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Plant architecture is an important agronomic trait that affects crop yield. Here, we report that a gene involved in programmed cell death, OsPDCD5, negatively regulates plant architecture and grain yield in rice. We used the CRISPR/Cas9 system to introduce loss-of-function mutations into OsPDCD5 in 11 rice cultivars. Targeted mutagenesis of OsPDCD5 enhanced grain yield and improved plant architecture by increasing plant height and optimizing panicle type and grain shape. Transcriptome analysis showed that OsPDCD5 knockout affected auxin biosynthesis, as well as the gibberellin and cytokinin biosynthesis and signaling pathways. OsPDCD5 interacted directly with OsAGAP, and OsAGAP positively regulated plant architecture and grain yield in rice. Collectively, these findings demonstrate that OsPDCD5 is a promising candidate gene for breeding super rice cultivars with increased yield potential and superior quality.
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Proteínas Reguladoras de la Apoptosis/metabolismo , Grano Comestible/crecimiento & desarrollo , Proteínas de Plantas/metabolismo , Estructuras de las Plantas/crecimiento & desarrollo , Proteínas Reguladoras de la Apoptosis/genética , Citocininas/metabolismo , Grano Comestible/genética , Regulación de la Expresión Génica de las Plantas , Giberelinas/metabolismo , Ácidos Indolacéticos/metabolismo , Mutagénesis , Oryza/genética , Oryza/crecimiento & desarrollo , Fitomejoramiento , Proteínas de Plantas/genética , Estructuras de las Plantas/genética , Plantas Modificadas Genéticamente , Unión Proteica , Transducción de Señal/genéticaRESUMEN
Spikelet number per panicle (SNP) is one of the most important yield components in rice. Rice ENHANCING BIOMASS AND SPIKELET NUMBER (OsEBS), a gene involved in improved SNP and yield, has been cloned from an accession of Dongxiang wild rice. However, the mechanism of OsEBS increasing rice SNP is poorly understood. In this study, the RNA-Seq technology was used to analyze the transcriptome of wildtype Guichao 2 and OsEBS over-expression line B102 at the heading stage, and analysis of the evolution of OsEBS was also conducted. A total of 5369 differentially expressed genes (DEGs) were identified between Guichao2 and B102, most of which were down-regulated in B102. Analysis of the expression of endogenous hormone-related genes revealed that 63 auxin-related genes were significantly down-regulated in B102. Gene Ontogeny (GO) enrichment analysis showed that the 63 DEGs were mainly enriched in eight GO terms, including auxin-activated signaling pathway, auxin polar transport, auxin transport, basipetal auxin transport, and amino acid transmembrane transport, most of which were directly or indirectly related to polar auxin transport. Kyoto Encyclopedia of Genes and Genomes (KEGG) metabolic pathway analysis further verified that the down-regulated genes related to polar auxin transport had important effects on increased SNP. Analysis of the evolution of OsEBS found that OsEBS was involved in the differentiation of indica and japonica, and the differentiation of OsEBS supported the multi-origin model of rice domestication. Indica (XI) subspecies harbored higher nucleotide diversity than japonica (GJ) subspecies in the OsEBS region, and XI experienced strong balancing selection during evolution, while selection in GJ was neutral. The degree of genetic differentiation between GJ and Bas subspecies was the smallest, while it was the highest between GJ and Aus. Phylogenetic analysis of the Hsp70 family in O. sativa, Brachypodium distachyon, and Arabidopsis thaliana indicated that changes in the sequences of OsEBS were accelerated during evolution. Accelerated evolution and domain loss in OsEBS resulted in neofunctionalization. The results obtained from this study provide an important theoretical basis for high-yield rice breeding.
