Identification and validation of stable reference genes for RT-qPCR analyses of Kobresia littledalei seedlings.

BACKGROUND: Kobreisa littledalei, belonging to the Cyperaceae family is the first Kobresia species with a reference genome and the most dominant species in Qinghai-Tibet Plateau alpine meadows. It has several resistance genes which could be used to breed improved crop varieties. Reverse Transcription Quantitative Real-Time Polymerase Chain Reaction (RT-qPCR) is a popular and accurate gene expression analysis method. Its reliability depends on the expression levels of reference genes, which vary by species, tissues and environments. However, K.littledalei lacks a stable and normalized reference gene for RT-qPCR analysis. RESULTS: The stability of 13 potential reference genes was tested and the stable reference genes were selected for RT-qPCR normalization for the expression analysis in the different tissues of K. littledalei under two abiotic stresses (salt and drought) and two hormonal treatments (abscisic acid (ABA) and gibberellin (GA)). Five algorithms were used to assess the stability of putative reference genes. The results showed a variation amongst the methods, and the same reference genes showed tissue expression differences under the same conditions. The stability of combining two reference genes was better than a single one. The expression levels of ACTIN were stable in leaves and stems under normal conditions, in leaves under drought stress and in roots under ABA treatment. The expression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) expression was stable in the roots under the control conditions and salt stress and in stems exposed to drought stress. Expression levels of superoxide dismutase (SOD) were stable in stems of ABA-treated plants and in the roots under drought stress. Moreover, RPL6 expression was stable in the leaves and stems under salt stress and in the stems of the GA-treated plants. EF1-alpha expression was stable in leaves under ABA and GA treatments. The expression levels of 28 S were stable in the roots under GA treatment. In general, ACTIN and GAPDH could be employed as housekeeping genes for K. littledalei under different treatments. CONCLUSION: This study identified the best RT-qPCR reference genes for different K. littledalei tissues under five experimental conditions. ACTIN and GAPDH genes can be employed as the ideal housekeeping genes for expression analysis under different conditions. This is the first study to investigate the stable reference genes for normalized gene expression analysis of K. littledalei under different conditions. The results could aid molecular biology and gene function research on Kobresia and other related species.

Insights into the Impact of Trans-Zeatin Overproduction-Engineered Sinorhizobium meliloti on Alfalfa (Medicago sativa L.) Tolerance to Drought Stress.

Plant growth-promoting rhizobacteria have been shown to enhance plant tolerance to drought stress through various mechanisms. However, there is limited research on improving drought resistance in alfalfa by genetically modifying PGPR to produce increased levels of cytokinins. Herein, we employed synthetic biology approaches to engineer two novel strains of Sinorhizobium meliloti capable of overproducing trans-Zeatin and investigated their potential in enhancing drought tolerance in alfalfa. Our results demonstrate that alfalfa plants inoculated with these engineered S. meliloti strains exhibited reduced wilting and yellowing while maintaining higher relative water content under drought conditions. The engineered S. meliloti-induced tZ activated the activity of antioxidant enzymes and the accumulation of osmolytes. Additionally, the increased endogenous tZ content in plants alleviated the impact of drought stress on the alfalfa photosynthetic rate. However, under nondrought conditions, inoculation with the engineered S. meliloti strains had no significant effect on alfalfa biomass and nodule formation.

Overexpression of the alfalfa (Medicago sativa) gene, MsKMS1, negatively regulates seed germination in transgenic tobacco (Nicotiana tabacum).

Soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE)-associated proteins are a class of transmembrane proteins involved in intracellular trafficking pathways. However, the functions of many SNARE domain-containing proteins remain unclear. We have previously identified a SNARE-associated gene in alfalfa (Medicago sativa ) KILLING ME SLOWLY1 (MsKMS1 ), which is involved in various abiotic stresses. In this study, we investigated the function of MsKMS1 in the seed germination of transgenic tobacco (Nicotiana tabacum ). Phylogenetic analysis showed that MsKMS1 was homologous to the SNARE-associated or MAPR component-related proteins of other plants. Germination assays revealed that MsKMS1 negatively regulated seed germination under normal, D-mannitol and abscisic acid-induced stress conditions, yet MsKMS1 -overexpression could confer enhanced heat tolerance in transgenic tobacco. The suppressive effect on germination in MsKMS1 -overexpression lines was associated with higher abscisic acid and salicylic acid contents in seeds. This was accompanied by the upregulation of abscisic acid biosynthetic genes (ZEP and NCED ) and the downregulation of gibberellin biosynthetic genes (GA20ox2 and GA20ox3 ). Taken together, these results suggested that MsKMS1 negatively regulated seed germination by increasing abscisic acid and salicylic acid contents through the expression of genes related to abscisic acid and gibberellin biosynthesis. In addition, MsKMS1 could improve heat tolerance during the germination of transgenic tobacco seeds.

