Transcription factor NtNAC56 regulates jasmonic acid-induced leaf senescence in tobacco.

Leaf senescence is a vital aspect of plant physiology and stress responses and is induced by endogenous factors and environmental cues. The plant-specific NAC (NAM, ATAF1/2, CUC2) transcription factor family influences growth, development, and stress responses in Arabidopsis (Arabidopsis thaliana) and other species. However, the roles of NACs in tobacco (Nicotiana tabacum) leaf senescence are still unclear. Here, we report that NtNAC56 regulates leaf senescence in tobacco. Transgenic plants overexpressing NtNAC56 (NtNAC56-OE) showed induction of senescence-related genes and exhibited early senescence and lower chlorophyll content compared to wild-type (WT) plants and the Ntnac56-19 mutant. In addition, root development and seed germination were inhibited in the NtNAC56-OE lines. Transmission electron microscopy observations accompanied by physiological and biochemical assays revealed that NtNAC56 overexpression triggers chloroplast degradation and reactive oxygen species accumulation in tobacco leaves. Transcriptome analysis demonstrated that NtNAC56 activates leaf senescence-related genes and jasmonic acid (JA) biosynthesis pathway genes. In addition, the JA content of NtNAC56-OE plants was higher than in WT plants, and JA treatment induced NtNAC56 expression. We performed DNA affinity purification sequencing to identify direct targets of NtNAC56, among which we focused on LIPOXYGENASE 5 (NtLOX5), a key gene in JA biosynthesis. A dual-luciferase reporter assay and a yeast one-hybrid assay confirmed that NtNAC56 directly binds to the TTTCTT motif in the NtLOX5 promoter. Our results reveal a mechanism whereby NtNAC56 regulates JA-induced leaf senescence in tobacco and provide a strategy for genetically manipulating leaf senescence and plant growth.

Coplanar Conformational Structure of π-Conjugated Polymers for Optoelectronic Applications.

Hierarchical structure of conjugated polymers is critical to dominating their optoelectronic properties and applications. Compared to nonplanar conformational segments, coplanar conformational segments of conjugated polymers (CPs) demonstrate favorable properties for applications as a semiconductor. Herein, recent developments in the coplanar conformational structure of CPs for optoelectronic devices are summarized. First, this review comprehensively summarizes the unique properties of planar conformational structures. Second, the characteristics of the coplanar conformation in terms of optoelectrical properties and other polymer physics characteristics are emphasized. Five primary characterization methods for investigating the complanate backbone structures are illustrated, providing a systematical toolbox for studying this specific conformation. Third, internal and external conditions for inducing the coplanar conformational structure are presented, offering guidelines for designing this conformation. Fourth, the optoelectronic applications of this segment, such as light-emitting diodes, solar cells, and field-effect transistors, are briefly summarized. Finally, a conclusion and outlook for the coplanar conformational segment regarding molecular design and applications are provided.

LESION MIMIC MUTANT 1 confers basal resistance to Sclerotinia sclerotiorum in rapeseed via a salicylic acid-dependent pathway.

Rapeseed (Brassica napus) is a major edible oilseed crop consumed worldwide. However, its yield is seriously affected by infection from the broad-spectrum non-obligate pathogen Sclerotinia sclerotiorum due to a lack of highly resistant germplasm. Here, we identified a Sclerotinia-resistant and light-dependent lesion mimic mutant from an ethyl methanesulfonate-mutagenized population of the rapeseed inbred Zhongshuang 11 (ZS11) named lesion mimic mutant 1 (lmm1). The phenotype of lmm1 is controlled by a single recessive gene, named LESION MIMIC MUTANT 1 (LMM1), which mapped onto chromosome C04 by bulked segregant analysis within a 2.71-Mb interval. Histochemical analysis indicated that H2O2 strongly accumulated and cell death occurred around the lesion mimic spots. Among 877 differentially expressed genes (DEGs) between ZS11 and lmm1 leaves, 188 DEGs were enriched in the defense response, including 95 DEGs involved in systemic acquired resistance, which is consistent with the higher salicylic acid levels in lmm1. Combining bulked segregant analysis and transcriptome analysis, we identified a significantly up-regulated gene, BnaC4.PR2, which encodes ß-1,3-glucanase, as the candidate gene for LMM1. Overexpression of BnaC4.PR2 may induce a reactive oxygen species burst to trigger partial cell death and systemic acquired resistance. Our study provides a new genetic resource for S. sclerotiorum resistance as well as new insights into disease resistance breeding in B. napus.

