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Hepatic fibrosis is a chronic liver disease that lacks effective pharmacotherapeutic treatments. As part of the disease's mechanism, hepatic stellate cells (HSCs) are activated by damage-related stimuli to secrete excessive extracellular matrix, leading to collagen deposition. Currently, the drug delivery system that targets HSCs in the treatment of liver fibrosis remains an urgent challenge due to the poor controllability of drug release. Since the level of reactive oxygen species (ROS) increases sharply in activated HSCs (aHSCs), we designed ROS-responsive micelles for the HSC-specific delivery of a traditional Chinese medicine, resveratrol (RES), for treatment of liver fibrosis. The micelles were prepared by the ROS-responsive amphiphilic block copolymer poly(l-methionine

Strigolactones (SLs) constitute a class of plant hormones that regulate many aspects of plant development, including repressing tillering in rice (Oryza sativa). However, how SL pathways are regulated is still poorly understood. Here, we describe a rice mutant dwarf and high tillering1 (dht1), which exhibits pleiotropic phenotypes (such as dwarfism and increased tiller numbers) similar to those of mutants defective in SL signaling. We show that DHT1 encodes a monocotyledon-specific hnRNP-like protein that acts as a previously unrecognized intron splicing factor for many precursor mRNAs (pre-mRNAs), including for the SL receptor gene D14. We find that the dht1 (DHT1I232F) mutant protein is impaired in its stability and RNA binding activity, causing defective splicing of D14 pre-mRNA and reduced D14 expression, and consequently leading to the SL signaling-defective phenotypes. Overall, our findings deepen our understanding of the functional diversification of hnRNP-like proteins and establish a connection between posttranscriptional splicing and SL signaling in the regulation of plant development.

Vesicular trafficking plays critical roles in cell expansion in yeast and mammals, but information linking vesicular trafficking and cell expansion in plants is limited. Here, we isolated and characterized a rice (Oryza sativa) mutant, decreased plant height 1-1 (dph1-1), which exhibited a wide spectrum of developmental phenotypes, including reduced plant height and smaller panicles and grains. Cytological analysis revealed that limited cell expansion was responsible for the dph1-1 mutant phenotype compared to the wild-type. Map-based cloning revealed that DPH1 encodes a plant-specific protein, OsSCD2, which is homologous to Arabidopsis (Arabidopsis thaliana) STOMATAL CYTOKINESIS DEFECTIVE2 (SCD2). Subcellular localization revealed that OsSCD2 is associated with clathrin. Confocal microscopy showed that the dph1-1 mutant has defective endocytosis and post-Golgi trafficking. Biochemical and confocal data indicated that OsSCD2 physically interacts with OsSCD1 and that they are associated with intracellular structures that colocalize with microtubules. Furthermore, we found that cellulose synthesis was affected in the dph1-1 mutant, evidenced by reduced cellulose synthase gene accumulation at the transcript and protein levels, most likely resulting from an impaired localization pattern. Our results suggest that OsSCD2 is involved in clathrin-related vesicular trafficking with an important role in maintaining plant growth in rice.

Endosperm as the storage organ of starch and protein in cereal crops largely determines grain yield and quality. Despite the fact that several pentatricopeptide repeat (PPR) proteins required for endosperm development have been identified in rice, the molecular mechanisms of many P-type PPR proteins in endosperm development remains unclear. Here, we isolated a rice floury endosperm mutant ppr5 that developed small starch grains and an abnormal aleurone layer, accompanied by decreased starch, protein, and amylose contents. Map-based cloning combined with a complementation test demonstrated that PPR5 encodes a P-type PPR protein that is localized to the mitochondria. The mutation in PPR5 caused reduced splicing efficiency of mitochondrial NADH dehydrogenase 4 (nad4) gene intron 3 and reduced complex I assembly and activity. Loss of PPR5 function greatly up-regulated expression of alternative oxidases (AOXs), reduced ATP production, and affected mitochondrial morphology. We demonstrate that PPR5, as a P-type PPR protein, is required for mitochondrial function and endosperm development by controlling the cis-splicing of mitochondrial nad4 intron 3.

