RESUMEN
Rice (Oryza sativa) is a cereal crop with a starchy endosperm. Starch is composed of amylose and amylopectin. Amylose content (AC) is the principal determinant of rice quality, but varieties with similar ACs can still vary substantially in their quality. In this study, we analyzed the total AC (TAC) and its constituent fractions, the hot water-soluble amylose content (SAC) and hot water-insoluble amylose content (IAC), in two sets of related chromosome segment substitution lines of rice with a common genetic background grown in two years. We searched for quantitative trait loci (QTLs) associated with SAC, IAC, and TAC and identified one common QTL (qSAC-6, qIAC-6, and qTAC-6) on chromosome 6. Map-based cloning revealed that the gene underlying the trait associated with this common QTL is Waxy (Wx). An analysis of the colors of soluble and insoluble starch-iodine complexes and their λmax values (wavelengths at the positions of their peak absorbance values) as well as gel permeation chromatography revealed that Wx is responsible for the biosynthesis of amylose, comprising a large proportion of the soluble fractions of the SAC. Wx is also involved in the biosynthesis of long chains of amylopectin, comprising the hot water-insoluble fractions of the IAC. These findings highlight the pleiotropic effects of Wx on the SAC and IAC. This pleiotropy indicates that these traits have a positive genetic correlation. Therefore, further studies of rice quality should use rice varieties with the same Wx genotype to eliminate the pleiotropic effects of this gene, allowing the independent relationship between the SAC or IAC and rice quality to be elucidated through a multiple correlation analysis. These findings are applicable to other valuable cereal crops as well.
Asunto(s)
Amilosa , Oryza , Proteínas de Plantas , Sitios de Carácter Cuantitativo , Solubilidad , Oryza/genética , Oryza/metabolismo , Amilosa/metabolismo , Amilosa/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Agua/química , Grano Comestible/genética , Grano Comestible/metabolismo , Pleiotropía Genética , Calor , Mapeo Cromosómico , Almidón Sintasa/genética , Almidón Sintasa/metabolismoRESUMEN
Starch is a significant ingredient of the seed endosperm with commercial importance in food and industry. Crop varieties with glutinous (waxy) grain characteristics, i.e. starch with high amylopectin and low amylose, hold longstanding cultural importance in some world regions and unique properties for industrial manufacture. The waxy character in many crop species is regulated by a single gene known as GBSSI (or waxy), which encodes the enzyme Granule Bound Starch Synthase1 with null or reduced activity. Several allelic variants of the waxy gene that contribute to varying levels of amylose content have been reported in different crop plants. Phylogenetic analysis of protein sequences and the genomic DNA encoding GBSSI of major cereals and recently sequenced millets and pseudo-cereals have shown that GBSSI orthologs form distinct clusters, each representing a separate crop lineage. With the rapidly increasing demand for waxy starch in food and non-food applications, conventional crop breeding techniques and modern crop improvement technologies such as gene silencing and genome editing have been deployed to develop new waxy crop cultivars. The advances in research on waxy alleles across different crops have unveiled new possibilities for modifying the synthesis of amylose and amylopectin starch, leading to the potential creation of customized crops in the future. This article presents molecular lines of evidence on the emergence of waxy genes in various crops, including their genesis and evolution, molecular structure, comparative analysis and breeding innovations.