Maternal High-Fat Diet Multigenerationally Programs HPA Function and Behaviors in Male Rat Offspring.

Maternal environmental factors have been demonstrated to exert significant influences on the health of offspring. The hypothalamic-pituitary-adrenal (HPA) axis is an important neuroendocrine stress system that can be influenced by early life challenges. Our previous research has revealed that the consumption of a high-fat diet (HFD) by pregnant and lactating rats leads to the programming of HPA axis activity in male offspring of the first generation (referred to as F1HFD/C). The present study aimed to investigate whether the observed remodeling of the HPA axis could be inherited by second-generation male offspring (referred to as F2HFD/C), following maternal HFD exposure. The results showed that F2HFD/C rats exhibited enhanced basal HPA axis activity, similar to their F1HFD/C ancestors. Moreover, F2HFD/C rats displayed exacerbated corticosterone responses to restraint and lipopolysaccharide-induced stress, but not to insulin-induced hypoglycemia stress. Furthermore, maternal HFD exposure significantly aggravated depression-like behavior in the F2 generation subjected to chronic unpredictable mild stress. To investigate the role of central calcitonin gene-related peptide (CGRP) signaling in maternal diet-induced programming of the HPA axis across generations, we conducted central infusion of αCGRP8-37, a CGRP receptor antagonist, in F2HFD/C rats. The results demonstrated that αCGRP8-37 attenuated depression-like behaviors and reduced the hyperresponsiveness of the HPA axis to restraint stress in these rats. Therefore, central CGRP signaling may contribute to maternal diet-induced programming of HPA axis across generations. In conclusion, our study provides evidence that maternal HFD consumption can lead to multigenerational programming of the HPA axis and behaviors in adult male descendants.

Comparative transcriptome and co-expression network analysis revealed the genes associated with senescence and polygalacturonase activity involved in pod shattering of rapeseed.

BACKGROUND: The pod shattering (PS) trait negatively affects the crop yield in rapeseed especially under dry conditions. To better understand the trait and cultivate higher resistance varieties, it's necessary to identify key genes and unravel the PS mechanism thoroughly. RESULTS: In this study, we conducted a comparative transcriptome analysis between two materials significantly different in silique shatter resistance lignin deposition and polygalacturonase (PG) activity. Here, we identified 10,973 differentially expressed genes at six pod developmental stages. We found that the late pod development stages might be crucial in preparing the pods for upcoming shattering events. GO enrichment results from K-means clustering and weighed gene correlation network analysis (WGCNA) both revealed senescence-associated genes play an important role in PS. Two hub genes Bna.A05ABI5 and Bna.C03ERF/AP2-3 were selected from the MEyellow module, which possibly regulate the PS through senescence-related mechanisms. Further investigation found that senescence-associated transcription factor Bna.A05ABI5 upregulated the expression of SAG2 and ERF/AP2 to control the shattering process. In addition, the upregulation of Bna.C03ERF/AP2-3 is possibly involved in the transcription of downstream SHP1/2 and LEA proteins to trigger the shattering mechanism. We also analyzed the PS marker genes and found Bna.C07SHP1/2 and Bna.PG1/2 were significantly upregulated in susceptible accession. Furthermore, the role of auxin transport by Bna.WAG2 was also observed, which could reduce the PG activity to enhance the PS resistance through the cell wall loosening process. CONCLUSION: Based on comparative transcriptome evaluation, this study delivers insights into the regulatory mechanism primarily underlying the variation of PS in rapeseed. Taken together, these results provide a better understanding to increase the yield of rapeseed by reducing the PS through better engineered crops.

qPrimerDB: A Powerful and User-Friendly Database for qPCR Primer Design.

