Discrete Miktoarm Star Block Copolymers with Tailored Molecular Architecture.

Molecular architecture is a critical factor in regulating phase behaviors of the block copolymer and prompting the formation of unconventional nanostructures. This work meticulously designed a library of isomeric miktoarm star polymers with an architectural evolution from the linear-branched block copolymer to the miktoarm star block copolymer and to the star-like block copolymer (i.e., 3AB → 3(AB1)B2 → 3(AB)). All of the polymers have precise chemical composition and uniform chain length, eliminating inherent molecular uncertainties such as chain length distribution or architectural defects. The self-assembly behaviors were systematically studied and compared. Gradually increasing the relative length of the branched B1 block regulates the ratio between the bridge and loop configuration and effectively releases packing frustration in the formation of the spherical or cylindrical structures, leading to a substantial deflection of phase boundaries. Complex structures, such as Frank-Kasper phases, were captured at a surprisingly higher volume fraction. Rationally regulating the molecular architecture offers rich possibilities to tune the packing symmetry of block copolymers.

Interfacial Behaviors of Giant Amphiphilic Molecules Composed of Hydrophobic Isobutyl POSS and Hydrophilic POSS Bearing Carboxylic Acid Groups at the Air-Water Interface.

The behavior of giant amphiphilic molecules at the air-water interface has become a subject of concern to researchers. Small changes in the molecular structure can cause obvious differences in the molecular arrangement and interfacial properties of the monolayer. In this study, we have systematically investigated the interfacial behaviors of the giant amphiphilic molecules with different number of hydrophobic BPOSS blocks and one hydrophilic ACPOSS block ((BPOSS)n-ACPOSS (n = 1-5)) at the air-water interface by the surface pressure-area (π-A) isotherm, Brewster angle microscopy (BAM), compression modulus measurement, and hysteresis measurement. We found that both the number of BPOSS blocks and the compression rate can significantly influence the interfacial behaviors of giant molecules. The π-A isotherms of giant molecules (BPOSS)n-ACPOSS (n = 2-5) exhibit a "cusp" phenomenon which can be attributed to the transition from monolayer to multilayer. However, the cusp is dramatically different from the "collapse" of the monolayer studied in other molecular systems, which is highly dependent on the compression rate of the monolayer. In addition, the compression modulus and hysteresis measurements reveal that the number of BPOSS blocks of (BPOSS)n-ACPOSS plays an important role in the static elasticity, stability, and reversibility of the Langmuir films.

Local Chain Feature Mandated Self-Assembly of Block Copolymers.

This work demonstrates an effective and robust approach to regulate phase behaviors of a block copolymer by programming local features into otherwise homogeneous linear chains. A library of sequence-defined, isomeric block copolymers with globally the same composition but locally different side chain patterns were elaborately designed and prepared through an iterative convergent growth method. The precise chemical structure and uniform chain length rule out all inherent molecular defects associated with statistical distribution. The local features are found to exert surprisingly pronounced impacts on the self-assembly process, which have yet to be well recognized. While other molecular parameters remain essentially the same, simply rearranging a few methylene units among the alkyl side chains leads to strikingly different phase behaviors, bringing about (i) a rich diversity of nanostructures across hexagonally packed cylinders, Frank-Kasper A15 phase, Frank-Kasper σ phase, dodecagonal quasicrystals, and disordered state; (ii) a significant change of lattice dimension; and (iii) a substantial shift of order-to-disorder transition temperature (up to 40 °C). Different from the commonly observed enthalpy-dominated cases, the frustration due to the divergence between the native molecular geometry originating from side chain distribution and the local packing environment mandated by lattice symmetry is believed to play a pivotal role. Engineering the local chain feature introduces another level of structural complexity, opening up a new and effective pathway for modulating phase transition without changing the chemistry or composition.

A reference-grade genome assembly for Astragalus mongholicus and insights into the biosynthesis and high accumulation of triterpenoids and flavonoids in its roots.

