Antifreeze Polysaccharides from Wheat Bran: The Structural Characterization and Antifreeze Mechanism.

Exploring a novel natural cryoprotectant and understanding its antifreeze mechanism allows the rational design of future sustainable antifreeze analogues. In this study, various antifreeze polysaccharides were isolated from wheat bran, and the antifreeze activity was comparatively studied in relation to the molecular structure. The antifreeze mechanism was further revealed based on the interactions of polysaccharides and water molecules through dynamic simulation analysis. The antifreeze polysaccharides showed distinct ice recrystallization inhibition activity, and structural analysis suggested that the polysaccharides were arabinoxylan, featuring a xylan backbone with a majority of Araf and minor fractions of Manp, Galp, and Glcp involved in the side chain. The antifreeze arabinoxylan, characterized by lower molecular weight, less branching, and more flexible conformation, could weaken the hydrogen bonding of the surrounding water molecules more evidently, thus retarding the transformation of water molecules into the ordered ice structure.

Spermidine alleviates oxidative damage and enhances phenolic compounds accumulation in barley seedlings under UV-B stress.

BACKGROUND: Ultraviolet B (UV-B) radiation can enhance the accumulation of phenolic compounds (PCs) in barley seedling, although this may result in severe oxidative damage. In the present study, the role of spermidine in alleviating oxidative damage and regulating synthesis of PCs in barley seedlings under UV-B stress was investigated. RESULTS: Exogenous spermidine increased the length and fresh weight as well as PCs contents of barley seedlings under UV-B stress. Application of dicyclohexylamine, an inhibitor of endogenous spermidine synthesis, significantly inhibited the growth and PC accumulation of barley seedlings under UV-B stress, although this inhibitory effect can be alleviated by exogenous spermidine. Exogenous spermidine increased the contents of vanillic acid, syringic acid, protocatechuic acid and p-coumaric acid in barley seedlings under UV-B stress by 20-200% through enhancing the activities of enzymes related to synthesis of these acids. In addition, exogenous spermidine enhanced activities and gene expression of antioxidant enzymes in barley seedlings under UV-B stress, including peroxidase, glutathione reductase and glutathione S-transferase. CONCLUSION: Spermidine can alleviate oxidative damage of barley seedlings under UV-B stress and enhance the accumulation of PCs. © 2022 Society of Chemical Industry.

Effect of Amino Acids on Folates Accumulation in Wheat Seedlings during Germination under Red Light Radiation.

Deficiency of folates can cause various health problems, and germination is a potential way to enrich folates in grain-based food materials. In the present study, the effects of six amino acids (phenylalanine, tyrosine, tryptophan, glutamate, γ-aminobutyric acid, and p-aminobenzoic acid) on folate accumulation during wheat germination under red light radiation were investigated, and an optimized combination of amino acids for promoting folate enrichment was established. The results showed that applying phenylalanine, tyrosine, tryptophan, glutamate, or p-aminobenzoic acid to wheat seedlings during germination can significantly increase the content of total folates through activating the synthesis of the precursors for folate synthesis (pterin and p-aminobenzoic acid) or condensation of these two moieties. Meanwhile, up-regulation of corresponding genes was observed by measuring their expressions to investigate the mechanism for promoting the accumulation of folates. The highest content of folates (ca. 417 µg/100 g DW) was observed when the germinated wheat was cultured with a mixture of 1.5 mM phenylalanine, 0.5 mM tyrosine, 0.5 mM tryptophan, 0.75 mM p-aminobenzoic acid, and 0.5 mM glutamic acid, which was 50% higher than the control seedlings. This study established a promising and practical approach to enhance the accumulation of folates in wheat seedlings.

The mechanism of freeze-thawing induced accumulation of γ-aminobutyric acid in germinated soybean.

