Characterization of the closely related Arabidopsis thaliana ß-ketoacyl-CoA synthases KCS3, KCS12 and KCS19.

The cuticle, consisting of cuticular wax and cutin, is a lipid membrane that seals the plant surface against environmental stress. ß-Ketoacyl-CoA synthases (KCSs) are condensing enzymes catalyzing crucial reactions elongating hydrocarbon chains into precursors for various cuticular wax components. Although many KCS genes were well characterized in various species, the functions of the closely related Arabidopsis KCS3, KCS12, KCS19 enzymes remained unclear. Here, we found KCS3 preferentially expressed in growing organs, especially in guard cells. kcs3 mutants and kcs3kcs12 double mutants displayed sepal fusion phenotypes, suggesting defects in cuticle formation. The mutants had decreased amounts of wax components with relatively short hydrocarbon chains in the developing organs but increased levels of wax compounds in mature organs. In contrast, kcs19 mutants showed seed fusion phenotypes and altered chain length distributions in seed suberin. Taken together, our results show that KCS12 and KCS3 share redundant functions in flower development, while KCS19 is involved in seed coat formation. All three condensing enzymes are involved in the elongation of C>18 hydrocarbon chains in young, actively expanding tissues.

Tetracosanoic acids produced by 3-ketoacyl-CoA synthase 17 are required for synthesizing seed coat suberin in Arabidopsis.

Very long-chain fatty acids (VLCFAs) are precursors for the synthesis of membrane lipids, cuticular waxes, suberins, and storage oils in plants. 3-Ketoacyl CoA synthase (KCS) catalyzes the condensation of C2 units from malonyl-CoA to acyl-CoA, the first rate-limiting step in VLCFA synthesis. In this study, we revealed that Arabidopsis KCS17 catalyzes the elongation of C22-C24 VLCFAs required for synthesizing seed coat suberin. Histochemical analysis of Arabidopsis plants expressing GUS (ß-glucuronidase) under the control of the KCS17 promoter revealed predominant GUS expression in seed coats, petals, stigma, and developing pollen. The expression of KCS17:eYFP (enhanced yellow fluorescent protein) driven by the KCS17 promoter was observed in the outer integument1 of Arabidopsis seed coats. The KCS17:eYFP signal was detected in the endoplasmic reticulum of tobacco epidermal cells. The levels of C22 VLCFAs and their derivatives, primary alcohols, α,ω-alkane diols, ω-hydroxy fatty acids, and α,ω-dicarboxylic acids increased by ~2-fold, but those of C24 VLCFAs, ω-hydroxy fatty acids, and α,ω-dicarboxylic acids were reduced by half in kcs17-1 and kcs17-2 seed coats relative to the wild type (WT). The seed coat of kcs17 displayed decreased autofluorescence under UV and increased permeability to tetrazolium salt compared with the WT. Seed germination and seedling establishment of kcs17 were more delayed by salt and osmotic stress treatments than the WT. KCS17 formed homo- and hetero-interactions with KCR1, PAS2, and ECR, but not with PAS1. Therefore, KCS17-mediated VLCFA synthesis is required for suberin layer formation in Arabidopsis seed coats.

Microencapsulation of phenolic compounds extracted from soybean seed coats by spray-drying.

