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.

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.

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.

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.

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.

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.

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.

桑叶粉及桑叶提取物的功效和应用现状

桑叶为药食同源性食品,具有多种药理作用。近年来,随着人们对健康饮食需求的不断增加,桑叶粉及桑叶提取物作为一种新型的保健食品,在食品药品加工中的应用越来越广泛。通过文献综述和案例分析,介绍了桑叶粉及桑叶提取物的加工工艺、营养价值、功能特点和在食品药品加工中的应用现状及研究进展,展望了桑叶粉及桑叶提取物的未来发展方向,以期为进一步推动桑叶产业的发展,促进其在食品药品加工中的应用提供参考。

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.

桑葚果实、桑葚干酒和桑葚利口酒中酚类化合物的高效液相色谱串联电喷雾二级质谱分析及其抗氧化活性比较

为了解桑葚果酒和桑葚果实在酚类化合物的种类、含量及抗氧化活性之间的差异,该研究以桑葚果实、桑葚干酒和桑葚利口酒为试材,采用高效液相色谱串联电喷雾二级质谱(high performance liquid chromatography-electron spray ionization-mass spectrometry/mass spectrometry,HPLC-ESI-MS/MS)对酚类化合物进行定性定量分析。HPLC-ESI-MS/MS分析结果表明,在桑葚果实、桑葚干酒和桑葚利口酒中均检测到花青素-3-葡萄糖苷和花青素-3-芸香糖苷,而花葵素-3-芸香糖苷和4-丙酮酸-花青素-3-葡萄糖苷分别只在桑葚干酒和桑葚利口酒中被检测到。同时,桑葚干酒和桑葚利口酒中花青素-3-葡萄糖苷和花青素-3-芸香糖苷的含量均显著低于桑葚果实。此外,在桑葚果实中鉴定的8种非花色苷多酚中,仅有4种存在于桑葚干酒和桑葚利口酒中。相较于桑葚果实,桑葚干酒和利口酒中原儿茶素、二氢槲皮素和槲皮素的含量显著升高。桑葚果实DPPH自由基清除能力介于桑葚干酒和利口酒之间。综上,桑葚果酒和桑葚果实中酚类化合物的种类、含量及抗氧化活性之间差异显著。

一种优化的桑树花芽石蜡切片制作方法

针对常规石蜡切片方法无法获得完整桑树花芽切片的问题,以“无籽大十”品种桑树花芽为供试材料,在常规石蜡切片技术基础上,对石蜡切片样品制作过程中固定、软化、脱水、透明及染色等环节进行了优化。结果表明,使用15%氢氟酸软化后,采用优化后的系列脱水、透明等步骤进行制片,可以使切片连续性、完整性得到明显提高。通过切片显微观察,可见细胞轮廓明显、裂痕较少、结构较完整,染色较清晰,并且对发育后期的花芽效果更为明显。优化后的制样流程解决了常规制片出现浸蜡不彻底、组织变脆、蜡带不连续、内部结构不完整等一系列问题。该研究可以从解剖学上为桑树花芽发育形态和花芽分化机制研究提供技术参考。

不同比例桑枝叶粉复合饲料对肉鸡血液生化指标及胆固醇含量的影响

桑树是在我国广泛栽种且兼具药食两用价值的木本植物,其营养丰富产量又高。随着机械化栽培生产技术升级和应用推广,桑多元化应用前景十分广阔。为探究机械化收获的杂交桑枝叶粉制备复合饲料对肉鸡生产的影响,在等氮等能的基础上,按基础日粮含0%(对照组)、2%(低剂量组)、4%(中剂量组)、6%(高剂量组)的不同比例配制桑枝叶粉复合饲料,经连续8周的饲养观察,测定肉鸡生长性能、血液理化指标,分析与脂质代谢相关基因的表达特征。结果显示,与对照组相比,桑枝叶粉复合饲料组鸡生长状态、觅食表现和增重无明显变化,但肉鸡的血清总蛋白含量、抗氧化能力、淋巴细胞增殖活性和血清溶菌酶活性显著升高;同时,桑枝叶粉饲养组肉鸡的血清谷丙转氨酶(ALT)活性、谷草转氨酶(AST)活性、尿素氮(UREA)含量和胆固醇含量降低,其中4%桑枝叶粉复合饲料组表现尤为明显。经定量PCR检测表明,内源性胆固醇含量的变化可能与HMGCR、SREBP-2等蛋白基因水平的表达受到抑制,以及LDLR外运蛋白基因的表达量增加有关。综上所述,桑枝叶粉可改善肉鸡体内的物质代谢,提高肝肾及抗氧化和免疫保护功能,促进胆固醇脂质代谢,具有较好的市场应用潜力。