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Oryza , RNA-Seq , Oryza/genética , Filogenia , Fitomejoramiento , Perfilación de la Expresión Génica , TranscriptomaRESUMEN
Amylose content (AC) is the main factor determining the palatability, viscosity, transparency, and digestibility of rice (Oryza sativa) grains. AC in rice grains is mainly controlled by different alleles of the Waxy (Wx) gene. The AP2/EREBP transcription factor OsEBP89 interacts with the MYC-like protein OsBP5 to synergistically regulate the expression of Wx. Here, we determined that the GLYCOGEN SYNTHASE KINASE 5 (OsGSK5, also named SHAGGY-like kinase 41 [OsSK41]) inhibits the transcriptional activation activity of OsEBP89 in rice grains during amylose biosynthesis. The loss of OsSK41 function enhanced Wx expression and increased AC in rice grains. By contrast, the loss of function of OsEBP89 reduced Wx expression and decreased AC in rice grains. OsSK41 interacts with OsEBP89 and phosphorylates four of its sites (Thr-28, Thr-30, Ser-238, and Thr-257), which makes OsEBP89 unstable and attenuates its interaction with OsBP5. Wx promoter activity was relatively weak when regulated by the phosphomimic variant OsEBP89E -OsBP5 but relatively strong when regulated by the nonphosphorylatable variant OsEBP89A -OsBP5. Therefore, OsSK41-mediated phosphorylation of OsEBP89 represents an additional layer of complexity in the regulation of amylose biosynthesis during rice grain development. In addition, our findings provide four possible sites for regulating rice grain AC via precise gene editing.
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Endospermo , Oryza , Endospermo/metabolismo , Amilosa/metabolismo , Oryza/metabolismo , Regiones Promotoras Genéticas , Grano Comestible/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMEN
Auxin is an important phytohormone in plants, and auxin signaling pathways in rice play key roles in regulating its growth, development, and productivity. To investigate how rice grain yield traits are regulated by auxin signaling pathways and to facilitate their application in rice improvement, we validated the functional relationships among regulatory genes such as OsIAA10, OsSK41, and OsARF21 that are involved in one of the auxin (OsIAA10) signaling pathways. We assessed the phenotypic effects of these genes on several grain yield traits across two environments using knockout and/or overexpression transgenic lines. Based on the results, we constructed a model that showed how grain yield traits were regulated by OsIAA10 and OsTIR1, OsAFB2, and OsSK41 and OsmiR393 in the OsSK41-OsIAA10-OsARF module and by OsARF21 in the transcriptional regulation of downstream auxin response genes in the OsSK41-OsIAA10-OsARF module. The population genomic analyses revealed rich genetic diversity and the presence of major functional alleles at most of these loci in rice populations. The strong differentiation of many major alleles between Xian/indica and Geng/japonica subspecies and/or among modern varieties and landraces suggested that they contributed to improved productivity during evolution and breeding. We identified several important aspects associated with the genetic and molecular bases of rice grain and yield traits that were regulated by auxin signaling pathways. We also suggested rice auxin response factor (OsARF) activators as candidate target genes for improving specific target traits by overexpression and/or editing subspecies-specific alleles and by searching and pyramiding the 'best' gene allelic combinations at multiple regulatory genes in auxin signaling pathways in rice breeding programs.
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Oryza , Oryza/metabolismo , Fitomejoramiento , Fenotipo , Ácidos Indolacéticos/metabolismo , Grano Comestible/genética , Grano Comestible/metabolismoRESUMEN
BACKGROUND: Rice eating quality largely dictates consumer preference, and the demand for new rice varieties with excellent eating quality from farmers is increasing. Identification of factors contributing to eating quality is helpful for developing high-quality rice varieties. RESULTS: Two groups of rice with different apparent amylose content (AACs) were used in this study. One group contained four varieties with low AACs (8.8-9.4%), whereas the other contained four traditional varieties with medium AACs (17.2-17.5%). The physicochemical properties, starch fine structure and crystallinity and storage protein composition of the two groups were analyzed. We found that, in both groups, the rice varieties with high eating quality had more short-chain amylopectin, lower glutelin and prolamin content, and a higher albumin content. In addition, the low-AAC varieties produced opaque endosperms, which may result from an increased number of pores in the center of starch granules. CONCLUSIONS: Both the fine structure of starch and the storage protein composition were closely related to rice eating quality. In both groups, short branch-chain amylopectin, short-chain amylopectin [degree of polymerization (DP) 6-12], and albumin had positive effects on eating quality. By contrast, long branch-chain amylopectin, long-chain amylopectin (DP 35-60), glutelin and prolamin had adverse effects on eating quality of rice. © 2020 Society of Chemical Industry.
