Characterization of mulberry leaf instant tea and evaluation of its hypolipidemia effect via regulation of intestinal microbiota

The purpose of this study was to characterize mulberry leaf instant tea(MLIT)powder prepared from the'Longsang No.1'(Morus abla L.cv.Longsang 1)mulberry leaves in Heilongjiang Province(China)and assess its obesity-preventing/relieving effects.A total of 174 compounds including quercetin,chlorogenic acid,1-deoxyecomycin(1-DNJ)related to antihyperlipidemia effects were identified from the MLIT powder.MLIT treatment reversed the Lee's index,fat coefficient,and serum biochemical parameters in both the obesity relieving and obesity preventing mice fed with high-fat diet.In the obesity relieving experiment,the relative abundance of Desulfovibrio in mouse feces decreased after both 0.5%and 1%MLIT treatments.In obesity preventing experiments,mouse with different amount of MLIT treatments showed increased relative abundance of Akkermansia,Bifidobacterium and Lactobacillus,while Deferribacteres,Desulfobacterota decreased.The beneficial bacteria in the intestinal tract of mice treated with MLIT increased.This study proved that MLIT had antihyperlipidemia potential via modulating intestinal microbiota in mice.

Green Synthesis of Iron(Ⅱ,Ⅲ)-polyphenol Nanoparticles and Their Adsorption of Malachite Green

Three kinds of iron nanoparticles(FeNPs)were prepared via green route based on pomegranate(PG),green tea(GT),and mulberry(ML)extracts under ambient conditions.The obtained materials were characterized by scanning electron microscopy(SEM),transmission electronic microscopy(TEM),X-ray energy-dispersive spectrometer(EDS),X-ray diffraction(XRD),fourier transform infrared spectroscopy(FTIR),and X-ray photoelectron spectroscopy(XPS)techniques.The experimental results show that FeNPs were in the form of amorphous iron(Ⅱ,Ⅲ)-polyphenol complex with different dispersity and morphologies.GT-Fe has the smallest size range of 25-35 nm,PG-Fe has a moderate size-distribution of 30-40 nm,while ML-Fe formed a tuberous net-type with a sheeting structure.PG-Fe displays the highest removal efficiency of 90.2%in 20 min towards cationic dye of malachite green(16.6%by ML-Fe and 69.3%by GT-Fe),which is attributed to its highest polyphenol content,lowest zeta potential,as well as the most Fe^(2+)on the surface of FeNPs.The removal mechanism was mainly induced by electrostatic adsorption based on pH and zeta potential tests.

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.

Response Mechanisms to Flooding Stress in Mulberry Revealed by Multi-Omics Analysis

Abiotic stress,including flooding,seriously affects the normal growth and development of plants.Mulberry(Morus alba),a species known for its flood resistance,is cultivated worldwide for economic purposes.The transcriptomic analysis has identified numerous differentially expressed genes(DEGs)involved in submergence tolerance in mulberry plants.However,a comprehensive analyses of metabolite types and changes under flooding stress in mulberry remain unreported.A non-targeted metabolomic analysis utilizing liquid chromatographytandem mass spectrometry(LC-MS/MS)was conducted to further investigate the effects of flooding stress on mulberry.A total of 1,169 metabolites were identified,with 331 differentially accumulated metabolites(DAMs)exhibiting up-regulation in response to flooding stress and 314 displaying down-regulation.Pathway enrichment analysis identified significant modifications in many metabolic pathways due to flooding stress,including amino acid biosynthesis and metabolism and flavonoid biosynthesis.DAMs and DEGs are significantly enriched in the Kyoto Encyclopedia of Genes and Genomes(KEGG)pathways for amino acid,phenylpropanoid and flavonoid synthesis.Furthermore,metabolites such as methyl jasmonate,sucrose,and D-mannose 6-phosphate accumulated in mulberry leaves post-flooding stress.Therefore,genes and metabolites associated with these KEGG pathways are likely to exert a significant influence on mulberry flood tolerance.This study makes a substantial contribution to the comprehension of the underlying mechanisms implicated in the adaptation of mulberry plants to submergence.

