Phylogenomics of mulberries (Morus, Moraceae) inferred from plastomes and single copy nuclear genes.

Mulberries (genus Morus), belonging to the order Rosales, family Moraceae, are important woody plants due to their economic values in sericulture, as well as for nutritional benefits and medicinal values. However, the taxonomy and phylogeny of Morus, especially for the Asian species, remains challenging due to its wide geographical distribution, morphological plasticity, and interspecific hybridization. To better understand the evolutionary history of Morus, we combined plastomes and a large-scale nuclear gene analyses to investigate their phylogenetic relationships. We assembled the plastomes and screened 211 single-copy nuclear genes from 13 Morus species and related taxa. The plastomes of Morus species were relatively conserved in terms of genome size, gene content, synteny, IR boundary and codon usage. Using nuclear data, our results elucidated identical topologies based on coalescent and concatenation methods. The genus Morus was supported as monophyletic, with M. notabilis as the first diverging lineage and the two North American Morus species, M. microphylla and M. rubra, as sister to the other Asian species. In the Asian Morus species, interspecific relationships were completely resolved. However, cyto-nuclear discordances and gene tree-species tree conflicts were detected in the phylogenies of Morus, with multiple evidences supporting hybridization/introgression as the main cause of discordances between nuclear and plastid phylogenies, while gene tree-species tree conflicts were mainly caused by ILS.

Chemometric evaluation of functional components and anti-quorum sensing activity of mulberry leaves from Indian cultivars: a potential contribution to the food industry.

BACKGROUND: Potential use of many native, easily available vegetal materials for human consumption and value addition is not well recognized. Mulberry, being a traditional industrial crop rich in nutrients and nutraceuticals can be of great importance for the food industry. However, mulberry leaves are mainly being utilized in sericulture and are not exploited for their functional components. Thus, the selection of promising mulberry cultivars, rich in bioactive compounds, like resveratrol and 1-deoxynojirimycin, increase their potential use in functional foods. RESULTS: Chlorogenic acid, myricetin and kaempferol were the major polyphenols present in the nine selected cultivars, in the range 0.001-0.086, 0.003-0.079 and 0.003-0.163 g kg-1 fresh weight (FW), respectively. Protocatechuic acid, epicatechin and rutin were predominantly present in cultivars V-1, G-2 and ML (0.103, 0.080 and 0.121 g kg-1 FW, respectively). Similarly, resveratrol and 1-deoxynojirimycin were highest in cultivars ML and K-2 (0.078 and 0.079 g kg-1 FW, respectively). Leaf extracts of cultivars G-2 and ML were able to effectively inhibit the violacein production with 64.08% and 70.04%, respectively at the concentration of 6 mg mL-1 presumably due to a higher content of polyphenols. Chemometric evaluation of chromatographic data showed the intraspecific variability and secondary metabolite co-existence in different cultivars. CONCLUSIONS: Considering phytoconstituents, cultivars G-2, ML, K-2 and V-1 could contribute efficiently to the rational utilization of mulberry in agro-food industries. Furthermore, cultivars G-2 and ML leaves can be a new source of quorum sensing inhibitory agents. © 2021 Society of Chemical Industry.

Application of economic plant for remediation of cadmium contaminated soils: Three mulberry (Moms alba L.) varieties cultivated in two polluted fields.

In order to study the role of mulberry (Moms alba L) as an economic crop for remediation of cadmium (Cd) contaminated soil, the transport of Cd from mulberry to silkworm were investigated. Three varieties of mulberry (Yuesang-11, Nongsang-14, and Qiangsang-1) with three planting densities were cultivated in two heavy metal-contaminated fields named Dongkou in Shaoyang city and Linxiang in Yueyang city in Hunan province respectively. The both field soils were contaminated by heavy metals, especially by Cd. The potential risks of heavy metals in Linxiang's soil were higher than those in Dongkou's because of higher concentrations of Cd. Since the promotion of Cd concentrations in aerial parts (stem, branch and leaf) resulted from the increase of planting density, the method of high planting density is beneficial to improve the efficiency of the remediation of Cd contaminated soil. The percentages of average Cd contents of mulberry in Dongkou accounted for 44%, 20%, 18% and 16% in roots, stems, branches and leaves respectively, while the Cd contents were 38%, 27%, 19% and 16% distributed in roots, stems, branches and leaves respectively. Mulberry leaves from contaminated soils was applied in food source of silkworms in this study. Although there is Cd uptake occurred in silkworm growth and its products (cocoons and chrysalis), Cd contents in cocoons are lower than the national standard (100 µg*kg-1) for textile industry of China. Therefore, mulberry can be regarded as an economical crop to control soil contamination with Cd.