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Oryza/química , Proteínas de Almacenamiento de Semillas/química , Almidón/química , Amilopectina/química , Amilopectina/metabolismo , Amilosa/química , Amilosa/metabolismo , Análisis de los Alimentos , Humanos , Oryza/clasificación , Oryza/metabolismo , Semillas/química , Semillas/clasificación , Semillas/metabolismo , Almidón/metabolismoRESUMEN
Grain number per panicle is a major component of rice yield that is typically controlled by many quantitative trait loci (QTLs). The identification of genes controlling grain number per panicle in rice would be valuable for the breeding of high-yielding rice. The Oryza glaberrima chromosome segment substitution line 9IL188 had significantly smaller panicles compared with the recurrent parent 9311. QTL analysis in an F2 population derived from a cross between 9IL188 and 9311 revealed that qgnp7(t), a major QTL located on the short arm of chromosome 7, was responsible for this phenotypic variation. Fine mapping was conducted using a large F3 population containing 2250 individuals that were derived from the F2 heterozygous plants. Additionally, plant height, panicle length, and grain number per panicle of the key F4 recombinant families were examined. Through two-step substitution mapping, qgnp7(t) was finally localized to a 41 kb interval in which eight annotated genes were identified according to available sequence annotation databases. Phenotypic evaluation of near isogenic lines (NIL-qgnp7 and NIL-qGNP7) indicated that qgnp7(t) has pleiotropic effects on rice plant architecture and panicle structure. In addition, yield estimation of NILs indicated that qGNP7(t) derived from 9311 is the favorable allele. Our results provide a foundation for isolating qgnp7(t). Markers flanking this QTL will be a useful tool for the marker-assisted selection of favorable alleles in O. glaberrima improvement programs.
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Docosahexaenoic acid (DHA), an essential omega-3 fatty acid, offers significant health benefits but faces challenges such as distinct odor, oxidation susceptibility, and limited intestinal permeability, hindering its broad application. Microencapsulation, widely employed, enhances DHA performance by facilitating controlled release, digestion, and absorption in the gastrointestinal tract. Despite extensive studies on DHA microcapsules and related delivery systems, understanding the mechanisms governing encapsulated DHA release, digestion, and absorption, particularly regarding the influence of wall materials and DHA sources, remains limited. This review starts with an overview of current techniques commonly applied for DHA microencapsulation. It then proceeds to outline up-to-date advances in the release, digestion and absorption of DHA microcapsules, highlighting the roles of wall materials and DHA sources. Importantly, it proposes strategies for overcoming challenges and exploiting opportunities to enhance the bioavailability of DHA microcapsules. Notably, spray drying dominates DHA microencapsulation (over 90 % usage), while complex coacervation shows promise for future applications. The combination of proteins and carbohydrates or phospholipids as wall material exhibits potential in controlling release and digestion of DHA microcapsules. The source of DHA, particularly algal oil, demonstrates higher lipid digestibility and absorptivity of free fatty acids (FFAs) than fish oil. Future advancements in DHA microcapsule development include formulation redesign (e.g., using plant proteins as wall material and algal oil as DHA source), technique optimization (such as co-microencapsulation and pre-digestion), and creation of advanced in vitro systems for assessing DHA digestion and absorption kinetics.