Heading date (or flowering time) is one of the most important agronomic traits in rice, influencing its regional adaptability and crop yield. Many major-effect genes for rice heading date have been identified, but in practice they are difficult to be used for rice molecular breeding because of their dramatic effects on heading date. Genes with minor effects on heading date, which are more desirable for fine-tuning flowering time without significant yield penalty, were seldom reported. In this study, we identified a new minor-effect heading date repressor, Delayed Heading Date 4 (DHD4). The dhd4 mutant shows a slightly earlier flowering phenotype without a notable yield penalty compared with wild-type plants under natural long-day conditions. DHD4 encodes a CONSTANS-like transcription factor localized in the nucleus. Molecular, biochemical, and genetic assays show that DHD4 can compete with 14-3-3 to interact with OsFD1, thus affecting the formation of the Hd3a-14-3-3-OsFD1 tri-protein FAC complex, resulting in reduced expression of OsMADS14 and OsMADS15, and ultimately delaying flowering. Taken together, these results shed new light on the regulation of flowering time in rice and provide a promising target for fine-tuning flowering time to improve the regional adaptability of rice.

Summary Thioredoxins (TRXs) occur in plant chloroplasts as complex disulphide oxidoreductases. Although many biological processes are regulated by thioredoxins, the regulatory mechanism of chloroplast TRXs are largely unknown.Here we report a rice white panicle2 mutant caused by a mutation in the thioredoxin z gene, an orthologue of AtTRX z in Arabidopsis. white panicle2 (wp2) seedlings exhibited a high‐temperature‐sensitive albinic phenotype.We found that plastid multiple organellar RNA editing factors (MORFs) were the regulatory targets of thioredoxin z. We showed that OsTRX z protein physically interacts with OsMORFs in a redox‐dependent manner and that the redox state of a conserved cysteine in the MORF box is essential for MORF–MORF interactions. wp2 and OsTRX z knockout lines show reduced editing efficiencies in many plastidial‐encoded genes especially under high‐temperature conditions. An Arabidopsis trx z mutant also exhibited significantly reduced chloroplast RNA editing.Our combined results suggest that thioredoxin z regulates chloroplast RNA editing in plants by controlling the redox state of MORFs.

This paper concerns a disassembly line balancing problem (DLBP) in remanufacturing that aims to allocate a set of tasks to workstations to disassemble a product. We consider two objectives in the same time, i.e., minimising the number of workstations required and minimising the operating costs. A common approach to such problems is to covert the multiple objectives into a single one and solve the resulting problem with either exact or heuristic methods. However, the appropriate weights must be determined a priori, yet the results provide little insight on the trade-off between competing objectives. Moreover, DLBP problems are proven NP-complete and thus the solvable instances by exact methods are limited. To this end, we formulate the problem into a multi-objective dynamic program and prove the monotonicity property of both objective functions. A backward recursive algorithm is developed to efficiently generate all the non-dominated solutions. The numerical results show that our proposal is more efficient than alternative exact algorithms proposed in the literature and can handle much larger problem instances.

Summary Heading date is a key agronomic trait affecting crop yield. In rice, Early heading date 1 (Ehd1) is an important B‐type response regulator in determination of heading date. Although many regulatory factors of Ehd1 expression have been functionally characterized, the direct regulators of Ehd1 largely remain to be identified. Here, we identified a new regulator of Ehd1, OsRE1, that directly binds to the A‐box motif in the Ehd1 promoter. Osre1 confers an early heading phenotype due to elevated expression levels of Ehd1. OsRE1 is a nucleus‐localized bZIP transcription factor with a diurnal rhythmic expression pattern. Furthermore, we identified an OsRE1‐interacting protein, OsRIP1, and demonstrated that OsRIP1 can repress the transcript expression of Ehd1 in an OsRE1‐dependent manner. Our genetic data showed that OsRE1 and OsRIP1 may function upstream of Ehd1 in regulating heading date. Together, our results suggest that OsRE1 functions cooperatively with OsRIP1 to regulate heading date through finely modulating the expression of Ehd1. In addition, OsRE1 and OsRIP1 are two minor heading date regulators, which are more desirable for fine‐tuning heading date to improve rice regional adaptability.

A partial disassembly line balancing problem under uncertainty is studied in this paper, which concerns the allocation of a sequence of tasks to workstations such that the overall profit is maximised. We consider the processing time uncertainty and develop robust solutions to accommodate it. The problem is formulated as a non-linear robust integer program, which is then converted into an equivalent linear program. Due to the intractability of such problems, the exact algorithms are only applicable to small-scale instances. We develop an improved migrating birds optimisation algorithm. Two enhancement techniques are proposed. The first one finds the optimal number of tasks to be performed for each sequence rather than random selection used in the literature; while the second one exploits the specific problem structure to construct effective neighbourhoods. The numerical results show the strong performance of our proposal compared to CPLEX and the improved gravitational search algorithm (IGSA), especially for large-scale problems. Moreover, the enhancement due to the proposed techniques is obvious across all instances considered.