Real-time quantitative polymerase chain reaction (qPCR) is a powerful tool for analyzing and quantifying gene expression, and its primer design is the first and most important step. In order to improve the efficiency and effectiveness of primer design, we designed a database qPrimerDB, based on thermodynamics gene-specific for multispecies qPCR primers design.In this chapter, we explained the working principle of the database and detailed the step-by-step practical steps with examples. The valuable and time-saving qPrimerDB database is publicly accessible at http://biodb.swu.edu.cn/qprimerdb and will be routinely updated.

Stereoisomer-Independent Stable Blue Emission in Axial Chiral Difluorenol.

Bulky conjugated molecules with high stability are the prerequisite for the overall improvement of performance in wide-bandgap semiconductors. Herein, a chiral difluorenol, 2,2'-(9,9'-spirobi[fluorene]-2,2'-diyl)bis(9-(4-(octyloxy)phenyl)-9H-fluoren-9-ol) (DOHSBF), is set as a desirable model to reveal the stereoisomeric effects of wide-bandgap molecules toward controlling photophysical behavior and improving thermal and optical stability. Three diastereomers are obtained and elucidated by NMR spectra. Interestingly, the effect of modifying the stereo-centers is not observed on optical properties in solutions, pristine films, or post-treated film states. All three diastereomers as well as the mixture exhibit excellent spectral stability without undesirable green emission. Therefore, this stereoisomer-independent blue-emitting difluorenol will be a promising candidate for next-generation wide-bandgap semiconductors that would have extensive application in organic photonics.

A Bio-Inspired Molecular Design Strategy toward 2D Organic Semiconductor Crystals with Superior Integrated Optoelectronic Properties.

Inspired by the 2D bilayer lipid membranes in nature, a unique supramolecular "push-pull" synergetic strategy toward self-assembled 2D organic crystals (2DOCs) is proposed in this work, which can effectively suppress the interlayer 3D stacking while maintaining the assembly of the intralayer for 2D growth. For this purpose, a model molecule PF-Py consisting of a planar supramolecular "attractor" and a nonplanar steric "repellor" is designed for the solution self-assembly process. Well-defined 2DOCs including crystal nanosheets and millimeter-sized crystal films with layered amphiphile-like packing are obtained, which is analogical to the cell membranes of living organisms. Thanks to the special packing mode, the 2DOCs have fascinating integrated photoelectric property, with high mobility of 7.8 × 10-2 cm2 V-1 s-1 , high crystalline state photoluminescence quantum yield of 55%, and superior deep-blue laser characteristics with a low threshold of 5.51 µJ cm-2 . This supramolecular synergetic strategy advances the design of 2D organic semiconductor crystals for high performance optoelectronics.

Genome-wide association study and transcriptome comparison reveal novel QTL and candidate genes that control petal size in rapeseed.

Petal size determines the value of ornamental plants, and thus their economic value. However, the molecular mechanisms controlling petal size remain unclear in most non-model species. To identify quantitative trait loci and candidate genes controlling petal size in rapeseed (Brassica napus), we performed a genome-wide association study (GWAS) using data from 588 accessions over three consecutive years. We detected 16 significant single nucleotide polymorphisms (SNPs) associated with petal size, with the most significant SNPs located on chromosomes A05 and C06. A combination of GWAS and transcriptomic sequencing based on two accessions with contrasting differences in petal size identified 52 differentially expressed genes (DEGs) that may control petal size variation in rapeseed. In particular, the rapeseed gene BnaA05.RAP2.2, homologous to Arabidopsis RAP2.2, may be critical to the negative control of petal size through the ethylene signaling pathway. In addition, a comparison of petal epidermal cells indicated that petal size differences between the two contrasting accessions were determined mainly by differences in cell number. Finally, we propose a model for the control of petal size in rapeseed through ethylene and cytokinin signaling pathways. Our results provide insights into the genetic mechanisms regulating petal size in flowering plants.

Unveiling the Effects of Interchain Hydrogen Bonds on Solution Gelation and Mechanical Properties of Diarylfluorene-Based Semiconductor Polymers.