Genome assembly and multi-omic analyses reveal the mechanisms underlying flower color formation in Torenia fournieri.

Torenia fournieri Lind. is an ornamental plant that is popular for its numerous flowers and variety of colors. However, its genomic evolutionary history and the genetic and metabolic bases of flower color formation remain poorly understood. Here, we report the first T. fournieri reference genome, which was resolved to the chromosome scale and was 164.4 Mb in size. Phylogenetic analyses clarified relationships with other plant species, and a comparative genomic analysis indicated that the shared ancestor of T. fournieri and Antirrhinum majus underwent a whole genome duplication event. Joint transcriptomic and metabolomic analyses identified many metabolites related to pelargonidin, peonidin, and naringenin production in rose (TfR)-colored flowers. Samples with blue (TfB) and deep blue (TfD) colors contained numerous derivatives of petunidin, cyanidin, quercetin, and malvidin; differences in the abundances of these metabolites and expression levels of the associated genes were hypothesized to be responsible for variety-specific differences in flower color. Furthermore, the genes encoding flavonoid 3-hydroxylase, anthocyanin synthase, and anthocyanin reductase were differentially expressed between flowers of different colors. Overall, we successfully identified key genes and metabolites involved in T. fournieri flower color formation. The data provided by the chromosome-scale genome assembly establish a basis for understanding the differentiation of this species and will facilitate future genetic studies and genomic-assisted breeding.

Healing the Buried Interface by a Plant-Derived Green Passivator for Carbon-Based CsPbIBr2 Perovskite Solar Cells.

Perovskite solar cells (PSCs) have attracted extensive attention in photovoltaic applications owing to their superior efficiency, and the buried interface plays a significant role in determining the efficiency and stability of PSCs. Herein, a plant-derived small molecule, ergothioneine (ET), is adopted to heal the defective buried interface of CsPbIBr2-based PSC to improve power conversion efficiency (PCE). Because of the strong interaction between Lewis base groups (-C═O and -C═S) in ET and uncoordinated Pb2+ in the perovskite film from the theoretical simulations and experimental results, the defect density of the CsPbIBr2 perovskite film is significantly reduced, and therefore, the nonradiative recombination in the corresponding device is simultaneously suppressed. Consequently, the target device achieves a high PCE of 11.13% with an open-circuit voltage (VOC) of 1.325 V for hole-free, carbon-based CsPbIBr2 PSCs and 14.56% with a VOC of 1.308 V for CsPbI2Br PSCs. Furthermore, because of the increased ion migration energy, the detrimental phase segregation in this mixed-halide perovskite is weakened, delivering excellent long-term stability for the unencapsulated device in ambient conditions over 70 days with a 96% retention rate of initial efficiency.

Integrated physiological, metabolomic, and transcriptomic analyses elucidate the regulation mechanisms of lignin synthesis under osmotic stress in alfalfa leaf (Medicago sativa L.).

Alfalfa, an essential forage crop known for its high yield, nutritional value, and strong adaptability, has been widely cultivated worldwide. The yield and quality of alfalfa are frequently jeopardized due to environmental degradation. Lignin, a constituent of the cell wall, enhances plant resistance to abiotic stress, which often causes osmotic stress in plant cells. However, how lignin responds to osmotic stress in leaves remains unclear. This study explored the effects of osmotic stress on lignin accumulation and the contents of intermediate metabolites involved in lignin synthesis in alfalfa leaves. Osmotic stress caused an increase in lignin accumulation and the alteration of core enzyme activities and gene expression in the phenylpropanoid pathway. We identified five hub genes (CSE, CCR, CADa, CADb, and POD) and thirty edge genes (including WRKYs, MYBs, and UBPs) by integrating transcriptome and metabolome analyses. In addition, ABA and ethylene signaling induced by osmotic stress regulated lignin biosynthesis in a contradictory way. These findings contribute to a new theoretical foundation for the breeding of high-quality and resistant alfalfa varieties.