The microgrid technology, which can dispatch power independently, is an effective way to increase the efficiency of energy utilization meanwhile develop and utilize the clean and renewable energy. However, the power generation of a single microgrid is unstable, because it is greatly affected by the external environment. Therefore, the development and application of the multi-microgrid system have gradually drawn various countries' attention. In order to minimize the operating cost and gaseous pollutant emission of the multi-microgrid system, which is composed of renewable energies and electric vehicles and so on, this paper builds a 24 hours day-ahead multi-objective complex constrained optimization model, using interval optimization to handle uncertainties of renewable energies. In view of the model characteristics, the metaheuristic strategies about initialization and repair of solution are designed. Furthermore, the fuzzy membership degree and Chebyshev function are used in parallel to decompose the multi-objective optimization problem, thus a multi-objective evolutionary algorithm based on hybrid decomposition (MOEA/HD) is constructed. Finally, the effectiveness of the metaheuristic strategies can be verified by analyzing the simulation results in this paper. Moreover, the results prove that the MOEA/HD is more efficient, which can get a higher-quality Pareto optimal solution set when compared to other algorithms.

The transient elevation of cytoplasmic calcium is essential for pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI). However, the calcium channels responsible for this process have remained unknown. Here, we show that rice CDS1 (CELL DEATH and SUSCEPTIBLE to BLAST 1) encoding OsCNGC9, a cyclic nucleotide-gated channel protein, positively regulates the resistance to rice blast disease. We show that OsCNGC9 mediates PAMP-induced Ca(2+) influx and that this event is critical for PAMPs-triggered ROS burst and induction of PTI-related defense gene expression. We further show that a PTI-related receptor-like cytoplasmic kinase OsRLCK185 physically interacts with and phosphorylates OsCNGC9 to activate its channel activity. Our results suggest a signaling cascade linking pattern recognition to calcium channel activation, which is required for initiation of PTI and disease resistance in rice.

OsVIN2, a vacuolar invertase, affects grain size and yield by altering sugar composition, transport, and starch accumulation in rice. Grain size, a major determinant of rice yield, is influenced by many developmental and environmental factors. Sugar metabolism plays vital roles in plant development. However, the way in which sugar metabolism affects rice grain size remains largely elusive. In this study, we characterized the small grain-size rice mutant sgs1. Histological analyses showed that reduced spikelet hull and endosperm size results from decreased cell size rather than cell number. Map-based cloning and complementation tests revealed that a DaiZ7 transposon insertion in a vacuolar invertase gene OsVIN2 is responsible for the mutant phenotype. Subcellular distribution and biochemical analysis indicated that OsVIN2 is located in the vacuolar lumen, and that its sucrose hydrolysis activity is maintained under acidic conditions. Furthermore, an altered sugar content with increased sucrose and decreased hexose levels, as well as changes in invertase and sucrose synthase activities, sugar transport gene expression, and starch constitution in sgs1 implies that OsVIN2 affects sucrose metabolism, including sugar composition, transport, and conversion from the source to the sink organs. Collectively, OsVIN2 is involved in sugar metabolism, and thus regulates grain size; our findings provide insights into grain development and also suggest a potential strategy to improve grain quality and yield in rice.

Endosperm, the major storage organ in cereal grains, determines grain yield and quality. Despite the fact that a role for P-type pentatricopeptide repeat (PPR) proteins in the regulation of endosperm development has emerged, molecular functions of many P-type PPR proteins remain obscure. Here, we report a rice endosperm defective mutant, floury endosperm10 (flo10), which developed smaller starch grains in starchy endosperm and abnormal cells in the aleurone layer. Map-based cloning and rescued experiments showed that FLO10 encodes a P-type PPR protein with 26 PPR motifs, which is localized to mitochondria. Loss of function of FLO10 affected the trans-splicing of the mitochondrial nad1 intron 1, which was accompanied by the increased accumulation of the nad1 exon 1 and exons 2-5 precursors. The failed formation of mature nad1 led to a dramatically decreased assembly and activity of complex I, reduced ATP production, and changed mitochondrial morphology. In addition, loss of function of FLO10 significantly induced an alternative respiratory pathway involving alternative oxidase. These results reveal that FLO10 plays an important role in the maintenance of mitochondrial function and endosperm development through its effect on the trans-splicing of the mitochondrial nad1 intron 1 in rice.

