A new alkylene dihydrofuran glycoside with antioxidation activity from the root bark of Morus alba L.

A new alkylene dihydrofuran glycoside（1） was isolated from the root bark ofMorus alba L.,along with moracin M-3′-O-β-D-glucopyranoside （2）,and moracin M-6,3′-di-O-β-D-glucopyranoside（3）.Compound 1 was identified as 2-methylene-3-methoxy-2, 5-dihydrofuran-4-O-β-D-glucopyranoside on the basis of chemical and spectroscopic data including 1D and 2D NMR spectral analysis.In addition,the antioxidant activity of 1 was evaluated using the 1,1-diphenyl-2-picrylhydrazyl（DPPH） and 2,2′-azinobis- 3-ethylbenzothiazoline-6-sulphonic acid（ABTS） assay.The IC_（50） values were 2.49 and 0.45 mg/mL,respectively.

Correlation between Different Morus alba L. Leaf Nodes and the Change in Leaf Size or Mass

This paper aimed to explore the correlation between Morus alba L. leaf node and its size or mass change and provide a theoretical basis for the development of scientific Morus alba L. cultivation technology. During 2016-2017,the correlation between Morus alba L. leaf node and its size or mass change was investigated and researched. Results showed that through the correlation coefficient analysis,there was a weakly positive correlation between the leaf node and some indicators such as( leaf length ÷ leaf width) and petiole length,there was a weakly negative correlation between the leaf node and some indicators such as leaf width,( leaf length + leaf width),( leaf length × leaf width) and leaf mass,and there was a weakly negative correlation between the leaf node and the other indicators; there was a highly positive correlation between the leaf mass and some indicators such as Morus alba L. leaf length,leaf width,( leaf length + leaf width) and( leaf length × leaf width),there was a significantly positive correlation between the leaf mass and some indicators such as petiole transverse diameter in width and petiole mass,and there was a weakly positive correlation between the leaf mass and some indicators such as petiole transverse diameter in thickness. In terms of significant level of difference,it was found that there was no significant difference in the correlation coefficients between the leaf nodes on the Morus alba L. shoots and some indicators such as leaf length,leaf width,( leaf length + leaf width),( leaf length × leaf width) and leaf mass; there was an extremely significant difference in the correlation coefficients between the leaf mass and some indicators such as Morus alba L. leaf length,leaf width,( leaf length + leaf width),( leaf length × leaf width),petiole transverse diameter in width and petiole mass; there was no significant difference in the correlation coefficients between the leaf mass and the other indicators. The regression analysis was performed on the leaf node and leaf length,leaf width,( leaf length + leaf width),( leaf length × leaf width),( leaf length ÷ leaf width),petiole length,petiole diameter in width,petiole diameter in thickness,petiole mass and leaf mass. Significance F values were0. 4864,0. 1995,0. 2888,0. 3004,0. 2350,0. 2407,0. 8925,0. 5227,0. 7022 and 0. 2000,respectively,that is,there was an extremely significant difference between Morus alba L. leaf node and petiole diameter in width,there was a significant difference between Morus alba L.leaf node and petiole mass,and there was no significant difference between Morus alba L. leaf node and the other indicators. The comparative analysis of quadratic curve regression equation and linear regression equation was performed on some indicators such as leaf length,and it was found that the R value between the leaf node and petiole diameter in width was in line with the logarithmic curve regression equation,and the other R values of quadratic curve regression equation was larger than the other R values of linear regression equation,indicating that different Morus alba L. leaf nodes,leaf length,leaf mass and other indicators were in line with the quadratic curve regression equation. It was concluded that by investigating the leaf node on the Morus alba L. shoots and regression equation,we could predict the leaf size,leaf mass and other indicators,and we could take the leaf node on the Morus alba L. shoots as a main factor affecting the Morus alba L. leaf size and mass.