Genome-wide identification and expression analysis of the glycosyl hydrolase family 1 genes in Medicago sativa revealed their potential roles in response to multiple abiotic stresses.

Glycoside hydrolase family 1 (GH1) ß-glucosidases (BGLUs), are encoded by a large number of genes, which participate in the development and stress response of plants, particularly under biotic and abiotic stresses through the activation of phytohormones. However, there are few studies systematically analyzing stress or hormone-responsive BGLU genes in alfalfa. In this study, a total of 179 BGLU genes of the glycoside hydrolase family 1 were identified in the genome of alfalfa, and then were classified into five distinct clusters. Sequence alignments revealed several conserved and unique motifs among these MsBGLU proteins. Many cis-acting elements related to abiotic stresses and phytohormones were identified in the promoter of some MsBGLUs. Moreover, RNA-seq and RT-qPCR analyses showed that these MsBGLU genes exhibited distinct expression patterns in response to different abiotic stress and hormonal treatments. In summary, this study suggests that MsBGLU genes play crucial roles in response to various abiotic stresses and hormonal responses, and provides candidate genes for stress tolerance breeding in alfalfa.

A 3-Ketoacyl-CoA Synthase 10 (KCS10) Homologue from Alfalfa Enhances Drought Tolerance by Regulating Cuticular Wax Biosynthesis.

Cuticular wax, forming the first line of defense against adverse environmental stresses, comprises very long-chain fatty acids (VLCFAs) and their derivatives. 3-Ketoacyl-CoA synthase (KCS) is a rate-limiting enzyme for VLCFA biosynthesis. In this study, we isolated KCS10, a KCS gene from alfalfa, and analyzed the effect of gene expression on wax production and drought stress in transgenic plants. MsKCS10 overexpression increased compact platelet-like crystal deposition and promoted primary alcohol biosynthesis through acyl reduction pathways in alfalfa leaves. Overexpression of MsKCS10 induced the formation of coiled-rodlet-like crystals and increased n-alkane content through decarbonylation pathways in tobacco and tomato fruits. Overexpression of MsKCS10 enhanced drought tolerance by limiting nonstomatal water loss, improving photosynthesis, and maintaining osmotic potential under drought stress in transgenic tobacco. In summary, MsKCS10 plays an important role in wax biosynthesis, wax crystal morphology, and drought tolerance, although the mechanisms are different among the plant species. MsKCS10 can be targeted in future breeding programs to improve drought tolerance in plants.

Osmotic stress-induced lignin synthesis is regulated at multiple levels in alfalfa (Medicago sativa L.).

Alfalfa is an important forage crop. Yield and quality are frequently threatened by extreme environments such as drought and salt stress. As a component of the cell wall, lignin plays an important role in the abiotic stress response, the mechanisms of which have not been well clarified. In this study, we combined physiological, transcriptional, and metabolic analyses to reveal the changes in lignin content in alfalfa under mannitol-induced osmotic stress. Osmotic stress enhanced lignin accumulation by increasing G and S units, which was associated with increases in enzyme activities and decreases in 8 intermediate metabolites. Upon combined analysis of the transcriptome and metabolome, we identified five key structural genes and several coexpressed transcription factors, such as MYB and WRKY, which may play a core role in regulating lignin content and composition under osmotic stress. In addition, lignin synthesis was positively regulated by ABA but negatively regulated by ethylene under osmotic stress. These results provide new insight into the regulatory mechanism of lignin synthesis under abiotic stress.

A dual-mode fluorescent probe based on carbon dots for detecting solution polarity.

