Morus Species Research Articles

Antitumor progression potential of morusin suppressing STAT3 and NFκB in human hepatoma SK-Hep1 cells

Morusin is a prenylated flavonoid that has been isolated from the root bark of the mulberry tree (Morus species, Moraceae), a Chinese traditional medicine. It has been synthesized by our laboratory from commercially available phloroglucinol, and has demonstrated to possess antitumor effects of cell lines including A549, MCF-7, and MDA-MB-231. In this study, at non-cytotoxic concentrations, morusin altered invasive morphology and suppressed cell-matrix adhesion, cell motility and cell invasion in SK-Hep1 cells. Morusin also increased the expression of E-cadherin, an epithelial cell junction protein, decreased the expression of vimentin, a mesecnchymal marker, and α2-, α6-, β1- integrin, which regulated cancer attachment and migration. In addition, morusin reduced the activity of matrix metalloproteinase-2 and 9 (MMP-2 and MMP-9), which were involved in extracellular matrix (ECM) degradation and promoting cancer cell invasion. Furthermore, morusin suppressed the signal transducer and activator of transcription 3 (STAT3) and nuclear factor-κB (NFκB) signaling pathways, which modulate the protein expression involved in the invasion process. Finally, morusin decreased the lung colonization of the SK-Hep1 cells in the nude mice. These results indicate morusin possesses antitumor progression potential through suppressing STAT3 and NFκB.

Anthocyanin composition of different wild and cultivated berry species

A broad range of anthocyanins (glycosides of cyanidin, pelargonidin, peonidin, delphinidin, malvidin, and petunidin) was identified and quantified in the fruit of 24 wild and cultivated berry species using HPLC-DAD-MS2. The anthocyanin composition in species of Ribes, Rubus, Vaccinium, and Fragaria genus as well as in less known species of Crataegus, Morus, Amelanchier, Sorbus, Sambucus and Aronia genus was determined. Cyanidin was the most commonly occurring anthocyanidin, meanwhile malvidin glycosides were only detected in blueberries. Glycosides of pelargonidin were detected in raspberries, strawberry, black mulberry and chokeberry. Peonidin glycosides were identified in hawthorn, black currant and gooseberry as well as in blueberry species. The richest species in the content of peonidin glycosides were blueberries. Delphinidin glycosides were the prevailing form of anthocyanins in black currant and bilberry. The highest total anthocyanin content was determined in dark colored fruit of cultivated elderberry and bilberry whereas light-colored dog rose and Chinese hawthorn fruit had the lowest anthocyanin content. The composition of anthocyanidin glycosides did not differ between the fruit of wild growing and cultivated species, but their contents were generally different.

Morus species through centuries in pharmacy and as food

The use of various Morus species in pharmacy, as well as in traditional medicine is well known worldwide. In Serbia there is a proverb 'Health comes in through the mouth', i.e. through food. So, the aim of the present review paper is the construction of a mosaic from many known uses of Morus species which have been in use through centuries and in different cultures as a remedy, but also as an important nutrient.

Cytogenetical Studies in Some Triploid Mulberry Varieties (Moraceae)

Most Morus species are diploid, having 28 chromosomes, but a few species, specifically, M. tiliafolia Makino, Morus cathyana and M. nigra L., are higher polyploidy. In these species, the chromosome number varies from 2n=28 to 2n=308, with ploidy level from x to 22x. In the present study, three diploid mulberry varieties, namely, V1, S13 and S34 have been analyzed for detailed meiotic studies. Based on chromosome configuration and other meiotic behaviour, x=14 has been considered as the basic number of the genus. Meiosis shows marginal differences. Various meiotic abnormalities, such as the occurrence of one large chromosome at metaphase I, unequal separation at anaphase I, the presence of laggards and precocious movement of chromosomes at anaphase II, and two micronuclei at tetrad, have been observed. The low pollen fertility and seed sets may be due to irregular chromosomal pairing and separation rather than due to genetic or physiological causes. © 2013 The Japan Mendel Society.

