Extracting Total Anthocyanin from Purple Sweet Potato Using an Effective Ultrasound-Assisted Compound Enzymatic Extraction Technology.
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ABSTRACT:
SUBMITTER: Wang F
PROVIDER: S-EPMC9317032 | biostudies-literature |
REPOSITORIES: biostudies-literature
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Project description:Sweet potato anthocyanins are water-soluble pigments with many physiological functions. Previous research on anthocyanin accumulation in sweet potato has focused on the roots, but the accumulation progress in the leaves is still unclear. Two purple sweet potato cultivars (Fushu No. 23 and Fushu No. 317) with large quantities of anthocyanin in the leaves were investigated. Anthocyanin composition and content were assessed with ultra-performance liquid chromatography diode-array detection (UPLC-DAD) and ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS), and the expressions of genes were detected by qRT-PCR. The two cultivars contained nine cyanidin anthocyanins and nine peonidin anthocyanins with an acylation modification. The acylation modification of anthocyanins in sweet potato leaves primarily included caffeoyl, p-coumaryl, feruloyl, and p-hydroxy benzoyl. We identified three anthocyanin compounds in sweet potato leaves for the first time: cyanidin 3-p-coumarylsophoroside-5-glucoside, peonidin 3-p-coumarylsophoroside-5-glucoside, and cyanidin 3-caffeoyl-p-coumarylsophoroside-5-glucoside. The anthocyanidin biosynthesis downstream structural genes DFR4, F3H1, anthocyanin synthase (ANS), and UDP-glucose flavonoid 3-O-glucosyltransferase (UFGT3), as well as the transcription factor MYB1, were found to be vital regulatory genes during the accumulation of anthocyanins in sweet potato leaves. The composition of anthocyanins (nine cyanidin-based anthocyanins and nine peonidin-based anthocyanins) in all sweet potato leaves were the same, but the quantity of anthocyanins in leaves of sweet potato varied by cultivar and differed from anthocyanin levels in the roots of sweet potatoes. The anthocyanidin biosynthesis structural genes and transcription factor together regulated and controlled the anthocyandin biosynthesis in sweet potato leaves.
| S-EPMC6832942 | biostudies-literature
Project description:Given the rise of morbidity and mortality caused by Klebsiella pneumoniae (KP), the increasing number of strains resistant to antibiotics, and the emergence of hypervirulent Klebsiella pneumonia, treatment of KP infection becomes difficult; thus, novel drugs are necessary for treatment. Anthocyanins, or natural flavonoids, have an extensive effect against bacterial infection. However, few studies on anti-KP are identified. Here, we evaluated the therapeutic effect of purple sweet potato anthocyanins (PSPAs) on KP, containing 98.7% delphinidin 3-sambubioside. Results showed that KP-infected mice after PSPAs treatment manifested decreased mortality, weakened lung injury, dampened inflammatory responses, and reduced bacterial systemic dissemination in vivo. In Vitro, PSPAs significantly suppressed pyroptosis and restricted NLRP3 inflammasome activation in alveolar macrophages infected with KP. As for the mechanism, PSPAs promote mitophagy by recruiting Parkin to the mitochondria. PSPAs-conferred mitophagy increased mitochondrial membrane potential and decreased mitochondrial reactive oxygen species and mitochondrial DNA, resulting in impaired NLRP3 inflammasome activation. In addition, the promotion of mitophagy by PSPAs required the Nrf2 signaling pathway. Collectively, these findings suggest that PSPAs are a potential option for the treatment of KP infection.
| S-EPMC8583717 | biostudies-literature
Project description:Purple-fleshed sweet potato is good for health due to rich anthocyanins in tubers. Although the anthocyanin biosynthetic pathway is well understood in up-ground organs of plants, the knowledge on anthocyanin biosynthesis in underground tubers is limited. In the present study, we isolated and functionally characterized a root-preferential gene encoding dihydrokaempferol reductase (IbDHKR) from purple-fleshed sweet potato. IbDHKR showed highly similarity with the reported dihydroflavonol reductases in other plant species at the sequence levels and the NADPH-binding motif and the substrate-binding domain were also found in IbDHKR. The tissue profile showed that IbDHKR was expressed in all the tested organs, but with much higher level in tuber roots. The expression level of IbDHKR was consistent with the anthocyanin content in sweet potato organs, suggesting that tuber roots were the main organs to synthesize anthocyanins. The recombinant 44 kD IbDHKR was purified and fed by three different dihydroflavonol substrates including dihydrokaempferol (DHK), dihydroquerctin, and dihydromyrecetin. The substrate feeding assay indicated that only DHK could be accepted as substrate by IbDHKR, which was reduced to leucopelargonidin confirmed by LC-MS. Finally, IbDHKR was overexpressed in transgenic tobacco. The IbDHKR-overexpression tobacco corolla was more highly pigmented and contained higher level of anthocyanins than the wild-type tobacco corolla. In summary, IbDHKR was a root-preferential gene involved in anthocyanin biosynthesis and its encoding protein, specifically catalyzing DHK reduction to yield leucopelargonidin, was a candidate gene for engineering anthocyanin biosynthetic pathway.