Identification of Mulberry Bacterial Blight Caused by Klebsiella oxytoca in Bazhong,Sichuan,China

To provide a scientific basis for controlling mulberry bacterial blight in Bazhong,Sichuan,China(BSC),this study aimed to isolate and purify pathogenic bacteria from diseased branches of mulberry trees in the region and to clarify their taxonomic status using morphological observation,physiological and biochemical detection,molecular-level identification,and the construction of a phylogenetic tree.A total of 218 bacterial strains were isolated from samples of diseased mulberry branches.Of these,7 strains were identified as pathogenic bacteria based on pathogenicity tests conducted in accordance with Koch’s postulates.Preliminary findings from the analysis of the 16S rRNA sequence indicated that the 7 pathogenic bacteria are members of Klebsiella spp.Morphological observation revealed that the pathogenic bacteria were oval-shaped and had capsules but no spores.They could secrete pectinase,cellulase,and protease and were able to utilize D-glucose,D-mannose,D-maltose,and D-Cellobiose.The 7 strains of pathogenic bacteria exhibited the highest homology with Klebsiella oxytoca.This study identifies Klebsiella oxytoca as the causative agent of mulberry bacterial blight in BSC,laying the foundation for the prevention and control of this pathogen and further investigation into its pathogenic mechanism.

Synergistic effects of three traditional herbs green tea,mulberry leaf and corn silk on glucose uptake level of L6 myoblasts and the hypoglycemic mechanism

Background:Green tea,mulberry leaf and corn silk are traditional herbs used in the prevention and treatment of diabetes in China for a long time,but their synergistic hypoglycemic effects and mechanisms remain unclear.Methods:The effective components of green tea,mulberry leaf and corn silk were extracted and enriched.Mixture design of experiments was used to study the influences of different combinations on the cell viability and glucose uptake level of L6 myoblasts,so as to determine the optimal synergistic hypoglycemic combination.The possible hypoglycemic mechanism of the optimal synergistic combination was explored by cytotoxicity assay,glucose uptake assay,and western blot.Results:Three polyphenol enrichment fractions of the herbs,30%ethanol elution fraction of green tea(GT),50%ethanol elution fraction of mulberry leaf(ML)and 60%ethanol elution fraction of corn silk(CS)were obtained.The antioxidant activities of GT-30%,ML-50%and CS-60%were superior to those of crude extracts,and showed strong potential inα-amylase andα-glucosidase inhibition activities.The optimal synergistic combination of crude extracts G7(crude extract of green tea:crude extract of mulberry leaf:crude extract of corn silk=1:5:3),polyphenol enrichment fractions R3(GT-30%:ML-50%:CS-60%=1:7:1)and monomers X2(epigallocatechin gallate:morusin:formononetin=3:1:2)were selected,respectively.G7,R3,and X2 showed promoting effects on the cell viability and glucose uptake of L6 myoblasts within the detected concentration range.In addition,G7,R3,and X2 could increase the expression levels of p-PI3K/PI3K and p-Akt/Akt in L6 myoblasts,and promote the translocation of Glut4,but G7 and R3 showed more significant effects.Conclusion:The synergistic hypoglycemic effects of green tea,mulberry leaf and corn silk had the characteristics of multiple-components and multiple-targets with p-PI3K/PI3K,p-Akt/Akt and the translocation of Glut4 signal pathways involved.The three traditional herbs might have the potential to be combined used for the prevention and treatment of diabetes based on the synergistic hypoglycemic effects.

Quality Evaluation of Fermented Mulberry Wine

[Objectives]This study to conducted to clarify various nutritional components and the composition and contents of aromatic substances in mulberry wine prepared by fermentation,and promote the quality improvement and product development of mulberry wine.[Methods]Mulberry wine was prepared with mulberry Shengguo 1 as the material using Saccharomyces cerevisiae,and its contents of total sugars,total phenols,total acids,resveratrol,protein,dry extract,ethanol content,Fe,methanol,sulfur dioxide,volatile acids,total flavonoids and proanthocyanidins were determined.Aromatic substances were extracted by n-hexane,and their components and relative contents were determined by gas chromatography-mass spectrometry(GC-MS).[Results]The physical and chemical indexes of the mulberry wine prepared by fermentation were in accordance with the national standards,and it was rich in resveratrol(10.5±0.6 mg/L),protein(381±11 mg/L),total flavonoids(406±18 mg/L)and proanthocyanidins(855±22 mg/L),which were 8.95,19.49,3.76 and 1.63 times higher than those of grape wine,respectively.The aromatic substances were mainly composed of ethyl formate(9.9%),ethyl acetate(7.6%),acetal(6.2%),propanol(5.7%),isobutanol(5.6%)and isoamyl acetate(5.3%).[Conclusions]The results of this study demonstrated that mulberry wine was rich in active substances,which suggested that mulberry wine has great potential in the fruit wine market.