Characterization of Morus species in respect to micro, macro, and toxic elements

This study examines the mineral composition of the extracts of the fruits, leaves and roots of white mulberry (Morus alba L.) and black mulberry (Morus nigra L.) grown in Serbia. All extract samples of white and black mulberry were analyzed for the content of micro (B, Co, Cr, Cu, Fe, Li, Mn, Ni, Se, Sr, Zn), macro (Ca, Mg, Na), and toxic metals (Al, As, Cd, Hg, Pb) by inductively coupled plasma optical emission spectrometry (ICP-OES). The study revealed that parts of the plant had statistically significant impact on the levels of the examined elements among the two Morus species. All extracts contained high amounts of Ca, Mg, Na, B, Cu, Fe, Mn and Zn. The studies showed that in the most of extracts dominant macro element was Mg (591- 1942 ?g/g of dry extract), while dominant microelements were Zn, B, Cu in all extracts, except for the black mulberry leaves, whose extract was most abundant in Fe (143 ?g/g of dry extract). The highest content of micro and macro elements was detected in the extract of black mulberry leaves. This work contributes to the knowledge of the nutritional properties of Morus species. The obtained results may be useful in the evaluation of new dietary and food products.

Phenols, Flavonoids, and Antioxidant and Antibacterial Activity of Leaves and Stem Bark of Morus Species

Leaves and stem bark composition from Morus species (M. alba var. alba, M. alba var. rosa and M. rubra) was evaluated for its antioxidant and antimicrobial activities in order to enhance its therapeutic uses. Total phenolics and flavonoids contents were estimated in hydromethanolic and aqueous extracts. Results showed highest content in M. rubra leaves aqueous extract (1129 mg gallic acid equivalent/100 g dry weight and 816 mg gallic acid equivalent/100 g dry weight, respectively). Using 2,2-diphenyl-1-picrylhydrazyl assay, the highest antioxidant activity was obtained in aqueous stem barks extracts. M. alba var. alba has an IC50 of 2.84 mg/ml and the IC50 value of M. rubra was the highest (4.78 mg/ml). ABTS.+ (2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid)) method and reducing power assay were used to confirm the results from the 2,2-diphenyl-1-picrylhydrazyl test. All extracts expressed considerable free radical-scavenging properties. Hydromethanolic and aqueous extracts were tested against Staphylococcus aureus, Enterococcus feacalis, Staphylococcus epidermis, Escherichia coli, and Salmonella Typhimurium bacteria. Hydromethanolic stem bark extracts have the highest antimicrobial activities, and it may be a good antimicrobial agent for human gastrointestinal infections. This plant could be used as an additive to foods and also as a possible source to obtain new and effective herbal medicines to treat infections of multi-drug resistant strains of microorganisms.

English

Genetic stability of Morus alba, Morus indica, Morus laevigata (indigenous collection) and Morus species (exotic collection) have been studied in in vitro regenerated plants of mulberry (fresh, before and after cryopreservation) using random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) markers. This study examined the genetic stability of cryopreserved dormant buds of Morusgermplasm that were stored in liquid nitrogen using two-step freezing, then rewarmed and regrown. Dormant buds of mulberry collected during winter period were found suitable for the cryopreservation in liquid nitrogen. In the present study, the plants were regenerated directly from dormant buds (before and after cryopreservation) without intermediary callus phase. These regenerants thus bear low risk of genetic instability. Both the single primer amplification reaction (SPAR) markers showed reproducible and well resolved banding patterns in mulberry germplasm, in which RAPD marker generated a total of 201 bands based on 15 primers; however, ISSR markers were given 145 bands using 11 primers. Both markers showed monomorphic banding patterns and did not reveal any polymorphism among the mother plant and in vitro regenerants before and after cryopreservation, suggesting that cryopreservation, using two-step freezing, does not affect genetic stability of mulberry germplasm. The transitory nature of these polymorphisms should be carefully considered when monitoring for genetic stability. Key words: Cryopreservation, Genetic stability, in vitro culture, ISSR, mulberry, RAPD.