The intrinsically rigid and limited strain of most conjugated polymers has encouraged us to optimize the extensible properties of conjugated polymers. Herein, learning from the hydrogen bonds in glucose, which were facilitated to the toughness enhancement of cellulose, we introduced interchain hydrogen bonds to polydiarylfluorene by amide-containing side chains. Through tuning the copolymerization ratio, we systematically investigated their influence on the hierarchical condensed structures, rheology behavior, tensile performances, and optoelectronic properties of conjugated polymers. Compared to the reference copolymers with a low ratio of amide units, copolymers with 30% and 40% amide units present a feature of the shear-thinning process that resembled the non-Newtonian fluid, which was enabled by the interchain dynamic hydrogen bonds. Besides, we developed a practical and universal method for measuring the intrinsic mechanical properties of conjugated polymers. We demonstrated the significant impact of hydrogen bonds in solution gelation, material crystallization, and thin film stretchability. Impressively, the breaking elongation for P4 was even up to ~30%, which confirmed the partially enhanced film ductility and toughness due to the increased amide groups. Furthermore, polymer light-emitting devices (PLEDs) based on these copolymers presented comparable performances and stable electroluminescence (EL). Thin films of these copolymers also exhibited random laser emission with the threshold as low as 0.52 µJ/cm2, suggesting the wide prospective application in the field of flexible optoelectronic devices.

Comparative transcriptome and metabolomic profiling reveal the complex mechanisms underlying the developmental dynamics of tobacco leaves.

Although the leaf is the most important photosynthetic organ in most plants, many of the molecular mechanisms underlying leaf developmental dynamics remain to be explored. To better understand the transcriptional regulatory mechanisms involved in leaf development, we conducted comparative transcriptomic and metabolomic analysis of leaves from seven positions on tobacco (Nicotiana tabacum) plants. A total of 35,622 unique differentially expressed genes and 79 metabolites were identified. A time-series expression analysis detected two interesting transcriptional profiles, one comprising 10,197 genes that displayed continual up-regulation during leaf development and another comprising 4696 genes that displayed continual down-regulation. Combining these data with co-expression network results identified four important regulatory networks involved in photorespiration and the tricarboxylic acid cycle; these networks may regulate carbon/nitrogen balance during leaf development. We also found that the transcription factor NtGATA5 acts as a hub associated with C and N metabolism and chloroplast development during leaf development through regulation of phytohormones. Furthermore, we investigated the transcriptional dynamics of genes involved in the auxin, cytokinin, and jasmonic acid biosynthesis and signaling pathways during tobacco leaf development. Overall, our study greatly expands the understanding of the regulatory network controlling developmental dynamics in plant leaves.

Enhanced emission in organic nanocrystals via asymmetrical design of spirocyclic aromatic hydrocarbons.

Two spirocyclic aromatic hydrocarbon derivatives were prepared to clarify the molecular geometry effects on the regulation of the crystalline morphologies and photophysical behaviors of organic nanocrystals. Due to the different structural symmetry of a spiro-center, distinguishing nanocrystal morphologies with unique crystallization-enhanced/quenched emission was achieved.

Hierarchical Uniform Supramolecular Conjugated Spherulites with Suppression of Defect Emission.

Easily processed, well-defined, and hierarchical uniform artificial architectures with intrinsic strong crystalline emission properties are necessary for a range of light-emitting optoelectronic devices. Herein, we designed and prepared ordered supramolecular spherulites, comprising planar conformational molecules as primary structures and multiple hydrogen bonds as physical cross-links. Compared with serious aggregation-induced fluorescence quenching (up to 70%), these highly ordered architectures exhibited unique and robust crystalline emission with a high PLQY of 55%, which was much higher than those of other terfluorenes. The primary reasons for the high PLQY are the uniform exciton energetic landscape created in the planar conformation and the highly ordered molecular packing in spherulite. Meanwhile, minimal residual defect (green-band) emissions are effectively suppressed in our oriented crystalline framework, whereas the strong and stable blue light radiations are promoted. These findings may confirm that supramolecular ordered artificial architectures may offer higher control and tunability for optoelectronic applications.