This study aimed to characterize and microencapsulate soybean seed coats phenolic compounds by spray-drying, evaluating physicochemical properties and storage stability. Different extraction methodologies were used to obtain crude extract (SCE), ethyl acetate fraction, water fraction, and bound phenolic extract. Extraction yield, total phenolic and flavonoid contents, and antioxidant capacity were determined. HPLC-electrospray ionization source-MS/MS analysis was performed on SCE. Microencapsulation by spray-drying of SCE incorporating 10%, 20%, and 30% maltodextrin (MD) was carried out. Drying yield (DY), encapsulation efficiency (EE), moisture, morphology and particle size, dry, and aqueous storage stability were evaluated on the microcapsules. SCE had 7.79 g/100 g polyphenolic compounds (mainly isoflavones and phenolic acids) with antioxidant activity. Purification process by solvent partitioning allowed an increase of phenolic content and antioxidant activity. Microcapsules with 30% MD exhibited the highest DY, EE, and stability. Microencapsulated polyphenolic compounds from soybean seed coats can be used as functional ingredients in food products. PRACTICAL APPLICATION: Soybean seed coat is a usually discarded agro-industrial by-product, which presents antioxidant compounds of interest to human health. These compounds are prone to oxidation due to their chemical structure; therefore, microencapsulation is a viable and reproducible solution to overcome stability-related limitations. Microencapsulation of soybean seed coats polyphenols is an alternative which protects and extends the stability of phenolic compounds that could be potentially incorporated into food products as a natural additive with antioxidant properties.

Mass Spectrometry-Based Untargeted Metabolomics Reveals the Importance of Glycosylated Flavones in Patterned Lentil Seed Coats.

Lentil seed coats are rich in antioxidant polyphenols that are important for plant defense and have potential as valorized byproducts. Although biochemical differences among lentil seed coat colors have been previously studied, differences among seed coat patterns remain largely unexplored. This study used mass spectrometry-based untargeted metabolomics to investigate polyphenol differences among lentil seed coat patterns to search for biochemical pathways potentially responsible for seed coat pattern differences. Comparing patterned with non-patterned green lentil seed coats, 28 significantly upregulated metabolites were found in patterned seed coats; 19 of them were identified as flavones. Flavones were virtually absent in non-patterned seed coats, thereby strongly suggesting a blockage in their flavone biosynthetic pathway. Although the black pattern is not readily discernible on black seed coats, many of the same flavones found in green marbled seed coats were also found in black seed coats, indicating that black seed coats likely have a marbled pattern.

An update on nutritional profile, phytochemical compounds, health benefits, and potential applications in the food industry of pulses seed coats: A comprehensive review.

Pulses, as a sustainable source of nutrients, are an important choice for human diets, but vast quantities of seed coats generated in pulses processing are usually discarded or used as low-value ruminant feed. It has been demonstrated that pulses seed coats are excellent sources of dietary nutrients and phytochemicals with potential health benefits. With growing interest in the sustainable use of resources and the circular economy, utilization of pulses seed coats to recover these valuable components is a core objective for their valorization and an important step toward agricultural sustainability. This review comprehensively provides a comprehensive insight on the nutritional and phytochemical profiles presented in pulses seed coats and their health benefits obtained from the findings of in vitro and in vivo studies. Furthermore, in the food industry, pulses seed coats can be acted as potential food ingredients with nutritional, antioxidant and antimicrobial characteristics or as the matrix or active components of films for food packaging and edible coatings. A better understanding of pulses seed coats may provide a reference for increasing the overall added value and realizing the pulses' sustainable diets.

The walnut shell network: 3D visualisation of symplastic and apoplastic transport routes in sclerenchyma tissue.

MAIN CONCLUSION: High symplastic connectivity via pits was linked to the lignification of the developing walnut shell. With maturation, this network lessened, whereas apoplastic intercellular space remained and became relevant for shell drying. The shell of the walnut (Juglans regia) sclerifies within several weeks. This fast secondary cell wall thickening and lignification of the shell tissue might need metabolites from the supporting husk tissue. To reveal the transport capacity of the walnut shell tissue and its connection to the husk, we visualised the symplastic and apoplastic transport routes during shell development by serial block face-SEM and 3D reconstruction. We found an extensive network of pit channels connecting the cells within the shell tissue, but even more towards the husk tissue. Each pit channel ended in a pit field, which was occupied by multiple plasmodesmata passing through the middle lamella. During shell development, secondary cell wall formation progressed towards the interior of the cell, leaving active pit channels open. In contrast, pit channels, which had no plasmodesmata connection to a neighbouring cell, got filled by cellulose layers from the inner cell wall lamellae. A comparison with other nut species showed that an extended network during sclerification seemed to be linked to high cell wall lignification and that the connectivity between cells got reduced with maturation. In contrast, intercellular spaces between cells remained unchanged during the entire sclerification process, allowing air and water to flow through the walnut shell tissue when mature. The connectivity between inner tissue and environment was essential during shell drying in the last month of nut development to avoid mould formation. The findings highlight how connectivity and transport work in developing walnut shell tissue and how finally in the mature state these structures influence shell mechanics, permeability, conservation and germination.