Asunto(s)
Productos Agrícolas , Almidón Sintasa , Amilopectina/metabolismo , Amilopectina/genética , Amilosa/metabolismo , Amilosa/genética , Productos Agrícolas/genética , Genotipo , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Almidón/metabolismo , Almidón/genética , Almidón/biosíntesis , Almidón Sintasa/genética , Almidón Sintasa/metabolismoRESUMEN
Soluble starch synthases (SSs) play important roles in the synthesis of cassava starch. However, the expression characteristics of the cassava SSs genes have not been elucidated. In this study, the MeSSIII-1 gene and its promoter, from SC8 cassava cultivars, were respectively isolated by PCR amplification. MeSSIII-1 protein was localized to the chloroplasts. qRT-PCR analysis revealed that the MeSSIII-1 gene was expressed in almost all tissues tested, and the expression in mature leaves was 18.9 times more than that in tuber roots. MeSSIII-1 expression was induced by methyljasmonate (MeJA), abscisic acid (ABA), and ethylene (ET) hormones in cassava. MeSSIII-1 expression patterns were further confirmed in proMeSSIII-1 transgenic cassava. The promoter deletion analysis showed that the -264 bp to -1 bp MeSSIII-1 promoter has basal activity. The range from -1228 bp to -987 bp and -488 bp to -264 bp significantly enhance promoter activity. The regions from -987 bp to -747 bp and -747 bp to -488 bp have repressive activity. These findings will provide an important reference for research on the potential function and transcriptional regulation mechanisms of the MeSSIII-1 gene and for further in-depth exploration of the regulatory network of its internal functional elements.
Asunto(s)
Regulación de la Expresión Génica de las Plantas , Manihot , Proteínas de Plantas , Plantas Modificadas Genéticamente , Regiones Promotoras Genéticas , Manihot/genética , Manihot/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente/genética , Almidón Sintasa/genética , Almidón Sintasa/metabolismo , Ácido Abscísico/farmacología , Ácido Abscísico/metabolismo , Etilenos/metabolismoRESUMEN
Starch structure is often characterized by the chain-length distribution (CLD) of the linear molecules formed by breaking each branch-point. More information can be obtained by expanding into a second dimension: in the present case, the total undebranched-molecule size. This enables answers to questions unobtainable by considering only one variable. The questions considered here are: (i) are the events independent which control total size and CLD, and (ii) do ultra-long amylopectin (AP) chains exist (these chains cannot be distinguished from amylose chains using simple size separation). This was applied here to characterize the structures of one normal (RS01) wheat and two high-amylose (AM) mutant wheats (an SBEIIa knockout and an SBEIIa and SBEIIb knockout). Absolute ethanol was used to precipitate collected fractions, then size-exclusion chromatography for total molecular size and for the size of branches. The SBEIIa and SBEIIb mutations significantly increased AM and IC contents and chain length. The 2D plots indicated the presence of small but significant amounts of long-chain amylopectin, and the asymmetry of these plots shows that the corresponding mechanisms share some causal effects. These results could be used to develop plants producing improved starches, because different ranges of the chain-length distribution contribute independently to functional properties.
Asunto(s)
Amilopectina , Amilosa , Almidón Sintasa , Triticum , Triticum/metabolismo , Triticum/química , Triticum/genética , Amilopectina/química , Amilopectina/biosíntesis , Amilosa/química , Amilosa/biosíntesis , Almidón Sintasa/genética , Almidón Sintasa/metabolismo , Almidón Sintasa/química , Almidón/química , Almidón/biosíntesis , Almidón/metabolismo , Mutación , Proteínas de Plantas/química , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMEN
Amylose biosynthesis is strictly associated with granule-bound starch synthase I (GBSSI) encoded by the Waxy gene. Mutagenesis of single bases in the Waxy gene, which induced by CRISPR/Cas9 genome editing, caused absence of intact GBSSI protein in grain of the edited line. The amylose and amylopectin contents of waxy mutants were zero and 31.73%, while those in the wild type were 33.50% and 39.00%, respectively. The absence of GBSSI protein led to increase in soluble sugar content to 37.30% compared with only 10.0% in the wild type. Sucrose and ß-glucan, were 39.16% and 35.40% higher in waxy mutants than in the wild type, respectively. Transcriptome analysis identified differences between the wild type and waxy mutants that could partly explain the reduction in amylose and amylopectin contents and the increase in soluble sugar, sucrose and ß-glucan contents. This waxy flour, which showed lower final viscosity and setback, and higher breakdown, could provide more option for food processing.