Draft genome sequence of the mulberry tree Morus notabilis

Human utilization of the mulberry–silkworm interaction started at least 5,000 years ago and greatly influenced world history through the Silk Road. Complementing the silkworm genome sequence, here we describe the genome of a mulberry species Morus notabilis. In the 330-Mb genome assembly, we identify 128 Mb of repetitive sequences and 29,338 genes, 60.8% of which are supported by transcriptome sequencing. Mulberry gene sequences appear to evolve ~3 times faster than other Rosales, perhaps facilitating the species’ spread worldwide. The mulberry tree is among a few eudicots but several Rosales that have not preserved genome duplications in more than 100 million years; however, a neopolyploid series found in the mulberry tree and several others suggest that new duplications may confer benefits. Five predicted mulberry miRNAs are found in the haemolymph and silk glands of the silkworm, suggesting interactions at molecular levels in the plant–herbivore relationship. The identification and analyses of mulberry genes involved in diversifying selection, resistance and protease inhibitor expressed in the laticifers will accelerate the improvement of mulberry plants.

A field study on solar-induced chlorophyll fluorescence and pigment parameters along a vertical canopy gradient of four tree species in an urban environment

To better understand the potential uses of vegetation indices based on the sun-induced upward and downward chlorophyll fluorescence at leaf and at canopy scales, a field study was carried out in the city of Valencia (Spain). Fluorescence yield (FY) indices were derived for trees at different traffic intensity locations and at three canopy heights. This allowed investigating within-tree and between-tree variations of FY indices for four tree species. Several FY indices showed a significant (p<0.05) and important effect of tree location for the species Morus alba (white mulberry) and Phoenix canariensis (Canary Island date palm). The upward FY parameters of M. alba, and the upward to downward ratios at 687 and 741nm for both species, were significantly related to tree location. It was found that not the total chlorophyll (Chl) content, but rather the Chl a/b ratio showed the strongest correlations with several of the indices applied. Chl a/b was lowest at the bottom level of the highest traffic intensity location for both species due to an increased Chl b, indicating a larger light harvesting complex related to Photosystem II (LHCII) as a response to limiting light. The leaf deposits from traffic observed at this sampling location possibly led to a shading effect, resulting further in an adaptive response of the photosynthetic system and subsequent difference of FY indices. This study therefore indicated the importance of the size of LHCII on the fluorescence emission, observed under different traffic generated pollution conditions.

Quantitative determination of 1-deoxynojirimycin in mulberry leaves from 132 varieties

Mulberry 1-deoxynojirimycin (DNJ) shows high α-glucosidase inhibitory, antioxidant, antimicrobial, and anti-inflammatory activity. This is a widely used functional food constituent in China, Japan, Korea, Thailand and many other Asian countries. In this study, the DNJ content of mulberry leaves from 132 varieties of nine Morus species was determined. The degree of maturity of the mulberry leaves affected the DNJ content significantly. The younger leaves contained higher DNJ concentrations than did the older leaves. DNJ concentrations in mature leaves varied among these 132 mulberry varieties from 0.1341 to 1.472mg/g of dry leaves. DNJ content in leaves was variety dependent and had no obvious relationship with either the species or their place of origin.

Natural Antioxidants Effect of Mulberry Fruits (Morus nigra and Morus alba L.) On Lipids Profile and Oxidative Stress in Hypercholestrolemic Rats

