endogenous hormone analysis revealed that the gibberellic acid (GA4) content significantly increased in the transgenic tomato plants, whereas the auxin (IAA) and abscisic acid (ABA) contents exhibited differential changes at different developmental stages. Further transcriptome analysis of the transgenic plants revealed that BR6OX1 overexpression significantly affects plant hormone signaling pathways, particularly by causing significant changes in the expression of genes related to hormone synthesis and signaling, such as those encoding auxin, gibberellin, abscisic acid, and ethylene. These results suggest that the BR6OX1 gene may influence fruit set by regulating the balance of plant hormones. This study provides an important theoretical basis for developing seedless blueberries and lays the foundation for the application of this gene in the improvement of fruit quality.
Carbon starvation is one of the proposed mechanisms of drought-induced plant mortality. However, it has not been implicated in drought mortality as much as hydraulic failure. We tested the role of carbon on responses to drought by limiting stem photosynthesis and increasing tissue non-structural carbohydrates (NSC) in saplings of the tropical tree Calophyllum longifolium Willd. (Calophyllaceae). We first artificially increased [NSC] by exposing half of the saplings to 2000 µmol mol-1 of CO2, while the other half remained at ambient [CO2] for six weeks. Following CO2 treatments, there were no significant differences in predawn leaf water potential (Ψpd), leaf photosynthetic rate, and stem re-assimilation rate between elevated and ambient [CO2] treatments, whereas stem re-assimilation percentage (percentage of dark respiration rate that is re-assimilated) was greater in ambient [CO2]. Exposure to elevated [CO2] produced higher starch and total [NSC] in the tissues of those plants. We then covered the stems of half of the plants from each CO2 treatment to block stem photosynthesis and applied a drought treatment to all plants. Light exclusion reduced stem photosynthesis and most traits responded similarly to drought across the four treatment levels. Drought decreased soluble sugar concentration and increased starch concentration with minimal effects of prior [CO2] treatment. Despite initial differences in starch and total [NSC] between the two [CO2] treatment levels, and physiological responses to light exclusion, leaf and plant mortality occurred at the same pace. Our results demonstrate that stem photosynthesis does not contribute to drought survival in saplings of C. longifolium.
Salt stress is an abiotic stressor that adversely affects the growth and productivity of caraway, an important aromatic plant. A randomized complete split-plot design was used to determine the nanoparticles of zinc oxide (ZnO NPs) effects at levels of 0, 0.2, and 0.4 g L-1 on fruit yield, chemical and biochemical composition, and essential oil (EO) productivity of caraway plants subjected to saline irrigation water containing 0, 1, 2, 3, and 4 g L-1 sodium chloride (NaCl). The results indicated that increasing NaCl concentrations significantly reduced yield traits, relative water content, salinity tolerance index, leaf pigment concentrations, N, P, K, and Zn concentrations, fruits' total proteins and total carbohydrates and percentage and yield/plant of essential oil (EO) in comparison to the untreated plants. Conversely, proline content, Na% and Cl%, and the activities of catalase, peroxidase, superoxide dismutase, and polyphenol oxidase increased with increasing NaCl concentrations relative to the control. Foliar application of ZnO NPs significantly increased the parameters relative to the untreated control except proline content, Na%, and Cl%, which were in comparison to respective control significantly reduced. Moreover, utilization of ZnO NPs positively affected the traits mentioned above under all NaCl concentrations tested compared to treatments without ZnO NPs. Different combinations of NaCl and ZnO NPs concentrations had varying effects on EO composition. A total of 32 compounds were identified across all treatment combinations, with the highest number (14) observed in the control. The major EO compounds were carvone (up to 49.71%) in the 2 g L-1 NaCl + 0.2 g L-1 ZnO NPs treatment, limonene (up to 28.68%) in the 2 g L-1 NaCl + 0.4 g L-1 ZnO NPs treatment, and D-limonene (up to 25.95%) in the 4 g L-1 NaCl + 0 g L-1 ZnO NPs treatment. These results suggest that the application of ZnO NPs may provide a sustainable approach to caraway cultivation under saline conditions.