BACKGROUND: γ-Aminobutyric acid (GABA) is a non-protein amino acid with several functions in the human body. Although freeze-thawing could effectively accumulate GABA in soybean sprouts, the mechanism has not been revealed. The mechanism by which freeze-thawing enhances GABA accumulation in germinated soybean was revealed by evaluating GABA content, the activity of related synthesis enzymes, and the microstructure of the tissues and cells of sprouts. The germinated soybeans were treated at different temperatures (from -196 °C to 25 °C) for 12 h and then thawed at 25 °C for 6 h. RESULTS: The results showed that GABA content in frozen soybean sprouts did not change significantly before thawing. After thawing, the GABA content of sprouts increased by 83.9% and 82.9% when treated by liquid nitrogen flash freeze at - 80 °C for 12 h compared with the control (4 °C treatment for 12 h). The results indicated that GABA formation mainly occurred during thawing. However, glutamate decarboxylase (GAD), diamine oxidase (DAO), and aminoaldehyde dehydrogenase (AMADH) activity decreased during thawing. Based on the malonaldehyde (MDA) content and microstructure of sprouts, it was suggested that freezing at lower temperatures (< -20 °C) maintained the integrity of the cell structure, while the tissues and cell membranes were broken during thawing. CONCLUSION: These results could provide evidence for the hypothesis that GABA formation resulted from full contact between enzymes and substrates during thawing, rather than the contribution of higher enzyme activity. © 2019 Society of Chemical Industry.

NaCl stress on physio-biochemical metabolism and antioxidant capacity in germinated hulless barley (Hordeum vulgare L.).

BACKGROUND: Hulless barley generally grows in barren fields, where soil salinization is serious. However, only a few studies have been carried out investigating germinated hulless barley under salt stress. In the present study, the effect of NaCl stress on the physio-biochemical metabolism and antioxidant capacity of germinated hulless barley was investigated. RESULTS: NaCl stress inhibited seedling growth and caused oxidative damage, although it enhanced the accumulation of phenolic compounds and antioxidant capacity. The highest contents of total phenolic and main phenolic acids (vanillic acid, p-coumaric acid, ferulic acid and sinapic acid) were found with 60 mmol L-1 NaCl treatment, whereas 120 mmol L-1 NaCl inhibited the synthesis of phenolic components. Gene expression of phenylalanine ammonia lyase (PAL), cinnamic acid 4-hydroxylase (C4H), 4-coumarate coenzyme A ligase (4CL), p-coumaric acid 3-hdroxylase (C3H) and caffeic acid/5-hydroxyferulic acid O-methyltransferase (COMT), which participated in the synthesis of phenolic compounds, was up-regulated by NaCl stress, as were the enzyme activities of PAL, C4H and 4CL. NaCl treatment also enhanced the antioxidant enzyme activities of germinated hulless barley. CONCLUSION: NaCl stress inhibited seedlings growth and caused oxidative damage. Simultaneously, the antioxidant system of germinated hulless barley was enhanced. The results of the present study provide a theoretical basis with respect to the growth of hulless barley under salt stress. © 2018 Society of Chemical Industry.

Role of Ca2+ in phenolic compound metabolism of barley (Hordeum vulgare L.) sprouts under NaCl stress.