Development of Morus alba Reinforced Poly-Lactic Acid with Elevated Mechanical and Thermal Properties

This research investigates the mechanical and thermal properties of Morus alba combined with polylactic acid in comparison with other natural fibers. The study uses three different fiber and PLA compositions - 20%, 30%, and 40% respectively - to produce composite materials. In addition, another composite with the same fiber volume is treated with a 4% NaOH solution to improve mechanical properties. The composites are processed by twin-screw extrusion, granulation, and injection molding. Tensile strength measurements of raw fibers and NaOH-treated fibers were carried out using a single-fiber tensile test with a gauge length of 40 mm. It was observed that the NaOH surface treatment increases the resistance against tensile loading and exhibited improved properties for raw fiber strands. The diameter of the fibers was measured using optical microscopy. During this research, flexural tests, impact tests, differential scanning calorimetry (DSC), and heat deflection temperature measurements (HDT) were conducted to evaluate the mechanical and thermal properties of the developed composite samples. The results indicate that the mechanical properties of NaOH-treated Morus alba-reinforced polylactic acid outperform both virgin PLA samples and untreated Morus alba samples.

The gap-free genome of mulberry elucidates the architecture and evolution of polycentric chromosomes

Mulberry is a fundamental component of the global sericulture industry,and its positive impact on our health and the environment cannot be overstated.However,the mulberry reference genomes reported previously remained unassembled or unplaced sequences.Here,we report the assembly and analysis of the telomere-to-telomere gap-free reference genome of the mulberry species,Morus notabilis,which has emerged as an important reference in mulberry gene function research and genetic improvement.The mulberry gap-free reference genome produced here provides an unprecedented opportunity for us to study the structure and function of centromeres.Our results revealed that all mulberry centromeric regions share conserved centromeric satellite repeats with different copies.Strikingly,we found that M.notabilis is a species with polycentric chromosomes and the only reported polycentric chromosome species up to now.We propose a compelling model that explains the formation mechanism of new centromeres and addresses the unsolved scientific question of the chromosome fusion-fission cycle in mulberry species.Our study sheds light on the functional genomics,chromosome evolution,and genetic improvement of mulberry species.

Potential of Thai Bast Fibers for Injection Molded PLA Composites

Thailand has a huge variability of bast fiber plants,some of which have been little researched regarding their applicability in composites.Bast fiber(bundle)s from different species were investigated and incorporated into a polylactide(PLA)matrix by injection molding.Hemp and kenaf were used as well-studied fibers,while roselle,Fryxell and paper mulberry are less extensively characterized.Tensile strength,tensile modulus and interfacial shear strength(IFSS)of single fiber(bundle)s were highest for hemp,followed by kenaf,roselle,Fryxell and paper mulberry.Despite the lower tensile strength and IFSS of paper mulberry,the highest tensile strength was achieved for the paper mulberry/PLA composite followed by hemp/PLA.Scanning electron microscope(SEM)analyses showed that the single cells in paper mulberry fiber bundles,in contrast to the other fiber types investigated,were only partially bonded to each other,which explains the lower strength of the fiber bundles.The higher aspect ratio of fiber(bundle)s of paper mulberry in the PLA composite can explain the good composite characteristics.Apart from hemp,paper mulberry shows the best reinforcing effect in the PLA matrix and offers interesting potential for composite applications.Compared to neat PLA,the tensile strength could be increased by 24%and the tensile modulus by 54%.

Transcriptome Profiling of the Salt-Stress Response in Paper Mulberry

Paper mulberry is a high-quality woody feed resource plant with high crude protein content.It is widely distributed in China and has excellent characteristics of salt and alkali tolerance.Paper mulberry has ecological and economic importance.Salt stress has become a critical factor with the increasing degree of soil salinity that restricts plant growth.In the saline-alkali environments,transcriptome expression is altered leading to phenotypic defects in most plants.However,the regulatory mechanism related to paper mulberry’s salt-stress(SS)response is not clearly understood.In the present study de novo transcriptomic assembly was performed,and gene expression levels were measured between different SS and natural conditions(25℃)as a control for paper mulberry plants.According to the results of our study,under NaCl stress conditions,the differential gene expression was observed in the leaves of paper mulberry compared with the control.A total of 2126 differentially expressed unigenes were observed.Among these unigenes the expression of 812 DEGs was up-regulated and the expression of 1,314 DEGs was down-regulated.Additionally,The GO and KEGG analyses regarding differentially expressed unigenes(DEUs)revealed that the observed critical transcriptomic alterations under salt stress(SS)conditions were associated with primary and secondary metabolism,photosynthesis,and plant hormone signaling pathways.Further investigations such as gene function studies regarding the unigenes depicting altered expression under salt stress conditions in paper mulberry will help understand the mechanism of salt tolerance,and this information can be utilized in paper mulberry breeding and improvement programs.

