Physiological and Transcriptome Analysis Illuminates the Molecular Mechanisms of the Drought Resistance Improved by Alginate Oligosaccharides in Triticum aestivum L.

Alginate oligosaccharides(AOS)enhance drought resistance in wheat(Triticum aestivum L.),but the definite mechanisms remain largely unknown.The physiological and transcriptome responses of wheat seedlings treated with AOS were analyzed under drought stress simulated with polyethylene glycol-6000.The results showed that AOS promoted the growth of wheat seedlings and reduced oxidative damage by improving peroxidase and superoxide dismutase activities under drought stress.A total of 10,064 and 15,208 differentially expressed unigenes(DEGs)obtained from the AOS treatment and control samples at 24 and 72 h after dehydration,respectively,were mainly enriched in the biosynthesis of secondary metabolites(phenylpropanoid biosynthesis,flavonoid biosynthesis),carbohydrate metabolism(starch and sucrose metabolism,carbon fixation in photosynthetic organisms),lipid metabolism(fatty acid elongation,biosynthesis of unsaturated fatty acids,alpha-linolenic acid metabolism,cutin,suberine and wax biosynthesis),and signaling transduction pathways.The up-regulated genes were related to,for example,chlorophyll a-b binding protein,amylosynthease,phosphotransferase,peroxidase,phenylalanine ammonia lyase,flavone synthase,glutathione synthetase.Signaling molecules(including MAPK,plant hormones,H_(2)O_(2) and calcium)and transcription factors(mainly including NAC,MYB,MYB-related,WRKY,bZIP family members)were involved in the AOS-induced wheat drought resistance.The results obtained in this study help underpin the mechanisms of wheat drought resistance improved by AOS,and provides a theoretical basis for the application of AOS as an environmentally sustainable biological method to improve drought resistance in agriculture.

Enzymatically prepared alginate oligosaccharides improve broiler chicken growth performance by modulating the gut microbiota and growth hormone signals

Background Alginate oligosaccharide(AOS)holds great potential as a novel feed supplement in farm animals.However,the effects of AOS on chicken health and the underlying mechanisms are not fully understood.This study aimed to optimize the enzymatic preparation of AOS by using bacterial alginate lyases expressed in yeast,investigate the effects of the prepared AOS on the growth performance and gut health of broiler chickens,and reveal the underlying mechanisms.Results Five alginate lyases from bacteria were cloned into Pichia pastoris GS115 and the alginate lyase PDE9 was expressed at relatively high yield,activity and stability in P.pastoris.Animal trials were carried out using 3201-day-old male Arbor Acres broilers(four groups;8 replicates/group×10 chicks/replicate)receiving either a basal diet or the same diet supplemented with 100,200 and 400 mg/kg PDE9-prepared AOS for 42 d.The results showed that dietary supplementation of 200 mg/kg AOS displayed the highest activity in promoting the birds’ADG and ADFI(P<0.05).AOS ameliorated the intestinal morphology,absorption function and barrier function,as indicated by the enhanced(P<0.05)intestinal villus height,maltase activity,and the expression of PEPT,SGLT1,ZNT1,and occludin.AOS also increased serum insulin-like growth factor-1,ghrelin(P<0.05),and growth hormone(P<0.1).Moreover,the concentrations of acetate,isobutyrate,isovalerate,valerate,and total SCFAs in cecum of birds fed AOS were significantly higher than the control birds(P<0.05).Metagenomic analysis indicated that AOS modulated the chicken gut microbiota structure,function,and microbial interactions and promoted the growth of SCFAs-producing bacteria,for example,Dorea sp.002160985;SCFAs,especially acetate,were found positively correlated with the chicken growth performance and growth-related hormone signals(P<0.05).We further verified that AOS can be utilized by Dorea sp.to grow and to produce acetate in vitro.Conclusions We demonstrated that the enzymatically produced AOS effectively promoted broiler chicken growth performance by modulating the chicken gut microbiota structure and function.For the first time,we established the connections among AOS,chicken gut microbiota/SCFAs,growth hormone signals and chicken growth performance.