Fluorescent probes always attract huge attention. In particular, carbon dots, due to their unique biocompatibility and variable fluorescence characteristics, can be used in multiple fields and were fulled of expectation by researchers. Since the advent of the dual-mode carbon dots probe, which greatly improved the accuracy of quantitative detection, there are higher hopes for dual-mode carbon dots probes. Herein, we have successfully developed a new dual-mode fluorescent carbon dots probe based on 1,10-phenanthroline (Ph-CDs). Ph-CDs detect the object to be measured based on both down-conversion luminescence and up-conversion luminescence at the same time, different from the reported dual-mode fluorescent probes which are based on the wavelength and intensity changes in down-conversion luminescence. As-prepared Ph-CDs have good linear relationships with the polarity of solvents in down-conversion luminescence (R2 = 0.9909) and up-conversion luminescence (R2 = 0.9374), respectively. Hence, Ph-CDs provide a new in-depth insight into the design of fluorescent probes with dual-mode detection and they can give more accurate, reliable and convenient detection results.

Round-comb Fe2O3@SnO2 heterostructures enable efficient light harvesting and charge extraction for high-performance all-inorganic perovskite solar cells.

The precise design of an electron transport layer (ETL) to improve the light-harvesting and quality of perovskite (PVK) film plays a crucial role in the photovoltaic performance of n-i-p perovskite solar cells (PSCs). In this work, a novel three-dimensional (3D) round-comb Fe2O3@SnO2 heterostructure composites with high conductivity and electron mobility induced by its Type-II band alignment and matched lattice spacing is prepared and employed as an efficient mesoporous ETL for all-inorganic CsPbBr3 PSCs. Arising from the multiple light scattering sites provided by the 3D round-comb structure, the diffuse reflectance of Fe2O3@SnO2 composites is increased to improve the light absorption of the deposited PVK film. Besides, the mesoporous Fe2O3@SnO2 ETL affords not only more active surface for sufficient exposure to the CsPbBr3 precursor solution but also a wettable surface to reduce the barrier for heterogeneous nucleation, which realizes the regulated growth of a high-quality PVK film with less undesired defect. Hence, both the light-harvesting capability, the photoelectrons transport and extraction are improved, and the charge recombination is restrained, delivering an optimized power conversion efficiency (PCE) of 10.23 % with a high short-circuit current density of 7.88 mA cm-2 for the c-TiO2/Fe2O3@SnO2 ETL based all-inorganic CsPbBr3 PSCs. Moreover, under lasting erosion at 25 °C and 85 % RH for 30 days and light-soaking (AM 1.5G) for 480 h in air atmosphere, the unencapsulated-device shows superiorly persistent durability.

Construction of Ti3C2Tx MXene wrapped urchin-like CuCo2S4 microspheres for high-performance asymmetric supercapacitors.

Copper cobalt sulfide (CuCo2S4) nanomaterials are regarded as promising electrode materials for high-performance supercapacitors due to their abundant redox states and considerable theoretical capacities. However, the intrinsic poor electrical conductivity, sluggish reaction kinetics and insufficient number of electroactive sites of these materials are huge barriers to realize their practical applications. In this study, a facile two-step strategy to engineer a hierarchical 3D porous CuCo2S4/MXene composite electrode is presented for enhanced storage properties. This well-constructed CuCo2S4/MXene composite not only provides abundant active sites for the faradaic reaction, but also offers more efficient pathways for rapid electron/ion transport and restricts the volumetric expansion during the charge/discharge process. When evaluated in a 3 M KOH electrolyte, the CuCo2S4/MXene-3 electrode exhibits a specific capacity of 1351.6 C g-1 at 1 A g-1 while retaining excellent cycling stability (95.2% capacity retention at 6 A g-1 after 10 000 cycles). Additionally, the solid-state asymmetric supercapacitor (ASC) CuCo2S4/MXene//AC device displays an energy density of 78.1 W h kg-1 and a power density of 800.7 W kg-1. Two ASC devices connected in series can illuminate a blue LED indicator for more than 20 min, demonstrating promising prospects for practical applications. These electrochemical properties indicate that the high-performance CuCo2S4/MXene composites are promising electrode materials for advanced asymmetric supercapacitors.

Interface engineering of Fe2P@CoMnP4 heterostructured nanoarrays for efficient and stable overall water splitting.