Asunto(s)
Amilosa , Edición Génica , Hordeum , Proteínas de Plantas , Almidón Sintasa , Amilosa/metabolismo , Hordeum/genética , Hordeum/metabolismo , Edición Génica/métodos , Almidón Sintasa/genética , Almidón Sintasa/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Sistemas CRISPR-Cas , Amilopectina/metabolismo , Sacarosa/metabolismo , Azúcares/metabolismo , Regulación de la Expresión Génica de las Plantas , Mutación , beta-Glucanos/metabolismo , Plantas Modificadas Genéticamente , SolubilidadRESUMEN
Transitory starch is an important carbon source in leaves, and its biosynthesis and metabolism are closely related to grain quality and yield. The molecular mechanisms controlling leaf transitory starch biosynthesis and degradation and their effects on rice (Oryza sativa) quality and yield remain unclear. Here, we show that OsLESV and OsESV1, the rice orthologs of AtLESV and AtESV1, are associated with transitory starch biosynthesis in rice. The total starch and amylose contents in leaves and endosperms are significantly reduced, and the final grain quality and yield are compromised in oslesv and osesv1 single and oslesv esv1 double mutants. Furthermore, we found that OsLESV and OsESV1 bind to starch, and this binding depends on a highly conserved C-terminal tryptophan-rich region that acts as a starch-binding domain. Importantly, OsLESV and OsESV1 also interact with the key enzymes of starch biosynthesis, granule-bound starch synthase I (GBSSI), GBSSII, and pyruvate orthophosphote dikiase (PPDKB), to maintain their protein stability and activity. OsLESV and OsESV1 also facilitate the targeting of GBSSI and GBSSII from plastid stroma to starch granules. Overexpression of GBSSI, GBSSII, and PPDKB can partly rescue the phenotypic defects of the oslesv and osesv1 mutants. Thus, we demonstrate that OsLESV and OsESV1 play a key role in regulating the biosynthesis of both leaf transitory starch and endosperm storage starch in rice. These findings deepen our understanding of the molecular mechanisms underlying transitory starch biosynthesis in rice leaves and reveal how the transitory starch metabolism affects rice grain quality and yield, providing useful information for the genetic improvement of rice grain quality and yield.
Asunto(s)
Grano Comestible , Oryza , Proteínas de Plantas , Almidón Sintasa , Almidón , Oryza/genética , Oryza/metabolismo , Almidón/metabolismo , Almidón/biosíntesis , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Almidón Sintasa/genética , Almidón Sintasa/metabolismo , Grano Comestible/metabolismo , Grano Comestible/genética , Hojas de la Planta/metabolismo , Hojas de la Planta/genética , Amilosa/metabolismo , Amilosa/biosíntesis , Regulación de la Expresión Génica de las PlantasRESUMEN
The starch synthase (SS) plays important roles in regulating plant growth and development and responding to adversity stresses. Although the SS family has been studied in many crops, it has not been fully identified in sweet potato and its two related species. In the present study, eight SSs were identified from Ipomoea batatas (I. batata), Ipomoea trifida (I. trifida), and Ipomoea trlioba (I. trlioba), respectively. According to the phylogenetic relationships, they were divided into five subgroups. The protein properties, chromosomal location, phylogenetic relationships, gene structure, cis-elements in the promoter, and interaction network of these proteins were also analyzed; stress expression patterns were systematically analyzed; and real-time polymerase chain reaction (qRT-PCR) analysis was performed. Ipomoea batatas starch synthase (IbSSs) were highly expressed in tuber roots, especially Ipomoea batatas starch synthase 1 (IbSS1) and Ipomoea batatas starch synthase 6 (IbSS6), which may play an important role in root development and starch biosynthesis. At the same time, the SS genes respond to potassium deficiency, hormones, cold, heat, salt, and drought stress. This study offers fresh perspectives for enhancing knowledge about the roles of SSs and potential genes to enhance productivity, starch levels, and resistance to environmental stresses in sweet potatoes.
