Insights into tissue-specific anthocyanin accumulation in Japanese plum (Prunus salicina L.) fruits: A comparative study of three cultivars.

In the present study, three matured Japanese plum cultivars with different colored peel and flesh were selected to mine the key transcription factors regulating anthocyanin formation in tissues. Results showed that PsMYB10 was correlated with structural genes C4H, F3H, and ANS. PsMYB6 could positively regulate C4H (r = 0.732) and accumulated anthocyanins in Sanhua plum's flesh. Sanhua plum has the highest phenolic and anthocyanin contents (10.24 ± 0.37 gallic acid equivalent mg g-1 dry weight (DW) and 68.95 ± 1.03 µg g-1 DW), resulting itself superior biological activity as 367.1 ± 42.9 Trolox equivalent mg g-1 DW in oxygen radical absorbance capacity value and 72.79 ± 4.34 quercetin equivalent mg g-1 DW in cellular antioxidant activity value. The present work provides new insights into the regulatory mechanism of tissue-specific anthocyanin biosynthesis, confirming the pivotal role of anthocyanins in the biological activity of plums, providing essential support for the development of horticultural products enriched with anthocyanins.

Comparative Analysis of Phytochemical Profiles and Antioxidant Activities between Sweet and Sour Wampee (Clausena lansium) Fruits.

As a local medicine and food, wampee fruit, with abundant bioactive compounds, is loved by local residents in Southern China. Titratable acid (TA), total sugar (TS), and total phenolic and flavonoid contents were detected, and phytochemical profiles and cellular antioxidant activities were analyzed by the HPLC and CAA (cellular antioxidant activity) assay in five sweet wampee varieties and five sour wampee varieties. Results showed that the average TS/TA ratio of sweet wampee varieties was 29 times higher than sour wampee varieties, while TA content was 19 times lower than sour wampee varieties. There were much lower levels of total phenolics, flavonoids, and antioxidant activities in sweet wampee varieties than those in sour wampee varieties. Eight phytochemicals were detected in sour wampee varieties, including syringin, rutin, benzoic acid, 2-methoxycinnamic acid, kaempferol, hesperetin, nobiletin, and tangeretin, while just four of them were detected in sweet wampee varieties. Syringin was the only one that was detected in all the sour wampee varieties and was not detected in all sweet wampee varieties. Correlation analysis showed significant positive correlations between TA with phenolics, flavonoids, and total and cellular (PBS wash) antioxidant activities, while there were significant negative correlations between TS/TA with phenolic and cellular (no PBS wash) antioxidant activities. This suggested that the content of titratable acid in wampee fruit might have some relationship with the contents of phenolics and flavonoids. Sour wampee varieties should be paid much attention by breeders for their high phytochemical contents and antioxidant activities for cultivating germplasms with high health care efficacy.

Phytochemical Profiles and Cellular Antioxidant Activities in Chestnut (Castanea mollissima BL.) Kernels of Five Different Cultivars.

In this study, the phytochemical profiles, total and cellular antioxidant activities of five different Chinese chestnut (Castanea mollissima BL.) cultivars were analyzed. Phenolics, flavonoids as well as phytochemical compounds in five cultivars of chestnut kernels were determined. Results showed that the free forms played a dominant role in total phenolics, flavonoids and antioxidant activities of all five cultivars of chestnut kernels. The cultivar 'Fyou' showed the highest total and free phenolic contents, 'Heguoyihao' showed the highest total and free flavonoids contents, and 'Chushuhong' showed the highest total and cellular antioxidant activities. Eight phenolic compounds were detected, and chlorogenic acid, gallic acid, and quercetin were shown as three predominant components in all five cultivars. These results provide valuable information which may be a guidance for selection of good chestnut variety to be used as functional food.

The Combination of Hot Air and Chitosan Treatments on Phytochemical Changes during Postharvest Storage of 'Sanhua' Plum Fruits.

Plum fruits would become putrid quickly after harvest. In order to prolong postharvest life, 'Sanhua' plum fruits were treated by hot air combined with a chitosan coating, and stored at low temperature. Fruit firmness, total soluble solids, total phytochemical contents were evaluated along with total antioxidant activities and phytochemical components. Results showed that hot air treatment delayed softening process of plum fruit. The total phenolics and flavonoids accumulated and antioxidant activities increased in both control and treatment samples during storage. These values in the samples treated with hot air and chitosan were all higher than control and hot air treatments. Phytochemicals of epicatechin, cyanidin, pelargonidin, and hesperetin were all upregulated by hot air and chitosan treatment, especially epicatechin. This suggested that chitosan might play an important role in regulating phytochemical profiles of 'Sanhua' plum fruits during storage.

Dynamic Changes of Ascorbic Acid, Phenolics Biosynthesis and Antioxidant Activities in Mung Beans (Vigna radiata) until Maturation.