Study on Photosynthetic Characteristics of Hybrid Mulberry in Autumn under Herbaceous Cultivation

[Objectives]This study was conducted to clarify the physiological mechanism of growth of hybrid mulberry after autumn cutting in herbaceous cultivation.[Methods]The net photosynthetic rate(Pn),stomatal conductance(Gs),intercellular CO_(2) concentration(Ci)and transpiration rate(Tr)of leaves were measured under different treatment conditions using a Li-6400XT portable photosynthetic measurement system.[Results]After harvest of mulberry shoots in autumn,leaf Pn decreased with the extension of branch and leaf growth time,while Gs,Ci and Tr showed a trend of first increasing and then decreasing.The Pn was affected by factors such as leaf positions,mulberry varieties,cutting,and fertilization,which was manifested by the 6 th to 10 th mature leaves>the 2 nd to 4 th tender leaves,‘Nongsang 14’>hybrid mulberry,intermediate cut>uncut,and normal fertilization>no fertilization,all showing significant differences(P<0.05).Combined with the results of Gs,Ci and Tr measurements,it was found that the changes in leaf Pn were mainly related to non-stomatal factors.Timely cutting and harvesting during summer and autumn could significantly improve the photosynthetic rate of mulberry leaf,which was beneficial for extending the late autumn growth period of hybrid mulberry under herbaceous cultivation.[Conclusions]This study provides a theoretical reference for mulberry shoot harvesting techniques in summer and autumn.

Flower Characteristics and Resource Evaluation of Fruit Mulberry

[Objectives]In order to study the flower characteristics of mulberry and evaluate the resources of fruit mulberry.[Methods]Firstly,the resources of fruit mulberry were collected,and four varieties of fruit mulberry(‘Dashi’mulberry,‘Changguo’milk mulberry,‘agate’mulberry and‘Xiangjin’milk mulberry)were collected.Then,the flower characteristics,fruit quality,and adaptability of fruit mulberry were studied.[Results]Mulberry is a dioecious plant,and the inflorescences are lurocatkin.The male flowers will fall off automatically,while the female flowers will develop into fruit.The four varieties of fruit mulberry have good adaptability in Shanghai,and their fruits have their own characteristics.At the same time,in order to use the resources of fruit mulberry,mulberry fruit picking activities were carried out for many times.[Conclusions]This study provides a reference for the cultivation,collection,utilization and evaluation of mulberry.

Identification of 13th-14th Century Chinese Handmade Paper Fibers Collected in Yuan Dynasty Paper Currencies

The earliest paper currencies in existence in China were handcrafted during the Yuan dynasty.These currencies were scientifically excavated from different ruins or tombs,whereas scientific analyses of the papers are rare.This study used optical and scanning electron microscopy to examine the fibers collected in Yuan dynasty paper currencies in conjunction with the Herzberg staining method.Despite differences in circulation period,paper fibers in both Zhi Yuan Tong Xing Bao Chao(two Guan)and Zhong Tong Yuan Bao Jiao Chao(one Guan and 500 Wen,issued in Zhi Zheng period)were identified as similar papermaking materials,bast fibers of mulberry bark.The results indicate that mulberry bark,a durable papermaking material used since ancient times,was mainly utilized as a raw material in these Yuan dynasty paper currency.This fiber identification work solved the critical problem of papermaking material in the Yuan dynasty paper currency and provided important information for conserving these precious cultural relics.

Polyphenol content and antioxidant activity in mulberry powder treated at high temperature

The aim of this study was to investigate changes of polyphenols content and the antioxidant activity of mulberry fruit powder after heating at high temperature.The mulberry fruit powder was heated by baking method at different temperature and time.The DPPH scavenging ability,the OH^(•-) scavenging ability and the O_(2)^(•-) scavenging ability were used to evaluate the changes in the antioxidant activity of the mulberry fruit powder after the heat treatment.The results showed that 74.87%of the polyphenols in the mulberry powder were lost after heating at 150℃for 5 min,and almost all the polyphenols were lost with the increase of the heating time and temperature.The antioxidant experiment showed that the DPPH scavenging rate,OH^(•-) scavenging rate and O_(2)^(•-) scavenging rate of mulberry fruit powder were 68.90%,39.15%and 14.09%,respectively after treatment at 150℃ for 5 min.Mulberry fruit powder still has some potential to be added to baked goods as a functional ingredient.