Mulberry belongs to the Morus genus of the Moraceae family. There are 24 species of Morus, with at least 100 known varieties. Mulberry leaves, bark and branches have long been used in Chinese medicine. In most European countries mulberries are grown for fruit production. Mulberry are a good source of polyphenols, especially anthocyanins and micronutrients. In epidemiologic al and clinical studies, these constituents have been associated with improved cardiovascular risk profiles. Human intervention studies using (either fresh, or as juice, or freeze-dried) , or purified anthocyanin extracts have demonstrated significant improvements in LDL oxidation, lipid peroxidation, total plasma antioxidant capacity, dyslipidemia and glucose metabolism. Benefits were seen in healthy subjects and in those with existing metabolic risk factors. The present study was performed to determine the natural antioxidants content of black and white mulberry fruits (Morus nigra and Morus alba L.) in Egypt and study the effects of different levels different levels of black and white mulberry fruits on lipid profile and oxidative stress in hypercholestrolemic rats. Forty adult male albino rats Sprague Dawley strain with an average weight of (100±120 g) randomly divided into 8 groups (each of 5 rats), Group 1: Control negative, rats fed on basal diet, Group 2: Control positive, rats fed on hypercholesterolemic diet composed of basal diet +2% cholesterol. Others groups (3-8), rats fed on hypercholestrolemic diet supplemented by different levels of black or white mulberry dry fruits (2.5, 5 and 10%) for four weeks. Results observed that rats fed on hypercholestrolemic diet without supplementation (control positive) had significant increase in malonaldhyde (MDA), nitric oxide (NO), total cholesterol, triglycerides (T.G), low density lipoprotein (LDL) and very low density lipoprotein (VLDL) while it ha d significant decrease in total antioxidant capacity (TAC) and high density lipoprotein (HDL) comparing with rats fed on basal diet (control negative). Moreover, The group administrated with Morus nigra L. 10% had lowest value in malonaldhyde (MDA) and nitric oxide (NO) followed by the group administrated with Morus alba L. 10%. In addition, all hypercholestrolemic groups administrated with different levels (2.5, 5 and 10%) of black and white mulberry fruits (Morus nigra and Morus alba L.) had significant increase in total antioxidant capacity (TAC). The group administrated with Morus nigra L. 10% had highest value in total antioxidant capacity (TAC) followed by the group administrated with Morus alba L. 10%. In addition, all hypercholestrolemic groups administrated with different levels (2.5, 5 and 10%) of black and white mulberry fruits (Morus nigra and Morus alba L.) had significant decrease in total cholesterol, triglycerides (T.G), low density lipoprotein (LDL) and very low density lipoprotein (VLDL). and significant increase in high density lipoprotein (HDL),comparing with rats fed on hypercholestrolemic diet (control positive). It concluded that, consumption of black and white mulberry fruits (Morus nigra and Morus alba L.) my modify the risk of hypercholesterolemia and it have more potential as a health supplement rich in natural antioxidants. Moreover, mulberries are emerging as a dietary source of multiple compounds and nutrients, including anthocyanins, flavonoids, vitamins and fiber, that reduce CVD risk. While limited epidemiological data inversely associate consumption of berries with inflammation and CVD, these conclusions need to b e strengthened in future case-control or cohort studies investigating the long-term health benefits of berries in specific populations.

Morus alba and active compound oxyresveratrol exert anti-inflammatory activity via inhibition of leukocyte migration involving MEK/ERK signaling.

BackgroundMorus alba has long been used in traditional Chinese medicine to treat inflammatory diseases; however, the scientific basis for such usage and the mechanism of action are not well understood. This study investigated the action of M. alba on leukocyte migration, one key step in inflammation.MethodsGas chromatography-mass spectrometry (GC-MS) and cluster analyses of supercritical CO2 extracts of three Morus species were performed for chemotaxonomy-aided plant authentication. Phytochemistry and CXCR4-mediated chemotaxis assays were used to characterize the chemical and biological properties of M. alba and its active compound, oxyresveratrol. fluorescence-activated cell sorting (FACS) and Western blot analyses were conducted to determine the mode of action of oxyresveratrol.ResultsChemotaxonomy was used to help authenticate M. alba. Chemotaxis-based isolation identified oxyresveratrol as an active component in M. alba. Phytochemical and chemotaxis assays showed that the crude extract, ethyl acetate fraction and oxyresveratrol from M. alba suppressed cell migration of Jurkat T cells in response to SDF-1. Mechanistic study indicated that oxyresveratrol diminished CXCR4-mediated T-cell migration via inhibition of the MEK/ERK signaling cascade.ConclusionsA combination of GC-MS and cluster analysis techniques are applicable for authentication of the Morus species. Anti-inflammatory benefits of M. alba and its active compound, oxyresveratrol, may involve the inhibition of CXCR-4-mediated chemotaxis and MEK/ERK pathway in T and other immune cells.

Antioxidant Rich Morus alba leaf Extract Induces Apoptosis in Human Colon and Breast Cancer Cells by the Downregulation of Nitric Oxide Produced by Inducible Nitric Oxide Synthase

Morus species had been used widely in the traditional medicines for various diseases. In this study we report the in vitro antiproliferative activity of the methanol extract of Morus alba. The extract is capable of inducing cytotoxicity in human colon cancer (HCT-15) cells (IC50 = 13.8 μg/ml) and breast cancer (MCF-7) cells (IC50 = 9.2 μg/ml), resulted in significant morphological changes of the cells, fragmentation of DNA, and caspase-3 activation- characteristics of apoptosis. It downregulated the amount of nitric oxide (NO) produced as a result of inducible nitric oxide (iNOS) activation. The HPLC analysis of the extract showed epicatechin (20%), myricetin (10%), quercetin hydrate (12%), luteolin (12%), and kaempferol (6%) as the major active components and ascorbic acid, gallic acid, pelargonidine, and p-coumaric acid as the minor components. To the best of our knowledge, this is the first report showing the downregulation of iNOS and induction of apoptosis by M. alba extract.

