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Bioscience Reports Oct 2022Seed vigor and longevity are important agronomic attributes, as they are essentially associated with crop yield and thus the global economy. Seed longevity is a measure... (Review)
Review
Seed vigor and longevity are important agronomic attributes, as they are essentially associated with crop yield and thus the global economy. Seed longevity is a measure of seed viability and the most essential property in gene bank management since it affects regeneration of seed recycling. Reduced seed life or storability is a serious issue in seed storage since germplasm conservation and agricultural enhancement initiatives rely on it. The irreversible and ongoing process of seed deterioration comprises a complex gene regulatory network and altered metabolism that results in membrane damage, DNA integrity loss, mitochondrial dysregulation, protein damage, and disrupted antioxidative machinery. Carbohydrates and/or sugars, primarily raffinose family oligosaccharides (RFOs), have emerged as feasible components for boosting or increasing seed vigor and longevity in recent years. RFOs are known to perform diverse functions in plants, including abiotic and biotic stress tolerance, besides being involved in regulating seed germination, desiccation tolerance, vigor, and longevity. We emphasized and analyzed the potential impact of RFOs on seed vigor and longevity in this review. Here, we comprehensively reviewed the molecular mechanisms involved in seed longevity, RFO metabolism, and how RFO content is critical and linked with seed vigor and longevity. Further molecular basis, biotechnological approaches, and CRISPR/Cas applications have been discussed briefly for the improvement of seed attributes and ultimately crop production. Likewise, we suggest advancements, challenges, and future possibilities in this area.
Topics: Longevity; Oligosaccharides; Raffinose; Seeds; Sugars
PubMed: 36149314
DOI: 10.1042/BSR20220198 -
Molecules (Basel, Switzerland) Sep 2022α-Galactosidase (EC 3.2.1.22) refers to a group of enzymes that hydrolyze oligosaccharides containing α-galactoside-banded glycosides, such as stachyose, raffinose,...
α-Galactosidase (EC 3.2.1.22) refers to a group of enzymes that hydrolyze oligosaccharides containing α-galactoside-banded glycosides, such as stachyose, raffinose, and verbascose. These enzymes also possess great potential for application in sugar production, and in the feed and pharmaceutical industries. In this study, a strain of (WHPC005) that produces α-galactosidase was identified from the soil of Western Hunan, China. It was determined that the optimal temperature and pH for this α-galactosidase were 45 °C and 5.5, respectively. The activity of α-galactosidase was inhibited by K, Al, Fe, fructose, sucrose, lactose, galactose, SDS, EDTA, NaCl, and (NH)SO, and enhanced by Ca, Fe, Mn, Zn, glucose, and raffinose. The optimal inducer was raffinose, and the optimal induction concentration was 30 μmol/L. The α-galactosidase gene was cloned using random fragment cloning methods. Sequence analysis demonstrated that the open reading frame of the α-galactosidase gene was 1230 bp, which encodes a putative protein of 409 amino acids in length. Bioinformatics analysis showed that the isoelectric point and molecular weight of this α-galactosidase were 4.84 and 47.40 kD, respectively. Random coils, alpha helixes, and beta turns were observed in its secondary structure, and conserved regions were found in the tertiary structure of this α-galactosidase. Therefore, this α-galactosidase-producing bacterial strain has the potential for application in the feed industry.
Topics: Amino Acids; Carnobacteriaceae; Edetic Acid; Fructose; Galactose; Galactosides; Glucose; Glycosides; Hydrogen-Ion Concentration; Kinetics; Lactose; Oligosaccharides; Raffinose; Sodium Chloride; Soil; Substrate Specificity; Sucrose; alpha-Galactosidase
PubMed: 36144675
DOI: 10.3390/molecules27185942 -
The Plant Journal : For Cell and... Oct 2019Raffinose is thought to play an important role in plant tolerance of abiotic stress. We report here that maize HEAT SHOCK FACTOR A2 (ZmHSFA2) and HEAT SHOCK BINDING...