江西桑树病害种类调查与鉴定

至今为止,江西的桑树病害尚未见系统的调查与报道。为了探明我省桑树病害种类及主要病害的发生情况,为防治提供依据,笔者于1991～1993年对赣中、南、北地区部分桑园进行了普查,采集的标本经鉴定有以下47种,其中叶部病害18种,枝干病害22种,根部病害2种,全株性病害5种,江西曾记载的桑树病害中,笔者已鉴定到11种,此外,还查到江西未曾记载的桑树病害34种;国内曾记载的75种桑树病害中,笔者已查到37种,此外。

Effects of Nitrogen Amount and Nitrogen Form on 1-deoxynojimycin Content in Mulberry Leaf

[Objective] Effects of nitrogen amount and nitrogen form on 1-deoxynojimycin（DNJ）content in mulberry leaf was explored in order to provide a reference for improving DNJ content in mulberry leaf and its medicinal value.[Method] Effects of nitrogen amount（NO-3-N and NH＋4-N）and nitrogen form（NO-3-N/NH＋4-N）on DNJ content in mulberry leaf were studied by changing nitrogen amount and nitrogen form in water culture solution.[Result] DNJ contents in mulberry leaf both increased at first and then decreased with the increase of nitrogen amount,furthermore compared with NH＋4-N,NO-3-N was more beneficial to the accumulation of DNJ;DNJ content in mulberry leaf obviously increased at first and then decreased with the decrease of NO-3-N/NH＋4-N,among them when its rate was 25/75,DNJ content reached the highest.[Conclusion] Proper nitrogen amount and nitrogen form could effectively improve the DNJ content in mulberry leaf.

表面活性剂辅助提取桑叶总黄酮的工艺研究

[目的]研究表面活性剂辅助提取桑叶中总黄酮的工艺条件。[方法]在单因素考察的基础上,利用正交试验优选最佳提取工艺条件。[结果]最佳提取工艺条件为:Tween 80的浓度为1.5%,料液比1∶40(g/ml),提取时间120 min,提取温度80℃,提取2次;在此条件下,总黄酮平均提取率为1.742%。[结论]表面活性剂辅助提取法是一种新型高效的提取方法,可用于桑叶中总黄酮的提取。

Induced Expression of Cytochrome P450 CYP305 B1V1 Gene in Different Tissues of Wild Mulberry Silkworm(Bombyx mandarina)

[ Objective] The aim of this study was to investigate effects of various inducers on the expression of cytochrome P450 CYP305 B1 V1 Gene in different tissues of wild mulberry silkworm. [ Method] Referring to the mRNA sequence of CYP305 B1 V1 Gene published in GenBank for wild mulberry silkworm, one pair of primers was designed, and the expression of cytochrome P450 CYP305 B1 V1 Gene in different tissues of wild mulberry silkworm treated by NaF, rutin, cypermethrin and ecdysone was also analyzed by the semi - quantitative RT - PCR. Furthermore, homology comparison and phylogenetic analysis for amino acid sequences of this gene were studied. [ Result] Rutin, cypermethrin and NaF had effects on the expression of P450 CYP305 B1 V1 Gene in different tissues of wild mulberry silkworm, while ecdysone had no significant effect. Homology comparison for amino acids indicated that the amino acid sequence of this gene was the most similar to that of CYP305 B1 gene in Bombyx mori with 100% amino acid identity, and highly similar to those of Tribolium casmneum CYP305A1, Apis mellifera CYP305A1, Drosophi- la melanogaster CYP305A1, Anopheles gambiae CYP305A2and Culex pipiens quinquefasciatus CYP2LI. [ Conclusion] CYP305 B1 V1 Gene of wild mulberry silkworm is likely to mainly take part in the metabolism of exogenous compounds, which is of great significance for revealing the function of cytochrome P450 and the metabolic mechanism of different drugs.

Effects of Exogenous Hormones on Subculture Multiplication and Root Induction of Tissuecultured Seedlings of Mulberry

Objective The purpose of this experiment was to investigate the effects of exogenous hormone factors on subculture multiplication and root induction of tis-suecultured seedlings in mulberry. [Method] The aseptic seedlings of Guisangyou 12 were used as the materials and the comparative experiment on subculture multi-plication and root induction used different kinds,concentrations and combinations of exogenous hormones. [Result] The hormone combination of 0.1 mg/L IBA＋1.5 mg/L 6-BA＋0.03 mg/L TDZ showed the best effects to bud multiplication, and the multiple of bud multiplication was 5.28. The effects of NAA was better than IBA and IAA on root induction. The hormone combination of 2.0 mg/L NAA＋1.0 mg/L PP333 was most suitable to the root induction, in which the rooting rate arrived to 100%, the root number was 7.01 and the root length was 1.38 cm on average. [Conclusion] The results wil provide some technical reference for large-scale propagation of mul-berry seedlings in vitro.