Alginate oligosaccharides preparation, biological activities and their application in livestock and poultry

Alginate oligosaccharides(AOS), belonging to the class of functional marine oligosaccharides, are low-molecular polymers linked by β-1,4-mannuronic acid(M) and α-1,4-guluronic acid(G), which could be classically obtained by enzymatic hydrolysis of alginate. With low viscosity and good water solubility, as well as anti-oxidant, immune regulation, anti-bacterial and antiinflammatory activities, AOS have been widely used in medical science and functional food, green agriculture and other fields. As new bio-feed additives, AOS have broad potential applications in animal husbandry. In this review, the sources of alginate, chemical structure and preparation methods of AOS, and their biological activities and application in livestock and poultry are summarized. We expect this review could contribute to lay a foundation of application and further research for AOS in livestock and poultry.

Response of Salmonella enterica serovar Typhimurium to alginate oligosaccharides fermented with fecal inoculum:integrated transcriptomic and metabolomic analyses

Alginate oligosaccharides(AOS),extracted from marine brown algae,are a common functional feed additive;however,it remains unclear whether they modulate the gut microbiota and microbial metabolites.The response of Salmonella enterica serovar Typhimurium,a common poultry pathogen,to AOS fermented with chicken fecal inocula was investigated using metabolomic and transcriptomic analyses.Single-strain cultivation tests showed that AOS did not directly inhibit the growth of S.Typhimurium.However,when AOS were fermented by chicken fecal microbiota,the supernatant of fermented AOS(F-AOS)exhibited remarkable antibacterial activity against S.Typhimurium,decreasing the abundance ratio of S.Typhimurium in the fecal microbiota from 18.94 to 2.94%.Transcriptomic analyses showed that the 855 diferentially expressed genes induced by F-AOS were mainly enriched in porphyrin and chlorophyll metabolism,oxidative phosphorylation,and Salmonella infection-related pathways.RT-qPCR confrmed that F-AOS downregulated key genes involved in fagellar assembly and the type III secretory system of S.Typhimurium,indicating metabolites in F-AOS can infuence the growth and metabolism of S.Typhimurium.Metabolomic analyses showed that 205 microbial metabolites were signifcantly altered in F-AOS.Among them,the increase in indolelactic acid and 3-indolepropionic acid levels were further confrmed using HPLC.This study provides a new perspective for the application of AOS as a feed additive against pathogenic intestinal bacteria.

Alginate oligosaccharide-mediated butyrate-HIF-1α axis improves skin aging in mice

The“gut-skin”axis has been proved and is considered as a novel therapy for the prevention of skin aging.The antioxidant efficacy of oligomannonic acid(MAOS)makes it an intriguing target for use to improve skin aging.The present study further explored whereby MAOS-mediated gut-skin axis balance prevented skin aging in mice.The data indicated the skin aging phenotypes,oxidative stress,skin mitochondrial dysfunction,and intestinal dysbiosis(especially the butyrate and HIF-1a levels decreased)in aging mice.Similarly,fecal microbiota transplantation(FMT)from aging mice rebuild the aging-like phenotypes.Further,we demonstrated MAOS-mediated colonic butyrate-HIF-1a axis homeostasis promoted the entry of butyrate into the skin,upregulated mitophagy level and ultimately improving skin aging via HDAC3/PHD/HIF-1a/mitophagy loop in skin of mice.Overall,our study offered a better insights of the effectiveness of alginate oligosaccharides(AOS),promised to become a personalized targeted therapeutic agents,on gut-skin axis disorder inducing skin aging.

Synergistic effects of alginate oligosaccharide and cyanidin-3-O-glucoside on the amelioration of intestinal barrier function in mice