Raffinose is thought to play an important role in plant tolerance of abiotic stress. We report here that maize HEAT SHOCK FACTOR A2 (ZmHSFA2) and HEAT SHOCK BINDING PROTEIN 2 (ZmHSBP2) physically interact with each other and antagonistically modulate expression of GALACTINOL SYNTHASE2 (ZmGOLS2) and raffinose biosynthesis in transformed maize protoplasts and Arabidopsis plants. Overexpression of ZmHSFA2 in Arabidopsis increased the expression of Arabidopsis AtGOLS1, AtGOLS2 and AtRS5 (RAFFINOSE SYNTHASE), increased the raffinose content in leaves and enhanced plant heat stress tolerance. Contrary to ZmHSFA2, overexpression of ZmHSBP2 in Arabidopsis decreased expression of AtGOLS1, AtGOLS2 and AtRS5, decreased the raffinose content in leaves and reduced plant heat stress tolerance. ZmHSFA2 and ZmHSBP2 also interact with their Arabidopsis counterparts AtHSBP and AtHSFA2 as determined using bimolecular fluorescence complementation assays. Furthermore, endogenous ZmHSBP2 and Rluc, controlled by the ZmHSBP2 promoter, are transcriptionally activated by ZmHSFA2 and inhibited by ZmHSBP2 in maize protoplasts. These findings provide insights into the transcriptional regulation of raffinose biosynthetic genes, and the tolerance their product confers to plant heat stress.
Topics: Arabidopsis; Arabidopsis Proteins; Galactosyltransferases; Gene Expression Regulation, Plant; Heat Shock Transcription Factors; Heat-Shock Response; Plant Proteins; Plants, Genetically Modified; Raffinose; Stress, Physiological; Thermotolerance; Zea mays
PubMed: 31180156
DOI: 10.1111/tpj.14434 -
The Journal of Biological Chemistry Jun 2020Raffinose and its precursor galactinol accumulate in plant leaves during abiotic stress. RAFFINOSE SYNTHASE (RAFS) catalyzes raffinose formation by transferring a...
Raffinose and its precursor galactinol accumulate in plant leaves during abiotic stress. RAFFINOSE SYNTHASE (RAFS) catalyzes raffinose formation by transferring a galactosyl group of galactinol to sucrose. However, whether RAFS contributes to plant drought tolerance and, if so, by what mechanism remains unclear. In this study, we report that expression of from maize (or corn, ) () is induced by drought, heat, cold, and salinity stresses. We found that mutant maize plants completely lack raffinose and hyper-accumulate galactinol and are more sensitive to drought stress than the corresponding null-segregant (NS) plants. This indicated that ZmRAFS and its product raffinose contribute to plant drought tolerance. overexpression in enhanced drought stress tolerance by increasing -inositol levels via ZmRAFS-mediated galactinol hydrolysis in the leaves due to sucrose insufficiency in leaf cells and also enhanced raffinose synthesis in the seeds. Supplementation of sucrose to detached leaves converted ZmRAFS from hydrolyzing galactinol to synthesizing raffinose. Taken together, we demonstrate that ZmRAFS enhances plant drought tolerance through either raffinose synthesis or galactinol hydrolysis, depending on sucrose availability in plant cells. These results provide new avenues to improve plant drought stress tolerance through manipulation of the raffinose anabolic pathway.
Topics: Arabidopsis; Disaccharides; Droughts; Galactosyltransferases; Hydrolysis; Mutation; Raffinose; Stress, Physiological; Substrate Specificity; Zea mays
PubMed: 32366461
DOI: 10.1074/jbc.RA120.013948 -
Applied and Environmental Microbiology Jan 2019Herein, we present a biocatalytic method to produce raffinose and stachyose using sucrose as the substrate. An multienzyme system was developed using five enzymes,...