Enhanced adsorption of copper ions by phosphoric acid-modified Paeonia ostii seed coats.

Novel adsorbent, phosphoric acid-modified Paeonia ostii seed coats (PA-PSC) were successfully prepared by low-temperature pyrolysis to effectively remove Cu(II) from aqueous solution. The results revealed that equilibrium adsorption capacity (qe) of PA-PSC for Cu(II) was notably enhanced up to 4-folds compared with the raw PSC. FT-IR and XPS analyses suggested that the adsorption of Cu(II) by PA-PSC was primarily ascribed to electrostatic forces and complexing effects. Besides, equilibrium and kinetic studies demonstrated that Freundlich and pseudo-second-order models were the actually fairly good approximations of Cu(II) adsorption. Thermodynamic analysis revealed that the adsorption of Cu(II) onto PA-PSC was a chemical, endothermic, and spontaneous process. Lastly, reusability study further confirmed the applicability of PA-PSC as a promising adsorbent for removing Cu(II) from aqueous solution.

Assessment of the use of cooked Araucaria angustifolia seed coats extract as food for brine shrimp.

The use of vegetable waste and its screening for potential cytotoxicity is of utmost importance to ensure its safe use in the feed industry for fish and other animals. We evaluated aqueous and ethanolic extracts of cooked Araucaria angustifolia seed coats. The Stiasny index for the aqueous and ethanolic extracts was 2.87% ± 0.03% and 60.53% ± 4.79%, respectively. Condensed tannins were 11-fold higher in the ethanolic extract than the aqueous extract. The flavonoid and polyphenol contents were 1.7- and 1.8-fold higher in the ethanolic extract than in the aqueous extract, respectively. The 36 h EC50 for brine shrimp hatchability was 300.32 µg/mL for the aqueous extract, and 76.60 µg/mL for the ethanolic extract. The 24 h LC50 was 1405.96 µg/mL for the aqueous extract, and it was 356.32 µg/mL for the ethanolic extract. The aqueous extract was nontoxic to A. salina nauplii, and therefore, it can be used as a possible food additive in fish feed. The results also demonstrated that the different solvents used in the extraction affected the yield and the total phenolic, total flavonoid, and condensed tannin content. Further in vivo and cell line cytotoxicity testing is recommended to substantiate these findings.

[Simultaneous rapid detection of ten stilbenes in serum of mice by UPLC-MS/MS].

Stilbenes is a class of natural polyphenols with 1,2-diphenylethylene as the skeleton structure which have structural and active diversity. However, there are fewer studies on their metabolic process, which limits the in-depth research and development of such components. An UPLC-MS/MS method simultaneously determining contents of ten stilbenes was firstly established in this study and applied to study the ten stilbenes of peony seed coats in the serum of C57 mice.Piceatannol was the internal standard, and methanol was used for protein precipitation, UPLC-MS/MS with negative ion mode was used for analysis, and the method was validated.The serum samples were collected and detected after mice being oral administered with 800 mg·kg~(-1) peony seed coat extracts for 8 weeks. The results showed that suffruticosol A, suffruticosol B, suffruticosol C, trans-ε-viniferin, cis-gnetin H, trans-suffruticosol D and trans-gnetin H were detected in serum samples, and the highest is suffruticosol A. The method is simple and quick with high specificity and sensitivity, and it is suitable for quantitative determination of ten stilbenes in the serum of mice.