Emerging evidence shows that dietary oligosaccharides are important prebiotics that can improve intestinal flora,while dietary polyphenols can act directly on intestinal cells.However,information about their synergistic effects on gut health is still limited.In this study,alginate oligosaccharide(AOS)and cyanidin-3-O-glucoside(C3G)were selected as a common marine plant oligosaccharide and terrestrial plant polyphenol,respectively,to study their effects on intestinal health.The results show that,in comparison to their individual applications,the combination of AOS and C3G(mass ratio,3:1)displayed a stronger ability to up-regulate the expression of tight junction proteins,while enhanced intestinal epithelial barrier was also observed and levels of mucin-2 andβ-defensins were simultaneously increased in the intestinal mucus.Interestingly,the secretion of immunoglobulin A and immune-related cytokines were approximately doubled by the AOS+C3G mixture.In addition,the AOS+C3G mixture was found to be more conducive to the positive transformation of intestinal flora,which stimulated the growth of beneficial bacteria Akkermansia,Lachnospiraceae and Feacalibaculum while inhibiting the growth of harmful bacteria Helicobacter and Turicibacter.The data generated herein thus suggests that dietary oligosaccharides and dietary polyphenols may be more beneficial to intestinal health when applied in combination than their individual effects alone.

Alginate oligosaccharide-induced intestinal morphology, barrier function and epithelium apoptosis modifications have beneficial effects on the growth performance of weaned pigs

Background: Alginate oligosaccharide(AOS), produced from alginate by alginate lyase-mediated depolymerisation, is a potential substitute for antibiotics and possesses growth-enhancing effects. Nevertheless, the mechanisms by which AOS regulates porcine growth remain to be elucidated. Therefore, we investigated the AOS-mediated changes in the growth performance of weaned pigs by determining the intestinal morphology, barrier function,as well as epithelium apoptosis.Methods: Twenty-four weaned pigs were distributed into two groups(n = 12) and received either a basal diet(control group) or the same diet supplemented with 100 mg/kg AOS. On d 15, D-xylose(0.1 g/kg body weight)was orally administrated to eight randomly selected pigs per treatment, and their serum and intestinal mucosa samples were collected 1 h later.Results: Our results showed that inclusion of AOS in the diet for 2 wk increased(P < 0.05) the average daily body weight gain in weaned pigs. Notably, AOS supplementation ameliorated the intestinal morphology and barrier function, as suggested by the enhanced(P < 0.05) intestinal villus height, secretory immunoglobulin A content and goblet cell counts. Compared to the control group, AOS ingestion both decreased(P < 0.05) the total apoptotic percentage and increased(P < 0.05) the proportion of S phase in the intestinal epithelial cells. Furthermore, AOS not only up-regulated(P < 0.05) the B-cell lymphoma-2(BCL2) transcriptional level but also down-regulated(P < 0.05) the B-cell lymphoma-2-associated X protein(BAX), cysteinyl aspartate-specific proteinase-3(caspase-3) and caspase-9 transcriptional levels in the small intestine.Conclusions: In summary, this study provides evidence that supplemental AOS beneficially affects the growth performance of weaned pigs, which may result from the improved intestinal morphology and barrier function,as well as the inhibited enterocyte death, through reducing apoptosis via mitochondria-dependent apoptosis.

Alginate oligosaccharide improves resistance to postharvest decay and quality in kiwifruit(Actinidia deliciosa cv. Bruno)

Kiwifruit is an extremely perishable fruit;postharvest disease and senescence during storage can reduce the fruit quality,resulting in economic loss.Considerable research effort has focused on identifying safe and cost-effective ways to preserve fresh kiwifruit.To this end,the present study investigated the effects of alginate oligosaccharide(AOS)soaking treatment on postharvest quality and disease in the‘Bruno’variety of kiwifruit.The involved physiological mechanisms were further explored.The results showed that AOS did not inhibit the growth of Botrytis cinerea in vitro,the causal agent of gray mold in kiwifruit,but reduced the incidence of gray mold and diameter of lesions of kiwifruit during storage.Kiwifruit treated with 50 mg·L-1 AOS showed a higher degree of firmness and lower soluble solid content than control fruit treated with distilled water.Moreover,AOS treatment inhibited the activity of polygalacturonase and pectinesterase,while enhancing the activity of polyphenoloxidase,l-phenylalanine ammonia lyase andβ-1,3-glucanase related to pathogen defense,and also improved total antioxidant capacity determined by the DPPH,FRAP,and ABTS methods in kiwifruit.These results indicate that 50 mg·L-1 AOS can confer disease resistance in kiwifruit during storage.