Asunto(s)
Regulación de la Expresión Génica de las Plantas , Ipomoea batatas , Filogenia , Proteínas de Plantas , Almidón Sintasa , Almidón Sintasa/genética , Almidón Sintasa/metabolismo , Ipomoea batatas/genética , Ipomoea batatas/crecimiento & desarrollo , Ipomoea batatas/enzimología , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Estrés Fisiológico/genética , Familia de Multigenes , Genoma de Planta/genética , Ipomoea/genéticaRESUMEN
Potato starch with mutations in starch branching enzyme genes (SBEI, SBEII) and granule-bound starch synthase gene (GBSS) was characterized for molecular and thermal properties. Mutations in GBSS were here stacked to a previously developed SBEI and SBEII mutation line. Additionally, mutations in the GBSS gene alone were induced in the wild-type variety for comparison. The parental line with mutations in the SBE genes showed a â¼ 40 % increase in amylose content compared with the wild-type. Mutations in GBSS-SBEI-SBEII produced non-waxy, low-amylose lines compared with the wild-type. An exception was a line with one remaining GBSS wild-type allele, which displayed â¼80 % higher amylose content than wild-type. Stacked mutations in GBSS in the SBEI-SBEII parental line caused alterations in amylopectin chain length distribution and building block size categories of whole starch. Correlations between size categories of building blocks and unit chains of amylopectin were observed. Starch in GBSS-SBEI-SBEII mutational lines had elevated peak temperature of gelatinization, which was positively correlated with large building blocks.
Asunto(s)
Enzima Ramificadora de 1,4-alfa-Glucano , Solanum tuberosum , Almidón Sintasa , Amilopectina/química , Almidón Sintasa/genética , Almidón Sintasa/metabolismo , Amilosa , Solanum tuberosum/metabolismo , Estructura Molecular , Almidón/química , Mutación , Enzima Ramificadora de 1,4-alfa-Glucano/químicaRESUMEN
Head rice yield (HRY) measures rice milling quality and determines final grain yield and commercial value. Here, we report that two major quantitative trait loci for milling quality in rice, qMq-1 and qMq-2, represent allelic variants of Waxylv/Waxyb (hereafter Wx) encoding Granule-Bound Starch Synthase I (GBSSI) and Alkali Spreading Value ALKc/ALKb encoding Soluble Starch Synthase IIa (SSIIa), respectively. Complementation and overexpression transgenic lines in indica and japonica backgrounds confirmed that Wx and ALK coordinately regulate HRY by affecting amylose content, the number of amylopectin branches, amyloplast size, and thus grain filling and hardness. The transcription factor OsDOF18 acts upstream of Wx and ALK by activating their transcription. Furthermore, rice accessions with Wxb and ALKb alleles showed improved HRY over those with Wxlv and ALKc. Our study not only reveals the novel molecular mechanism underlying the formation of HRY but also provides a strategy for breeding rice cultivars with improved HRY.
Asunto(s)
Alelos , Oryza , Proteínas de Plantas , Oryza/genética , Oryza/crecimiento & desarrollo , Oryza/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Sitios de Carácter Cuantitativo/genética , Regulación de la Expresión Génica de las Plantas , Plantas Modificadas Genéticamente/genética , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Almidón Sintasa/genética , Almidón Sintasa/metabolismoRESUMEN
Amylose content (AC) is a significant quality trait in starchy crops, affecting their processing and application by the food and non-food industries. Therefore, fine-tuning AC in these crops has become a focus for breeders. Granule-bound starch synthase (GBSS) is the core enzyme that directly determines the AC levels. Several excellent reviews have summarized key progress in various aspects of GBSS research in recent years, but they mostly focus on cereals. Herein, we provide an in-depth review of GBSS research in monocots and dicots, focusing on the molecular characteristics, evolutionary relationships, expression patterns, molecular regulation mechanisms, and applications. We also discuss future challenges and directions for controlling AC in starchy crops, and found simultaneously increasing both the PTST and GBSS gene expression levels may be an effective strategy to increase amylose content.