BACKGROUND: The literature on the role of calcium ion (Ca2+ ) in relation to phenolic compounds metabolism and related enzymes activities remains controversial. It is still unclear whether Ca2+ affects phenolic compounds content of barley sprouts. This study investigated the role and function of Ca2+ in phenolic compound metabolism of barley (Hordeum vulgare L.) sprouts under sodium chloride (NaCl) stress. RESULTS: Calcium chloride (CaCl2 ) significantly improved total calcium and calmodulin (CaM) contents as well as Ca2+ concentration, and enhanced phenolic compound accumulation by regulating the gene, protein expression and the activities of enzymes related to phenolics biosynthesis. Specifically, CaCl2 significantly increased the activities of phenylalanine ammonia lyase (PAL), cinnamic acid 4-hydroxylase (C4H), 4-coumarate coenzyme A ligase (4CL) and ferulic acid 5-hydroxylase (F5H) by up-regulating the corresponding protein expression. The activity of p-coumaric acid 3-hydroxylase (C3H) decreased during germination while caffeic acid O-methyltransferase (COMT) increased initially and then decreased, which was consistent with the changes in gene and protein expression under CaCl2 treatment. Conversely, lanthanum(III) chloride (LaCl3 ), ethylene glycol tetraacetic acid (EGTA) and 2-aminoethoxydiphenyl borate (2-APB) induced opposite effects. Decreased calcium and CaM contents and Ca2+ concentration were observed, and fluctuation change of relevant gene and protein expressions and PAL, C4H, 4CL, C3H, COMT and F5H activitives were also detected. CONCLUSION: Calcium ion played an important role for mediating NaCl stress-induced phenolics accumulation in barley sprouts. It required both Ca2+ influx and release from apoplast and intracellular stores, respectively. © 2019 Society of Chemical Industry.

Polyamines regulating phytic acid degradation in mung bean sprouts.

BACKGROUND: Polyamines are essentially involved in cell division and differentiation. Transport of polyamines is adenosine triphosphate (ATP)-dependent, while phytic acid is the major reserve of phosphate essential to the energy-producing machinery of cells. Thus polyamines might enhance phytic acid degradation during mung bean germination. In this study, different polyamines (putrescine (Put), spermidine (Spd) and spermine (Spm)) and dicyclohexylamine (DCHA, an inhibitor of Spd synthesis) were applied to investigate the function of polyamines on phytic acid degradation. RESULTS: Spd exhibited the best effect at the same concentration. Simultaneously, exogenous Spd improved sprout growth and enhanced the accumulation of gibberellin acid 3 (GA3 ), indole-3-acetic acid (IAA), abscisic acid (ABA) and cytokinin (CTK). This must be due to the increased endogenous polyamine contents. Apart from dramatically reducing phytic acid content, Spd resulted in the up-regulation of PA, PAP, MIPP and ALP transcript levels and the enhancement of phytase and acid phosphatase activities. However, DCHA application caused the opposite results, because it decreased endogenous polyamine contents. Furthermore, Spd alleviated the DCHA-induced inhibitory effect to some extent. CONCLUSION: Overall, polyamines, especially Spd, could accelerate phytic acid degradation in mung bean sprouts by inducing the synthesis of endogenous polyamines and phytohormones and enhancing the growth of sprouts. © 2017 Society of Chemical Industry.

Ca2+ influxes and transmembrane transport are essential for phytic acid degradation in mung bean sprouts.

BACKGROUND: Phytic acid is considered as an antinutrient. Ca2+ addition during germination has been proved to be an effective method for reducing phytic acid content in seeds. In this study, mung bean sprouts were treated with LaCl3 (La), verapamil (VP), ruthenium red (RR), and CaCl2 to explore the effect of Ca2+ influxes on phytic acid degradation. RESULTS: CaCl2 (6 mmol L-1 ) significantly improved extracellular and intracellular calcium precipitates and calcium content, elevated phytase and acid phosphatase activity, and further enhanced phytic acid degradation. Conversely, La, VP, or RR induced the opposite results. Among them, RR exhibited the most significant inhibitory effect. Decreased PA, PAP, MIPP, and ALP gene expression after VP or RR treatment was also observed. Enhanced or weakened extracellular Ca2+ influx or intracellular Ca2+ efflux was detected with increased or decreased calcium precipitates distributed in different compartments. However, CaCl2 addition differentially reversed the inhibitory effects of all channel blockers. CONCLUSION: CaCl2 enhanced Ca2+ influxes and accumulation in cells, which contributed to the regulation of phytic acid degradation. This study demonstrates that calcium channels play an essential role in mediating phytic acid degradation in mung bean sprouts, and both extracellular and intracellular Ca2+ -regulation were involved in phytic acid degradation. © 2017 Society of Chemical Industry.