Cisgenic introduction of Rvi15 or FB_MR5 into 'Gala' generated single-copy lines with ≤ 201 bp vector remnants and high disease resistance, supporting cisgenesis as a powerful tool for commercial apple improvement. Cisgenesis can be used to transfer disease resistance genes from wild apple germplasm into established apple cultivars, reducing their susceptibility to diseases. This approach allows combining disease resistance and fruit quality traits without compromising the desirable characteristics of the original cultivar. In our study, we used the binary plasmid vector pMF1 to create three apple scab-resistant cisgenic 'Gala' lines with the apple scab resistance gene Rvi15, and three fire blight-resistant cisgenic 'Gala' lines with the fire blight resistance gene FB_MR5. Quantitative PCR revealed that five of six cisgenic lines had a single integrated T-DNA copy, and targeted locus amplification enabled the identification and characterization of insertion sites in five lines. All characterized lines showed T-DNA trimming at the borders, with a total length of non-endogenous sequences at their respective insertion sites ranging from 135 to 201 base pairs. The presence of such residual vector sequences can influence the regulatory status of cisgenic plants, as some countries may classify these lines as genetically modified plants. All cisgenic lines exhibited increased disease resistance compared to the untransformed control genotype 'Gala'. However, the achieved resistances are based on single genes, and it will be necessary to develop combinations of different resistance genes for more durable resistance. Resistance, durability, and evolving regulatory frameworks are crucial factors that together will determine the success of the cisgenic approach in achieving both high fruit quality and resistance for sustainable apple production.
Plants interact with a vast variety of microbes that inhabit both above- and belowground tissues. Through their effect on host physiology and growth, plant-microbe interactions define the success of a plant's life cycle. A key aspect of these interactions is the requirement for highly cell-type-specific responses from the plant, be it to form symbiotic structures in certain cells or to mount a highly localised immune response. There has been long-standing interest in uncovering the cell-specific transcriptomic changes that underpin these processes to better understand the establishment, functioning, and regulation of plant-microbe interactions. The recent optimisation of single-cell and spatial transcriptomics for plants now allows us to investigate these interactions in unprecedented detail. Here, we discuss how single-cell technologies can help unravel the many mysteries of plant-microbe interactions. We focus on the key lessons we have learned from recent single-cell studies in the field and highlight the current limitations of single-cell technologies. We also offer promising avenues for future exploration and conclude by suggesting experimental and bioinformatic considerations to maximise insights from past and future studies and help make the most of this new single-cell era in the field of plant-microbe interactions.
Cadmium (Cd) contamination of agricultural soils disrupts plant signaling networks, impairing nutrient communication, photosynthetic efficiency, and stress responses. Microbial inoculants offer eco-biotechnological solutions by modulating signal perception and transduction under heavy metal stress. This field study investigated the role of Acinetobacter schindleri strain SR-5-1 in influencing pea (Pisum sativum L.) signaling pathways under Cd toxicity. Plants exposed to environmentally relevant Cd concentrations (250 and 500 µM) exhibited disrupted chlorophyll biosynthesis, elevated oxidative stress markers, and impaired nutrient signaling. Inoculation with SR-5-1 restored chlorophyll levels, enhanced ROS-scavenging enzyme activities, and reduced lipid peroxidation, indicating microbial effects on oxidative signaling cascades. Importantly, the inoculated plants accumulated less Cd in the roots and leaves, reflecting microbial mediation of ion transporter activity and rhizosphere detoxification. SR-5-1 also improved nitrogen, iron, zinc, potassium, and magnesium acquisition, highlighting its role in nutrient uptake and homeostasis. These findings demonstrate that SR-5-1 functions as a bio-communicator, alleviating xenobiotic stress by modulating ROS and nutrient signaling pathways. The study underscores the ecological relevance of microbial inoculants in supporting integrative plant communication and resilience, positioning SR-5-1 as a promising bioresource for signaling-driven sustainable agriculture in Cd-affected soils.