| S-EPMC5329058 | biostudies-literature
Project description:BackgroundAnthocyanins, which have important biological functions and have a beneficial effect on human health, notably account for pigmentation in purple-fleshed sweet potato tuberous roots. Individual regulatory factors of anthocyanin biosynthesis have been identified; however, the regulatory network of anthocyanin biosynthesis in purple-fleshed sweet potato is unclear.ResultsWe functionally determined that IbMYB340 cotransformed with IbbHLH2 in tobacco and strawberry receptacles induced anthocyanin accumulation, and the addition of IbNAC56a or IbNAC56b caused increased pigmentation. Furthermore, we confirmed the interaction of IbMYB340 with IbbHLH2 and IbNAC56a or IbNAC56b via yeast two-hybrid and firefly luciferase complementation assays; these proteins could form a MYB340-bHLH2-NAC56a or MYB340-bHLH2-NAC56b transcriptional complex to regulate anthocyanin biosynthesis by binding to the IbANS promoter rather than the IbUFGT promoter. Furthermore, it was found by a transient expression system in tobacco leaves that IbMYB44 could decrease anthocyanin accumulation. Moreover, the interaction of IbMYB44 with IbMYB340 and IbNAC56a or IbNAC56b was verified. This result suggested that IbMYB44 acts as a repressor of anthocyanin in sweet potato.ConclusionsThe repressor IbMYB44 affected anthocyanin biosynthesis by competitively inhibiting the IbMYB340-IbbHLH2-IbNAC56a or IbMYB340-IbbHLH2-IbNAC56b regulatory complex formation. Overall, the present study proposed a novel regulatory network whereby several vital TFs play key roles in regulating anthocyanin biosynthesis, and it provides strong insight into the potential mechanism underlying anthocyanin biosynthesis in sweet potato tuberous roots with purple color.
| S-EPMC7275474 | biostudies-literature
Project description:The purple sweet potato, Ipomoea batatas, belongs to the family Convolvulaceae. It is one of the most widely consumed tubers in Asia and is found in many dishes. Many people with diabetes eat purple sweet potato tubers to help reduce blood glucose in China.To predict the ultrasonic conditions for getting the optimal in vitro antioxidant and antiglycated activity of ultrasonic extracted polysaccharides from purple sweet potato (I. batatas) tubers, the artificial neural network (ANN) regression models was used in this study.The antioxidant activity of polysaccharides was quantified by evaluating the hydroxyl radical scavenging effect after ultrasonic extraction, and the data were used in conjunction with optimized extraction conditions to train the predictive ANN models.The following conditions were predicted to yield optimal hydroxyl scavenging activity: 200 W, 22°C, and 40 min. In contrast, conditions of 230 W, 22°C, and 50 min yielded the greatest inhibitory effect on albumin nonenzymatic glycosylation. The accuracy and predictive ability of the models ranged from good to excellent, as indicated by R2 values ranging from 0.953 to 0.998.The results of the present study showed that ANN predictive models are useful in ultrasonic processing, which can rapidly and accurately predict the optimum extraction conditions for polysaccharides based on their antioxidant and antiglycated activities. In addition, the results of the present study suggest that the consumption of sweet potatoes may help reduce free radicals in the body and prevent or treat diabetes.Ultrasonic extraction conditions were simulated and optimized using artificial neural networkBioactivities showed nonlinear relationship with ultrasonic conditionsThe optimal extraction conditions were 200 W, 22°C, and 40 min for the highest antioxidant capacityThe optimal extraction conditions were 230 W, 22°C, and 50 min for the highest antiglycated effect. Abbreviations used: IBP: Polysaccharide of Ipomoea batatas; RSM: response surface methodology; ANN: Artificial neural network; BSA: Bovine serum albumin.
