Synthesis of chitosan oligosaccharide selenium and its antitumor activity

Background:Polysaccharides have various biological activities;the complexation of polysaccharides with trace element ions can produce synergistic effects,improving the original biological activities of sugars and trace elements.Methods:The preparation process of chitosan oligosaccharide selenium(COSSe)was optimized by the response surface method,followed by a detailed analysis of the resultant compound’s characteristics.The anti-cancer activity of COSSe was studied using the human ovarian cancer cell line SKOV3 as a cell model.Results:The prepared COSSe response surface was well predicted,indicating successful chitosan oligosaccharide binding with selenium.Response surface method analyses identified the optimal synthesis conditions for COSSe:the reaction time of 5.08 h,the reaction temperature of 71.8°C,and mass ratio(Na2SeO3:chitosan oligosaccharide)of 1.02.Under the optimal conditions,the final product,the selenium content,reached 1.302%.The results of cell experiments showed that COSSe significantly inhibited SKOV3 proliferation in a concentration-dependent manner.RNA-seq results showed that chitosan oligosaccharide and COSSe significantly modulated the expression of genes’DNA metabolic processes and cell cycle in SKOV3 cells.Gene enrichment analysis showed the inhibition of the cell cycle,and the results of flow cytometry showed that SKOV3 cells increased in the S phase and decreased in the G2/M phase,with a noted suppression in the protein expression of cyclin-dependent kinase 2(CDK2)and cyclin A1(CCNA1).Conclusion:COSSe has a stronger effect than chitosan oligosaccharide,leading to the arrest of the cell cycle in the S phase.Thus,COSSe may be an effective candidate for the treatment of ovarian cancer.

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.

In situ visualization of the cellular uptake and sub-cellular distribution of mussel oligosaccharides

Unlike chemosynthetic drugs designed for specific molecular and disease targets,active small-molecule natural products typically have a wide range of bioactivities and multiple targets,necessitating extensive screening and development.To address this issue,we propose a strategy for the direct in situ microdynamic examination of potential drug candidates to rapidly identify their effects and mechanisms of action.As a proof-of-concept,we investigated the behavior of mussel oligosaccharide(MOS-1)by tracking the subcellular dynamics of fluorescently labeled MOS-1 in cultured cells.We recorded the entire dynamic process of the localization of fluorescein isothiocyanate(FITC)-MOS-1 to the lysosomes and visualized the distribution of the drug within the cell.Remarkably,lysosomes containing FITC-MOS-1 actively recruited lipid droplets,leading to fusion events and increased cellular lipid consumption.These drug behaviors confirmed MOS-1 is a candidate for the treatment of lipid-related diseases.Furthermore,in a high-fat HepG2 cell model and in high-fat diet-fed apolipoprotein E(ApoE)^(-/-)mice,MOS-1 significantly promoted triglyceride degradation,reduced lipid droplet accumulation,lowered serum triglyceride levels,and mitigated liver damage and steatosis.Overall,our work supports the prioritization of in situ visual monitoring of drug location and distribution in subcellular compartments during the drug development phase,as this methodology contributes to the rapid identification of drug indications.Collectively,this methodology is significant for the screening and development of selective small-molecule drugs,and is expected to expedite the identification of candidate molecules with medicinal effects.

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.

Structural elucidation of mulberry leaf oligosaccharide and its selective promotion of gut microbiota to alleviate type 2 diabetes mellitus

Two oligosaccharide fractions(MLO 2-1 and 2-2)were purified from enzymatic hydrolysate of mulberry leaf polysaccharide.The results of simulated digestion showed that MLO 2-2 was a digestible oligosaccharide,which could be degraded by human digestive juice;while MLO 2-1 possessed the non-digestible property in the upper gastrointestinal tract and performed the function by regulating the gut microbiota.Hence,MLO 2-1 was selected for the further analysis.The structure of MLO 2-1 was elucidated as follow:α-T-Glcp-(1→3)-β-Glcp-(1→5)-α-Araf-(1→5)-α-Araf-1→5)-α-Araf-(1→3)-α-(2-OAc)-Glcp-1.The in vitro fecal fermentation results showed that MLO 2-1 could modulate the composition of gut microbiota.Meanwhile,MLO 2-1 was effectively metabolized by fecal bacteria to produce lactate and short chain fatty acids,especially acetate and butyrate.The specific metabolic pathways of MLO 2-1 by gut microbiota were further illuminated.Gut microbiota analysis revealed that MLO 2-1 selectively promoted the growth of Ligilactobacillus murinus,a commensal bacterium presented a reduced level in T2DM mice.Animal experiments indicated that MLO 2-1 and L.murinus exhibited hypoglycemic activities.These results demonstrated that MLO 2-1 might alleviate T2DM by selectively accelerating the proliferation of L.murinus.