EMS Induced Morphometric Biomass and Phytochemical Variations in Morus Species (Genotype RFS135)

EMS Induced Morphometric Biomass and Phytochemical Variations in Morus Species (Genotype RFS135)

EMS Induced Morphometric Biomass and Phytochemical Variations in Morus Species (Genotype RFS135)

Aims: In the present investigation mutations were induced by EMS in rain fed mulberry genotype RFS135 and evolved mutants were evaluated for crop improvement by morphometric and phytochemical studies. Study Design: Cross-sectional study. Place and Duration of Study: Department of sericulture, Jnanabharathi, Bangalore University, Bangalore-560056, Karnataka, India, between June 2006 and July 2010. Methodology: The active bud sprouts of vegetative cuttings of RFS135 in multiple sets were treated for twelve hours intermittently (every one hour) with three different concentrations (0.1%, 0.3% and 0.5%).Further M1V1 and M1V2 generation clones were evaluated for biomass, nutritive and morpho-metric characters. Results: The results revealed that concentrations of 0.1% and 0.3% EMS treatment were effective in significantly altering the morpho-metric characters, biomass yield and phyto chemical constituents. The significant variation in the morpho-metric characters such as height of the plant, number of branches, stem girth, number of leaves per plant and Research Article American Journal of Experimental Agriculture, 3(1): 43-55, 2013 44 increased biomass was recorded among the M1V2 clones of 0.1% EMS treatment (p=0.0001) and 0.3% EMS treatment (p=0.0006). Further significant improvement was recorded in nutritive parameters such as proteins, reducing sugars, minerals and moisture content. Moisture retention capacity and Chlorophyll contents were also found to be high in mutant clones recovered from 0.1% and 0.3% EMS treatments. Conclusion: This suggests that EMS at lower concentrations below 0.5% can be safely used for crop improvement in mulberry and can also be tried in other vegetative propagated crops.

Antidepressant Activities of the Methanol Extract, Petroleum Ether and Ethyl Acetate Fractions of &amp;lt;i&amp;gt;Morus mesozygia&amp;lt;/i&amp;gt; Stem Bark

Morus mesozygia Stapf (Moraceae) is the only Morus species indigenous to Tropical Africa. It is believed in traditional medicine that the plant exhibits several medicinal properties. The antidepressant-like activities of the crude methanol extract, petroleum ether and ethyl acetate fractions were investigated using the Forced Swimming Test (FST) and Tail Suspension Test (TST) models. The petroleum ether fraction at a dose of 140 mg/kg was the most effective in both the FST and TST when compared to the activity of the reference drug Imipramine (250 mg/kg) by reducing the immobility duration of the animals by 66.8% and 49.4% respectively.

Macrophage activating activity of pyrrole alkaloids from Morus alba fruits

Ethnopharmacological relevanceThe fruits of Morus alba have been traditionally used as a tonic to enhance immune responses. Materials and methodsThe macrophage activating constituents of Morus alba fruits were purified using various column chromatography techniques. The structures of isolated compounds were determined on the basis of spectroscopic data interpretation such as 1D and 2D NMR analysis. The macrophage activating activities of isolated compounds were evaluated by measuring the production of nitric oxide, TNF-α and IL-12 in RAW 264.7 cells. The phagocytic activity was also evaluated. ResultsFive pyrrole alkaloids, 5-(hydroxymethyl)-1H-pyrrole-2-carboxaldehyde (1), 2-formyl-1H-pyrrole-1-butanoic acid (2), 2-formyl-5-(hydroxymethyl)-1H-pyrrole-1-butanoic acid (3), 2-formyl-5-(methoxymethyl)-1H-pyrrole-1-butanoic acid (4) and Morrole A (5) were isolated from the fruits of Morus alba. Morrole A (5) is first reported in nature and other pyrrole alkaloids (1–4) are first reported from Morus species. Among the isolated compounds, compounds 3 and 4 significantly activated macrophage activity by the enhancement of nitric oxide, TNF-α and IL-12 production, and the stimulation of phagocytic activity in RAW 264.7 cells. ConclusionPyrrole alkaloids, including a new compound, were isolated from Morus alba fruits. These compounds activated macrophage activity in RAW 264.7 cells.