Ubiquitination regulates pattern-triggered immune responses in plants, but the underlying mechanisms remain incompletely understood. In Arabidopsis, the E3 ubiquitin ligase PUB4 has been implicated in growth, development, and stress responses, and is known to interact with and be phosphorylated by CERK1, resulting in the positive regulation of chitin-triggered immunity. In this study, we identified ubiquitinated proteins enriched from chitin-treated microsomal fractions and discovered the receptor kinase FERONIA as a potential target of PUB4. FERONIA interacted with PUB4 both in vivo and in vitro, as evidenced by yeast two-hybrid, BiFC, and co-immunoprecipitation assays. An in vitro ubiquitination assay demonstrated that PUB4 ubiquitinates the cytoplasmic domain of FERONIA. Moreover, transient expression assays in wild-type and pub4-2 mutant protoplasts revealed that PUB4 mediates chitin-induced FERONIA degradation. Unlike PUB4, which positively regulates chitin-induced defense responses, FERONIA negatively regulates them. For example, fer4 mutants exhibited elevated basal and chitin-induced ROS and callose levels. This suggests that FERONIA suppresses immune signaling under steady-state conditions, and that PUB4-mediated degradation of FERONIA relieves this inhibition. These findings clearly reveal a novel mechanism by which PUB4 regulates chitin-induced immunity driven by CERK1 through targeted degradation of FERONIA. This highlights the balance between positive and negative regulators of plant immune signaling.
In higher plants, ferredoxin (Fd) is present as distinct isoproteins of photosynthetic type (LFd) and non-photosynthetic type (RFd), which exhibit differential function despite their similarity in the 3D structures. We previously showed that pH-dependency of electron transfer activity with ferredoxin-NADP+ reductase (FNR) was opposite between LFd and RFd, which was explained by the opposite pH-dependent profile of Km for the two Fds, and that the differences of C-terminal residues and 78th residue between the two Fds were partly responsible for the different pH dependency. In this study, we further investigated the determinants responsible for the different pH dependency between the LFd and RFd. Site-directed mutants of Fd, substituted at the residues on the interface with FNR, were prepared. Kinetic analyses using the single site-directed mutants and their combined, multiple mutants showed that combination of the substitutions of Fd residues at 61, 63, 78th and C-terminal region located at the interface with FNR conferred opposite pH-dependency, which indicated that these residues are the enough determinant for their opposite pH dependency of LFd and RFd in the electron transfer reaction with FNR.
Microtubules (MTs) are crucial for cell division, growth, development and morphogenesis in plants. Cotton fibres are single-celled trichomes that originate from the epidermal cells of the ovule, making them an excellent model for studying plant cell differentiation and rapid elongation. However, the roles of MTs in cotton fibre development remain incompletely understood. In this study, we identified GhTTLL12, a tubulin-tyrosine ligase-like protein 12, as a positive regulator of fibre initiation and elongation via Gh-Gb introgression analysis. GhTTLL12 was preferentially expressed during the rapid elongation stage of cotton fibres. Overexpression of GhTTLL12 enhanced plant height, root length, fibre cell protrusion number and fibre length in cotton. Conversely, CRISPR/Cas9-mediated knockout of GhTTLL12 led to opposite phenotypes, thereby significantly reducing fibre quality. MT co-sedimentation and immunofluorescence assays demonstrated that GhTTLL12 binds directly to MTs and promotes their assembly while facilitating the formation of transverse MT arrays in elongating fibres. Further investigation of the molecular mechanisms revealed that after GhTTLL12 is recruited into the nucleus by GhTUB8, it promotes mitosis in ovule epidermal cells upon activation by GhMML3, increasing the number of fibre cell protrusions. GhMYB86, a negative regulator of cotton fibre elongation, represses GhTTLL12 transcription in the nucleus, leading to attenuated mitotic activity. Cytoplasmic GhTTLL12 modulates fibre cell elongation by regulating MT assembly and ordered arrangement. Collectively, our findings define a GhMML3/GhMYB86-GhTTLL12-GhTUB8 regulatory module that links stage-specific transcriptional regulation to MT remodelling during cotton fibre development, providing new insights into the improvement of fibre quality and yield.