Asunto(s)
Almidón Sintasa , Almidón Sintasa/genética , Almidón Sintasa/metabolismo , Amilosa , AlmidónRESUMEN
A chalky endosperm mutant (GM03) induced from an indica rice GLA4 was used to investigate the functional gene in starch biosynthesis. Bulked segregant analysis and sanger sequencing determined that a novel mutation in soluble starch synthase IIIa (SSIIIa) is responsible for the chalky phenotype in GM03. Complementary test by transforming the active SSIIIa gene driven by its native promoter to GM03 recovered the phenotype to its wildtype. The expression of SSIIIa was significantly decreased, while SSIIIa protein was not detected in GM03. The mutation of SSIIIa led to increased expression of most of starch synthesis related genes and elevated the levels of most of proteins in GM03. The CRISPR/Cas9 technology was used for targeted disruption of SSIIIa, and the mutant lines exhibited chalky endosperm which phenocopied the GM03. Additionally, the starch fine structure in the knockout mutant lines ss3a-1 and ss3a-2 was similar with the GM03, which showed increased amylose content, higher proportions of B1 and B2 chains, much lower proportions of B3 chains and decreased degree of crystallinity, leading to altered thermal properties with lower gelatinization temperature and enthalpy. Collectively, these results suggested that SSIIIa plays an important role in starch synthesis by elongating amylopectin long chains in rice.
Asunto(s)
Oryza , Almidón Sintasa , Amilopectina/metabolismo , Almidón Sintasa/genética , Almidón Sintasa/metabolismo , Oryza/genética , Oryza/metabolismo , Alelos , Almidón/metabolismo , Amilosa/metabolismoRESUMEN
Waxy (WX) and high-amylose (HA) wheat flours have interesting functional and/or nutritional characteristics, but low technological properties compared to regular wheat. Here a set of three wheat lines, having different amylose content but sharing the same varietal background, were compared to shed light on the role of the amylose/amylopectin ratio on the protein conformational changes that lead to gluten formation. Despite the absence of differences in their protein profile, as also confirmed by thiolomic approaches, both WX and HA lines developed a weaker gluten than the control sample. The altered amylose/amylopectin ratio exerts a matrix effect establishing a competition for water with proteins, leading to a different protein structure and three-dimensional organization of the gluten network. These results add a piece to the understanding of the molecular aspects that oversee matrix effects on gluten formation in wheat, which description can be helpful for a rational optimization of the transformation process.
Asunto(s)
Amilosa , Almidón Sintasa , Amilosa/química , Amilopectina/química , Almidón Sintasa/metabolismo , Glútenes/metabolismo , Triticum/química , Almidón/químicaRESUMEN
Amylose fraction of grain starch is correlated with a type of resistant starch with better nutritional quality. Granule-bound starch synthase I (GBSSI) is the known starch synthase, responsible for elongation of linear amylose chains. GBSSI expression, activity, and binding to starch and other proteins are the key factors that can affect amylose content. Previously, a QTL, qhams7A.1 carrying GBSSI mutant allele, was identified through QTL mapping using F2 population of the high amylose mutant line, 'TAC 75'. This high amylose mutant line has >2-fold higher amylose content than wild variety 'C 306'. In this study, we characterized this novel mutant allele, GBSSI.L539P. In vitro starch synthase activity of GBSSI.L539P showed improved activity than the wild type (GBSSI-wt). When expressed in yeast glycogen synthase mutants (Δgsy1gsy2), GBSSI-wt and GBSSI.L539P partially complemented the glycogen synthase (gsy1gsy2) activity in yeast. Structural analysis by circular dichroism (CD) and homology modelling showed no significant structural distortion in the mutant enzyme. Molecular docking studies suggested that the residue Leu539 is distant from the catalytic active site (ADP binding pocket) and had no detectable conformational changes in active site. Both wild and mutant enzymes were assayed for starch binding in vitro, and demonstrating higher affinity of the GBSSI.L539P mutant for starch than the wild type. The present study indicated that distant residue (L539P) influenced GBSSI activity by affecting its starch-binding ability. Therefore, it may be a potential molecular target for enhanced amylose content in grain.