P048 Suppression of proinflammatory cytokines by mulberry derivatives

Introduction The mulberry belongs to the Morus genus of the Moraceae family. There are 24 species of Morus, with at least 100 known varieties. Methods RAW 264.7 cells were pretreated with mulberry or mulberry derivatives for 2 h, and then stimulated with lipopolysaccharide (LPS). Gene expression of proinflammatory cytokines were determined by RT-PCR. The levels of NO were determined by measuring the amount of nitrate using Griess reagents, and absorption was measured at 570 nm. Intracellular ROS levels were evaluated using a DCFH-DA fluorescent probe by flow cytometry. Results We found that mulberry derivatives inhibited the gene expression of proinflammatory cytokines including IL-6, IL-1b, TNF-aand COX-2 in Raw264.7 cells stimulated with lipopolysaccharide (LPS). In addition, LPS-induced intracellular ROS and NO levels were decreased in Raw264.7 cells treated with mulberry derivatives compared with those in mulberry. Conclusion These data suggest that mulberry derivatives showed anti-inflammatory activity by reducing intracellular ROS and NO production via suppression of proinflammatory cytokine expression, and mulberry derivatives may be useful for the prevention and therapy of inflammation-related diseases.

Endozoochorous dispersal of alien and native plants by two palearctic avian frugivores with special emphasis on invasive giant goldenrod Solidago gigantea

Abstract The role of frugivorous birds in dispersal of the seeds of fleshy-fruited plants is well understood, but other groups of birds like granivores and omnivores can also act as vectors, especially in the process of non-standard dispersal. In this study we examined the role of two widely distributed birds (the Eurasian Blackbird, Turdus merula, and the Black Redstart, Phoenicurus ochruros) in seed dispersal. We analyzed the content of 279 droppings of the Black Redstart collected between July and October and 85 droppings of the Eurasian Blackbird collected during January. In total we found 366 seeds, representing 11 plant taxa, in the faecal samples; 93% of the seeds belonged to fleshy-fruited species. The most numerous were two native taxa, Sambucus racemosa/nigra, in summer and autumn, Crataegus monogyna in winter, and one alien species Morus alba, which is dispersed in summer. We also found seeds of four dry-fruited species; the most interesting of which was the presence of anemochorous seeds of Solidago gigantea (a highly invasive alien species). The seeds were found only in droppings of the Eurasian Blackbird. Considering the obtained results the need for further studies on bird-plant interactions should be a priority.

Free radical scavenging activity, total phenolic and flavonoid contents of mulberry (Morus spp. L., Moraceae) extracts

Mulberry (Morus spp. L., Moraceae) fruits, leaves, bark and branch have been used in traditional medicine as diuretic, hypoglycemic and hypotensive. The mechanism of their effects is correlated with the content of active components. Objective of this work was to evaluate and compare antioxidant properties of different extracts of two Morus species growing in Serbia: Morus alba L. (white mulberry) and Morus nigra L. (black mulberry). Potential antioxidant activity, content of antioxidant compounds (phenolics and flavonoids) and radical scavenging capacity, tested by DPPH method, were evaluated. The phenolic and flavonoid composition of different Morus extracts was determined by the HPLC method. The extracts prepared from fruits, leaves and roots of M. alba and M. nigra exhibited different characteristics. The highest extraction yield was achieved by M. alba leaves extraction (23.40%). M. nigra roots extract shown the highest total phenolics (186.30 mg CAE/g), while highest total flavonoids content (67.37 mg RE/g) was determined for M. nigra leaves extracts. In addition, black mulberry leaves extracts with the highest antioxidant activity had the highest phenolic acids contents. The dominant phenolic components in the samples were rutin and chlorogenic acid. All investigated mulberry dry extracts shown high content of phenolic compounds and significant antioxidant activity. This work contributes to knowledge of the antioxidant properties of Morus species. The obtained results may be useful in the evaluation of new dietary supplements and food products.