Gibberellic acid promoting phytic acid degradation in germinating soybean under calcium lactate treatment.

BACKGROUND: Phytic acid as a phosphorus storage vault provides phosphorus for plant development. It is an anti-nutritional factor for humans and some animals. However, its degradation products lower inositol phosphates have positive effects on human health. In this study, the effect of gibberellic acid (GA) on phytic acid degradation under calcium lactate (Ca) existence was investigated. RESULTS: The results showed that Ca + GA treatment promoted the growth status, hormone metabolism and phytic acid degradation in germinating soybean. At the same time, the availability of phosphorus, the activity of phytic acid degradation-associated enzyme and phosphoinositide-specific phospholipase C (PI-PLC) increased. However, the relative genes expression of phytic acid degradation-associated enzymes did not vary in accordance with their enzymes activity. CONCLUSION: The results revealed that GA could mediate the transport and function of calcium and a series of physiological and biochemical changes to regulate phytic acid degradation of soybean sprouts. © 2017 Society of Chemical Industry.

Cyclic ADP-ribose and IP3 mediate abscisic acid-induced isoflavone accumulation in soybean sprouts.

In this study, the roles of ABA-cADPR-Ca2+ and ABA-IP3-Ca2+ signaling pathways in UV-B-induced isoflavone accumulation in soybean sprouts were investigated. Results showed that abscisic acid (ABA) up regulated cyclic ADP-ribose (cADPR) and inositol 1,4,5-trisphosphate (IP3) levels in soybean sprouts under UV-B radiation. Furthermore, cADPR and IP3, as second messengers of UV-B-triggered ABA, induced isoflavone accumulation by up-regulating proteins and genes expression and activity of isoflavone biosynthetic-enzymes (chalcone synthase, CHS; isoflavone synthase, IFS). After Ca2+ was chelated by EGTA, isoflavone content decreased. Overall, ABA-induced cADPR and IP3 up regulated isoflavone accumulation which was mediated by Ca2+ signaling via enhancing the expression of proteins and genes participating in isoflavone biosynthesis in soybean sprouts under UV-B radiation.

Effects of ABA and CaCl2 on GABA accumulation in fava bean germinating under hypoxia-NaCl stress.

Effects of exogenous abscisic acid (ABA) and CaCl2 on γ-aminobutyric acid (GABA) accumulation of germinated fava bean under hypoxia-NaCl stress were investigated. Exogenous ABA resulted in the enhancement of glutamate decarboxylase (GAD) and diamine oxidase (DAO) activity as well as GABA content in cotyledon and shoot. CaCl2 increased both enzyme activities in shoot and GABA content in cotyledon and shoot. ABA downregulated GAD expression in cotyledon and radicle, while upregulated that in shoot; it also upregulated DAO expression in each organ. CaCl2 upregulated GAD expression in cotyledon, while downregulated that in radicle. However, it upregulated DAO expression in shoot, downregulated that in radicle. ABA inhibitor fluridon and ethylenediaminetetraacetic acid inhibited GAD and DAO activities significantly so that inhibited GABA accumulation through reducing ABA biosynthesis and chelating Ca(2+), respectively. However, they upregulated GAD and DAO expression in varying degrees. These results indicate that ABA and Ca(2+) participate in GABA biosynthesis in fava bean during germination under hypoxia-NaCl stress.

Cloning of genes related to aliphatic glucosinolate metabolism and the mechanism of sulforaphane accumulation in broccoli sprouts under jasmonic acid treatment.