| S-EPMC5551373 | biostudies-other
Project description:Dihydroflavonol-4-reductase (DFR) is a key enzyme in the catalysis of the stereospecific reduction of dihydroflavonols to leucoanthocyanidins in anthocyanin biosynthesis. In the purple sweet potato (Ipomoea batatas Lam.) cv. Ayamurasaki, expression of the IbDFR gene was strongly associated with anthocyanin accumulation in leaves, stems and roots. Overexpression of the IbDFR in Arabidopsis tt3 mutants fully complemented the pigmentation phenotype of the seed coat, cotyledon and hypocotyl. Downregulation of IbDFR expression in transgenic sweet potato (DFRi) using an RNAi approach dramatically reduced anthocyanin accumulation in young leaves, stems and storage roots. In contrast, the increase of flavonols quercetin-3-O-hexose-hexoside and quercetin-3-O-glucoside in the leaves and roots of DFRi plants is significant. Therefore, the metabolic pathway channeled greater flavonol influx in the DFRi plants when their anthocyanin and proanthocyanidin accumulation were decreased. These plants also displayed reduced antioxidant capacity compared to the wild type. After 24 h of cold treatment and 2 h recovery, the wild-type plants were almost fully restored to the initial phenotype compared to the slower recovery of DFRi plants, in which the levels of electrolyte leakage and hydrogen peroxide accumulation were dramatically increased. These results provide direct evidence of anthocyanins function in the protection against oxidative stress in the sweet potato. The molecular characterization of the IbDFR gene in the sweet potato not only confirms its important roles in flavonoid metabolism but also supports the protective function of anthocyanins of enhanced scavenging of reactive oxygen radicals in plants under stressful conditions.
| S-EPMC3817210 | biostudies-literature
Project description:In various plant species, many transcription factors (TFs), such as MYB, bHLH, and WD40, have been identified as regulators of anthocyanin biosynthesis in underground organs. However, the regulatory elements of anthocyanin biosynthesis in the tuberous roots of sweet potato have not been elucidated yet. Here, we selected the purple-fleshed sweet potato cultivar "Zhezi1" (ZZ P ) and its spontaneous yellow-fleshed mutant "Xinli" (XL Y ) to investigate the regulatory mechanism of the anthocyanin biosynthesis in the tuberous roots of sweet potato. By analyzing the IbMYB1 genotype in ZZ P and XL Y , we found that the IbMYB1-2, a MYB TF involved in anthocyanin biosynthesis, was missing in the XL Y genome, which might lead to an extreme decrease in anthocyanins in XL Y . A comparative transcriptome analysis of ZZ P and XL Y was conducted to find the TFs involved in anthocyanin biosynthesis in ZZ P and XL Y . The anthocyanin structural genes were significantly enriched among the differentially expressed genes. Moreover, one MYB activator (IbMYB1), one bHLH (IbbHLH2), three WRKY activator candidates (IbWRKY21, IbWRKY24, and IbWRKY44), and two MYB repressors (IbMYB27 and IbMYBx-ZZ) were highly expressed in ZZ P accompanied with anthocyanin structural genes. We also tested the expression of these TFs in six purple- and two orange-fleshed sweet potato cultivars. Interestingly, most of these TFs were significantly positively correlated with anthocyanin contents in these cultivars. The function of the anthocyanin biosynthesis repression of IbMYB27 and IbMYBx-ZZ was verified through transient co-transformation with IbMYB1 into tobacco leaves. Further functional verification of the above TFs was conducted by Y2H, BiFC, and dual-luciferase assays. These tests showed that the MYB-bHLH-WD40/MYB-bHLH-WD40-WRKY complex activated the promoter of anthocyanin structural gene IbDFR and promoters for IbWRKY44, IbMYB27, and IbMYBx-ZZ, indicating reinforcement and feedback regulation to maintain the level of anthocyanin accumulation in the tuberous roots of purple-fleshed sweet potato. These results may provide new insights into the regulatory mechanism of anthocyanin biosynthesis and accumulation in underground organs of sweet potatoes.