Selection and characterisation of lactic acid bacteria isolated from different origins for ensiling Robinia pseudoacacia and Morus alba L.leaves

The objective of this study was to isolate lactic acid bacteria（LAB） strains from different origins and to select the best strains for ensiling Robinia pseudoacacia（RB） and Morus alba L.（MB）leaves. The LAB strains were inoculated into the extracted liquid obtained from RB and MB leaves to evaluate the fermentation products. 11 LAB strains were selected for further experiments based on the highest products of lactic or acetic acid, including 1 strain of Weissella confusa, 2 of Lactobacillus reuteri and 8 of Lactobacillus plantarum.The API 50 CH fermentation experiment indicated that all of the selected 11 LAB strains utilised most of the carbohydrates. All the strains grew at temperatures between 10 and 45℃ and at a p H of 3.5 to 4.5; however, L. reuteri F7 and F8 tolerated a p H as low as 3.0. All 11 LAB strains showed antibacterial activity against Listeria monocytogens, Escherichia coil, Salmonella sp. and Acetobacter pasteurianus; however, after excluding the effect of organic acids, only F7 and F8 still exhibited antibacterial activity. The present study indicated that the selected 11 LAB strains could be used to prepare silages of RB and MB leaves, especially L. reuteri F7 and F8.

New flavanone-monoterpene hybrids as α-glucosidase inhibitors from the root bark of Morus alba

Objective The Morus alba root bark is a well-known Chinese herbal medicine called Sang-Bai-Pi and has often been used to relieve the hyperglycemic symptom of diabetes patients.The current work aims to further explore its bioactive constituents with α-glucosidase inhibitory activity for the potential treatment of diabetes.Methods A combination of different separating techniques including routine column chromatograph and HPLC especially on chiral columns were applied for the isolation of target molecules,while comprehensive spectroscopic experiments comprising MS,NMR,ECD,etc.were carried out to complete the structural assignment.The anti-hyperglycemic property of the isolates was evaluated by an in vitro α-glucosidase inhibitory bioassay.Results Two pairs of new flavanone-monoterpene hybrid enantiomers were isolated and identified,and an interesting phenomenon of mutual transformation between these cometabolites were detected,which resulted in their regio-isomerization and enantiomerization.The bioassay results revealed remarkable α-glucosidase inhibitory activity for these fascinating molecules.Conclusions The Morus alba root bark is a rich source of bioactive flavonoid derivatives and deserves further investigations to develop new potential chemotherapies for diabetes control and treatment.

Inhibition of Sanggenon G Isolated from Morus alba on the Metastasis of Cancer Cell

Objective An organic layer prepared from the cortex of Morus alba(Moraceae)was studied in order to identify the active compounds for heparinase.Methods Bioassay-guided fractionation resulted in the isolation of sanggenon G.Results The compound showed inhibitory activity with IC50 of 3.7μmol/L on heparinase in vitro as well as 24μmol/L in invasion assay using MDA-MB231 cells.Sanggenon G also had the moderate cytotoxicity at SW 620(colon)and ACHN(kidney)cancer cell lines with IC50 of 10.96 and 13.44μmol/L,respectively.Conclusion This is the first time that prenylated flavonoid sanggenon G is described as heparinase inhibitor.Besides,this flavonoid would be expected to be a metastasis inhibitor of cancer cells and also a valuable reagent to explore the mechanism of heparinase/heparanase-mediated metastasis.

A New Geranylated 2-Arylbenzofuran from Morus alba

Objective To study the chemical constituents from the root barks of Morus alba. Methods Various chromatographic techniques were used to separate the constituents and their structures were elucidated on the basis of extensive spectroscopic interpretation. Results Six compounds were isolated from the root barks of M. alba. The structure of new compound, named as albafuran D（1）, was established together with known compounds sanggenon H（2）, kuwanon E（3）, kuwanon U（4）, kuwanon F（5）, and benzokuwanon E（6）. Conclusion Compound 1 is a new geranylated 2-arylbenzofuran.