Comparison of the protective effect of black and white mulberry against ethyl carbamate-induced cytotoxicity and oxidative damage.

Increasing evidence indicates that crude extracts derived from mulberry confer protection against oxidative stress. However, the antioxidant capacity of mulberry extract among different cultivars remains elusive. The main objective of the present study was therefore to investigate the effect of black mulberry extract (BMB) and white mulberry extract (WMB) on ethyl carbamate (EC)-induced cytotoxicity and oxidative stress. This study showed that the contents of total phenolics, total flavonoids, total procyanidins, cyanidin-3-O-glucoside, cyanidin-3-O-rutinoside and pelargonidin-3-O-glucoside of BMB were higher than that of WMB. Moreover, our results showed that phenolics-abundant BMB was stronger than WMB in scavenging ABTS and DPPH free radicals. BMB was more effective in ameliorating EC-induced cytotoxicity by inhibiting excessive ROS generation, suppressing mitochondrial dysfunction and increasing GSH concentration in HepG2 cells than WMB. Taken together, our study revealed that BMB afforded better protection against EC-caused cytotoxicity than WMB.

[Identification of origin plant of Uygur medicine mulberry based on DNA barcode].

To provide molecular evidence for medical material identification, we analyzed the nucleotide sequence of ITS2, psbA-trnH gene in Morus genus plants and commercial products which were obtained from different places in Xinjiang. The sequence of ITS2 and psbA-trnH in fifty-one samples were amplified and sequenced, MEGA 6.0 was used to analyze the intra- and interspecific K-2P distances, neighbor-joining (NJ) tree was used to constructing clustering tree. ITS2 sequence analyzed results showed that there is no intra-specific variation among Morus alba, M. alba var. tatarica and M. nigra, but 13 variations sites were exist between M. alba and M. nigra and their inter-specific K-2P distances was 0.04, which indicated that there had significant variation in them. We didn't find informative variation sites between Morus genus plants and commercial products, and we also found that M. nigra can be distinguished from other two species by NJ Tree. PsbA-trnH analysis results showed there was only one variation site between M. alba and M. nigra, but insertion or deletion variation were remarkable evidence among M. alba, M. alba var. tatarica and M. Nigra. Inter-specific variation was accordance with intra-specific variation of commercial products. So ITS2 and psbA-trnH gene were important marker for M. alba, M. alba var. tatarica and M. nigra identification. This study provided important evidence for Uygur medicine identification and market supervision.

Comparison of Different Strains of Agrobacterium rhizogenes for Hairy Root Induction and Betulin and Betulinic Acid Production in Morus alba.

Three Agrobacterium rhizogenes strains were tested for their ability to transform the plant Mois alba L. and to induce production of the secondary metabolites betulin and betulinic acid. All the tested strains of A. rhizogenes (R1601, LBA9402 and R1000) were able to induce hairy root formation in leaf tissue explants. Strain LBA9402 had the highest rate of infection (92.7% ± 8.8%), whereas strain R1601 had the lowest rate (87.4% ± 9.3%). The highest number of hairy roots per explant (5.6 ±0.5) and the greatest root length (2.4 ± 0.2 mm) were obtained with strain LBA9402. We also evaluated dry weight (a measure of growth) and betulin and betulinic acid production in hairy roots and found that the highest growth (167.8 ± 14.5 mg/flask) occurred after infection with strain LBA9402. Furthermore the highest production of betulin (5.4 ± 0.4 mg/g dry weight) and betulinic acid (2.3 ± 0.2 mg/g dry weight) was noted using strain LBA9402. Among three elicitors, yeast extract showed the highest induction of betulin production (8.7 ± 0.4 mg/g) and silver nitrate induced the highest yield of betulinic acid (4.1 ± 0.2 mg/g). Our study showed that A. rhizogenes strain LBA9402 was the most effective of the three tested strains for production-of transformed root cultures and betulin and betulinic acid.

Evolutionary and functional analysis of mulberry type III polyketide synthases.