BACKGROUND: Cytochrome P450 79F1 (CYP79F1), cytochrome P450 83A1 (CYP83A1), UDP-glucosyltransferase 74B1 (UGT74B1), sulfotransferase 18 (ST5b) and flavin-containing monooxygenase GS-OX1 (FMOGS - OX1 ) are important enzymes in aliphatic glucosinolate biosynthesis. In this study, their full-length cDNA in broccoli was firstly cloned, then the mechanism of sulforaphane accumulation under jasmonic acid (JA) treatment was investigated. RESULTS: The full-length cDNA of CYP79F1, CYP83A1, UGT74B1, ST5b and FMOGS - OX1 comprised 1980, 1652, 1592, 1378 and 1623 bp respectively. The increase in aliphatic glucosinolate accumulation in broccoli sprouts treated with JA was associated with elevated expression of genes in the aliphatic glucosinolate biosynthetic pathway. Application of 100 µmol L(-1) JA increased myrosinase (MYR) activity but did not affect epithiospecifier protein (ESP) activity in broccoli sprouts, which was supported by the expression of MYR and ESP. Sulforaphane formation in 7-day-old sprouts treated with 100 µmol L(-1) JA was 3.36 and 1.30 times that in the control and 300 µmol L(-1) JA treatment respectively. CONCLUSION: JA enhanced the accumulation of aliphatic glucosinolates in broccoli sprouts via up-regulation of related gene expression. Broccoli sprouts treated with 100 µmol L(-1) JA showed higher sulforphane formation than those treated with 300 µmol L(-1) JA owing to the higher glucoraphanin content and myrosinase activity under 100 µmol L(-1) JA treatment. © 2016 Society of Chemical Industry.

Accumulation of γ-aminobutyric acid in soybean by hypoxia germination and freeze-thawing incubation.

BACKGROUND: γ-Aminobutyric acid (GABA) can be synthesised by the GABA shunt and polyamine degradation pathway in plants under hypoxia stress and lower temperature. The hypoxia germination freeze-thawing incubation was used as a new technique for accumulating more GABA in soybean. RESULTS: Results showed that glutamate decarboxylase (GAD) and diamine oxidase (DAO) activity as well as GABA content increased during germination within 24 h under hypoxia. However, the contents of dry matters and protein decreased. When the hypoxia-treated sprouts were frozen at -18 °C for 12 h and thawed at 25 °C for 6 h, GABA content increased drastically to 7.21-fold of the non-frozen sprouts. Subsequently, the freeze-thawing sprouts were ground into homogenates and incubated. GABA content was 14.20-fold of the only-soaked seeds when homogenates was incubated at 45 °C for 80 min within 400 µmol L(-1) pyridoxine (VB6) (pH 6.5). CONCLUSION: The hypoxia germination freeze-thawing incubation was an effective method for accumulating GABA in soybean. During incubation, DAO was more important for GABA formation in homogenate of germinated soybean compared with GAD.

Major nutrient compositions and functional properties of sorghum flour at 0-3 days of grain germination.

Germination is of importance to improving nutritional attributes of cereal grains for human consumption. The effect of germination time on major nutrient compositions and functional properties of sorghum flour was investigated in this study. Grains of Butanua, a new Sudanese sorghum cultivar, were germinated for 0, 1, 2, and 3 days to analyze their chemical and functional properties. The contents of starch, protein, oil, foaming stability, bulk density, and least gelation concentration of sorghum flour decreased, whereas oil absorption capacity, foaming capacity, and emulsion capacity and stability enhanced with an increase in germination time. Improved functional properties of sorghum flour by germination of the grains not only make it useful and suitable for various food processing formulations, but also improve the food product quality. This new finding will beneficially help develop innovative technologies, design new types of functional foods, and promote both sorghum production and relevant food processing industry in the future.

Effect of NaCl stress on health-promoting compounds and antioxidant activity in the sprouts of three broccoli cultivars.