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.

Therapeutic potential and mechanism of functional oligosaccharides in inflammatory bowel disease: a review

Inflammatory bowel disease(IBD)is characterized by recurrent attacks and long courses,and the number of patients has expanded rapidly year by year.Additionally,current conventional strategies exist serious adverse effects.In this case,it is an urgent issue to find out an effective and safe treatment.Functional oligosaccharides possess safe and excellent physiological activities,and have attracted enormous attention due to their great therapeutic potential for IBD.This review emphasizes the attenuating effects of distinct functional oligosaccharides on IBD and their structure,and summarizes the main mechanisms from the aspects of regulating intestinal fl ora structure,repairing intestinal barrier,modulating immune function and mediating related signaling pathways in order to reveal the relationship between functional oligosaccharides,immune regulation,intestinal epithelial cells,gut fl ora and IBD treatment.Oligosaccharides possess excellent protective effects on IBD,and can be considered as safe and functional ingredients in the health food and pharmaceutical industry.

Mannan oligosaccharides alleviate oxidative injury in the head kidney and spleen in grass carp(Ctenopharyngodon idella)via the Nrf2 signaling pathway after Aeromonas hydrophila infection

Background Mannan oligosaccharides(MOS)are recommended as aquaculture additives owing to their excellent antioxidant properties.In the present study,we examined the effects of dietary MOS on the head kidney and spleen of grass carp(Ctenopharyngodon idella)with Aeromonas hydrophila infection.Methods A total of 540 grass carp were used for the study.They were administered six gradient dosages of the MOS diet(0,200,400,600,800,and 1,000 mg/kg)for 60 d.Subsequently,we performed a 14-day Aeromonas hydrophila challenge experiment.The antioxidant capacity of the head kidney and spleen were examined using spectrophotometry,DNA fragmentation,qRT-PCR,and Western blotting.Results After infection with Aeromonas hydrophila,400-600 mg/kg MOS supplementation decreased the levels of reactive oxygen species,protein carbonyl,and malonaldehyde and increased the levels of anti-superoxide anion,antihydroxyl radical,and glutathione in the head kidney and spleen of grass carp.The activities of copper-zinc superoxide dismutase,manganese superoxide dismutase,catalase,glutathione S-transferase,glutathione reductase,and glutathione peroxidase were also enhanced by supplementation with 400-600 mg/kg MOS.Furthermore,the expression of most antioxidant enzymes and their corresponding genes increased significantly with supplementation of 200-800 mg/kg MOS.mRNA and protein levels of nuclear factor erythroid 2-related factor 2 also increased following supplementation with 400-600 mg/kg MOS.In addition,supplementation with 400-600 mg/kg MOS reduced excessive apoptosis by inhibiting the death receptor pathway and mitochondrial pathway processes.Conclusions Based on the quadratic regression analysis of the above biomarkers(reactive oxygen species,malondialdehyde,and protein carbonyl)of oxidative damage in the head kidney and spleen of on-growing grass carp,the recommended MOS supplementation is 575.21,557.58,531.86,597.35,570.16,and 553.80 mg/kg,respectively.Collectively,MOS supplementation could alleviate oxidative injury in the head kidney and spleen of grass carp infected with Aeromonas hydrophila.

Effects of Different Oligosaccharides on Performance and Availability of Nutrients in Broilers

One hundred and forty-four Avin broilers (1 d of age) were randomly divided into six groups, with three replicates of eight birds each to study on the effects of different oligosaccharides on performance and availability of nutrients in broilers. The control group(Group I)was fed with corn-soybean meal as basal diet, and the trial groups (Group II, III, IV,V and VI) were fed with basal diet plus 0.1% Manoligosaccharides(MOS), 0.3% Soybean-Oligosaccharides(SBOS), 0.3% Fructo-oligosaccharides(FOS), 0.3% α-Glucooligosaccharides(α-GOS) and 50 mg·L-1 Chlortetracycline(CTC), respectively. The results showed that the supplementation of oligosaccharides slightly improved daily gain and feed intake. SBOS supplementation improved, but MOS and α-GOS significantly decreased availability of energy. Oligosaccharides supplementation improved availability of energy, phosphorus, calcium, magnesium and iron, and significantly increased cholesterol content of fecal, and did not affect on availability of protein and cholesterol contents in serum and muscle. Availability of phosphorus of broilers fed with FOS was much higher than that with α-GOS. SBOS supplementation remarkably increased availability of iron.