Polyphenolic Profiles of Yellow Bean Seed Coats and Their Relationship with Iron Bioavailability.

Previous work with Caco-2 cell cultures has shown that individual polyphenols can either promote or inhibit iron uptake. This investigation was designed to characterize the relationship between iron bioavailability and seed coat polyphenol composition in a panel of 14 yellow beans representing five market classes with the potential for fast cooking time and high iron content. The study included two white and two red mottled bean lines, which represent high and low iron bioavailability capacity in dry beans, respectively. Polyphenols were measured quantitatively by high-performance liquid chromatography-mass spectrometry (HPLC-MS)/UV and iron bioavailability of seed coat extracts was measured in Caco-2 assays. Thirteen of the yellow bean seed types contained high concentrations (up to 35.3 ± 2.7 µmol/g) of kaempferol 3-glucoside (k 3-g), a known promoter of iron uptake. A general association between the ratio of promoting to inhibiting polyphenols (P/I) and iron uptake was observed. The presence of iron uptake inhibiting condensed tannins proportionately countered the promotional effects of kaempferol compounds. Unidentified factors present in seed coats other than polyphenols also appeared to affect iron uptake.

Polyphenol profile comparisons of seed coats of five pulse crops using a semi-quantitative liquid chromatography-mass spectrometric method.

INTRODUCTION: Pulse crops are nutritious and therefore widely grown. Pulse seed coats are typically discarded, despite their high content of polyphenols that are known for their antioxidant properties and health benefits. A better understanding of polyphenol diversity and biochemical pathways will ultimately provide insight into how polyphenols are linked to health benefits, which will help to better utilise these seed coats. OBJECTIVES: To explore polyphenol profiles among seed coats of diverse genotypes of five pulse crops using a targeted liquid chromatography mass spectrometry (LC-MS) method. METHODS: Four genotypes of each of common bean, chickpea, pea, lentil and faba bean seed coats were selected for analysis. Following extraction, polyphenols were quantified using LC-MS. RESULTS: An LC-MS method was developed to quantify 98 polyphenols from 13 different classes in 30 min. The low-tannin seed coats had the lowest concentrations of all polyphenols. Chickpea and pea seed coats had the most similar polyphenolic profiles. The black common bean showed the most diverse seed coat polyphenol profile, including several anthocyanins not detected in any of the other seed coats. CONCLUSION: The LC-MS method reported herein was used to show polyphenol diversity within several polyphenol classes among the pulse crop seed coats. Detected in all seed coats, flavonols and hydroxybenzoic acids appear well-conserved in the edible Fabaceae. The presence of anthocyanins, flavan-3-ols and proanthocyanins in the coloured seed coats suggests that unique divergent branches were introduced in the flavonoid biosynthetic pathway, possibly in response to environmental stressors.

Simultaneous rapid detection of ten stilbenes in serum of mice by UPLC-MS/MS / 中国中药杂志

Stilbenes is a class of natural polyphenols with 1,2-diphenylethylene as the skeleton structure which have structural and active diversity. However, there are fewer studies on their metabolic process, which limits the in-depth research and development of such components. An UPLC-MS/MS method simultaneously determining contents of ten stilbenes was firstly established in this study and applied to study the ten stilbenes of peony seed coats in the serum of C57 mice.Piceatannol was the internal standard, and methanol was used for protein precipitation, UPLC-MS/MS with negative ion mode was used for analysis, and the method was validated.The serum samples were collected and detected after mice being oral administered with 800 mg·kg~(-1) peony seed coat extracts for 8 weeks. The results showed that suffruticosol A, suffruticosol B, suffruticosol C, trans-ε-viniferin, cis-gnetin H, trans-suffruticosol D and trans-gnetin H were detected in serum samples, and the highest is suffruticosol A. The method is simple and quick with high specificity and sensitivity, and it is suitable for quantitative determination of ten stilbenes in the serum of mice.