Electrocatalytic water splitting to generate high-quality hydrogen is an attractive renewable energy storage technology; however, it is still far from becoming a real-world application. In this study, we developed an effective and stable nickel foam-supported Fe2P@CoMnP4 heterostructure electrocatalyst for overall water splitting. As expected, the as-obtained Fe2P@CoMnP4/NF electrocatalyst exhibits superb bifunctional catalytic activity and only requires extremely low overpotentials of 53 and 249 mV to achieve a current density of 10 mA cm-2 for the hydrogen and oxygen evolution reactions, respectively. Moreover, a two-electrode electrolyzer assembled using Fe2P@CoMnP4/NF as electrodes operates at the low cell voltage of 1.54 V at 10 mA cm-2, showing excellent long-term stability for 140 h. Theoretical calculations indicate that the surface electronic structure is effectively adjusted by the generated heterointerfaces between the Fe2P and CoMnP4 in a two-phase matrix, resulting in a Gibbs free energy of hydrogen adsorption close to zero and high intrinsic activity. This innovative strategy is a valuable route for producing low-cost high-performance bifunctional electrocatalysts for water splitting.

Iron nanoparticles induced the growth and physio-chemical changes in Kobresia capillifolia seedlings.

Iron nanoparticles (NPs) priming is known to affect the seed germination and seedling growth in many plants. However, whether it has an important role in stimulating the growth of perennial Qinghai-Tibet Plateau plants remains unclear. In this study, the effects of seed priming with different concentrations of nFe2O3 and FeCl3 (10, 50, 100, 500, and 1000 mg L-1) on seed germination, plant growth, photosystem, antioxidant enzyme activities, root morphology, and biomass distribution of Kobresia capillifolia were evaluated under laboratory conditions. The results showed that compared with treatment materials, concentration had more significant effects on K. capillifolia development. There was no significant impact on germination rate were discovered under all treatments, but decreased the seed mildew rate at 100 mg L-1 nFe2O3. Compare with control, Fe-based priming significantly decreased root biomass. All Fe-based treatments increased rubisco activity of leaves, and significantly enhanced Pn at ranged from 10 to 100 mg L-1. Meanwhile, chlorophyll contents were decreased, the chloroplasts were swollen, and thylakoids were disorganized under all Fe treatments. Iron-based priming significantly enhanced SOD, POD, and CAT activities in Kobresia roots. In conclusion, the thick cuticle-covered seed coat of K. capillifolia postponed the penetration of FeNPs into seeds, so FeNPs priming had a weak impact on seed germination. The sustainable release of Fe ions from FeNPs and the uptake of Fe ions by roots affected the physiology, biochemistry and morphology of K. capillifolia. The findings of this study provide an in-depth understanding of how FeNPs impact the alpine meadow plant, K. capillifolia.

Enhanced Gas Sensitivity of Au-Decorated Flowery WSe2 Nanostructures.

With the continuous improvement in material life, people are paying more and more attention to air quality; therefore, it is critical to design efficient and stable gas sensor devices. In this work, a flowery WSe2 nanostructure and its nanocomposite (Au@WSe2) decorated with Au nanoparticles were fabricated by the hydrothermal method. The performance of a resistive sensor with flowery WSe2 and Au@WSe2 sensors was evaluated by detecting volatile organic compounds such as ethanol, isoamylol, n-butyl alcohol, isopropanol, isobutanol and n-propanol. The results show that Au-nanoparticle-decorated flowery WSe2 can decrease the optimal working temperature from 215 °C to 205 °C and significantly enhance the response of flowery WSe2. The response values to isoamylol are the highest (as high as 44.5) at a low gas concentration (100 ppm), while the response values to ethanol are the highest (as high as 178.5) at a high gas concentration (1000 ppm) among the six different alcohols. Moreover, the response is steady and repeatable. The results demonstrate that the Au@WSe2 substrate has good responsiveness and selectivity, which makes it a promising candidate for gas detection.

Identification of Competing Endogenous RNAs (ceRNAs) Network Associated with Drought Tolerance in Medicago truncatula with Rhizobium Symbiosis.