Astragalus membranaceus var. mongholicus (AMM), a member of the Leguminosae, is one of the most important medicinal plants worldwide. The dried roots of AMM have a wide range of pharmacological effects and are a traditional Chinese medicine. Here, we report the first chromosome-level reference genome of AMM, comprising nine pseudochromosomes with a total size of 1.47 Gb and 27 868 protein-encoding genes. Comparative genomic analysis reveals that AMM has not experienced an independent whole-genome duplication (WGD) event after the WGD event shared by the Papilionoideae species. Analysis of long terminal repeat retrotransposons suggests a recent burst of these elements at approximately 0.13 million years ago, which may explain the large size of the AMM genome. Multiple gene families involved in the biosynthesis of triterpenoids and flavonoids were expanded, and our data indicate that tandem duplication has been the main driver for expansion of these families. Among the expanded families, the phenylalanine ammonia-lyase gene family was primarily expressed in the roots of AMM, suggesting their roles in the biosynthesis of phenylpropanoid compounds. The functional versatility of 2,3-oxidosqualene cyclase genes in cluster III may play a critical role in the diversification of triterpenoids in AMM. Our findings provide novel insights into triterpenoid and flavonoid biosynthesis and can facilitate future research on the genetics and medical applications of AMM.

Regioisomeric Giant Triblock Molecules: Role of the Linker.

In this study, polyhedral oligomeric silsesquioxane (POSS) based giant triblock molecules with precisely defined regio-configuration are modularly prepared through highly efficient coupling reactions. The length of the linker connecting neighboring nanoparticles is elaborately designed to regulate the geometric constraints. The triblock molecules adopt a folded packing during phase separation, and the regio-configuration imparts direct influence on the self-assembly behaviors. The ortho-isomers form periodic structures with a larger domain size, larger interfacial curvature, and enhanced phase stability. The regio-effect is closely related to the length and symmetry of the linker. As the linker extends, the neighboring particles gradually decouple, and the regio-effect diminishes. The symmetry of the linker shows an even more profound impact. This work quantitatively scrutinized the role of the linker, opening an avenue for engineering the assembled structures with molecular precision.

Hyperspectral imaging with machine learning for non-destructive classification of Astragalus membranaceus var. mongholicus, Astragalus membranaceus, and similar seeds.

The roots of Astragalus membranaceus var. mongholicus (AMM) and A. membranaceus (AM) are widely used in traditional Chinese medicine. Although AMM has higher yields and accounts for a larger market share, its cultivation is fraught with challenges, including mixed germplasm resources and widespread adulteration of commercial seeds. Current methods for distinguishing Astragalus seeds from similar (SM) seeds are time-consuming, laborious, and destructive. To establish a non-destructive method, AMM, AM, and SM seeds were collected from various production areas. Machine vision and hyperspectral imaging (HSI) were used to collect morphological data and spectral data of each seed batch, which was used to establish discriminant models through various algorithms. Several preprocessing methods based on hyperspectral data were compared, including multiplicative scatter correction (MSC), standard normal variable (SNV), and first derivative (FD). Then selection methods for identifying informative features in the above data were compared, including successive projections algorithm (SPA), uninformative variable elimination (UVE), and competitive adaptive reweighted sampling (CARS). The results showed that support vector machine (SVM) modeling of machine vision data could distinguish Astragalus seeds from SM with >99% accuracy, but could not satisfactorily distinguish AMM seeds from AM. The FD-UVE-SVM model based on hyperspectral data reached 100.0% accuracy in the validation set. Another 90 seeds were tested, and the recognition accuracy was 100.0%, supporting the stability of the model. In summary, HSI data can be applied to discriminate among the seeds of AMM, AM, and SM non-destructively and with high accuracy, which can drive standardization in the Astragalus production industry.

Seed Vigour and Morphological and Physiological Characteristics of Epimedium brevicornu Maxim: In Different Stages of Seed Development.

Epimedium brevicornu Maxim is a traditional Chinese medicinal plant with important value for curing several diseases, including liver cancer. Seed germination, field seedling emergence, and morphological and physiological traits were measured in developing seeds of E. brevicornu, which were collected at 7, 14, 21, 28, and 35 days after flowering. The results showed that with the fruit pericarp changing from lime green to dark red, the seed volume increased. Furthermore, the dry mass of seeds gradually increased from 0.011 g at 7 d to 0.275 g at 35 d, which was a significantly positive correlation with seed vigour (r = 0.980). The soluble protein content initially increased and then decreased to 11.09 mg/g and presented a maximum at 28 d; however, the soluble sugar content gradually declined to a minimum of 30.45 mg/g at 35 d, which was also significantly negatively correlated with seed vigour (r = -0.915). Furthermore, the unsaturated fatty acids (oleic acid and linoleic acid) increase with seed development. Abscisic acid (ABA) reached a maximum value of 18.45 ng/g at 28 d, and gibberellin (GA3), 3-Indoleacetic acid (IAA) and zeatin-riboside (ZR) initially increased and then decreased. These results suggest that the vigour of E. brevicornu seeds is closely associated with their stage of development, with the highest vigour observed at 28~35 d after flowering.