Herein, we present a biocatalytic method to produce raffinose and stachyose using sucrose as the substrate. An multienzyme system was developed using five enzymes, namely, sucrose synthase (SUS), UDP-glucose 4-epimerase (GalE), galactinol synthase (GS), raffinose synthase (RS), and stachyose synthase (STS), and two intermedia, namely, UDP and inositol, which can be recycled. This reaction system produced 11.1 mM raffinose using purified enzymes under optimal reaction conditions and substrate concentrations. Thereafter, a stepwise cascade reaction strategy was employed to circumvent the instability of RS and STS in this system, and a 4.2-fold increase in raffinose production was observed. The enzymatic cascade reactions were then conducted using cell extracts to avoid the need for enzyme purification and supplementation with UDP. Such modification further increased raffinose production to 86.6 mM and enabled the synthesis of 61.1 mM stachyose. The UDP turnover number reached 337. Finally, inositol in the reaction system was recycled five times, and 255.8 mM raffinose (128.9 g/liter) was obtained. Soybean oligosaccharides (SBOS) have elicited considerable attention because of their potential applications in the pharmaceutical, cosmetics, and food industries. This study demonstrates an alternative method to produce raffinose and stachyose, which are the major bioactive components of SBOS, from sucrose via an enzyme system. High concentrations of galactinol, raffinose, and stachyose were synthesized with the aid of a stepwise cascade reaction process, which can successfully address the issue of mismatched enzyme characteristics of an metabolic engineering platform. The biocatalytic approach presented in this work may enable the synthesis of other valuable galactosyl oligosaccharides, such as verbascose and higher homologs, which are difficult to obtain through plant extraction.
Topics: Arabidopsis; Bacterial Proteins; Escherichia coli; Multienzyme Complexes; Oligosaccharides; Plant Proteins; Raffinose; Sucrose
PubMed: 30389762
DOI: 10.1128/AEM.02306-18 -
Journal of the Academy of Nutrition and... Dec 2023Fiber intake may lower digestive tract cancer risk, possibly by modulating the composition of gut microbiota. However, no data are available about the role of specific...
BACKGROUND
Fiber intake may lower digestive tract cancer risk, possibly by modulating the composition of gut microbiota. However, no data are available about the role of specific fiber fractions with prebiotic activity (e.g., inulin-type fructans (ITFs), fructo-oligosaccharides (FOSs) and galactooligosaccharides (GOSs)) on the risk lower digestive tract cancers.
OBJECTIVE
The objective was to assess the association between prebiotic intake and the risk of cancers of the upper digestive tract and stomach.
DESIGN
Within the PrebiotiCa study, data were derived from a network of Italian case-control studies conducted between 1992 and 2009. Participants' usual diet was assessed using a food frequency questionnaire. ITFs, and selected FOSs (nystose, kestose, and 1F-β-fructofuranosylnystose) and GOSs (raffinose and stachyose) were quantified in several food products via laboratory analyses. Participants' prebiotic intake was calculated by multiplying food frequency questionnaire intake by the prebiotic content of each food item.
PARTICIPANTS/SETTING
Cases were patients admitted to major hospitals with incident histologically confirmed cancers; there were 946 cases of cancer of the oral cavity/pharynx, 198 of the nasopharynx, 304 of the esophagus, 230 of the stomach. More than 4,000 patients admitted to the same hospitals for acute nonneoplastic and not diet-related conditions were selected as control subjects.
MAIN OUTCOME MEASURES
The outcomes were oral and pharyngeal, nasopharyngeal, esophageal, and stomach cancers.
STATISTICAL ANALYSES PERFORMED
The odds ratios and corresponding 95% CIs of the various cancers were derived using logistic regression models adjusted for major confounders and energy intake.
RESULTS
No association was observed between intake of prebiotics and risk of cancers of the oral cavity and pharynx, nasopharynx, and esophagus. High raffinose intake reduced stomach cancer risk (odds ratio for the third vs the first tertile 0.6, 95% CI 0.3 to 0.9); no other prebiotic was associated with stomach cancer.