Photophysical Identification of Three Kinds of Low-Energy Green Band Defects in Wide-Bandgap Polyfluorenes.

Blue-light-emitting semiconductors based on polyfluorenes often exhibit an undesired green emission band. In this report, three well-defined oligofluorenes corresponding to three types of "defects" attributed to aggregation, keto formation, and chain entanglement, respectively, are systemically investigated to unveil the origins of the green emission band in fluorene-based materials. First, the optical properties of defect molecules in different states are studied. The defect associated with aggregation is absent in dilute solutions and in films doped at 0.01 wt % with poly(methyl methacrylate). Second, the dependence of the emission spectra on the solvent was monitored to compare the effects of the "keto-" and "chain-entanglement defect" molecules. The green emission of keto defects exhibited a strong dependence on solvent polarity, whereas this cannot be observed in case of chain-entanglement defect. Third, energy transfer between poly[4-(octyloxy)-9,9-diphenylfluoren-2,7-diyl]- co-[5-(octyloxy)-9,9-diphenyl-fluoren-2,7-diyl] and the keto or chain-entanglement defect molecules is illustrated. Compared to those of the chain-entanglement defect, the spectra of the keto defect molecule (1:10-3) show signs of defect emission at lower proportions. These investigations not only provide insight into the photophysics of oligofluorenes but also supply a new strategy to explore defects in semiconductor polymers, which will aid in the development of effective approaches to obtain stable, pure blue organic light-emitting diodes based on polyfluorenes.

Supramolecular steric hindrance effect on morphologies and photophysical behaviors of spirocyclic aromatic hydrocarbon nanocrystals.

Three pyrene-based spirocyclic aromatic hydrocarbons (Py-SAHs) were prepared to clarify the roles of molecular segments in regulating the morphologies and photophysical properties of organic microcrystals. Due to the different supramolecular steric hindrance (SSH) effect between bulky groups and pyrene rings, distinct nanocrystal morphologies with unique photoluminescence properties were realized.

An eco-friendly water-assisted polyol method to enhance the aspect ratio of silver nanowires.

With water as an eco-friendly heterogeneous nucleation accelerator, silver nanowires (Ag NWs) have been successfully prepared with a high aspect ratio (>1600). The Ag NW-based film exhibits a low sheet resistance of 8.1 Ω sq-1 with a transparency of 81.9% at 550 nm, showing the potential application of electrode materials in polymer solar cells.

Ultrastable Supramolecular Self-Encapsulated Wide-Bandgap Conjugated Polymers for Large-Area and Flexible Electroluminescent Devices.

Controlling chain behavior through smart molecular design provides the potential to develop ultrastable and efficient deep-blue light-emitting conjugated polymers (LCPs). Herein, a novel supramolecular self-encapsulation strategy is proposed to construct a robust ultrastable conjugated polydiarylfluorene (PHDPF-Cz) via precisely preventing excitons from interchain cross-transfer/coupling and contamination from external trace H2 O/O2 . PHDPF-Cz consists of a mainchain backbone where the diphenyl groups localize at the 9-position as steric bulk moieties, and carbazole (Cz) units localize at the 4-position as supramolecular π-stacked synthon with the dual functionalities of self-assembly capability and hole-transport facility. The synergistic effect of the steric bulk groups and π-stacked carbazoles affords PHDPF-Cz as an ultrastable property, including spectral, morphological stability, and storage stability. In addition, PHDPF-Cz spin-coated gelation films also show thickness-insensitive deep-blue emission with respect to the reference polymers, which are suitable to construct solution-processed large-scale optoelectronic devices with higher reproducibility. High-quality and uniform deep-blue emission is observed in large-area solution-processed films. The electroluminescence shows high-quality deep-blue intrachain emission with a CIE (0.16, 0.12) and a very narrow full width at half-maximum of 32 nm. Finally, large-area and flexible polymer light-emitting devices with a single-molecular excitonic behavior are also fabricated. The supramolecular self-encapsulation design provides an effective strategy to construct ultrastable LCPs for optoelectronic applications.