Bacterial wilt caused by Ralstonia pseudosolanacearum poses a significant threat to turmeric (Curcuma longa L.) production in many growing regions. Although plant-associated microbial communities may contribute to disease suppression, the ecological roles of rhizosphere and endosphere microbiomes in turmeric are still underexplored. Here, we characterized rhizosphere and endosphere bacterial communities using 16S rRNA gene amplicon sequencing and evaluated their antagonistic activity against bacterial wilt pathogen of turmeric R. pseudosolanacearum strain RalsTur1. Microbiome profiling revealed compartment-specific patterns in turmeric-associated bacterial communities, with rhizosphere communities strongly structured by geographic location and endosphere communities comparatively stable across field sites. The endosphere core microbiome was dominated by bacterial members in the family Enterobacteriaceae, particularly Enterobacter, along with Pseudomonas, whereas rhizosphere communities included diverse taxa such as Bacillus and members of the Allorhizobium-Neorhizobium-Pararhizobium-Rhizobium group. Interbacterial competition assays showed that several turmeric-associated isolates reduced RalsTur1 populations in vitro. However, in planta assays using tissue-cultured turmeric plants revealed that only the endosphere-derived bacterial community, including Chryseobacterium gleum (ED4), Pseudomonas laurentiana (ED4-21), and Pantoea sp. (WEH1), significantly reduced pathogen populations, resulting in a two-log reduction in pathogen abundance. These findings suggest that colonization within plant tissues may contribute to suppression of vascular pathogens and highlight endophytic bacteria as candidates for further investigation in microbiome-based management of bacterial wilt.
Plants restrict microbial entry into their leaves by closing their stomata upon recognition of conserved microbe-associated molecular patterns (MAMPs). The hydathode pore is a stomata-like opening on leaf margins which are thought to lack MAMP recognition and to be an entry point for microbial pathogens. Here, we observe marginal hydathode pore closure in response to abscisic acid, and the MAMPs chitin and flg22 in Arabidopsis thaliana leaves. Hydathode pore closure occurs within 3-9 hours of chitin exposure and pores reopen after 12 hours. Under conditions when hydathode pores are open, external fluids enter approximately 80% of the marginal hydathodes within a leaf. After hydathode pores close in response to ABA, chitin or flg22, external fluids accumulate in under 20% of hydathodes with in a leaf. MAMP-induced hydathode restriction was similar for dye or fluorescent Pseudomonas syringae pv. tomato bacteria and was dependent on pattern recognition receptors including CERK1 for chitin and FLS2 for flg22. Chitin-induced hydathode limitation was also dependent on the NADPH oxidase RBOHF and was partially hampered in rbohD, lyk4 lyk5, bak1-5 bkk1, slac1-3 slah3-1 knockout mutants. Together, this work indicates that MAMP recognition regulates entry into the hydathode and induces transient closure of the hydathode pores.
Biomass-derived carbon quantum dots (BCDs) have attracted considerable research attention as a novel category of sustainable fluorescent nanomaterials, attributed to their adjustable photoluminescence, superior biocompatibility, and eco-friendly synthesis methods. BCDs are made from renewable biomass sources like plants, algae, animal byproducts, and microorganisms, following the principles of green chemistry. This makes them much better for the environment and cheaper to make than carbon dots made in the traditional way. The field still has three big problems, though: the luminescence mechanism is still not well understood, with different pathways like carbon-core and surface-state emissions not having a single theoretical framework; optical modulation strategies are still not well developed, with quantum yields often falling below 30% and poor batch-to-batch consistency making it hard to standardize applications; and not enough is known about in vivo metabolic pathways and long-term toxicity to allow for systematic toxicological evaluation and clinical translation. The complex luminescence mechanisms, including carbon-core-state, surface-state, molecular-state, and cross-linked enhanced emission (CEE), are thoroughly examined to clarify the structure-property relationships that dictate their optical behavior. By using controlled synthesis and surface modification techniques, BCDs can be made to emit light in the visible to near-infrared (NIR) range. This makes them perfect for use in different types of multimodal imaging, such as single-photon, multi-photon, and photoacoustic bioimaging. Their natural ability to emit light, along with their low toxicity to cells and high stability in light, makes it possible to see cells and tissues in high detail. We talk more about the problems we are having right now with standardizing synthesis and controlling optical properties with precision. Future research should concentrate on refining reaction conditions and clarifying luminescence mechanisms to promote the clinical application of BCDs as next-generation, sustainable imaging agents.