Combinatorial Enzyme Approach to Convert Wheat Insoluble Arabinoxylan to Bioactive Oligosaccharides

Combinatorial enzyme technology was applied for the conversion of wheat insoluble arabinoxylan to oligosaccharide structural variants. The digestive products were fractionated by Bio-Gel P4 column and screened for bioactivity. One fraction pool was observed to exhibit antimicrobial property resulting in the suppression of cell growth of the test organism ATCC 8739 E. coli. It has a MIC value of 1.5% (w/v, 35°C, 20 hr) and could be useful as a new source of prebiotics or preservatives. The present results further confirm the science and useful application of combinatorial enzyme approach.

Laminarin and Laminarin Oligosaccharides Originating from Brown Algae:Preparation,Biological Activities,and Potential Applications

Brown algae is one of the three major types of marine algae and includes approximately 2000 species.It is widely dis-tributed in various seas around the world.Brown algae contain a plethora of active substances,such as polysaccharides,polyphe-nols,omega-3 fatty acids,and carotenoids.Laminarin,a type of storage carbohydrate found abundantly in brown algae,is mainly formed by glucose monomers linked byβ-1,3-glucosidic bonds and partialβ-1,6-glucosidic bonds.Laminarin and laminarin oligo-saccharides,which contain 2-10 saccharide units,have extensive biological activities,such as antitumor,antioxidant,anti-inflam-matory,and prebiotic properties.Moreover,both laminarin and laminarin oligosaccharides can be considered as ideal substrates for bioethanol production because they are composed of abundant glucose residues.Therefore,brown algae-derived laminarin and lami-narin oligosaccharides have various potential applications in the food,medicine,cosmetics,and bioenergy fields.This paper reviews the preparation methods of laminarin and laminarin oligosaccharides,as well as their biological activities and potential applications.

Effects of in ovo delivered xylo-and mannan-oligosaccharides on growth performance,intestinal immunity,cecal short-chain fatty acids,and cecal microbiota of broilers

Background:This study investigated a novel in ovo feeding strategy to determine the prebiotic effects of xylo-and mannan-oligosaccharides(XOS and MOS)differing in the degree of polymerization.A total of 192 fertilized eggs were divided into 6 treatment groups:i)normal saline control(NSC),ii)xylotriose(XOS3),iii)xylotetraose(XOS4),iv)mannotriose(MOS3),v)mannotetraose(MOS4),and vi)no injection control(NIC),each containing 4 replicate trays with 8 eggs per replicate.On d 17 of incubation,3 mg of oligosaccharides(except for controls)dissolved in 0.5 mL of 0.85%normal saline were injected into the amnion of Cobb 500 broilers eggs.After hatch,the chicks were raised for 28 d under standard husbandry practices and were fed a commercial broilers diet ad libitum,and samples were collected periodically.Results:The hatchability,growth performance,and relative weights of breast,drumstick,liver,and proventriculus were not different among the treatments(P>0.05).The XOS3 injection increased the total short-chain fatty acid production at d 28 compared with both control groups(P<0.05).The villus height to crypt depth ratio was significantly higher in the XOS4 group than both controls on the hatch day(P<0.01)but were not different among any treatments on d 7 and 28(P>0.05).On the hatch day,the expression level of the CD3 gene(a T cell marker)was increased by XOS3,while the IL-10 gene(a marker of anti-inflammatory cytokine)was reduced by MOS4(P<0.05)compared with both controls.Compared with both controls,XOS3 exhibited a trend of reduction for IL-10(P=0.074).No cytokines or lymphocyte markers were affected by the treatments on d 7(P>0.05),except XOS4 increased IL-4 compared with NSC(P<0.05).The broilers in the MOS4 group had higher operational taxonomic units(OTUs)and had more differentially abundant taxa,including order Lactobacillales and family Leuconostocaceae(P<0.05)than both controls on d 28.The predictive functional profiling indicated that the linoleic acid metabolism pathway was enriched in the cecal microbiota of the XOS3 group compared with both controls(P<0.05).Conclusion:The effects of these XOS and MOS on ileal mucosa and immunity are transient,but the effects on fermentation and cecal microbiota are prolonged,and further research is warranted to determine their use as a gut health promoter in poultry.