Effect of Molecular Architecture and Symmetry on Self-Assembly: A Quantitative Revisit Using Discrete ABA Triblock Copolymers.

The inherent statistical heterogeneities associated with chain length, composition, and architecture of synthetic block copolymers compromise the quantitative interpretation of their self-assembly process. This study scrutinizes the contribution of molecular architecture on phase behaviors using discrete ABA triblock copolymers with precise chemical structure and uniform chain length. A group of discrete triblock copolymers with varying composition and symmetry were modularly synthesized through a combination of iterative growth methods and efficient coupling reactions. The symmetric ABA triblock copolymers self-assemble into long-range ordered structures with expanded domain spacings and enhanced phase stability, compared with the diblock counterparts snipped at the middle point. By tuning the relative chain length of two end blocks, the molecular asymmetry reduces the packing frustration, and thus increases the order-to-disorder transition temperature and enlarges the domain sizes. This study would serve as a quantitative model system to correlate the experimental observations with the theoretical assessments and to provide quantitative understandings for the relationship between molecular architecture and self-assembly.

Bt Cry1Ab/2Ab toxins disrupt the structure of the gut bacterial community of Locusta migratoria through host immune responses.

The gut microbiota of insects plays a vital role in digestion, nutrient acquisition, metabolism of dietary toxins, pathogen immunity and maintenance of gut homeostasis. Bacillus thuringinensis (Bt) poisons target insects through its toxins that are activated in the insect gut. The effects of Bt toxins on gut microbiota of insects and their underlying mechanisms are not well understood. In this study, we found that Cry1Ab/2Ab toxins significantly changed the gut bacterial community's structure and reduced the total load of gut bacteria in the Locusta migratoria. In addition, Cry toxins significantly increased the level of reactive oxygen species (ROS) in the gut of locusts. Our results also showed that Cry1Ab/2Ab toxins induced the host gut's immune response by up-regulating of key genes in the Immune deficiency (IMD) and Toll pathway. RNA interference showed that knocking down Relish could narrow the difference in the load, diversity, and composition in gut bacteria caused by Cry toxins. Our findings suggest that Bt potentially influences the gut bacterial community of L. migratoria through host immune response.

Agricultural Jiaosu: An Eco-Friendly and Cost-Effective Control Strategy for Suppressing Fusarium Root Rot Disease in Astragalus membranaceus.

Root rot caused by the pathogenic fungi of the Fusarium genus poses a great threat to the yield and quality of medicinal plants. The application of Agricultural Jiaosu (AJ), which contains beneficial microbes and metabolites, represents a promising disease control strategy. However, the action-effect of AJ on Fusarium root rot disease remains unclear. In the present study, we evaluated the characteristics and antifungal activity of AJ fermented using waste leaves and stems of medicinal plants, and elucidated the mechanisms of AJ action by quantitative real-time PCR and redundancy analysis. The effects of AJ and antagonistic microbes isolated from it on disease suppression were further validated through a pot experiment. Our results indicate that the AJ was rich in beneficial microorganisms (Bacillus, Pseudomonas, and Lactobacillus), organic acids (acetic, formic, and butyric acids) and volatile organic compounds (alcohols and esters). It could effectively inhibit Fusarium oxysporum and the half-maximal inhibitory concentration (IC50) was 13.64%. The antifungal contribution rate of the microbial components of AJ reached 46.48%. Notably, the redundancy analysis revealed that the Bacillus and Pseudomonas genera occupied the main niche during the whole inhibition process. Moreover, the abundance of the Bacillus, Pseudomonas, and Lactobacillus genera were positively correlated with the pH-value, lactic, formic and butyric acids. The results showed that the combined effects of beneficial microbes and organic acid metabolites increased the efficacy of the AJ antifungal activity. The isolation and identification of AJ's antagonistic microbes detected 47 isolates that exhibited antagonistic activities against F. oxysporum in vitro. In particular, Bacillus subtilis and Bacillus velezensis presented the strongest antifungal activity. In the pot experiment, the application of AJ and these two Bacillus species significantly reduced the disease incidence of Fusarium root rot and promoted the growth of Astragalus. The present study provides a cost-effective method to control of Fusarium root rot disease, and establishes a whole-plant recycling pattern to promote the sustainable development of medicinal plant cultivation.