Drought, bringing the risks of agricultural production losses, is becoming a globally environmental stress. Previous results suggested that legumes with nodules exhibited superior drought tolerance compared with the non-nodule group. To investigate the molecular mechanism of rhizobium symbiosis impacting drought tolerance, transcriptome and sRNAome sequencing were performed to identify the potential mRNA-miRNA-ncRNA dynamic network. Our results revealed that seedlings with active nodules exhibited enhanced drought tolerance by reserving energy, synthesizing N-glycans, and medicating systemic acquired resistance due to the early effects of symbiotic nitrogen fixation (SNF) triggered in contrast to the drought susceptible with inactive nodules. The improved drought tolerance might be involved in the decreased expression levels of miRNA such as mtr_miR169l-5p, mtr_miR398b, and mtr_miR398c and its target genes in seedlings with active nodules. Based on the negative expression pattern between miRNA and its target genes, we constructed an mRNA-miR169l-ncRNA ceRNA network. During severe drought stress, the lncRNA alternative splicings TCONS_00049507 and TCONS_00049510 competitively interacted with mtr_miR169l-5p, which upregulated the expression of NUCLEAR FACTOR-Y (NF-Y) transcription factor subfamily NF-YA genes MtNF-YA2 and MtNF-YA3 to regulate their downstream drought-response genes. Our results emphasized the importance of SNF plants affecting drought tolerance. In conclusion, our work provides insight into ceRNA involvement in rhizobium symbiosis contributing to drought tolerance and provides molecular evidence for future study.

Proteomic Analysis of Alfalfa (Medicago sativa L.) Roots in Response to Rhizobium Nodulation and Salt Stress.

(1) Background: Alfalfa is an important legume forage throughout the world. Although alfalfa is considered moderately tolerant to salinity, its production and nitrogen-fixing activity are greatly limited by salt stress. (2) Methods: We examined the physiological changes and proteomic profiles of alfalfa with active nodules (NA) and without nodules (NN) under NaCl treatment. (3) Results: Our data suggested that NA roots showed upregulation of the pathways of abiotic and biotic stress responses (e.g., heat shock proteins and pathogenesis-related proteins), antioxidant enzyme synthesis, protein synthesis and degradation, cell wall degradation and modification, acid phosphatases, and porin transport when compared with NN plants under salt stress conditions. NA roots also upregulated the processes or proteins of lipid metabolism, heat shock proteins, protein degradation and folding, and cell cytoskeleton, downregulated the DNA and protein synthesis process, and vacuolar H+-ATPase proteins under salt stress. Besides, NA roots displayed a net H+ influx and low level of K+ efflux under salt stress, which may enhance the salt tolerance of NA plants. (4) Conclusions: The rhizobium symbiosis conferred the host plant salt tolerance by regulating a series of physiological processes to enhance stress response, improve antioxidant ability and energy use efficiency, and maintain ion homeostasis.

High-performance perovskite photodetectors based on CsPbBr3 microwire arrays: erratum.

We correct two errors in our publication [Appl. Opt.60, 8896 (2021)APOPAI0003-693510.1364/AO.437478].

Distinct rhizosphere soil responses to nitrogen in relation to microbial biomass and community composition at initial flowering stages of alfalfa cultivars.

In the long-term growth process, alfalfa rhizosphere forms specific microbiome to provide nutrition for its growth and development. However, the effects of different perennial alfalfa cultivars on changes in the rhizosphere soil characteristics and microbiome are not well understood. In this study, 12 perennial alfalfa cultivars were grown continuously for eight years. Rhizosphere samples were tested using Illumina sequencing of the 16S rRNA gene coupled with co-occurrence network analysis to explore the relationship between alfalfa (biomass and crude protein content), soil properties, and the microbial composition and diversity. Redundancy analysis showed SOC and pH had the greatest impact on the composition of the rhizosphere microbial community. Moreover, microbial diversity also contributes to microbial composition. Soil properties (AP, EC, SOC and pH) exhibited a significant positive correlation with soil bacterial communities, which was attributed to the differences between plant cultivars. Partial least squares path modeling (PLS-PM) revealed that microbial biomass and community composition rather than diversity, are the dominant determinants in the rhizosphere soil nitrogen content of perennial alfalfa. Our findings demonstrate that the soil microbial biomass and composition of rhizosphere bacterial communities are strongly affected by cultivar, driving the changes in soil nitrogen content, and variances in the selective capacities of plants.