Asunto(s)
Almidón Sintasa , Almidón Sintasa/genética , Almidón Sintasa/metabolismo , Amilosa/metabolismo , Triticum/metabolismo , Glucógeno Sintasa/metabolismo , Alelos , Simulación del Acoplamiento Molecular , Saccharomyces cerevisiae/metabolismo , AlmidónRESUMEN
Plant chloroplasts conduct photosynthesis to convert solar energy into sugars for the carbon source essential for cell living and growth during the day. One fraction of photosynthetic products is stored in chloroplasts by forming starch granules to continue the provision of carbon energy during the night. Currently, profiling the starch temporal pattern requires either: (i) sacrificing the leaves, or (ii) generating transgenic plants at the risk of changing the metabolisms by incorporating a genetically modified granule-bound starch synthase (GBSS). In this paper, we demonstrated a nondestructive method using two-photon fluorescence (TPF) and second-harmonic generation (SHG) imaging to quantify starch granules within chloroplasts of fresh intact leaves across a day-night cycle. We did so using two Arabidopsis lines having normal and excess starch contents: wild-type (Columbia-0) and starch excess 1 (sex1). The starch granules were visualized by SHG imaging, while the chloroplasts in mesophyll cells were visualized by TPF imaging. Our results provided micron scale spatial resolution of starch distribution within leaves and showed starch circadian patterns consistent with those profiled by enzymatic assays in previous studies. We demonstrated that TPF-SHG imaging is a potential tool for revealing the real-time heterogeneity of starch circadian rhythm in leaf cells, without the need for destructive sample preparation.
Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Microscopía de Generación del Segundo Armónico , Almidón Sintasa , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Carbono/metabolismo , Hojas de la Planta/metabolismo , Almidón/metabolismo , Almidón Sintasa/metabolismo , Azúcares/metabolismoRESUMEN
BACKGROUND: Starch, a vital plant-derived polysaccharide comprised of branched glucans, is essential in nutrition and many industrial applications. Starch is often modified post-extraction to alter its structure and enhance its functionality. Targeted metabolic engineering of crops to produce valuable and versatile starches requires knowledge of the relationships between starch biosynthesis, structure, and properties, but systematic studies to obtain this knowledge are difficult to conduct in plants. Here we used Saccharomyces cerevisiae as a testbed to dissect the functions of plant starch biosynthetic enzymes and create diverse starch-like polymers. RESULTS: We explored yeast promoters and terminators to tune the expression levels of the starch-biosynthesis machinery from Arabidopsis thaliana. We systematically modulated the expression of each starch synthase (SS) together with a branching enzyme (BE) in yeast. Protein quantification by parallel reaction monitoring (targeted proteomics) revealed unexpected effects of glucan biosynthesis on protein abundances but showed that the anticipated broad range of SS/BE enzyme ratios was maintained during the biosynthetic process. The different SS/BE ratios clearly influenced glucan structure and solubility: The higher the SS/BE ratio, the longer the glucan chains and the more glucans were partitioned into the insoluble fraction. This effect was irrespective of the SS isoform, demonstrating that the elongation/branching ratio controls glucan properties separate from enzyme specificity. CONCLUSIONS: Our results provide a quantitative framework for the in silico design of improved starch biosynthetic processes in plants. Our study also exemplifies a workflow for the rational tuning of a complex pathway in yeast, starting from the selection and evaluation of expression modules to multi-gene assembly and targeted protein monitoring during the biosynthetic process.