Effect of genetically modified maize expressing the Cry1Ab and EPSPS proteins on growth, development, and gut bacterial diversity of the non-target arthropod Locusta migratoria.

The widespread cultivation of genetically modified (GM) crops has raised concerns for their safety. Here, we evaluated the effects of a GM maize variety expressing the Cry1Ab (14.76 ± 0.87 µg/g FW) and EPSPS proteins (191.55 ± 15.69 µg/g FW) on the life-history traits and gut bacterial community of a non-target arthropod, Locusta migratoria, in the laboratory. We found that GM maize had no significant effect on the survival or body weight of different development stages of L. migratoria. The midgut and hindgut bacterial diversities and compositions were determined using high-throughput sequencing targeting the V3-V4 regions of the 16S rRNA. No significant changes were found in the species diversity or abundance between insects in the GM-fed treatment and the non-GM control. Furthermore, the concentration of Cry1Ab and EPSPS in the gut was determined after digestion of GM maize. Results showed that the contents of Cry1Ab/EPSPS rapidly decreased and were hard to detect after 72 h. Based on the parameters assessed, we can conclude that the GM maize variety examined has no significant adverse effect on L. migratoria.

Carbohydrate regulation response to cold during rhizome bud dormancy release in Polygonatum kingianum.

BACKGROUND: The rhizome of Polygonatum kingianum Coll. et Hemsl (P. kingianum) is a crucial traditional Chinese medicine, but severe bud dormancy occurs during early rhizome development. Low temperature is a positive factor affecting dormancy release, whereas the variation in carbohydrates during dormancy release has not been investigated systematically. Therefore, the sugar content, related metabolic pathways and gene co-expression were analysed to elucidate the regulatory mechanism of carbohydrates during dormancy release in the P. kingianum rhizome bud. RESULTS: During dormancy transition, starch and sucrose (Suc) exhibited opposing trends in the P. kingianum rhizome bud, representing a critical indicator of dormancy release. Galactose (Gal) and raffinose (Raf) were increased in content and synthesis. Glucose (Glc), cellulose (Cel), mannose (Man), arabinose (Ara), rhamnose (Rha) and stachyose (Sta) showed various changes, indicating their different roles in breaking rhizome bud dormancy in P. kingianum. At the beginning of dormancy release, Glc metabolism may be dominated by anaerobic oxidation (glycolysis followed by ethanol fermentation). After entering the S3 stage, the tricarboxylic acid cycle (TCA) and pentose phosphate pathway (PPP) were may be more active possibly. In the gene co-expression network comprising carbohydrates and hormones, HYD1 was identified as a hub gene, and numerous interactions centred on STS/SUS were also observed, suggesting the essential role of brassinosteroids (BRs), Raf and Suc in the regulatory network. CONCLUSION: We revealed cold-responsive genes related to carbohydrate metabolism, suggesting regulatory mechanisms of sugar during dormancy release in the P. kingianum rhizome bud. Additionally, gene co-expression analysis revealed possible interactions between sugar and hormone signalling, providing new insight into the dormancy release mechanism in P. kingianum rhizome buds.

Identification of ST1 reveals a selection involving hitchhiking of seed morphology and oil content during soybean domestication.