Health-promoting compounds, antioxidant and myrosinase activity in the sprouts of three broccoli cultivars under 40 mM, 80 mM and 160 mM NaCl were investigated. LangYan (LY) sprouts had the richest health-promoting compounds among the evaluated cultivars. Treatment of 40 mM and 80 mM NaCl significantly decreased the content of ascorbic acid and total phenolic as well as antioxidant activity, but did not affect glucoraphanin, sulforaphane and myrosinase activity compared to the control. However, 160 mM NaCl treatment significantly enhanced the level of total phenolic, glucoraphanin, sulforaphane, antioxidant and myrosinase activity, while significantly decreased ascorbic acid content. The results suggest that 160 mM NaCl treatment would enhance antioxidant activity and sulforaphane yield in broccoli sprouts. The health-promoting value of broccoli sprouts depends on plant genotype and could be affected by NaCl stress.

Sequence analysis of diamine oxidase gene from fava bean and its expression related to γ-aminobutyric acid accumulation in seeds germinating under hypoxia-NaCl stress.

BACKGROUND: γ-Aminobutyric acid (GABA) is synthesized via the polyamine degradation pathway in plants, with diamine oxidase (DAO) being the key enzyme. In this study the cDNA of DAO in fava bean was cloned and its expression in seeds germinating under hypoxia-NaCl stress was investigated. RESULTS: Fava bean DAO cDNA is 2199 bp long and contains 2025 bp of open reading frame that encodes 675 amino acid peptides with a calculated molecular weight of 76.31 kDa and a pI of 5.41. Hypoxia and hypoxia-NaCl stress enhanced DAO activity and resulted in GABA accumulation in germinating fava bean. However, DAO gene expression was down-regulated under hypoxia compared with non-stress condition, while its expression in the cotyledon and shoot was up-regulated under hypoxia-NaCl. In addition, DAO expression could be promoted to enhance GABA accumulation after increasing the stress intensity using NaCl. DAO gene expression was significantly inhibited by aminoguanidine treatment under hypoxia but increased under hypoxia-NaCl. CONCLUSION: Under hypoxia, GABA accumulation due to NaCl was mainly concentrated in the cotyledon. The GABA content increase under hypoxia did not result from DAO gene expression, but DAO existing in seeds was activated under hypoxia. DAO gene expression was up-regulated to enhance GABA accumulation after increasing the stress intensity.

Production of High-Quality Wheat Sprouts of Strong Antioxidant Capacity: Process Optimization and Regulation Mechanism of Red Light Treatment.

Light treatment is an innovative method to enhance the synthesis of secondary metabolites in plants and improve the quality of plant-based food ingredients. This study investigated the effects of red light treatment on the physiological and biochemical changes during wheat germination, aiming to produce high-quality wheat sprouts with strong antioxidant capacity. Using response surface methodology, the study optimized the conditions for phenolic accumulation in wheat sprouts under red light treatment and explored the molecular mechanisms behind the enhancement of total phenolic content (TPC) and quality. The results indicated that red light treatment significantly increased the TPC in wheat sprouts. The highest TPC, reaching 186.61 µg GAE/sprout, was observed when wheat sprouts were exposed to red light at an intensity of 412 µmol/m²/s for 18.2 h/d over four days. Compared to no light, red light treatment significantly increased the content of photosynthetic pigments (chlorophyll and carotenoids). Red light treatment notably heightened the levels of both free and bound phenolic in the germinating wheat. Red light treatment markedly boosted the activities and relative gene expression levels of enzymes related to phenolic biosynthesis, including phenylalanine ammonia-lyase, cinnamate-4-hydroxylase, and 4-coumarate-CoA ligase. Additionally, red light treatment enhanced the antioxidant capacity of wheat sprouts by improving the activity and gene expression of four key antioxidant enzymes, thereby promoting growth and germination. This research suggested that red light treatment is an effective strategy for stimulating total phenolic biosynthesis, enhancing antioxidant capacity, and producing highly nutritious wheat sprouts, thus laying the groundwork for developing total phenolic-enriched wheat sprouts as valuable food ingredients in the future.