Analgesic and anti-inflammatory effects of UP1304,a botanical composite containing standardized extracts of Curcuma longa and Morus alba

OBJECTIVE： Though the initial etiologies of arthritis are multifactorial, clinically, patients share the prime complaints of the disease, pain. Here the authors assessed the analgesic and anti-inflammatory effects of UP1304, a composite that contains a standardized blend of extracts from the rhizome of Curcuma /onga and the root bark of Morus a/ba, on rats with carrageenan-induced paw edema. METHODS： A plant library was screened for bradykinin receptor antagonists./n vivo, the anti- inflammatory and analgesic effects of the standardized composite, UP1304, were evaluated in rats with carrageenan-induced paw edema using oral dose ranges of 100-400 mg/kg. Ibuprofen, at a dose of 200 mg/kg, was used as a reference compound. In vitro, cycleoxygenase （COX） and lipoxygenase （LOX） inhibition assays were performed to evaluate the degree of inflammation. RESULTS： Statistically significant improvements in pain resistance and paw edema suppression were observed in animals treated with UP1304, when compared to vehicle-treated rats. Results from the highest dose of UP1304 （400 mg/kg） were similar to those achieved by ibuprofen treatment at 200 mg/kg. /n vitro, UP1304 showed dose-dependent inhibition of the enzymatic activities of COX and LOX. A half-maximal inhibitory concentration of 9.6 tJg/mL for bradykinin B1 inhibition was calculated for the organic extract of C./onga. Curcumin showed Ki values of 2.73 and 58 IJg/mL for bradykinin receptors B1 and B2, respectively. CONCLUSION： Data presented here suggest that UP1304, analgesic and anti-inflammatory agent of botanical origin, acted as a bradykinin receptor B1 and B2 antagonist, and inhibited COX and LOX enzyme activities. This compound should be considered for the management of symptoms associated with arthritis.

12个桑品种叶片营养与活性成分分析

为筛选不同用途的桑(Morus alba L.)品种,选取7份栽培桑和5份野生桑种质夏季成熟的叶片,分别测定其12项成分含量,比较栽培桑和野生桑各组分含量间的差异,并采用相关性分析、主成分分析法对叶质的性能进行综合评价。结果表明,栽培桑的粗蛋白、粗纤维、类黄酮和总酚含量显著高于野生桑,粗脂肪、野尻霉素、钙和硒含量显著低于野生桑,而总糖、生物碱、γ-氨基丁酸含量与野生桑无显著差异;主成分分析中提取的4个主成分累计贡献率达88.37%,以此计算桑叶品质的综合得分,结果显示“908”叶质综合得分最高,其次为广1号、小红皮、黑玉墨斗、伦教408、GM41和“731”,而野生桑主成分得分均为负值;相关性分析表明夏季叶质综合评价指标可简化为粗蛋白、总糖、总酚和粗脂肪含量4项指标。该夏季叶质评价体系模型可为桑树优良品种选育以及桑叶产品加工原料的选取提供一定的理论依据。

桑树光合作用转录组研究

研究高产桑品种鄂桑1号(E1)和对照桑品种湖桑32号(H32)光合特性及光合相关基因表达变化,为桑树高光效育种以及种质资源筛选提供理论依据。利用LI-6400XT型便携式光合测定仪测定桑树叶片气体交换和叶绿素荧光参数,同时测定叶绿素含量和1,5二磷酸核酮糖羧化酶(RUBP)活性,在桑树光合日变化曲线峰值和谷底取样进行转录组测序,比较不同桑品种光合相关基因表达的差异。对E1和H32的光合日变化测定结果显示,桑品种E1叶片的Pn峰值显著大于H32峰值(P<0.05),H32叶片Tr峰值显著大于E1峰值(P<0.05);不同品种桑树叶片Pn-PAR和Pn-Ci的响应中,CE、CSP为E大于H32,AQY、LCP、LSP、CCP为H32大于E1。E1在较弱光强下实现光合产物积累的能力强于H32,E1对CO2低浓度的利用效率优于H32;不同桑品种叶片叶绿素荧光参数中,E1叶片ΦPSⅡ和qP显著大于H32(P<0.05),H32的NPQ显著大于E1(P<0.05),表明H32叶片吸收的光能较多的以热能的形式进行耗散;H32的叶绿素a、叶绿素b和叶绿素含量均显著小于E1(P<0.05),E1的RUBP活性大于H32(P>0.05),表明E1光合能力优于H32。不同品种不同时段发现3356个差异表达基因(Differentially expressed genes,DEGs),代谢通路分析显示共有1136个DEGs注释到KEEG数据库,DEGs显著富集的光合作用相关代谢通路主要是光合作用固碳、碳代谢、卟啉和叶绿素代谢等3条光合相关代谢通路。在FC≥2且FDR<0.01的高差异表达基因中筛选功能描述与光合作用相关的新基因共10个。其中MulberrynewGene3646、MulberrynewGene1405、MulberrynewGene2419和MulberrynewGene320共4个DEGs可能是造成H32和E1净光合速率和产量差异的关键基因,可重点研究。