Seed morphology and quality of cultivated soybean (Glycine max) have changed dramatically during domestication from their wild relatives, but their relationship to selection is poorly understood. Here, we describe a semi-dominant locus, ST1 (Seed Thickness 1), affecting seed thickness and encoding a UDP-D-glucuronate 4-epimerase, which catalyses UDP-galacturonic acid production and promotes pectin biosynthesis. Interestingly, this morphological change concurrently boosted seed oil content, which, along with up-regulation of glycolysis biosynthesis modulated by ST1, enabled soybean to become a staple oil crop. Strikingly, ST1 and an inversion controlling seed coat colour formed part of a single selective sweep. Structural variation analysis of the region surrounding ST1 shows that the critical mutation in ST1 existed in earlier wild relatives of soybean and the region containing ST1 subsequently underwent an inversion, which was followed by successive selection for both traits through hitchhiking during selection for seed coat colour. Together, these results provide direct evidence that simultaneously variation for seed morphology and quality occurred earlier than variation for seed coat colour during soybean domestication. The identification of ST1 thus sheds light on a crucial phase of human empirical selection in soybeans and provides evidence that our ancestors improved soybean based on taste.

Precisely Encoding Geometric Features into Discrete Linear Polymer Chains for Robust Structural Engineering.

Molecular shape is an essential parameter that regulates the self-organization and recognition process, which has not yet been well appreciated and exploited in block polymers due to the lack of precise and efficient modulation methods. This work (i) develops a robust approach to break the intrinsic symmetry of linear polymers by introducing geometric features into otherwise homogeneous chains and (ii) quantitatively highlights the critical contribution of molecular geometry/architecture to the self-assembly behaviors. Iteratively connecting homologous monomers of different side chains according to pre-designed sequences generates discrete polymers with exact chemical structure, uniform chain length, and programmable side-chain gradient along the backbone, which transcribes into diverse shapes. The precise chemistry eliminates all the defects and heterogeneities, providing a delicate platform for fundamental inquiries into the role of molecular geometry. A rich collection of unconventional complex phases, including Frank-Kasper A15 and σ phases, as well as a dodecagonal quasicrystal phase, were captured in these rigorous single-component systems. The self-assembly behaviors are strikingly sensitive to subtle variations of geometry, such that simply migrating a few methylene units among the side chains would generate substantial differences in lattice size or phase stability, or even trigger a phase transition toward distinct structures. The phenomena can be rationalized with a geometric argument that nonuniform side chain distribution leads to conformational mismatch between two immiscible blocks, resulting in varied interfacial curvatures and distinct lattice symmetries. The profound contribution demonstrates that molecular geometry is an effective and robust parameter for structural engineering.

Discrete Giant Polymeric Chains Based on Nanosized Monomers.

As size-amplified analogues of canonical macromolecules, polymeric chains built up by "giant" monomers represent an experimental realization of the "beads-on-a-string" model at larger length scales, which could provide insights into fundamental principles of polymer science. In this work, we modularly constructed discrete giant polymeric chains using nanosized building blocks (polyhedral oligomeric silsesquioxane, POSS) as basic repeat units through an efficient and robust iterative exponential growth approach, with precise control on molecular parameters, including size, composition, regioconfiguration, and surface functionalities. Their chemical structures were fully characterized by nuclear magnetic resonance spectroscopy, size-exclusion chromatography, and matrix-assisted laser desorption ionization time-of-flight mass spectrometry. With elaborately designed amphiphilic block POSS chains and their analogues made of conventional monomers, the size effects were delicately studied and highlighted. Interesting assembly behaviors emerge as a result of distinct interactions and molecular dynamics. This category of molecules shares general self-assembly characteristics as the conventional counterparts in terms of phase transition and evolution. Meanwhile, it turns out that the monomer size has profound impacts on phase stability, as a trade-off between entropic and enthalpic contributions. It may open up a door for modular and programmable design of interesting materials with complex structures and diverse functions.

Precise Amphiphilic Giant Polymeric Chain Based on Nanosized Monomers with Exact Regio-Configuration.

Polymeric chains made of "giant" monomers at a larger length scale provide intriguing insights into the fundamental principles of polymer science. In this study, we modularly prepared a library of discrete amphiphilic polymeric chains using molecular nanoparticles as repeat units, with exact control of composition, chain length, surface property, and regio-configuration. These giant polymeric chains self-assembled into a rich collection of highly ordered phases. The precise chemical structure and uniform chain length eliminate all the inherent molecular "defects", while the nanosized monomer amplifies minute structural differences, providing an ideal platform for a systematic scrutiny of the self-assembly behaviors at a larger length scale. The compositional and regio-configurational contribution was carefully studied. The regio-regularity is found to have a direct and profound impact on the chain conformation, leading to a distinct molecular packing scheme and therefore shifting the phase boundaries. With increasing the length of the linker, the regio-constraint gradually diminishes, and the neighboring particles would eventually be decoupled.