Photophysical and Fluorescence Anisotropic Behavior of Polyfluorene ß-Conformation Films.

We demonstrate a systematic visualization of the unique photophysical and fluorescence anisotropic properties of polyfluorene coplanar conformation (ß-conformation) using time-resolved scanning confocal fluorescence imaging (FLIM) and fluorescence anisotropy imaging microscopy (FAIM) measurements. We observe inhomogeneous morphologies and fluorescence decay profiles at various micrometer-sized regions within all types of polyfluorene ß-conformational spin-coated films. Poly(9,9-dioctylfluorene-2,7-diyl) (PFO) and poly[4-(octyloxy)-9,9-diphenylfluoren-2,7-diyl]-co-[5-(octyloxy)-9,9-diphenylfluoren-2,7-diyl] (PODPF) ß-domains both have shorter lifetime than those of the glassy conformation for the longer effective conjugated length and rigid chain structures. Besides, ß-conformational regions have larger fluorescence anisotropy for the low molecular rotational motion and high chain orientation, while the low anisotropy in glassy conformational regions shows more rotational freedom of the chain and efficient energy migration from amorphous regions to ß-conformation as a whole. Finally, ultrastable ASE threshold in the PODPF ß-conformational films also confirms its potential application in organic lasers. In this regard, FLIM and FAIM measurements provide an effective platform to explore the fundamental photophysical process of conformational transitions in conjugated polymer.

qPrimerDB: a thermodynamics-based gene-specific qPCR primer database for 147 organisms.

Real-time quantitative polymerase chain reaction (qPCR) is one of the most important methods for analyzing the expression patterns of target genes. However, successful qPCR experiments rely heavily on the use of high-quality primers. Various qPCR primer databases have been developed to address this issue, but these databases target only a few important organisms. Here, we developed the qPrimerDB database, founded on an automatic gene-specific qPCR primer design and thermodynamics-based validation workflow. The qPrimerDB database is the most comprehensive qPCR primer database available to date, with a web front-end providing gene-specific and pre-computed primer pairs across 147 important organisms, including human, mouse, zebrafish, yeast, thale cress, rice and maize. In this database, we provide 3331426 of the best primer pairs for each gene, based on primer pair coverage, as well as 47760359 alternative gene-specific primer pairs, which can be conveniently batch downloaded. The specificity and efficiency was validated for qPCR primer pairs for 66 randomly selected genes, in six different organisms, through qPCR assays and gel electrophoresis. The qPrimerDB database represents a valuable, timesaving resource for gene expression analysis. This resource, which will be routinely updated, is publically accessible at http://biodb.swu.edu.cn/qprimerdb.

Genome-Wide Identification, Evolutionary and Expression Analyses of the GALACTINOL SYNTHASE Gene Family in Rapeseed and Tobacco.

Galactinol synthase (GolS) is a key enzyme in raffinose family oligosaccharide (RFO) biosynthesis. The finding that GolS accumulates in plants exposed to abiotic stresses indicates RFOs function in environmental adaptation. However, the evolutionary relationships and biological functions of GolS family in rapeseed (Brassica napus) and tobacco (Nicotiana tabacum) remain unclear. In this study, we identified 20 BnGolS and 9 NtGolS genes. Subcellular localization predictions showed that most of the proteins are localized to the cytoplasm. Phylogenetic analysis identified a lost event of an ancient GolS copy in the Solanaceae and an ancient duplication event leading to evolution of GolS4/7 in the Brassicaceae. The three-dimensional structures of two GolS proteins were conserved, with an important DxD motif for binding to UDP-galactose (uridine diphosphate-galactose) and inositol. Expression profile analysis indicated that BnGolS and NtGolS genes were expressed in most tissues and highly expressed in one or two specific tissues. Hormone treatments strongly induced the expression of most BnGolS genes and homologous genes in the same subfamilies exhibited divergent-induced expression. Our study provides a comprehensive evolutionary analysis of GolS genes among the Brassicaceae and Solanaceae as well as an insight into the biological function of GolS genes in hormone response in plants.