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Disponibilidad Biológica , Cápsulas , Digestión , Ácidos Docosahexaenoicos , Ácidos Docosahexaenoicos/química , Humanos , Composición de Medicamentos , Absorción Intestinal , Secado por PulverizaciónRESUMEN
Food and formulation characteristics are crucial factors affecting the gastrointestinal release and absorption kinetics of oral solid preparations. In the present study, the dynamic continuous release and bioaccessibility of metformin hydrochloride immediate-release (IR) and sustained-release (SR) tablets were investigated in the dynamic human stomach-intestine (DHSI-IV) system simulating fasted and fed states in healthy adults. Both tablet formulations (particularly IR tablet) exhibited a postponed release in the fed state compared to the fasted state. Correspondingly, the bioaccessible fraction of metformin from IR tablets in the presence of high-fat meal was significantly reduced to 76.2 % of the fasted state. However, the in vitro bioaccessibility was less impaired by food for SR tablets with a fed/fasted ratio of 95.5 %. A convolution-based approach was used to convert in vitro bioaccessibility results to plasma concentration data. The predicted plasma concentration curve showed good agreement with human data in terms of pharmacokinetic (PK) parameters. In the fasted state, the predicted Cmax, Tmax and AUC0-24h of IR tablets were 943.9 ± 25.7 ng/mL, 2.0 ± 0.4 h and 7090.7 ± 112.0 ng.h/mL, respectively, mirroring values observed in healthy subjects. Overall, the DHSI-IV system has demonstrated potential to assess and predict the impact of meal intake on the in vivo release and absorption behaviors of oral solid preparations.
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Ayuno , Metformina , Adulto , Humanos , Administración Oral , Tracto Gastrointestinal , Comprimidos , Preparaciones de Acción Retardada/farmacocinética , Estudios Cruzados , Disponibilidad Biológica , Área Bajo la CurvaRESUMEN
An incomplete augmented Lagrangian preconditioner, for the steady incompressible Navier-Stokes equations discretized by stable finite elements, is proposed. The eigenvalues of the preconditioned matrix are analyzed. Numerical experiments show that the incomplete augmented Lagrangian-based preconditioner proposed is very robust and performs quite well by the Picard linearization or the Newton linearization over a wide range of values of the viscosity on both uniform and stretched grids.
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Matemática/métodos , Modelos Lineales , ViscosidadRESUMEN
Intestinal permeability is a key factor affecting the bioavailability and physiological efficacy of docosahexaenoic acid (DHA) encapsulated in microcapsules. However, how the DHA microcapsules are transformed and the components absorbed across the small intestinal membrane has seldom been examined previously. In this study, an ex vivo absorption model based on the permeability of the rat small intestine was established to evaluate the intestinal absorption of DHA microcapsules with five formulations after gastrointestinal digestion in vitro. For pure glucose solutions, the apparent permeability coefficient (Papp) increased from 5.70 ± 0.60 × 10-6 cm s-1 at 5 mg mL-1 to 20.25 ± 0.88 × 10-6 cm s-1 at 30 mg mL-1 and decreased to 15.73 ± 0.91 × 10-6 cm s-1 at 100 mg mL-1. The Papp values obtained using the ex vivo model are comparable to those reported in the human jejunum. For algal oil DHA microcapsules with whey protein as the wall material (A-WP-DHA) after in vitro digestion, the Papp of glucose released was 3.81 × 10-6 cm s-1 with an absorption ratio of 59.55% in the ex vivo model, significantly lower than that from the in vitro porcine casing model. The Papp and absorption ratio varied little among the in vitro dialysis models with different molecular weight cut-off values. A similar trend was observed for the absorption of amino acids. However, the absorption ratio (26.6%) was the highest in the ex vivo model for free fatty acids (FFAs) released from the microcapsules due to the rapid accumulation of compounds on the inner wall of the intestinal sac. In addition, the DHA microcapsules with algal oil as the DHA source (36.40%) exhibited a higher absorption ratio of FFAs than that from tuna oil (14.26%) in the ex vivo model. The wall material compositions seemed to have little effect on FFA absorption. The present study is practically meaningful for the future formulation of DHA microcapsules with enhanced absorption.