桑叶发酵浓缩液喷雾干燥工艺优化

目的研究桑叶发酵浓缩液喷雾干燥工艺条件。方法采用L^(9)(3^(4))正交设计试验法,以出粉率、含水量及1-脱氧野尻霉素(DNJ)含量为评价指标,考察发酵浓缩液相对密度、进风口温度、进料量、空气流量等对桑叶发酵浓缩液喷雾干燥工艺条件的影响。结果桑叶发酵浓缩液喷雾干燥工艺的优化组合条件为:发酵浓缩液的相对密度1.0,进风口温度190℃,进料量20 ml/min,空气流量600 L/h。结论所得产品颗粒均匀细腻,流动性较好,最大限度地保证了出粉率,提高了DNJ含量,为更好地利用桑叶资源提供了新思路和途径。

响应面法优化黑桑椹黄酮超声波提取工艺

为优化黑桑椹总黄酮超声波提取工艺,以库尔勒市阿瓦提乡采摘园种植的黑桑椹果实为原料,在单因素试验结果的基础上,分别选取乙醇体积分数、提取温度、提取时间、液料比4个因素开展Box-Behnken中心组合试验,利用Design-Expert 8.06软件对结果进行分析和优化。结果表明,黑桑椹中黄酮类物质的最佳提取工艺条件为:乙醇体积分数60%,提取温度70℃,超声时间40 min,液料比30∶1(mL/g)。在此工艺条件下,黑桑椹黄酮类物质提取量为9.565 mg/g。说明应用响应面法所得到的提取工艺参数可行性强、可靠性高。

桑葚发酵前后酚类组成变化及其抗氧化活性分析

目的:旨在分析桑葚饮料发酵前后酚类物质的组成变化和抗氧化功能差异。方法:利用毕赤酵母菌(Pichia kudriavzevii)和融合魏斯氏乳酸菌(Weissella confusa)通过二次发酵桑葚制备桑葚饮料,比较桑葚发酵前后酚类物质对DPPH、OH、ABTS+自由基的清除能力及还原力,并通过非靶向代谢组学技术对不同组别样品差异代谢物进行鉴定,进而探究发酵前后酚类物质组成变化与抗氧化性能之间的相关性。结果:发酵后桑葚酚类物质(1 mg/mL)对DPPH自由基清除率由发酵前的45.97%±2.23%提高到59.01%±3.59%(P<0.05);对OH自由基清除率由发酵前的68.68%±1.23%提高到78.55%±0.57%(P<0.05);发酵后的桑葚饮料中酚类物质的还原力显著提高(P<0.05);发酵前后的酚类化合物对ABTS+自由基清除能力无显著差异(P>0.05)。从发酵前后桑葚酚类物质中共筛选出46种差异代谢物,包括3种花色苷类、10种酚酸类、18种类黄酮及15种其它酚类物质,其中发酵后上调和下调的代谢物分别为26种和20种,共涉及4个相关代谢通路,包括植物次生代谢产物的生物合成、苯丙烷类生物合成、类黄酮生物合成和花色苷生物合成。58.70%的酚类物质与DPPH和OH自由基清除率呈正相关。结论:桑葚经发酵后酚类物质组成变化及含量的增加使得抗氧化能力得到明显上调。

玫瑰桑葚酒酿造工艺优化

以玫瑰花、桑葚为原料,在单因素试验的基础上,采用响应面法优化玫瑰桑葚酒的酿造工艺。结果表明,玫瑰桑葚酒的最佳酿造工艺条件为料液比3∶10(g/mL)、发酵时间为8 d、发酵温度为24℃、发酵初始pH值为3.8。此条件下发酵出的玫瑰桑葚酒酒体清亮,香气醇正,具有玫瑰花和桑葚独特的风味,感官评分为90,酒精度为10.9%vol;未检测出沙门氏菌、金黄色葡萄球菌,符合GB 2758—2012《食品安全国家标准发酵酒及其配制酒》要求。