CONCLUSIONS
The current study does not support a major role of prebiotic fibers on selected upper digestive tract cancers. The association between high raffinose intake and reduced stomach cancer risk needs further investigation.
Topics: Humans; Stomach Neoplasms; Risk Factors; Raffinose; Diet; Prebiotics
PubMed: 37468063
DOI: 10.1016/j.jand.2023.07.008 -
BMC Plant Biology Oct 2023Cassava is an ideal food security crop in marginal and drought environment. However, the post-harvest storage of cassava is urgent problem to be resolved. In this study,...
Cassava is an ideal food security crop in marginal and drought environment. However, the post-harvest storage of cassava is urgent problem to be resolved. In this study, the storage tolerant and non-tolerant cassava were screened by measuring the change of Peroxidase (POD), Superoxide dismutase (SOD), Catalase (CAT) and Malondialdehyde (MDA) in seven cultivars of cassava. Compared with other cultivars, the cultivar of SC14 showed the highest level of SOD, MDA and POD respectively at 0 day, 12 day and 9 day postharvest while exhibited lowest level of CAT at 0 day postharvest, indicating the strongest antioxidant capability and storage tolerance. In contrast, GR15231, termed as storage non-tolerance cultivars, showed lowest SOD and POD at 12 day and kept a relative high level of CAT at 12 day post-harvest. In addition, SC14 has higher level of starch and dry substance than GR15231. Mass spectrum was performed for SC14 and GR15231 to explore the key metabolites regulating the storage tolerance of cassava. The results showed that the expression of glutathione (reduced) and raffinose was significantly decreased at 12 day post-harvest both in tolerant SC14 and non-tolerant GR15231. Compared with GR15231, SC14 showed higher level of raffinose both at 0 and 12 day post-harvest, indicating that raffinose may be the potential metabolites protecting SC14 cultivar from deterioration post-harvest. Additionally, raffinose ratio of SC14a/SC14b was five times less than that of GR15231a/GR15231b, reflecting the slower degradation of raffinose in SC14 cultivar compared with GR15231 cultivar. In conclusion, the antioxidant microenvironment induced by reduced glutathione and higher level of raffinose in SC14 cultivar might be the promising metabolites to improve its antioxidant capacity and antibiosis and thus maintained the quality of Cassava root tubers.
Topics: Antioxidants; Manihot; Raffinose; Glutathione; Antibiosis; Superoxide Dismutase; Peroxidase; Peroxidases
PubMed: 37807058
DOI: 10.1186/s12870-023-04466-7 -
Poultry Science Jul 2020Soybean oligosaccharides have been previously shown to be associated with the production of major odor-causing compounds in broilers, although little is known about the...
Soybean oligosaccharides have been previously shown to be associated with the production of major odor-causing compounds in broilers, although little is known about the role of stachyose and raffinose, which are key components of soybean oligosaccharide, in broiler cecal microbiota and odor compound production. To this end, soybean oligosaccharide, stachyose, and raffinose were added to the birds' diets to investigate their effects on odor compound production and the microbial community characteristics of the cecum in broilers. A total of 300 one-day-old Arbor Acre broilers with similar initial live weight were randomly allocated into 5 dietary groups with 6 replicates of 10 birds. The diets included soybean meal (positive control), soybean meal-free (negative control), 0.6% soybean oligosaccharide, 0.6% stachyose, or 0.6% raffinose. After a 49-D feeding period, both ceca were aseptically removed postmortem, and the contents were collected and analyzed for skatole, indole, volatile fatty acids, and lactic acid by using high performance liquid chromatography. Bacterial communities were detected by using a high-throughput sequencing platform based on IlluminaMiSeq 2500. Levels of skatole and indole tended to be lower in the dietary supplementation of oligosaccharides. The lowest levels of skatole and indole were observed in the stachyose group (P < 0.05), while the highest levels were found in the negative control group (P < 0.05). Concentrations of acetic acid and propionic acid in the stachyose group were increased (P < 0.05) while those of butyric acid and lactic acid were decreased (P < 0.05) compared with the soybean oligosaccharide and raffinose groups. Firmicutes and Bacteroidetes were prevalent in all groups, the proportion of Bacteroidetes was slightly decreased in the stachyose group, and Verrucomicrobia was abundant in the raffinose group (P > 0.05). Bacterial genera Alistipes and Parabacteroides were comparably abundant in the stachyose group, while Bacteroides, Lactobacillus, and Akkermansia were more abundant in the negative control, stachyose, and raffinose groups, respectively. Collectively, these findings demonstrated that dietary oligosaccharide supplementation significantly reduced odor compound production by modulating the cecal microbial community. Compared with soybean oligosaccharide and raffinose, the addition of stachyose into diets may help improve gut fermentation and minimize odor compound generation in broilers.