Comparative Transcriptomics Analysis of Brassica napus L. during Seed Maturation Reveals Dynamic Changes in Gene Expression between Embryos and Seed Coats and Distinct Expression Profiles of Acyl-CoA-Binding Proteins for Lipid Accumulation.

Production of vegetable oils is a vital agricultural resource and oilseed rape (Brassica napus) is the third most important oil crop globally. Although the regulation of lipid biosynthesis in oilseeds is still not fully defined, the acyl-CoA-binding proteins (ACBPs) have been reported to be involved in such metabolism, including oil accumulation, in several plant species. In this study, progressive changes in gene expression in embryos and seed coats at different stages of seed development were comprehensively investigated by transcriptomic analyses in B. napus, revealing dynamic changes in the expression of genes involved in lipid biosynthesis. We show that genes encoding BnACBP proteins show distinct changes in expression at different developmental stages of seed development and show markedly different expression between embryos and seed coats. Both isoforms of the ankyrin-repeat BnACBP2 increased during the oil accumulation period of embryo development. By contrast, the expression of the three most abundant isoforms of the small molecular mass BnACBP6 in embryos showed progressive reduction, despite having the highest overall expression level. In seed coats, BnACBP3, BnACBP4 and BnACBP5 expression remained constant during development, whereas the two major isoforms of BnACBP6 increased, contrasting with the data from embryos. We conclude that genes related to fatty acid and triacylglycerol biosynthesis showing dynamic expression changes may regulate the lipid distribution in embryos and seed coats of B. napus and that BnACBP2 and BnACBP6 are potentially important for oil accumulation.

Instability of common beans during storage causes hardening: The role of glass transition phenomena.

Long-term storage of common beans leads to loss of cooking quality and an ill-defined solution, appropriate storage, is recommended. Therefore, the polymer science theory of glasses that hypothesizes stability of a system below its glass transition temperature (Tg) was applied to determine bean stability during storage in relation to cooking behavior. Since composition influences Tg, powders of cotyledons and seed coats in addition to whole beans were equilibrated above different saturated salt solutions in order to generate materials with different moisture contents. A thermal mechanical compression test which measures compressibility changes in a system upon reaching its glass-rubber transition temperature region was conducted to obtain the Tg. A Tg-moisture relation was established, whose relevance was confirmed by storage and cooking experiments which showed development of hard-to-cook in beans stored above Tg but not below it. Therefore, this relation constitutes a stability map for storage of common beans.

Cotyledon pectin molecular interconversions explain pectin solubilization during cooking of common beans (Phaseolus vulgaris).

Dynamics of pectin extractability in cotyledons and seed coats were explored for mechanistic insight into pectin changes due to aging and cooking of beans. In addition, changes in mineral distribution during cooking were determined in order to investigate their retention in the matrix. Pre-soaked fresh and aged beans were cooked in demineralized water for different times and the cotyledons, seed coats and cooking water were lyophilized. From cotyledon and seed coat powders, alcohol insoluble residue (AIR) was extracted and sequentially fractionated into water-, chelator- and sodium carbonate-extractable pectin (WEP, CEP and NEP, respectively). Characterization of pectin in AIR and pectin fractions revealed inherent structural differences between cotyledon and seed coat pectin with the latter exhibiting a lower degree of methylesterification (DM) and being more linear. Due to aging, WEP decreased whilst NEP substantially increased and the CEP fraction and DM of pectin in AIR did not change significantly, suggesting a more crucial role of increased covalent bonding than cation-mediated crosslinking in aging-induced hardening of beans. During cooking, some NEP was converted into WEP and no pectin depolymerization was observed from molar mass distribution profiles. Pectin changes due to aging and cooking of beans were more pronounced in the cotyledon compared to the seed coat. Whilst Ca2+, Fe2+ and Zn2+ were largely retained in the bean matrix during cooking, Mg2+ was largely leached from cotyledons into the cooking water. In conclusion, aging-induced hardening of beans and softening during cooking were found to be premised on interconversion of pectin fractions in cotyledons.