Thallium (Tl) is a toxic metal and priority pollutant, its soluble Tl levels in soil drive Tl accumulation in edible plants, posing health risks to gut microbiota via dietary exposure even at low doses. This study investigated Tl accumulation in sweet potatoes (4.45-32.87 µg/kg dry weight, 0.2442-1.7368 µg/kg wet weight) from soils (282.89-699.50 µg/kg) and its impact on a single pooled microbial community derived from fecal samples of three healthy adults (2 females, 1 male, 20-30 years) using an in vitro digestion-colon fermentation model. Low-dose Tl exposure drove significant, dose- and time-dependent genus-level restructuring of the pooled microbial community (Kruskal-Wallis, P = 0.001; PERMANOVA, P = 0.001, R2 = 0.783-0.980), without altering phylum-level alpha diversity (Kruskal-Wallis, P > 0.05), indicating compositional shifts rather than richness loss. Genus-level shifts included proliferation of harmful taxa (Escherichia_Shigella, Enterococcus) and reduction of beneficial taxa (Bacteroides, Prevotella, Akkermansia, Bifidobacterium, Blautia). Significant correlations (p < 0.05, 0.6883 < R2 < 0.9850) linked Tl content to Bacteroides, Prevotella, Escherichia_Shigella, and Enterococcus abundances. These findings demonstrate exposure-relevant microbiome shifts within this single pooled microbial community even at Tl concentrations below regulatory limits (300 µg/kg) via food chain transfer. However, as this in vitro model lacks host-microbe interactions (e.g., immune signaling, peristalsis) and the results reflect the response of one mixed inoculum from three donors rather than inter-individual variability, chronic in vivo studies are essential to validate these shifts and their metabolic and immune implications, informing soil-plant-human safety and public health strategies for low-dose dietary Tl exposure.
Climate change is shifting agriculture toward multifactorial abiotic stresses (drought, heat, and salinity). This study aims to characterize emergent, non-additive plant responses to combined stresses and to define the epigenetic and microbial frameworks that govern environmental memory and adaptive plasticity. We conducted a meta-synthesis of molecular and ecological studies, evaluating high-throughput data on DNA methylation, histone modifications, and ncRNA profiles. We further analyzed the plant holobiont to determine how rhizosphere and endosphere microbiota influence host stress imprinting. The analysis revealed that stress combinations trigger distinct transcriptomic and metabolic signatures, which are stabilized by an "epigenetic toolkit" such as RNA-directed DNA methylation and chromatin remodeling. Furthermore, plant-associated microbiota serve as an extrinsic regulatory layer, modulating host epigenetic states to prime plants for compound stress. While translational pathways such as epigenetic editing, CRISPR-mediated epigenome editing, and microbiome engineering show promise, their field-scale stability remains context-dependent. Building climate resilience requires a paradigm shift from traditional single-trait breeding toward multi-scale regulatory approaches. Harnessing the synergy between the plant epigenome and the microbiome enables the development of 'primed' crop varieties-an integrated strategy vital for safeguarding global food security amid intensifying environmental volatility.
Iron (Fe) and zinc (Zn) deficiencies severely threaten global human health. Thus, rice biofortification to enhance intrinsic Fe and Zn levels in grains represents an effective strategy to alleviate human Fe and Zn deficiencies. Several biofortification strategies have successfully increased Fe and Zn concentrations in the rice endosperm, with multigene approaches demonstrating synergistic micronutrient accumulation. To enhance Fe and Zn accumulation in rice endosperm, we designed a transformation construct (NYFN) in which OsNRAMP7 and OsNAS2 were driven by the 35S promoter, OsYSL2 by the OsSUT1 promoter, and OsFER2 by the endosperm-specific OsGluA2 promoter. Multi-year field trials were conducted at two locations to evaluate Fe and Zn accumulation in transgenic NYFN rice plants derived from Nipponbare (NB) and commercial cultivar Huaidao 5 (HD5) genetic backgrounds. The results showed that transgenic NB lines exhibited 10.93-14.72 μg/g DW Fe and 33.01-48.33 μg/g DW Zn in polished grains, representing 4.9- to 6.3-fold increases in Fe and 2- to 2.7-fold increases in Zn compared to the NB control. Polished grains of HD5 transformants contained 10.24-13.35 μg/g DW Fe and 32.17-50.33 μg/g DW Zn, corresponding to 4- to 7.4-fold elevations in Fe and 2- to 2.3-fold elevations in Zn relative to the HD5 control. X-ray fluorescence spectroscopy (µ-XRF) and Perls' Prussian blue staining analyses confirmed the significantly enhanced Fe and Zn accumulation in transgenic grains. Transgenic NB lines exhibited significant changes in certain agronomic traits, including reduced 1000-grain weight, grain size, and grain filling rate, whereas transgenic HD5 lines showed no significant agronomic differences relative to the wild type. Total grain weight per plant remained unchanged in both the transgenic NB and HD5 lines compared to the wild type. The results demonstrate that the NYFN strategy enables sustainable Fe and Zn biofortification, representing a promising solution to the global challenge of human Fe and Zn deficiency.