To better understand the regulatory mechanism of phenolics and ascorbic acid accumulation as well as antioxidant activities in mung beans during legume development, the gene expression profiles of 25 key-coding genes in ascorbic acid and phenolics metabolic pathways were analyzed. As well as the dynamitic changes of ascorbic acid, phenolic profiles and antioxidant activities with legume development were studied. The results indicated that gene expression profiles were closely related to the ascorbic acid and phenolics accumulation regularity during legume development. VrVTC2 and VrGME played important roles for ascorbic acid accumulation from 8 to 17 days after flowering (DAF). VrPAL and VrCHS exhibited positive correlations with daidzein and glycitin accumulation, and VrIFS had a strong positive correlation with glycitin biosynthesis. Antioxidant activities dramatically increased during mung bean maturing, which were significantly related to ascorbic acid and phenolics accumulation. Eight days after flowering was the essential stage for ascorbic acid and phenolics biosynthesis in mung beans.

Effect of Steaming Processing on Phenolic Profiles and Cellular Antioxidant Activities of Castanea mollissima.

The intention of this study was to investigate the effect of steaming processing on phenolic profiles and antioxidant activities in chestnuts. Steaming processing at different times and temperatures depicted diverse impacts on free and bound fractions. Though, bound phenolics were stable but long time steaming at higher temperatures tended to improve the levels of phenolics, flavonoids as well as antioxidant activities in chestnut kernels, by up to 60.11% of the original value. Seven phenolic compounds including ferulic acid, chlorogenic acid, gallic acid, vanillic acid, syringate, p-coumaric acid and quercetin were found to change during thermal processes. Significant relationships (p < 0.05) were identified between total phenolics and total antioxidant activities. However, the consistency of chlorogenic acid (p < 0.01) only with cellular antioxidant activity indicated poor bio-accessibility of the phytochemicals in chestnuts. However, this situation could be partly improved by steaming. Steaming could improve the cellular accessibility of free phytochemicals, particularly, increasing the bio-accessibility by 41.96%. This study provided valuable information on dynamic changes of phenolic profiles and antioxidant activity of chestnuts under a steaming process, which could offer possible guidance for the chestnut processing industry in the future.

Phytochemicals Accumulation in Sanhua Plum ( Prunus salicina L.) during Fruit Development and Their Potential Use as Antioxidants.

This study followed the flesh reddening of Sanhua plum from the surface to the center during fruit development. Five key stages were identified based on color changes during fruit ripening: full-green (FG), red-appeared, half-red, full-red, and purple-red (PR). Fruits were collected and analyzed for phytochemicals and antioxidant properties. Concurrently, the transcript levels of genes associated with phenolic, flavonoid, and anthocyanin production were investigated. The titratable acid content of Sanhua plum decreased during development, while total soluble sugar content increased. In addition, both the total phenolic content and total flavonoid content decreased during development, while anthocyanin content increased. The polyphenol oxidase activity peaked in the PR stage. The maximum antioxidant activity in vitro was observed in the FG stage, while cellular antioxidant activity peaked in the PR stage.

Dynamic effects of fermentation on phytochemical composition and antioxidant properties of wampee (Clausena lansium (Lour.) Skeel) leaves.

Variations in the phytochemical composition and antioxidant properties were studied in the wine of wampee leaves obtained at different stages of fermentation process. The highest concentrations of total phenolic and flavonoid contents were attained at Day 12 and Day 21 of fermentation, respectively. In addition, five phytochemical compounds including vanillic acid, p-coumaric acid, rutin, ferulic acid, and 7-hydroxycoumarin were identified and quantified by HPLC in fermented wampee products. The strongest antioxidant activity in wine was monitored on Day 12. Furthermore, total antioxidant activity was significantly correlated with vanillic acid, p-coumaric acid, ferulic acid, and 7-hydroxycoumarin compared with rutin. The obtained results suggested that 12-day fermentation could be an optimal process for excavation of applying wampee leaves into food and wine industries.

Impact of Leaf Development Stages on Polyphenolics Profile and Antioxidant Activity in Clausena lansium (Lour.) Skeels.

Clausena lansium (Lour.) Skeels, commonly known as "wampee," is an excellent food ingredient of medicinal value. Effects of leaf developmental stages on the composition of phenolics, flavonoids, and antioxidant activity were investigated. Phenolics composition was studied using HPLC-PAD, whereas antioxidant activity was estimated by oxygen radical absorbance capacity (ORAC) and cellular antioxidant activity (CAA) assays. Increase in bound flavonoids, quercetin, and cellular antioxidant activity was observed in bound and free fractions at different stages of leaf development. Predominantly, quercetin and ferulic acid contents were high in free and bound fractions of old leaves. In addition, phenolic components depicted highly significant positive association (p < 0.05) with antioxidant activity. Overall, old leaves of wampee have utility value similar to leaf buds, so they could be a more sustainable and economical source of bioactive compounds for commercial application in nutraceutical and pharmaceutical industries.

Transcriptome profiling reveals regulatory mechanisms underlying corolla senescence in petunia.