| S-EPMC9393619 | biostudies-literature
Project description:IntroductionThe potato (Solanum tuberosum) cultivar 'Xin Daping' is tetraploid with white skin and white flesh, while the cultivar 'Hei Meiren' is also tetraploid with purple skin and purple flesh. Comparative transcriptome analysis of white and purple cultivars was carried out using high-throughput RNA sequencing in order to further understand the mechanism of anthocyanin biosynthesis in potato.Methods and resultsBy aligning transcript reads to the recently published diploid potato genome and de novo assembly, 209 million paired-end Illumina RNA-seq reads from these tetraploid cultivars were assembled on to 60,930 transcripts, of which 27,754 (45.55%) are novel transcripts and 9393 alternative transcripts. Using a comparison of the RNA-sequence datasets, multiple versions of the genes encoding anthocyanin biosynthetic steps and regulatory transcription factors were identified. Other novel genes potentially involved in anthocyanin biosynthesis in potato tubers were also discovered. Real-time qPCR validation of candidate genes revealed good correlation with the transcriptome data. SNPs (Single Nucleotide Polymorphism) and indels were predicted and validated for the transcription factors MYB AN1 and bHLH1 and the biosynthetic gene anthocyanidin 3-O-glucosyltransferase (UFGT).ConclusionsThese results contribute to our understanding of the molecular mechanism of white and purple potato development, by identifying differential responses of biosynthetic gene family members together with the variation in structural genes and transcription factors in this highly heterozygous crop. This provides an excellent platform and resource for future genetic and functional genomic research.
| S-EPMC4459980 | biostudies-literature
Project description:Glycosylation contributes to the diversity and stability of anthocyanins in plants. The process is catalysed by various glucosyltransferases using different anthocyanidin aglycones and glycosyl donors. In this study, we found that an anthocyanidin 3-O-glucoside-2″-O-glucosyltransferase (3GGT) from purple sweet potato (Ipomoea batatas) catalyses the conversion of anthocyanidin 3-O-glucoside into anthocyanidin 3-O-sophoroside, which is functionally different from the 3GGT ortholog of Arabidopsis. Phylogenetic analysis indicated regioselectivity of 3GGT using uridine-5'-diphosphate (UDP)-xylose or UDP-glucose as the glycosyl is divergent between Convolvulaceae and Arabidopsis. Homology-based protein modeling and site-directed mutagenesis of Ib3GGT and At3GGT suggested that the Thr-138 of Ib3GGT is a key amino acid residue for UDP-glucose recognition and that it plays a major role in sugar-donor selectivity. Wild-type and ugt79b1 mutants (defective in UDP carbohydrate-dependent glycosyltransferases, UGTs) of Arabidopsis plants overexpressing Ib3GGT produced the new component cyanidin 3-O-sophoroside. Moreover, Ib3GGT expression was associated with anthocyanin accumulation in different tissues during I. batatas plant development and was regulated by the transcription factor IbMYB1. Localization assays for Ib3GGT showed that glycosyl extension occurs in the cytosol and not in the endoplasmic reticulum. This study therefore reveals the function of Ib3GGT in glycosyl extension of anthocyanins and demonstrates that Thr-138 is the key amino acid residue for UDP-glucose recognition.
| S-EPMC6255700 | biostudies-literature
Project description:BackgroundSpectral reflectance was evaluated for its usefulness as a nondestructive estimation of chlorophyll (Chl) content from three cultivars of sweet potato (Ipomoea batatas L.) with green, yellow, and purple leaves grown in a greenhouse for 22 days. While the green and yellow leaves contain variant amount of photosynthetic pigments without or with little level of anthocyanins, the purple leaves, except large amount of photosynthetic pigments, have high quantity of anthocyanins.ResultsFor green and yellow leaves, the reciprocal reflectance (R-1) and derived indices incorporating near infrared (NIR) reflectance, [(Rλ)-1 - (RNIR)-1] and [(RNIR/Rλ) - 1], in the green and red edge spectral ranges were shown to be strongly correlated (r2 = 0.8 ~ 0.9) with the chlorophyll content. The root mean square error (RMSE) of the chlorophyll content estimation using these indices was < 50 mg m-2. However, when purple leaves containing high levels of anthocyanins were included in the sample, R-1 in the green spectral range and the above-mentioned indices displayed much weaker correlations with the chlorophyll content. The RMSE of chlorophyll estimation using these indices in the green spectral range sharply increased to > 110 mg m-2 when the sample included purple leaves. The new index, [1 - (Rλ/RNIR)], was therefore inferred and developed to eliminate the distorting effect of anthocyanins on chlorophyll content estimation using reflectance in the green spectral range. For leaves with high levels of anthocyanins, the correlation between [1 - (Rλ/RNIR)] and the chlorophyll content remained strong (r2 = 0.8 ~ 0.9) in the green spectral range, and the RMSE was minimal.ConclusionThe reflectance index, [1 - (Rλ/RNIR)], therefore represents a new and useful parameter for estimating leaf chlorophyll content in leaves with any level of anthocyanins such as purple rice leaf.
| S-EPMC5432896 | biostudies-literature