Summary Starch is the most important form of energy storage in cereal crops. Many key enzymes involved in starch biosynthesis have been identified. However, the molecular mechanisms underlying the regulation of starch biosynthesis are largely unknown. In this study, we isolated a novel floury endosperm rice (Oryza sativa) mutant flo16 with defective starch grain (SG) formation. The amylose content and amylopectin structure were both altered in the flo16 mutant. Map‐based cloning and complementation tests demonstrated that FLO16 encodes a NAD‐dependent cytosolic malate dehydrogenase (CMDH). The ATP contents were decreased in the mutant, resulting in significant reductions in the activity of starch synthesis‐related enzymes. Our results indicated that FLO16 plays a critical role in redox homeostasis that is important for compound SG formation and subsequent starch biosynthesis in rice endosperm. Overexpression of FLO16 significantly improved grain weight, suggesting a possible application of FLO16 in rice breeding. These findings provide a novel insight into the regulation of starch synthesis and seed development in rice.

Tillering and panicle branching are important determinants of plant architecture and yield potential in rice (Oryza sativa). IDEAL PLANT ARCHITECTURE1 (IPA1) encodesSQUAMOSA PROMOTER BINDING PROTEIN-LIKE14, which acts as a key transcription factor regulating tiller outgrowth and panicle branching by directly activating the expression of O. sativa TEOSINTE BRANCHED1 (OsTB1) and O. sativa DENSE AND ERECT PANICLE1 (OsDEP1), thereby influencing grain yield in rice. Here, we report the identification of a rice mutant named shi1 that is characterized by dramatically reduced tiller number, enhanced culm strength, and increased panicle branch number. Map-based cloning revealed that O. sativa SHORT INTERNODES1 (OsSHI1) encodes a plant-specific transcription factor of the SHI family with a characteristic family-specific IGGH domain and a conserved zinc-finger DNA binding domain. Consistent with the mutant phenotype, OsSHI1 is predominantly expressed in axillary buds and young panicle, and its encoded protein is exclusively targeted to the nucleus. We show that OsSHI1 physically interacts with IPA1 both in vitro and in vivo. Moreover, OsSHI1 could bind directly to the promoter regions of both OsTB1 and OsDEP1 through a previously unrecognized cis-element (T/GCTCTAC motif). OsSHI1 repressed the transcriptional activation activity of IPA1 by affecting its DNA binding activity toward the promoters of both OsTB1 and OsDEP1, resulting in increased tiller number and diminished panicle size. Taken together, our results demonstrate that OsSHI1 regulates plant architecture through modulating the transcriptional activity of IPA1 and provide insight into the establishment of plant architecture in rice.

Summary Lesion mimic mutants that exhibit spontaneous hypersensitive response (HR)‐like necrotic lesions are ideal experimental systems for elucidating molecular mechanisms involved in plant cell death and defence responses. Here we report identification of a rice lesion mimic mutant, spotted leaf 35 (spl35), and cloning of the causal gene by TAIL‐PCR strategy. spl35 exhibited decreased chlorophyll content, higher accumulation of H2O2, up‐regulated expression of defence‐related marker genes, and enhanced resistance to both fungal and bacterial pathogens of rice. The SPL35 gene encodes a novel CUE (coupling of ubiquitin conjugation to ER degradation) domain‐containing protein that is predominantly localized in cytosol, ER and unknown punctate compartment(s). SPL35 is constitutively expressed in all organs, and both overexpression and knockdown of SPL35 cause the lesion mimic phenotype. SPL35 directly interacts with the E2 protein OsUBC5a and the coatomer subunit delta proteins Delta‐COP1 and Delta‐COP2 through the CUE domain, and down‐regulation of these interacting proteins also cause development of HR‐like lesions resembling those in spl35 and activation of defence responses, indicating that SPL35 may be involved in the ubiquitination and vesicular trafficking pathways. Our findings provide insight into a role of SPL35 in regulating cell death and defence response in plants.