Topics: Animal Feed; Animals; Cecum; Chickens; Diet; Dietary Supplements; Dose-Response Relationship, Drug; Gastrointestinal Contents; Gastrointestinal Microbiome; Odorants; Oligosaccharides; Raffinose; Random Allocation; Glycine max
PubMed: 32616249
DOI: 10.1016/j.psj.2020.03.034 -
International Journal of Molecular... Aug 2022The raffinose synthetase () and galactinol synthase () are two critical enzymes for raffinose biosynthesis, which play an important role in modulating plant growth and...
The raffinose synthetase () and galactinol synthase () are two critical enzymes for raffinose biosynthesis, which play an important role in modulating plant growth and in response to a variety of biotic or abiotic stresses. Here, we comprehensively analyzed the and gene families and their involvement in abiotic and biotic stresses responses at the genome-wide scale in kiwifruit. A total of 22 and 24 genes were identified in and genomes. Phylogenetic analysis showed that the and genes were clustered into four and six groups, respectively. Transcriptomic analysis revealed that abiotic stresses strongly induced some crucial genes members including and and their expression levels were further confirmed by qRT-PCR. The GUS staining of ::GUS transgenic plants revealed that the transcriptionlevel of was significantly increased by salt stress. Overexpression of in demonstrated that this gene enhanced the raffinose accumulation and the tolerance to salt stress. The co-expression networks analysis of hub transcription factors targeting key genes indicated that there was a strong correlation between and expression under salt stress. Furthermore, the yeast one-hybrid assays showed that could bind the promoter directly. These results may provide insights into the evolutionary and functional mechanisms of and genes in kiwifruit.
Topics: Actinidia; Arabidopsis; Galactosyltransferases; Gene Expression Regulation, Plant; Phylogeny; Plant Proteins; Raffinose; Stress, Physiological
PubMed: 36012101
DOI: 10.3390/ijms23168836 -
Scientific Reports Jun 2022Entrapment of glucose oxidase (GOx) within metallic gold converts this widely used enzyme into a general saccharide oxidase. The following sugar molecules were oxidized...
Entrapment of glucose oxidase (GOx) within metallic gold converts this widely used enzyme into a general saccharide oxidase. The following sugar molecules were oxidized by the entrapped enzyme (in addition to D-glucose): fructose, xylose, L-glucose, glucose-6-phosphate, sucrose, lactose, methylglucoside, and the tri-saccharide raffinose. With the exception of raffinose, none of these sugars have a natural specific oxidase. The origin of this generalization of activity is attributed to the strong protein-gold 3D interactions and to the strong interactions of the co-entrapped CTAB with both the gold, and the protein. It is proposed that these interactions induce conformational changes in the channel leading to the active site, which is located at the interface between the two units of the dimeric GOx protein. The observations are compatible with affecting the specific conformation change of pulling apart and opening this gate-keeper, rendering the active site accessible to a variety of substrates. The entrapment methodology was also found to increase the thermal stability of GOx up to 100 °C and to allow its convenient reuse, two features of practical importance.
Topics: Glucose; Glucose Oxidase; Gold; Oxidoreductases; Raffinose; Sugars
PubMed: 35739181
DOI: 10.1038/s41598-022-14957-6