Multiplex genome editing targeting soybean with ultra-low anti-nutritive oligosaccharides

Soybean is the primary source of plant protein for humans.Owing to the indigestibility of the raffinose family of oligosaccharides(RFO),raffinose and stachyose are considered anti-nutritive factors in soybean seeds.Low-RFO soybean cultivars are generated by mutagenesis of RFO biosynthesis genes,but the carbohydrate profiles invite further modification to lower RFOs.This study employed a pooled multiplex genome editing approach to target four seed-specifically expressed genes mediating RFO biosynthesis,encoding three raffinose synthases(RS2,RS3,and RS4)and one stachyose synthase.In T1progeny,rs2/rs3 and rs4/sts homozygous double mutants and a rs2/rs3/rs4/sts quadruple mutant(rfo-4m)were characterized.The rs2/rs3 mutant showed reduced raffinose and stachyose contents,but the rs4/sts mutant showed only reduced stachyose in seeds.The RFO contents in the rfo-4m mutant were almost eliminated.Metabolomic analysis showed that the mutation of four RFO biosynthesis genes led to a shift of metabolic profile in the seeds,including the accumulation of several oligosaccharides-related metabolites.These mutants could contribute to precision breeding of soybean cultivars for soy food production.

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.

Effectiveness of dietary xylo-oligosaccharides for broilers fed a conventional corn-soybean meal diet

An experiment was conducted to investigate the effect of dietary supplementation of xylo-oligosaccharides（XOS） on growth performance, meat quality, immune functions, duodenal morphology and intestinal microbial populations of broilers fed a conventional corn-soybean meal basal diet. A total of 450 1-day-old commercial Arbor Acres male broiler chicks were randomly allocated by bodyweight to 1 of 5 treatments with 6 replicate cages（15 broilers per cage） for each of 5 treatments in a completely randomized design. Chicks were fed the basal corn-soybean meal diets supplemented with 0, 25, 50, 75, or 100 mg of XOS kg–1 of diet, respectively, for an experimental duration of 42 days. The results showed that supplementation of XOS affected feed conversion rate（feed/gain, F/G） during days 22–42 and 1–42（P〈0.03）, drip loss in thigh muscle（P=0.02）, and duodenal crypt depth（P=0.005） on day 42, but had no effect（P〉0.05） on all other measured indices. The chicks fed the diet supplemented with 100 mg of XOS kg–1 had the lowest（P〈0.05） F/G and drip loss in thigh muscle. The drip loss in thigh muscle decreased linearly（P=0.003） as the supplemented XOS increased. Duodenal crypt depth decreased（P〈0.05） at the supplemental level of 75 mg of XOS kg–1. The results indicate that dietary supplementations of 75 and 100 mg of XOS kg–1 are beneficial to broilers fed a conventional corn-soybean meal diet.

Effect of Different Oligosaccharides on Immunity, and Cecal Microflora in Broilers

One hundred and forty-four Avin broilers (1 d old) were randomly divided into six treatments, with three replicates of eight birds each to investigate the effect of different oligosaccharides on index of immune organ, cecal weight and microflora in broilers. The control group (Group I) was fed with corn-soybean meal as basal diet, and the trial groups (group Ⅱ, Ⅲ, Ⅳ, Ⅴ and Ⅵ) were fed with basal diet plus 0.1% manoligosaccharides (MOS), 0.3% soybean-oligosaccharides (SBOS), 0.3% fructooligosaccharides (FOS), 0.3% α-glucooligosaccharides(α-GOS) and 0.03% chlortetracycline (CTC), respectively. The results showed that oligosaccharides supplementation significantly (P< 0.05) increased bifidobacteria concentration of young broilers and did not affect on bifidobacteria or escherichia coli concentration of the birds ( P >0.05). Cecal weights of 21 d old broilers fed with FOS and α-GOS were significantly(P <0.05) increased, higher than those with MOS and SBOS, but addition of oligosaccharides had no effect on the birds. The effect of oligosaccharides on the cecal length was the same as on the cecal weights. Oligosaccharides supplement did not affect on indexes of immune organs.

Effects of hyaluronan oligosaccharides on apoptosis ofhuman gingival fibroblasts

An?increased?content?of?low?molecular?weight (LMW)- HA?was?found?in?human?gingiva?during?the?initial phase of?periodontitis. A?number?of?functions?have?been?described?for LMW-HA?in?inflammatory?processes.?It?is?also?demonstrated?that?LMWHA?induces?apoptosis?in?chondrocytes?and?tumor?cells,?however, effects of?LMWHA on the apoptosis?of?gingival?fibroblasts?(GFs) remain?unclear. The?purpose?of?this?study?was?to?investigate?the effects?of HA?oligosaccharides?(HAoligo) on?the apoptosis of GFs. GFs were?isolated?from?palatal?gingival?tissues around?the upper?first?premolars of eight youth?(6 -10?years?old)?and eight adult?(26 -35?years?old) subjects.?Cultured?GFs?were?subjected?to?the?analyses,?ELISA?for?cell?proliferation,?quantitative real?time-PCR?for?gene?expression?of?Bcl-2,?and?caspase-3?assay?for?apoptotic potency. Proliferative?activity?of?cultured?GFs was increased?