Asunto(s)
Enzima Ramificadora de 1,4-alfa-Glucano , Arabidopsis , Almidón Sintasa , Enzima Ramificadora de 1,4-alfa-Glucano/metabolismo , Arabidopsis/metabolismo , Glucanos/química , Plantas/metabolismo , Isoformas de Proteínas , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Almidón/metabolismo , Almidón Sintasa/química , Almidón Sintasa/metabolismoRESUMEN
Ripening is the last, irreversible developmental stage during which fruit become palatable, thus promoting seed dispersal by frugivory. In Alisa Craig fruit, mRNAs with increasing m5C levels, such as STPK and WRKY 40, were identified as being involved in response to biotic and abiotic stresses. Furthermore, two mRNAs involved in cell wall metabolism, PG and EXP-B1, also presented increased m5C levels. In the Nr mutant, several m5C-modified mRNAs involved in fruit ripening, including those encoding WRKY and MADS-box proteins, were found. Targets of long non-coding RNAs and circular RNAs with different m5C sites were also found; these targets included 2-alkenal reductase, soluble starch synthase 1, WRKY, MADS-box, and F-box/ketch-repeat protein SKIP11. A combined analysis of changes in 5mC methylation and mRNA revealed many differentially expressed genes with differentially methylated regions encoding transcription factors and key enzymes related to ethylene biosynthesis and signal transduction; these included ERF084, EIN3, AP2/ERF, ACO5, ACS7, EIN3/4, EBF1, MADS-box, AP2/ERF, and ETR1. Taken together, our findings contribute to the global understanding of the mechanisms underlying fruit ripening, thereby providing new information for both fruit and post-harvest behavior.
Asunto(s)
Proteínas F-Box , Solanum lycopersicum , Almidón Sintasa , Solanum lycopersicum/genética , Solanum lycopersicum/metabolismo , Frutas/genética , Frutas/metabolismo , Regulación de la Expresión Génica de las Plantas/genética , Metilación , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , ARN Circular , Almidón Sintasa/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Proteínas F-Box/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo , Etilenos/metabolismo , ADN/metabolismo , ARN no Traducido/genética , ARN no Traducido/metabolismo , Oxidorreductasas/metabolismoRESUMEN
The starch branching enzyme IIb mutant (be2b) in rice significantly increases the resistant starch (RS) content and leads to reduced seed weight. However, the underlying metabolic mechanisms remain unclear. Proteomic analysis indicated that upregulation of starch synthase IIa (SSIIa) and SSIIIa and downregulation of BEI and SSI were possibly responsible for the decreased short amylopectin chains (DP 6-15) and increased longer chains (DP > 16) of be2b starch. The upregulation of granule-bound starch synthase led to increased amylose content (AC). These changes in the amylopectin structure and AC accounted for the increased RS content. α-Amylase 2A showed the strongest upregulation (up to 8.45-fold), indicating that the loss of BEIIb activity enhanced starch degradation. Upregulation of glycolysis-related proteins stimulated carbohydrate repartitioning through glycerate-3-phosphate and promoted the accumulation of tricarboxylic acid cycle intermediates, amino acids, and fatty acids. The unexpected carbohydrate partitioning and enhanced starch degradation resulted in the reduced seed weight in the be2b mutant.