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Ácidos Docosahexaenoicos , Diálisis Renal , Ratas , Humanos , Animales , Porcinos , Ácidos Docosahexaenoicos/química , Cápsulas , Absorción Intestinal , Intestino Delgado , GlucosaRESUMEN
A thorough understanding of the genetic basis of rice grain traits is critical for the improvement of rice (Oryza sativa L.) varieties. In this study, we generated an F2 population by crossing the large-grain japonica cultivar CW23 with Peiai 64 (PA64), an elite indica small-grain cultivar. Using QTL analysis, 17 QTLs for five grain traits were detected on four different chromosomes. Eight of the QTLs were newly-identified in this study. In particular, qGL3-1, a newly-identified grain length QTL with the highest LOD value and largest phenotypic variation, was fine-mapped to the 17 kb region of chromosome 3. A serine/threonine protein phosphatase gene encoding a repeat domain containing two Kelch motifs was identified as the unique candidate gene corresponding to this QTL. A comparison of PA64 and CW23 sequences revealed a single nucleotide substitution (CâA) at position 1092 in exon 10, resulting in replacement of Asp (D) in PA64 with Glu (E) in CW23 for the 364(th) amino acid. This variation is located at the D position of the conserved sequence motif AVLDT of the Kelch repeat. Genetic analysis of a near-isogenic line (NIL) for qGL3-1 revealed that the allele qGL3-1 from CW23 has an additive or partly dominant effect, and is suitable for use in molecular marker-assisted selection.
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Oryza/genética , Fosfoproteínas Fosfatasas/metabolismo , Sitios de Carácter Cuantitativo , Secuencia de Bases , Cromosomas de las Plantas , Cartilla de ADN , Fosfoproteínas Fosfatasas/genética , Reacción en Cadena de la PolimerasaRESUMEN
Microencapsulation is a promising technique to improve the bioavailability and mask the unpleasant smell of DHA oils. Yet, how the encapsulated DHA oils are 'released' and 'digested' within the gastrointestinal tract (GIT) and the effect of the wall material and source of DHA have been largely unknown. Here, two commercial DHA microcapsules from algae oil (A-DHA) and tuna oil (T-DHA) with 100% whey protein (WP) and 80% casein and 20% WP (C-WP) as wall materials were evaluated in vitro respectively. The release ratio was nearly linearly increased to 77.7% and 41.7% after the simulated gastric phase for T-DHA and A-DHA microcapsules, respectively. In contrast to A-DHA microcapsules for which the release of DHA approached equilibrium in the later intestinal phase, a decline in the release ratio was shown for T-DHA microcapsules perhaps due to the interaction of T-DHA with bile salts resulting in the formation of micelles. The more stable release behaviors might suggest a better performance of A-DHA coated by WP, which enables sustainable release during GIT digestion. This is supported by the better ability to resist gastric proteolysis for A-DHA microcapsules. Additionally, T-DHA (27.5%) showed a lower lipid digestibility than A-DHA (68.5%) in the end due to their structure difference. Significantly positive correlations were found for both microcapsules between DHA release ratio and protein hydrolysis. This study has provided quantitative information on the in vitro release and digestion of DHA microcapsules as influenced by the wall protein and DHA source. The findings are practically meaningful for future formulation of DHA microcapsules with controlled release rates at target sites of the GIT.
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Caseínas/química , Ácidos Docosahexaenoicos/química , Aceites de Pescado/química , Proteína de Suero de Leche/química , Reactores Biológicos , Cápsulas/química , Liberación de Fármacos , Lípidos/químicaRESUMEN
A dwarf mutant, designated LB4D, was obtained among the progeny of backcrosses to a wild rice introgression line. Genetic analysis of LB4D indicated that the dwarf phenotype was controlled by a single semidominant dwarfing gene, which was named LB4D. The mutants were categorized as dn-type dwarf mutants according to the pattern of internode reduction. In addition, gibberellin (GA) response tests showed that LB4D plants were neither deficient nor insensitive to GA. This study found that tiller formation by LB4D plants was decreased by 40% compared with the wild type, in contrast to other dominant dwarf mutants that have been identified, indicating that a different dwarfing mechanism might be involved in the LB4D dominant mutant. The reduction of plant height in F(1) plants ranged from 27.9% to 38.1% in different genetic backgrounds, showing that LB4D exerted a stronger dominant dwarfing effect. Using large F(2) and F(3) populations derived from a cross between heterozygous LB4D and the japonica cultivar Nipponbare, the LB4D gene was localized to a 46 kb region between the markers Indel 4 and Indel G on the short arm of chromosome 11, and four predicted genes were identified as candidates in the target region.