Induced Disease Resistance of Antarctic Bacteria B-3 Extracellular Oligosaccharide on Cucumber

[Objective] The induced disease resistance of Antarctic bacteria B-3 extracellular oligosaccharide on cucumber was studied.[Method] Taking the cucumber seedlings as experimental materials,the molecular weight,monosaccharide composition of Antarctic bacteria B-3 extracellular oligosaccharide were studied,its induced effect on the defense enzyme in cucumber leaves and the induced resistance against powdery mildew were also investigated.[Result] Through gel permeation chromatography,the molecular weight of B-3 oligosaccharides was determined to be 2 112 Da;B-3 oligosaccharides was composed by two monosaccharides including of mannose and glucose.The activities of chinitase,β-1,3-glucanse,phenylanine,ammonialyse（PAL）,superoxide dismutase（SOD）,and peroxidase（POD）in cucumber seedlings all increased compared with control when 0.3%,0.5% and 0.8% B-3 extracellular oligosaccharide were sprayed on the seedlings of cucumber for different times,respectively,which had the similar induction effect with 0.5% chitosan;at the same time,0.5% oligosaccharides could significantly reduce the disease index of cucumber powdery mildew,the control effect reached 24.49%.[Conclusion] B-3 oligosaccharides is expected to be developed as a new type of resistance elicitor.

Elicitation on Artemisinin Biosynthesis in Artemisia annua Hairy Roots by the Oligosaccharide Extract from the Endophytic Colletotrichum sp. B501

The oligosaccharide elicitor from the mycelial wall of an endophytic Colletotrichum sp. B501 promoted the production of artemisinin in Artemisia annua L. hairy root culture. When hairy roots of 22-day-old cultures (later growth phase) were exposed to the elicitor (20 mg/L) for 4 d, the maximum content of artemisinin reached 1.15 mg/g, a 64.29% increment over the control. The electron X-ray microanalysis disclosed the rapid accumulation of Ca 2+ in the elicited cortical cells of hairy root. The electronic microscope observation revealed the high electron density area in vacuole of elicited cells. During the first day of elicitation the peroxidase activity of hairy roots was improved sharply. Some cellular morphological changes including cell shrinkage, condensation of cytoplasm and nuclear fragmentation, coincident with the appearance of DNA ladders, were observed after the third day of elicitation. It was suggested that the oligosaccharide elicitor triggered the programmed cell death, which may provide the substance or chemical signal for artemisinin biosynthesis.

Cell Apoptosis Induced by Oligosaccharide in Suspension Cultures of Taxus chinensis

Taxol production of Taxus chinensis (Pilger) Rehd. var. mairei (Lemee et Lévl.) Cheng et L. K. Fu induced by oligosaccharide from Fusarium oxysporum f. vasinfectum (Atkinson) Snyder et Hansen was studied in suspension cultures, and it was found that oligosaccharide triggered cell apoptosis. Under transmission electron microscope the following morphological changes were observed: cell shrinkage, condensation of cytoplasm, nuclear fragmentation, and the increase of high electron density bodies in vacuole in great quantity. In oligosaccharide_treated cells, agarose gel electrophoresis revealed that DNA was digested into oligonucleosomal fragments that were times of 200 bp appearing as DNA ladders. Control cells were in normal physiological state, they were intact, abundant in organelle and with integral nucleus DNA, and the rate of taxol biosynthesis in these cells was very low. After the oligosaccharide to the culture system, the defense system of cells was elicited and the secondary metabolism was strengthened, i.e. phenolics were accumulated in the medium, the activity of polyphenol oxidase (PPO) was increased quickly and secondary wall of cells was thickened. The activity of L _phenylalanine ammonia lyase (PAL), the critical enzyme of the phenylpropanoid pathway, was increased promptly 1 h after elicitation. The rate of taxol production was improved sharply and the maximal taxol concentration at 72 h was six times that of control. Appearance of cell apoptosis was accompanied with the highest concentration of taxol in suspension cultures.