Confined Self-Assemblies of Chiral Block Copolymers in Thin Films.

Self-assembly of chiral block copolymers (BCPs*) can give rise to ordered chiral nanostructures, that is, a helical phase (H* phase), via chirality transfer from the molecular level to mesoscale. In the present work, we reported the self-assembly of BCPs* under one-dimensional spatial confinement. The morphological dependence of self-assembled BCPs* on the molecular weights and the film thickness was investigated. As chiral nanostructures, the H* phase can be formed in bulk, nonchiral nanostructures that were observed in the thin films. Also, the topology effect of self-assembly of BCPs* was examined. The self-assembly of BCPs* with a star-shaped topology exhibited a distinct morphology compared with that of linear BCPs*. The present work provides new insight into the chirality transfer of macromolecules under spatial confinement.

Discrete Block Copolymers with Diverse Architectures: Resolving Complex Spherical Phases with One Monomer Resolution.

This work describes the first rigorous example of a single-component block copolymer system forming unconventional spherical phases. A library of discrete block polymers with uniform chain length and diverse architectures were modularly prepared through a combination of a step-growth approach and highly efficient coupling reactions. The precise chemical structure eliminates all the molecular defects associated with molar weight, dispersity, and compositional ratio. Complex spherical phases, including the Frank-Kasper phase (A15 and σ) and quasicrystalline phase, were experimentally captured by meticulously tuning the composition and architectures. A phase portrait with unprecedented accuracy was mapped out (up to one monomer resolution), unraveling intriguing details of phase behaviors that have long been compromised by inherent molecular weight distribution. This study serves as a delicate model system to bridge the existing gaps between experimental observations and theoretical assessments and to provide insights into the formation and evolution of the unconventional spherical phases in soft matter systems.

Molecular Patchy Clusters with Controllable Symmetry Breaking for Structural Engineering.

Anisotropic patchy particles with molecular precision are exquisite building blocks for constructing diverse meso-structures of high complexity. In this research, a library of molecular patchy clusters consisting of a collection of functional polyhedral oligomeric silsesquioxane cages with exact regio-configuration and composition were prepared through a robust and modular approach. By meticulously tuning the composition, molecular symmetry, and other parameters, these patchy clusters could assemble into diverse nanostructures, including unconventional complex spherical phases (i.e., Frank-Kasper σ phase and dodecagonal quasicrystalline phase). As the size of the hydrophilic patch expands, a transition sequence from disorder to hexagonally packed cylinders and then to double gyroids was recorded, corresponding to a progressive decrease of interfacial curvature. On the other hand, regioisomers with the same composition but different regio-configuration adopt similar molecular packing but varied phase stability, as a result of the local self-sorting process to alleviate excess unfavorable interfacial contact. These precisely defined molecular patchy clusters provide a model system for a general understanding of the hierarchical structure formation and evolution based on anisotropic spherical building blocks at the nanoscale.

Characterizing rhizome bud dormancy in Polygonatum kingianum: Development of novel chill models and determination of dormancy release mechanisms by weighted correlation network analysis.

This study was conducted to explore specific chill models and the mechanisms underlying rhizome bud dormancy break in Polygonatum kingianum. Rhizome buds were subjected to various chilling temperatures for different duration and then transferred to warm conditions for germination and subsequent evaluation of their response to temperature and chilling requirements. A CUkingianum model was constructed to describe the contribution of low temperature to the chill unit, and it was suggested that 2.97°C was the optimum temperature and that 11.54°C was the upper limit for bud release. The CASkingianum model showed the relationship between chilling accumulation and sprouting percentage; therefore, rhizome bud development could be predicted through the model. Weighted correlation network analysis (WGCNA) of transcriptomic data of endo-, eco- and nondormant rhizome buds generated 33 gene modules, 6 of which were significantly related to bud sprouting percentage. In addition, 7 significantly matched transcription factors (TFs) were identified from the promoters of 17 "real" hub genes, and DAG2 was the best matched TF that bound to AAAG element to regulate gene expression. The current study is valuable for developing a highly efficient strategy for seedling cultivation and provides strong candidates for key genes related to rhizome bud dormancy in P. kingianum.