基于叶绿体matK基因序列的两份安康野生桑遗传特征分析

对陕西省安康市黄石滩、八仙镇2份野生桑叶绿体基因组进行PCR扩增,获得matK编码基因及其侧翼序列,扩增长度为1715 bp,ORF长度为1518 bp,2份序列仅存在1处单碱基差异。利用matK序列分析7份桑树(Morus alba L.)之间的遗传多样性,结果表明,不同品种桑树matK序列相似度较高,仅存在6个单位点碱基差异,定义3种单倍体,单倍型多样度为0.714,核苷酸多样度为0.00132,平均核苷酸差异为2.000。遗传距离分析表明,八仙镇桑树与印度桑(M.indica)、鲁桑(M.multicaulis)的遗传距离最近,黄石滩桑树与蒙桑(M.mongolica)、华桑(M.cathayana)的遗传距离最近。聚类分析将7份桑树聚为1支,且桑树品种之间的进化支长要明显短于其他科属之间的长度,表明桑树品种间的遗传多样性较低,经历较少进化事件。

基因编辑技术及其在桑树育种中的应用前景展望

综述了3种基因编辑技术的作用机制及CRISPR/Cas9基因编辑技术在植物育种中的应用。根据桑树(Morus alba L.)育种工作的现状,对CRISPR/Cas9基因编辑技术在桑树功能基因组学和遗传改良中的应用前景进行了展望,以期为桑树多元化种质创新提供借鉴和新思路。

α-Glucosidase inhibitors and antioxidants from root bark of Morusalba

Objective: To study the chemical constituents from root bark of Morus alba and their α-glucosidase inhibition and DPPH radical scavenging activities.Methods: The chemical constituents were isolated and purified by repeated column chromatographies on silica gel, Sephadex LH-20, and preparative HPLC. Their structures were elucidated by 1 D and 2 D NMR spectra and HR-ESI-MS.Results: Thirteen compounds 1–13 were isolated and identified. The bioactive assays revealed that compounds 1, 3 and 8 displayed strong α-glucosidase inhibitory activity with IC50 values of(147.1 ± 1.1),(314.1 ± 0.8), and(207.6 ± 0.1) μmol/L, respectively, which were stronger than the positive control of acarbose(418.6 ± 0.1 μmol/L). Compounds 10 and 11 displayed potent DPPH scavenging activity with EC50 values of(2.9 ± 0.1) and(5.0 ± 0.1) μmol/L [EC50 of positive control Vitamin C was(54.8 ± 0.1) μmol/L],respectively.Conclusion: To the best of our knowledge, this is the first report about the compounds 1, 3, and 8 of M.alba with α-glucosidase inhibitory effects.