The Dead Can Nurture: Novel Insights into the Function of Dead Organs Enclosing Embryos.

Plants have evolved a variety of dispersal units whereby the embryo is enclosed by various dead protective layers derived from maternal organs of the reproductive system including seed coats (integuments), pericarps (ovary wall, e.g., indehiscent dry fruits) as well as floral bracts (e.g., glumes) in grasses. Commonly, dead organs enclosing embryos (DOEEs) are assumed to provide a physical shield for embryo protection and means for dispersal in the ecosystem. In this review article, we highlight recent studies showing that DOEEs of various species across families also have the capability for long-term storage of various substances including active proteins (hydrolases and ROS detoxifying enzymes), nutrients and metabolites that have the potential to support the embryo during storage in the soil and assist in germination and seedling establishment. We discuss a possible role for DOEEs as natural coatings capable of "engineering" the seed microenvironment for the benefit of the embryo, the seedling and the growing plant.

Mechanistic insight into softening of Canadian wonder common beans (Phaseolus vulgaris) during cooking.

The relative contributions of cotyledons and seed coats towards hardening of common beans (Phaseolus vulgaris) were investigated and the rate-limiting process which controls bean softening during cooking was determined. Fresh or aged whole beans and cotyledons were soaked and cooked in demineralised water or 0.1 M NaHCO3 solution, and texture evolution, microstructure changes and thermal properties were studied. Fresh and aged whole beans cooked in demineralised water had significantly different softening rate constants and so did fresh and aged cotyledons. The comparable softening rate constants of aged whole beans and cotyledons indicated an insignificant role of the seed coat in hardening during storage. All samples cooked faster in 0.1 M NaHCO3 solution. Disintegration of cooked tissues followed by microscopic examination revealed a transition from cell breakage through a phase of cell breakage and separation to complete cell separation with increased cooking time wherefore texture decayed. Therefore, progressive solubilization of pectin in the middle lamella greatly promoted texture decay. While residual birefringence even after substantial cooking time suggested some molecular order of the starch, calorimetric analyses revealed complete starch gelatinisation before complete cell separation occurred. This implies an insignificant role of starch in texture decay during cooking but its hindered uncoiling into a viscous gel after gelatinisation due to the restricting cell wall could promote its retrogradation. Therefore, we suggest that the rate-determining process in bean softening relates to cell wall/middle lamella changes influencing pectin solubilization.

Characterization of Polyphenol Effects on Inhibition and Promotion of Iron Uptake by Caco-2 Cells.

Polyphenolic compounds present in the seed coat of common bean (Phaseolus vulgaris L.) are known to act collectively as inhibitors of iron bioavailability. Recent research identified specific polyphenols as being potent Fe uptake inhibitors. That research also identified other polyphenols as being promoters of Fe uptake. The present study extends that work using a Caco-2 cell model to characterize the effects of 43 additional polyphenols on Fe uptake. In addition, this study indicates that the inhibitory compounds have a more potent effect that outweighs the ability of promoting compounds to increase Fe uptake. For example, a ratio of 100:0 epicatechin (a promoter)/myricetin (an inhibitor) produced 78.5 ± 6.7 ng ferritin/mg protein, 90:10 yielded 27.4 ± 3.0, 50:50 yielded 3.42 ± 0.54, and 0:100 yielded 2.26 ± 0.25 ng ferritin/mg protein. A simulation of the relative concentrations of eight major polyphenols (four inhibitors, four promoters) present in a sample of black bean seed coats demonstrated that most of the inhibitory compounds would need to be removed to reduce the negative effect on Fe uptake. In vivo studies are now warranted to confirm the above in vitro effects. Such work would be significant as other bean color classes exist that are likely to have polyphenolic profiles that are more favorable to Fe bioavailability.