Ubiquitination and deubiquitination are reversible processes that play crucial roles in regulating organ size in plants. However, information linking deubiquitination and seed size in rice (Oryza sativa) is limited. Here, we characterized a dominant large-grain mutant, large grain1-D (lg1-D), with a 30.8% increase in seed width and a 34.5% increase in 1,000-grain weight relative to the wild type. The lg1-D mutant had more cells oriented in the lateral direction of the spikelet hull compared with the wild type. Map-based cloning showed that LG1 encodes a constitutively expressed ubiquitin-specific protease15 (OsUBP15) that possesses deubiquitination activity in vitro. Loss-of-function and down-regulated expression of OsUBP15 produced narrower and smaller grains than the control. A set of in vivo experiments indicated that the mutant Osubp15 had enhanced protein stability relative to wild-type OsUBP15. Further experiments verified that OsDA1 directly interacted with OsUBP15. Genetic data indicated that OsUBP15 and GRAIN WIDTH 2 (GW2) were not independent in regulating grain width and size. In summary, we identified OsUBP15 as a positive regulator of grain width and size in rice and provide a promising strategy for improvement of grain yield by pyramiding OsUBP15 and gw2.

Starch is a major storage substance in cereal grains, and starch biosynthesis is a complex process. In order to elucidate regulation of the starch biosynthesis pathway, we screened a series of rice (Oryza sativa L.) endosperm mutants. In this study, we identified a floury white-core endosperm mutant named floury endosperm12 (flo12). The flo12 mutant exhibited loosely packed starch granules and a lower thousand kernel weight compared to wild type. Semithin sections revealed that compound starch grains (SG) in flo12 interior endosperm cells were developed abnormally. Furthermore, amylose content was decreased, while total protein content was significantly increased in flo12 grains. Map-based cloning showed that FLO12 encodes rice alanine aminotransferase 1 (OsAlaAT1). OsAlaAT1 is highly expressed in developing endosperm. Subcellular localization showed that OsAlaAT1 is localized in the cytosol. Moreover, the expression of most starch synthesis-related genes was decreased, while most of the storage protein coding genes had elevated expression levels in the flo12 mutant. In addition, overexpression of the OsAlaAT1 gene increased grain weight. In brief, we demonstrated that OsAlaAT1 regulates carbon and nitrogen metabolism, which provides a new insight for the improvement of rice quality and yield.

This study deals with a sequence-dependent disassembly line balancing problem by considering the interactions among disassembly tasks, and a multi-objective mathematical model is establishe...

Anthocyanins cause purple, brown or red colors in various tissues of rice plants, but the specific determinant factors and regulatory systems for anthocyanin biosynthesis in almost all tissues remain largely unknown. In the present study, we mapped and isolated two complementary genes, OsC1 encoding a R2R3-MYB transcriptional factor and OsDFR encoding a dihydroflavonol 4-reductase, which are responsible for the purple coloration of apiculi and stigmas in indica cultivar Xieqingzao by the map-based cloning strategy. We also identified two tissue-specific pigmentation genes, OsPa for apiculi and OsPs for stigmas, by phylogenetic analysis of all anthocyanin biosynthesis-associated bHLH transcriptional factors in maize and rice, CRISPR/Cas9 knockout and transcriptional expression analysis. The OsC1, OsPa and OsPs proteins are all localized in the nucleus while the OsDFR protein is localized in the nucleus and cytoplasm, and the OsC1 and OsDFR genes are preferentially strongly expressed in both purple-colored tissues while the OsPa and OsPs genes are preferentially strongly expressed in apiculi and stigmas, respectively. OsC1 specifically interacts with OsPa or OsPs to activate OsDFR and other anthocyanin biosynthesis genes, resulting in purple-colored apiculi or stigmas. OsC1 itself does not produce color but can produce brown apiculi when functioning together with OsPa. Loss of function of OsDFR alone leads to brown apiculi and straw-white stigmas. Genotyping and phenotyping of a panel of 176 rice accessions revealed diverse genotypic combinations of OsC1, OsDFR, OsPa and OsPs that enable accurate prediction of their apiculus and stigma pigmentation phenotypes, thus validating the general applicability of the OsC1-OsDFR-OsPa and OsC1-OsDFR-OsPs models to natural populations. Our findings disclosed the biological functions of OsC1, OsPa and OsPs, and shed light on the specific regulatory systems of anthocyanin biosynthesis in apiculi and stigmas, a further step in understanding the regulatory network of anthocyanin biosynthesis in rice.

Additional file 15: Table S3. Amino acids sequences alignment of three bHLH transcription factors.