不同非生物胁迫对桑叶1-脱氧野尻霉素含量的影响

通过不同程度的干旱、淹水、盐处理等非生物胁迫,对桑叶的1-脱氧野尻霉素(DNJ)含量进行测定,结果表明,桑树(Morus alba L.)在一定程度的干旱、淹水和盐胁迫环境下,桑叶DNJ含量积累更高。本研究结果为桑树在消落带和盐碱地等地区更有效地开发桑树DNJ资源提供理论依据。

Oxidative Stressand Antioxidant Responses in Young Leaves of Mulberry Plants Grown Under Nitrogen, Phosphorus or Potassium Deficiency

The aim of this study was to associate the generation of reactive oxygen species （ROS） with Induced antloxidant responses and disturbed cellular redox environment in the nitrogen-（N）, phosphorus-（P）, or potassium-（K） deftcient mulberry （Morus alba L. var. Kanva-2） plants. The indicators of oxidative stress and cellular redox environment and antioxldant defense-related parameters were analyzed. Oeficlency of N, P or K suppressed growth, accelerated senescence, and decreased concentrations of chloroplastic pigments and glutathione. Lipid peroxidation and activities of superoxide dismutase, ascorbate peroxidase and glutathione reductase were also increased in these N, P, or K deprived plants. Concentration of hydrogen peroxide Increased in plants deficient in N or P. Oeficlency of N or P particularly altered the cellular redox environment as indicated by changes in the redox couples, namely ascorbic acid/total ascorbate decreased in P-, glutathione sulfydryl/total glutathione decreased in N-, and Increased in P-deficient plants. Activity staining of native gels for superoxide dismutase revealed Increased activity as Indicated by Increased intensity of bands, and induction of few new isoforms in P- and K-deficient plants. Oifferences in the patterns of superoxide dismutase isoforms and redox status （ascorbic acid/total ascorbate and glutathlone sulfydryl/total glutathione） Indicate that N-, P-, or K-deficiency altered antioxidant responses to varying extents in mulberry plants.

基于桑葚转录组测序的SSR位点分析

采用转录组测序技术对三个时期的桑葚进行转录组测序,建立桑葚的EST数据库。基于生物信息学方法分析桑葚转录组数据库的简单重复序列位点。从51895条Unigenes中共检索出23641条Unigenes含有44867个简单重复序列位点,共计252种基序类型。在所有基序类型中,以A/T与AT/AT型出现次数最多。单核苷酸基序类型与二核苷酸基序类型出现频率最高,共计75.69%。基序长度以10~20 bp为主,重复次数集中5~18次。设计27149对引物,随机选择20对引物,通过PCR验证,在4个桑树品种中展现出良好的重现性与通用性。桑葚转录组SSR位点类型丰富,具备开发出适宜于果桑选育的分子标记的潜力。

In vitro assessment of the effect of microencapsulation techniques on the stability,bioaccessibility and bioavailability of mulberry leaf bioactive compounds

Mulberry(Morus alba L.)leaf extract is well-known for its health-promoting features.However,food processing conditions affect its bioactive profile,which could be limited by encapsulation.Nevertheless,assessing the impact of encapsulating techniques on the digestibility of embedded biocompounds requires further knowledge.Hence,the aimed of this study was to assess the effects of carrier materials and drying techniques on(i)bioaccessibility and bioavailability indexes,(ii)stability of nutraceuticals,and(iii)changes in antioxidant activities of mulberry leaf extract powder by using an in vitro oral-gastrointestinal digestion.Digestion was performed out at 37°C in darkness by shaking encapsulated extract with human saliva during 2 min,followed by 2 h of incubation with a pepsin-HCl mixture,then mixed for 2 h with bile salts and pancreatin.After digestion the samples were acidified and HPLC assayed.A significant(p<0.05)decrease in nutraceuticals content was noted during the intestinal phase with more reduction in gamma-aminobutyric acid(34.91–51.14%)compared to that of flavonols(16.58–28.90%),1-deoxynojirimycin(17.56–20.42%)and phenolic acids(0.53–0.67%)in the gastric digesta.Although encapsulation was observed to enhance the bioaccessibility,negative effects were found in terms of the bioefficiency and bioavailability.Furthermore,encapsulation techniques mostly affected the ion reducing capacity than the radical scavenging capacity.The outcomes suggested that carriers had a major effect on the digestibility and antioxidative activity,whilst the drying techniques mostly affect the bioaccessibility and bioavailability of the flavonols.