The genetic regulatory mechanisms that govern natural corolla senescence in petunia are not well understood. To identify key genes and pathways that regulate the process, we performed a transcriptome analysis in petunia corolla at four developmental stages, including corolla fully opening without anther dehiscence (D0), corolla expansion, 2 days after anthesis (D2), corolla with initial signs of senescence (D4), and wilting corolla (D7). We identified large numbers of differentially expressed genes (DEGs), ranging from 4626 between the transition from D0 and D2, 1116 between D2 and D4, a transition to the onset of flower senescence, and 327 between D4 and D7, a developmental stage representing flower senescence. KEGG analysis showed that the auxin- and ethylene-related hormone biosynthesis and signaling transduction pathways were significantly activated during the flower development and highly upregulated at onset of flower senescence. Ethylene emission was detected at the D2 to D4 transition, followed by a large eruption at the D4 to D7 transition. Furthermore, large numbers of transcription factors (TFs) were activated over the course of senescence. Functional analysis by virus-induced gene silencing (VIGS) experiments demonstrated that inhibition of the expression of TFs, such as ethylene-related ERF, auxin-related ARF, bHLH, HB, and MADS-box, significantly extended or shortened flower longevity. Our data suggest that hormonal interaction between auxin and ethylene may play critical regulatory roles in the onset of natural corolla senescence in petunia.

Phenolic compounds, antioxidant activity, antiproliferative activity and bioaccessibility of Sea buckthorn (Hippophaë rhamnoides L.) berries as affected by in vitro digestion.

Phenolics, antioxidant and antiproliferative properties of Sea buckthorn berries were evaluated using a simulated in vitro digestion and compared with a chemical extraction method. Digested samples were subjected to antiproliferation evaluation against human liver, breast and colon cancer cells. Furthermore, the bioaccessibility of digested berries was evaluated using a Caco-2 cell culture model. Results revealed that after enzymatic digestion the phenolic compounds were quite different from the chemical extracts, more flavonoid aglycones were released, whereas less total phenolics, phenolic acids and flavonoid glycosides were detected. Although the extracellular antioxidant activity of the digesta was lower than that of extracts, the cellular antioxidant activity (CAA) and antiproliferative effects of berries were significantly enhanced by digestion. This was attributed to their higher flavonoid aglycone content and could be verified by testing individual active compounds, suggesting that the cellular uptake of samples might be improved, which was also certified by the Caco-2 cell uptake model. The digested samples showed an almost 5-fold cellular accumulative amount of isorhamnetin than pure isorhamnetin, which was attributed to the significant down regulation of the mRNA expression level of efflux transporters MRP2 and P-gp. This finding indicated that the digestion enhanced the bioaccessibility of bioactive compounds of berries.

A basic helix-loop-helix transcription factor, PhFBH4, regulates flower senescence by modulating ethylene biosynthesis pathway in petunia.

The basic helix-loop-helix (bHLH) transcription factors (TFs) play important roles in regulating multiple biological processes in plants. However, there are few reports about the function of bHLHs in flower senescence. In this study, a bHLH TF, PhFBH4, was found to be dramatically upregulated during flower senescence. Transcription of PhFBH4 is induced by plant hormones and abiotic stress treatments. Silencing of PhFBH4 using virus-induced gene silencing or an antisense approach extended flower longevity, while transgenic petunia flowers with an overexpression construct showed a reduction in flower lifespan. Abundance of transcripts of senescence-related genes (SAG12, SAG29) was significantly changed in petunia PhFBH4 transgenic flowers. Furthermore, silencing or overexpression of PhFBH4 reduced or increased, respectively, transcript abundances of important ethylene biosynthesis-related genes, ACS1 and ACO1, thereby influencing ethylene production. An electrophoretic mobility shift assay showed that the PhFBH4 protein physically interacted with the G-box cis-element in the promoter of ACS1, suggesting that ACS1 was a direct target of the PhFBH4 protein. In addition, ectopic expression of this gene altered plant development including plant height, internode length, and size of leaves and flowers, accompanied by alteration of transcript abundance of the gibberellin biosynthesis-related gene GA2OX3. Our results indicate that PhFBH4 plays an important role in regulating plant growth and development through modulating the ethylene biosynthesis pathway.

A Petunia homeodomain-leucine zipper protein, PhHD-Zip, plays an important role in flower senescence.

Flower senescence is initiated by developmental and environmental signals, and regulated by gene transcription. A homeodomain-leucine zipper transcription factor, PhHD-Zip, is up-regulated during petunia flower senescence. Virus-induced gene silencing of PhHD-Zip extended flower life by 20% both in unpollinated and pollinated flowers. Silencing PhHD-Zip also dramatically reduced ethylene production and the abundance of transcripts of genes involved in ethylene (ACS, ACO), and ABA (NCED) biosynthesis. Abundance of transcripts of senescence-related genes (SAG12, SAG29) was also dramatically reduced in the silenced flowers. Over-expression of PhHD-Zip accelerated petunia flower senescence. Furthermore, PhHD-Zip transcript abundance in petunia flowers was increased by application of hormones (ethylene, ABA) and abiotic stresses (dehydration, NaCl and cold). Our results suggest that PhHD-Zip plays an important role in regulating petunia flower senescence.