BACKGROUND: Type III polyketide synthases are important for the biosynthesis of flavonoids and various plant polyphenols. Mulberry plants have abundant polyphenols, but very little is known about the mulberry type III polyketide synthase genes. An analysis of these genes may provide new targets for genetic improvement to increase relevant secondary metabolites and enhance the plant tolerance to biotic and abiotic stresses. RESULTS: Eighteen genes encoding type III polyketide synthases were identified, including six chalcone synthases (CHS), ten stilbene synthases (STS), and two polyketide synthases (PKS). Functional characterization of four genes representing most of the MnCHS and MnSTS genes by coexpression with 4-Coumaroyl-CoA ligase in Escherichia coli indicated that their products were able to catalyze p-coumaroyl-CoA and malonyl-CoA to generate naringenin and resveratrol, respectively. Microsynteny analysis within mulberry indicated that segmental and tandem duplication events contributed to the expansion of the MnCHS family, while tandem duplications were mainly responsible for the generation of the MnSTS genes. Combining the evolution and expression analysis results of the mulberry type III PKS genes indicated that MnCHS and MnSTS genes evolved mainly under purifying selection to maintain their original functions, but transcriptional subfunctionalization occurred during long-term species evolution. Moreover, mulberry leaves can rapidly accumulated oxyresveratrol after UV-C irradiation, suggesting that resveratrol was converted to oxyresveratrol. CONCLUSIONS: Characterizing the functions and evolution of mulberry type III PKS genes is crucial for advancing our understanding of these genes and providing the basis for further studies on the biosynthesis of relevant secondary metabolites in mulberry plants.

Molecular cloning and expression analyses of RPS3a gene from mulberry under abiotic stresses and among different mulberry varieties.

A full-length cDNA sequence coding ribosomal protein S3a of mulberry tree, which we designated MmRPS3a (GenBank accession No. KR610331), was cloned based on mulberry expressed sequence tags. Sequence analysis showed that the MmRPS3a is 1089 bp long and contains a 80-bp 5'-UTR (untranslated region) and a 220-bp 3'-UTR. Its open reading frame consists of a 789-bp encoding 262 amino acids with a predicted molecular weight of 30.053 kDa and an isoelectric point of 9.84. Homology analysis revealed that MmRPS3a gene is highly conservative in mulberry and other species including Morus notabilis, Theobroma cacao, and Ricinus communis. Phylogenetic analysis based on MmRPS3a of other species showed that mulberry had a closer relationship with Prunus persica, Arabidopsis thaliana, Solanum tuberosum, Solanum lycopersicum, and Vitis vinifera. The results of quantitative PCR analysis showed that the transcriptional level of MmRPS3a mRNA changed significantly under the conditions of hypothermia, aridity, salt stress, and varieties of differing resistances.

Effect of Cultivar, Temperature, and Environmental Conditions on the Dynamic Change of Melatonin in Mulberry Fruit Development and Wine Fermentation.

High levels of melatonin have been reported in various foods but not in mulberry or its wine. This study investigated the dynamic changes of melatonin levels during mulberry fruit development and ethanol fermentation of 2 different colored mulberry cultivars ("Hongguo2Ë® Morus nigra, black and "BaiyuwangË® Morus alba, white) at 2 fermentation temperatures (16 and 25 °C). Our results showed that the melatonin level increased in the beginning of mulberry development but decreased in the end. The MnTDC gene expression level correlated with melatonin production, which implied that TDC may be the rate-limiting enzyme of the melatonin biosynthetic process in mulberries. During mulberry fermentation, the melatonin concentration increased rapidly in the beginning and then decreased gradually. Low temperature delayed the melatonin production during fermentation. A relatively high level of melatonin was found in "Hongguo2Ë® compared with "BaiyuwangË® during fruit development and fermentation. The variation of melatonin correlated with the ethanol production rate, suggesting that melatonin may participate in physiological regulation of Saccharomyces cerevisiae during the fermentation stage.

An Approach to Function Annotation for Proteins of Unknown Function (PUFs) in the Transcriptome of Indian Mulberry.

The modern sequencing technologies are generating large volumes of information at the transcriptome and genome level. Translation of this information into a biological meaning is far behind the race due to which a significant portion of proteins discovered remain as proteins of unknown function (PUFs). Attempts to uncover the functional significance of PUFs are limited due to lack of easy and high throughput functional annotation tools. Here, we report an approach to assign putative functions to PUFs, identified in the transcriptome of mulberry, a perennial tree commonly cultivated as host of silkworm. We utilized the mulberry PUFs generated from leaf tissues exposed to drought stress at whole plant level. A sequence and structure based computational analysis predicted the probable function of the PUFs. For rapid and easy annotation of PUFs, we developed an automated pipeline by integrating diverse bioinformatics tools, designated as PUFs Annotation Server (PUFAS), which also provides a web service API (Application Programming Interface) for a large-scale analysis up to a genome. The expression analysis of three selected PUFs annotated by the pipeline revealed abiotic stress responsiveness of the genes, and hence their potential role in stress acclimation pathways. The automated pipeline developed here could be extended to assign functions to PUFs from any organism in general. PUFAS web server is available at http://caps.ncbs.res.in/pufas/ and the web service is accessible at http://capservices.ncbs.res.in/help/pufas.