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Genes Dominantes/genética , Genes de Plantas/genética , Oryza/crecimiento & desarrollo , Oryza/genética , Mapeo Físico de Cromosoma/métodos , Proteínas de Plantas/genética , Emparejamiento Base/genética , China , Cruzamientos Genéticos , Inducción Enzimática/efectos de los fármacos , Marcadores Genéticos , Genotipo , Giberelinas/farmacología , Heterocigoto , Homocigoto , Mutación/genética , Oryza/citología , Oryza/enzimología , Fenotipo , Hojas de la Planta/efectos de los fármacos , Hojas de la Planta/crecimiento & desarrollo , Proteínas de Plantas/metabolismo , Recombinación Genética/efectos de los fármacos , Plantones/efectos de los fármacos , Plantones/enzimología , alfa-Amilasas/biosíntesisRESUMEN
Rice embryos are rich in high-quality protein, lipid, vitamins and minerals, representing the most important nutritional part of brown rice. However, the molecular mechanism of rice embryo development is poorly understood. In this study, two rice cultivars with contrasting embryo size (the giant embryo cultivar Dapeimi and the normal embryo cultivar 187R) were used to explore excellent genes controlling embryo size, and the developed near-isogenic lines (NILs) (NIL-D, which has the giant embryo phenotype, and its matching line, NIL-X) were used to explore transcript and metabolic properties in the earlier maturation stage of giant embryo development under natural conditions. The map-based cloning results demonstrated that Dapeimi is a novel allelic mutant of the rice GIANT EMBRYO (GE) gene, and the functional mutation site is a single cytosine deletion in the exon1. A total of 285 differentially accumulated metabolites (DAMs) and 677 differentially expressed genes (DEGs) were identified between NIL-D and NIL-X. The analysis of DAMs indicated that plants lacking GE mainly promoted energy metabolism, amino acid metabolism, and lipid metabolism pathways in the rice embryo. Pearson correlation coefficient showed that 300 pairs of gene-metabolites were highly correlated. Among them, OsZS_02G0528500 and OsZS_12G0013700 were considered to be key genes regulating L-Aspartic acid and L-Tryptophan content during rice giant embryo development, which are promising to be good candidate genes to improve rice nutrition. By analyzing rice embryo development through a combination of strategies, this research contributes to a greater understanding of the molecular mechanism of rice embryo development, and provides a theoretical foundation for breeding high-nutrition varieties.
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Grain size and shape are important determinants of grain weight and yield in rice. Here, we report a new major quantitative trait locus (QTL), qTGW3, that controls grain size and weight in rice. This locus, qTGW3, encodes OsSK41 (also known as OsGSK5), a member of the GLYCOGEN SYNTHASE KINASE 3/SHAGGY-like family. Rice near-isogenic lines carrying the loss-of-function allele of OsSK41 have increased grain length and weight. We demonstrate that OsSK41 interacts with and phosphorylates AUXIN RESPONSE FACTOR 4 (OsARF4). Co-expression of OsSK41 with OsARF4 increases the accumulation of OsARF4 in rice protoplasts. Loss of function of OsARF4 results in larger rice grains. RNA-sequencing analysis suggests that OsARF4 and OsSK41 repress the expression of a common set of downstream genes, including some auxin-responsive genes, during rice grain development. The loss-of-function form of OsSK41 at qTGW3 represents a rare allele that has not been extensively utilized in rice breeding. Suppression of OsSK41 function by either targeted gene editing or QTL pyramiding enhances rice grain size and weight. Thus, our study reveals the important role of OsSK41 in rice grain development and provides new candidate genes for genetic improvement of grain yield in rice and perhaps in other cereal crops.
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Glucógeno Sintasa Quinasa 3/genética , Oryza/genética , Sitios de Carácter Cuantitativo , Factores de Transcripción/metabolismo , Clonación Molecular , Grano Comestible , Regulación de la Expresión Génica de las Plantas , Oryza/anatomía & histología , Oryza/enzimología , FosforilaciónRESUMEN
A catalytic domino amination and oxidative coupling of in situ prepared gold acetylides has been developed for the synthesis of abnormal NHC (aNHC) gold complexes, and key vinylene digold intermediates are isolated and structurally characterized.