Antimicrobial Activities of Phenolic Extracts Derived from Seed Coats of Selected Soybean Varieties.

Soybean hulls or seed coats consist of complex carbohydrates, proteins, lipids, and polyphenols such as anthocyanidins, proanthocyanidins, and isoflavones. The polyphenolics in the seed coats give them various colors such as black, brown, green, yellow, or even a mottled appearance. In this study, the antimicrobial effects of phenolic extracts from the seed coats of different colored soybeans (yellow, dark brown, brown, and black) were evaluated against foodborne pathogens such as Salmonella Typhimurium, Escherichia coli O157:H7, and Campylobacter jejuni in broth-cultures as well as on chicken skin. The highest total phenolic content was observed for the phenolic extract from soybean variety (R07-1927) with black colored seed coat (74.1 ± 2.1 mg chlorogenic acid equivalent [CAE]/g extract) and was significantly different (P <0.0001) from the extract of the conventional soybean variety (R08-4004) with yellow colored seed coat (7.4 ± 1.2 mg CAE/g extract). The extract from black colored soybean produced reductions of 2.10 ± 0.08 to 2.20 ± 0.08-log CFU/mL for both E. coli O157:H7 and C. jejuni after 3 d when incubated in broth-culture having 4-log CFU/mL of bacteria, whereas a 6 d incubation was found to reduce S. Typhimurium and E. coli O157:H7 at 2.03 ± 0.05 and 3.3 ± 0.08-log CFU/mL, respectively. The extract also reduced S. Typhimurium and E. coli O157:H7 attached to chicken skin by 1.39 ± 0.03 and 1.24 ± 0.06-log CFU/g, respectively, upon incubation for 6 d. Soybean seed coat extracts may have a potency as antimicrobial agents to reduce foodborne bacteria contaminating poultry products.

Aspergillus awamori-fermented mung bean seed coats enhance the antioxidant and immune responses of weaned pigs.

The potential benefits of Aspergillus-fermented mung bean seed coats (FMSC) for weaned pigs remain unexplored. Both in vitro and in vivo experiments were employed to evaluate the potential of FMSC supplement on the growth, antioxidant and immune responses of weaned pigs. The total polyphenols and DPPH scavenging capability of ethanol extract of FMSC exhibited a greater (p < 0.01) increase than those of pre-fermentation. With the addition of the polyphenol of FMSC extract, an increase in phagocytosis by neutrophils and proliferation of peripheral blood mononuclear cells (PBMC) were found. However, these observations were significantly inhibited (p < 0.05) in those activated cells. Next, 96 weaned pigs were allotted with a randomized complete block design into four dietary treatments, including 0 (control), 600, 1200 or 1800 mg/kg FMSC in a corn-soya bean meal basal diet for a 35-day trial. The pigs were injected with swine enzootic pneumonia (SEP) vaccines at day 3 and day 21 respectively. The results showed that dietary treatment failed to affect growth performance or serum SEP titre. The diet supplemented with 600-1800 mg/kg FMSC decreased faecal lactoferrin on day 21 and increased plasma trolox equivalent antioxidant capacity (TEAC) and erythrocytes catalase activity, as well as decreased (p < 0.01) plasma malondialdehyde (MDA) concentration on day 35. Diet supplementation of 1800 mg/kg FMSC increased phagocytosis by neutrophils and PBMC proliferation induced by pokeweed mitogen (PWM). However, the polymorphonuclear leucocytes (PMN)-positive respiratory burst cells were decreased in the supplementation of 1200 or 1800 mg/kg FMSC respectively. In addition, the serum haptoglobin concentration was decreased in the supplementation with 1200 mg/kg FMSC. Taken together, FMSC enriches polyphenols with antioxidative and immune modulated properties. After feeding FMSC, an improvement in antioxidative capability and immunocompetence was found, implying that FMSC could provide as a feed additive at optimal level 1200 mg/kg for weaned pigs.