交配前后雌雄桑蚕蛾营养价值评价

为深入了解桑蚕蛾的营养物质成分,检测了交配前后9芙雌雄蚕蛾蛋白质、脂肪酸和氨基酸的组成和含量。通过粗蛋白、粗脂肪、不饱和脂肪酸含量、动脉粥样硬化指数(indices of atherogenicity,IA)、血栓形成指数(thrombogenicity,IT)以及氨基酸评分、化学评分、必需氨基酸指数(essential amino acid index,EAAI)、营养指数(nutrition index,NI)和生物价(biological value,BV)等指标评价了蚕蛾的营养价值。结果表明,蚕蛾氨基酸和不饱和脂肪酸含量丰富,交配前雌蛾蛋白质(21.6%,鲜重)、总氨基酸(46.94 g/100 g)、必需氨基酸含量(15.86%)以及EAAI(99.33)、NI(21.45)和BV(96.57)最高,同时含有丰富的甜味、药效和支链氨基酸,粗脂肪含量(7.11%,鲜重)低,多不饱和脂肪酸占比(78.18%)高,IA(0.21)和IT(0.14)低,是理想的蛋白源。交配后的雄蛾粗脂肪(24.46%,鲜重)、不饱和脂肪酸(44.31 g/100 g)、亚麻酸含量(19.37 g/100 g)最高,IA(0.28)和IT(0.22)指数较低,是提取不饱和脂肪酸的优质原料。该研究为桑蚕蛾在食品领域的开发应用提供了理论依据。

桑葚花青素的稳定性及降解动力学研究

桑葚花青素不稳定,外界因素的变化容易造成其降解。该试验以花青素保留率和颜色变化为评价指标,考察了光照、pH值、温度、添加剂、金属离子等因素对桑葚花青素稳定性的影响并探究其降解动力学。结果显示:室外光照加速了花青素的降解,k值由避光条件的1.80×10^(-2)升高至14.12×10^(-2),t_(1/2)由避光条件的38.5 h降至5.2 h。酸性环境(pH值1.00,3.00)下花青素的稳定性高,中性(pH值7.00)、碱性(pH值9.00)时花青素易发生降解,96 h后花青素的保留率分别降至83.25%、72.44%,颜色由酸性时的红色分别变为灰绿色、墨绿色。花青素在80,100℃分别放置10 h后ΔE^(*)分别变为3.4097,6.1928,红色褪去,花青素的保留率仅为65.39%、38.83%。酸度剂(柠檬酸)对花青素有明显的稳定和增色作用,防腐剂(山梨酸钾、苯甲酸钠)、抗氧化剂(抗坏血酸)加速了花青素的降解。Al^(3+)、Fe^(3+)、Fe^(2+)等金属离子的加入在一定程度上破坏了花青素的结构,颜色分别变为蓝紫色、黄褐色、蓝黑色。桑葚花青素的贮藏稳定性较差,适合低温、避光和酸性环境,并避免接触防腐剂、抗氧化剂等添加剂和铁、铝制品。该试验结果为花青素的贮藏、加工和应用提供了基础数据和理论依据。

基于改进YOLO v5的复杂环境下桑树枝干识别定位方法

为实现复杂自然环境下对桑树嫩叶处枝干的识别检测,改变当前桑叶采摘设备作业过程中依赖人工辅助定位的现状,解决识别目标姿态多样和环境复杂导致的低识别率问题,提出一种基于改进YOLO v5模型的桑树枝干识别模型(YOLO v5-mulberry),并结合深度相机构建定位系统。首先,在YOLO v5的骨干网络中加入CBAM(Convolutional block attention module)注意力机制,提高神经网络对桑树枝干的关注度;并增加小目标层使模型可检测4像素×4像素的目标,提高了模型检测小目标的性能;同时使用GIoU损失函数替换原始网络中的IoU损失函数,有效防止了预测框和真实框尺寸较小时无法正确反映预测框及真实框之间位置关系的情况;随后,完成深度图和彩色图的像素对齐,通过坐标系转换获取桑树枝干三维坐标。试验结果表明:YOLO v5-mulberry检测模型的平均精度均值为94.2%,较原模型提高16.9个百分点,置信度也提高12.1%;模型室外检测时应检测目标数53,实际检测目标数为48,检测率为90.57%;桑树嫩叶处枝干三维坐标识别定位系统的定位误差为(9.4985 mm,11.285 mm,19.11 mm),满足使用要求。该研究可实现桑树嫩叶处枝干的识别与定位,有助于推动桑叶智能化采摘机器人研究。