Asunto(s)
Enzima Ramificadora de 1,4-alfa-Glucano , Oryza , Almidón Sintasa , Enzima Ramificadora de 1,4-alfa-Glucano/metabolismo , Amilopectina/química , Amilosa/química , Oryza/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Proteómica , Almidón Resistente , Semillas/genética , Semillas/metabolismo , Almidón/química , Almidón Sintasa/genética , Almidón Sintasa/metabolismoRESUMEN
Starch synthase III plays a key role in starch biosynthesis and is highly expressed in developing wheat grains. To understand the contribution of SSIII to starch and grain properties, we developed wheat ssIIIa mutants in the elite cultivar Cadenza using in silico TILLING in a mutagenized population. SSIIIa protein was undetectable by immunoblot analysis in triple ssIIIa mutants carrying mutations in each homoeologous copy of ssIIIa (A, B and D). Loss of SSIIIa in triple mutants led to significant changes in starch phenotype including smaller A-type granules and altered granule morphology. Starch chain-length distributions of double and triple mutants indicated greater levels of amylose than sibling controls (33.8% of starch in triple mutants, and 29.3% in double mutants vs. 25.5% in sibling controls) and fewer long amylopectin chains. Wholemeal flour of triple mutants had more resistant starch (6.0% vs. 2.9% in sibling controls) and greater levels of non-starch polysaccharides; the grains appeared shrunken and weighed ~ 11% less than the sibling control which was partially explained by loss in starch content. Interestingly, our study revealed gene dosage effects which could be useful for fine-tuning starch properties in wheat breeding applications while minimizing impact on grain weight and quality.
Asunto(s)
Almidón Sintasa , Amilopectina/metabolismo , Pan , Grano Comestible/genética , Grano Comestible/metabolismo , Estructura Molecular , Fitomejoramiento , Almidón/metabolismo , Almidón Sintasa/metabolismo , Triticum/metabolismoRESUMEN
This review systematically documents the major different strategies of generating high-amylose (HAS) starch mutants aiming at providing high resistant starch, by engineering the starch biosynthesis metabolic pathways. We identify three main strategies based on a new representation of the starch structure: 'the building block backbone model': i) suppression of starch synthases for reduction of amylopectin (AP) side-chains; ii) suppression of starch branching enzymes (SBEs) for production of AM-like materials; and iii) suppression of debranching enzymes to restrain the transformation from over-branched pre-AP to more ordered AP. From a biosynthetic perspective, AM generated through the second strategy can be classified into two types: i) normal AM synthesized mainly by regular expression of granule-bound starch synthases, and ii) modified linear AP chains (AM-like material) synthesized by starch synthases due to the suppression of starch branching enzymes. The application of new breeding technologies, especially CRISPR, in the breeding of HAS crops is also reviewed.
Asunto(s)
Enzima Ramificadora de 1,4-alfa-Glucano , Almidón Sintasa , Enzima Ramificadora de 1,4-alfa-Glucano/genética , Enzima Ramificadora de 1,4-alfa-Glucano/metabolismo , Amilopectina/metabolismo , Amilosa/metabolismo , Vías Biosintéticas , Almidón/metabolismo , Almidón Sintasa/genética , Almidón Sintasa/metabolismoRESUMEN
BACKGROUND: An increased demand for food has mirrored the increasing global population. Obesity and diabetes are two disorders induced by poor eating choices. Consequently, there is an urgent need to develop modified foods that can ameliorate such illnesses. The objective of this study was to explore the effect of Waxy genes on the structural and functional properties of starch, with the aim of improving food quality. Wild-type tetraploid wheat was compared with three mutants with different Waxy gene combinations. RESULTS: The proportion of B-type granules was higher in the mutants than in the wild-type (Wx-AB), and there were significant changes in the starch granule size, number, and phenotype in the Wx free mutant (Wx-ab). The lowest branch chain length was observed in Wx-ab, whereas Wx-AB had the highest branch chain length of DP ≥ 37. Wx-ab had the highest degree of crystallinity. The crystallinity trend followed the order Wx-ab>Wx-Ab>Wx-aB>Wx-AB. The amount of slowly digestible starch (SDS) was higher in native, gelatinized, and retrograded starch in the mutant. The amount of retrograded starch was closer to gelatinized starch than to native starch. CONCLUSION: Waxy proteins make a substantial contribution to starch structure. A lack of waxy proteins reduced the unit chains markedly compared with the control. Waxy proteins significantly affected the smaller and longer chains of starch. The lines with differing waxy composition had different effects on food digestion. The Wx-AB in native starch and Wx-Ab in gelatinized starch can control obesity and diabetes by slow-digesting carbohydrates and high resistance to digestion. © 2022 Society of Chemical Industry.