Understanding crop root growth and distribution and soil water depletion under water deficit conditions is critical for developing sustainable production practices for cucumber (Cucumis sativus L.) in semi-arid regions like West Texas. Therefore, this study's main objective was to investigate the effect of deficit irrigation (DI) and biochar application on soil water depletion, root growth and distribution, and water productivity (WP) of cucumber. A two-year field study was conducted at Quaker Research Farm, Texas Tech University, Lubbock, TX. A split-plot design was used to randomize four irrigation levels I1 [100% crop evapotranspiration (ETc) throughout the growing season], I2 [80% ETc during early growth (crop establishment to mid-season), 60% ETc during late growth (mid-season to maturity)], I3 (60% ETc during early growth, 80% ETc during late growth), I4 (40% ETc throughout the growing season) and three biochar rates (0, 15, and 20 t/ha) in main-plots and sub-plots, respectively. Results showed a decrease in root length density (RLD) and root surface area density (RSAD) under DI treatments compared to control (I1). RLD was reduced by 24, 4, and 29%, whereas RSAD decreased by 24, 31, and 44% in I2, I3, and I4, respectively, compared to I1. There was greater soil water depletion under the severe DI treatment (I4) without enhanced root plasticity, with some occasional water storage in mild-DI treatments (I2 and I3). The crop water use decreased significantly by 17% in I2, with the least yield penalty of 14% compared to I1. The I2 was the most water productive treatment compared to other DI treatments. Although biochar showed some positive effects on RSAD, it had marginal effects on soil water depletion and water productivity. This study suggests DI strategy optimized soil water depletion by regulating root adaptations or compensatory responses to DI-induced mild to moderate water stress during the growing season, while improving WP for successful cucumber production. It is recommended to test biochar over a longer period (> 2 years) or at higher application rates to better understand its influence on cucumber production and WP in West Texas region.
1. Following the ban on prophylactic antibiotic use in feed for poultry, interest in natural alternatives has increased, particularly plant-derived extracts. In this study, the effects of Aronia melanocarpa fruit extract (AMF) on growth performance, serum biochemistry and caecal microbiota in broilers were evaluated.2. A total of 360 male Ross-308 broiler chicks were randomly allocated to one of the four treatment groups for 35 days. These included a control group (CON) receiving the basal diet and three treatment groups supplemented with 50 mg/kg (AMF1), 100 mg/kg (AMF2) or 200 mg/kg (AMF3) of AMF extract.3. Antioxidant capacity and phenolic profile were determined by DPPH and UHPLC-ESI-MS/MS analyses. Because the ethanol extract had the highest antioxidant activity (94.5%), it was used in the experiment. Chlorogenic acid, quercetin, hesperidin and cyanidin-3-O-glucoside were the major phenolic compounds identified.4. The AMF supplementation had no significant effect on growth performance (p > 0.05). In contrast, serum triglyceride (TG), phosphorus, sodium and iron concentrations exhibited statistically significant quadratic dose - response relationships (p < 0.05). Serum TG levels were significantly reduced in the AMF3 group, whereas serum iron levels were significantly increased in the AMF1 and AMF2 groups (p < 0.05). Phosphorus and sodium levels were found to be lower in the control group compared to the other groups (p < 0.091 and p < 0.063, respectively).5. The 16S rRNA sequencing showed that low and medium dose AMF applications supported caecal microbial diversity by increasing the relative abundance of beneficial bacteria such as Bacillus spp. and Lactobacillus jensenii. The high supplementation dose led to the dominance of the Morganella morganii population.6. In conclusion, AMF can modulate the gut microbiota in broiler chickens in a dose-dependent manner without negatively affecting growth performance. Moderate inclusion rates have the potential to positively impact gut health and metabolic balance by increasing beneficial bacteria populations.