Metabolomics Analysis of Different Quinoa Cultivars Based on UPLC-ZenoTOF-MS/MS and Investigation into Their Antioxidant Characteristics.

In recent years, quinoa, as a nutritious and sustainable food material, has gained increasing popularity worldwide. To investigate the diversity of nutritional characteristics among different quinoa cultivars and explore their potential health benefits, metabolites of five quinoa cultivars (QL-1, SJ-1, SJ-2, KL-1 and KL-2) were compared by non-targeted metabolomics analysis based on UPLC-ZenoTOF-MS/MS in this study. A total of 248 metabolites across 13 categories were identified. Although the metabolite compositions were generally similar among the different quinoa cultivars, significant variations existed in their respective metabolite contents. Among the identified metabolites, amino acids/peptides, nucleosides, saponins and phenolic acids were the most abundant. Notably, SJ-1 exhibited the most distinct metabolite profile when compared to the other cultivars. Amino acids/peptides and nucleosides were found to be crucial factors contributing to the unique metabolite profile of SJ-1. Collectively, these aforementioned metabolites accounted for a substantial 60% of the total metabolites observed in each quinoa variety. Additionally, a correlation between the DPPH radical scavenging activity and the free phenolic content of quinoa was observed. Variations in phenolic content resulted in different antioxidant capacities among the quinoa cultivars, and SJ-1 exhibited lower phenolic levels and weaker antioxidant activity than the others. These results can provide important information for the development of quinoa resources.

Refrigerated storage stimulates isoflavone and γ-aminobutyric acid accumulation in germinated soybeans.

This study aims to investigate the quality changes of germinated soybeans during refrigerated storage (4 °C), with an emphasis on the stimulatory effect of refrigeration on their special functional compounds. After germinating for two days, germinated soybeans were stored at 4 °C for seven days, while the germinated soybeans stored at 25 °C served as control group. The results showed that refrigerated storage significantly affected the physiological changes in germinated soybeans. The weight loss rate, browning rate, malondialdehyde (MDA) content and H2O2 content all decreased dramatically during refrigerated storage compared to the control group. The total phenolic and total flavonoid contents of germinated soybeans under refrigeration exhibited a trend of increasing and then decreasing over time. Additionally, during refrigerated storage, the total isoflavone content reached a peak of 8.72 g/kg on the fifth day, in which the content of daidzein and glycitin increased by 45% and 49% respectively, when compared with the control group. Moreover, the content of γ-aminobutyric acid (GABA) peaked on the first day, and kept a high level during storage. In which, the refrigerated group was 2.35-, 2.88-, 1.67-fold respectively after storage for three to seven days. These results indicated that refrigeration stimulated the biosynthesis of isoflavones and GABA in germinated soybeans during storage. More importantly, there was a sequential difference in the timing of the stimulation of the two functional components under refrigeration.

UV-B Radiation Exhibited Tissue-Specific Regulation of Isoflavone Biosynthesis in Soybean Cell Suspension Cultures.

Isoflavones, a class of substances with high biological activity, are abundant in soybeans. This study investigated isoflavone biosynthesis in soybean cell suspension cultures under UV-B radiation. UV-B radiation enhanced the transcription level and activity of key enzymes involved in isoflavone synthesis in cell suspension cultures. As a result, the isoflavone contents significantly increased by 19.80% and 91.21% in hypocotyl and cotyledon suspension cultures compared with the control, respectively. Meanwhile, a significant difference was observed in the composition of isoflavones between soybean hypocotyl and cotyledon suspension cultures. Genistin was only detected in hypocotyl suspension cultures, whereas glycitin, daidzein, and genistein accumulated in cotyledon suspension cultures. Therefore, UV-B radiation exhibited tissue-specific regulation of isoflavone biosynthesis in soybean cell suspension cultures. The combination of suspension cultures and abiotic stress provides a novel technological approach to isoflavone accumulation.