The complete chloroplast genome sequence of the mulberry Morus notabilis (Moreae).

The complete chloroplast genome of the mulberry Morus notabilis (Moreae) has been reconstructed from the whole-genome Illumina sequencing data. The circular genome is 158,680 bp in size, and comprises a pair of inverted repeat (IR) regions of 25,717 bp each, a large single-copy (LSC) region of 87,470 bp and a small single-copy (SSC) region of 19,776 bp. The total A+T content is 63.6%, while the corresponding values of the LSC, SSC and IR region are 65.9%, 70.7% and 57.1%, respectively. The chloroplast genome contains 129 genes, including 84 protein-coding genes (PCGs), eight ribosomal RNA (rRNA) genes and 37 transfer RNA (tRNA) genes. The maximum likelihood (ML) phylogenetic analysis revealed that M. notabilis was more related to its congeners than to the others.

The complete chloroplast genome sequence of Morus mongolica and a comparative analysis within the Fabidae clade.

The complete nucleotide sequence of the Morus mongolica chloroplast (cp) genome was reported and characterized in this study. The cp genome is a circular molecule of 158,459 bp containing a pair of 25,678 bp IR regions, separated by small and large single-copy regions of 19,736 and 87,363 bp, respectively. The number and relative positions of the 114 unique genes (80 PCGs, 30 tRNAs, and 4 rRNA genes) are almost identical to Morus indica cp genome. Further detailed comparative analyses revealed one hypervariable region, which is responsible for 88% of the total variation, and 64 indel events between two individuals. There are 78 simple sequence repeats (SSRs) in M. mongolica cp genome, in which 58 of them are mononucleotide repeats. Comparative analysis with M. indica cp genome indicated 22 SSRs with length polymorphisms and 1 SSR with nucleotide content polymorphism. The phylogenetic analysis of 60 PCGs from 62 cp genomes provided strong support for the monophyletic, single origin of Fabidae (N2-fixing) clade.

Comparison of Polyphenol Profile and Inhibitory Activities Against Oxidation and α-Glucosidase in Mulberry (Genus Morus) Cultivars from China.

UNLABELLED: Mulberry (genus Morus) is a significant source of polyphenols, which can promote positive effects on human health. China has various mulberry cultivars, however, many Chinese mulberry cultivars have been only minimally studied. To solve this lack of research, 8 mulberry cultivars (Da10, Tang10, Yueshen74, Yuefenshen, Longsang, Ningxia1hao, Taiwanguosang, and Baiyuwang) from 4 regions of China were assessed to determine their polyphenol profiles using HPLC-MS/MS and then tested for their antioxidant and anti-α-glucosidase activities in vitro. A total of 18 nonanthocyanins and 4 anthocyanins were quantified in mulberry cultivars; among these polyphenols, chlorogenic acid, quercetin 3-O-rutinoside, and cyanidin 3-O-glucoside were confirmed as the major phenolic acid, flavonol derivative, and anthocyanin, respectively. Two types of stilbene compounds, piceid, and piceatannol, were detected for the 1st time in all mulberry cultivars. Moreover, the methanolic extracts of different mulberry cultivars showed disparate antioxidant and α-glucosidase inhibitory activities, and this discrepancy was mainly attributed to varying the anthocyanin content. Based on our results, Taiwanguosang is proposed to be a good candidate suitable for further process due to its high level of anthocyanins. PRACTICAL APPLICATION: The polyphenols of mulberry cultivars are vital for human health and are relevant to the further development of mulberry-based products. China has a wide range of mulberry cultivar resources, and many of these cultivars have not yet been studied. Our research concentrated on the polyphenol profiles, antioxidant, and α-glucosidase inhibitory activities of various mulberry cultivars from different regions of China to provide basic information for mulberry cultivar selection and mulberry-based food production.

A holistic picture of Austronesian migrations revealed by phylogeography of Pacific paper mulberry.

The peopling of Remote Oceanic islands by Austronesian speakers is a fascinating and yet contentious part of human prehistory. Linguistic, archaeological, and genetic studies have shown the complex nature of the process in which different components that helped to shape Lapita culture in Near Oceania each have their own unique history. Important evidence points to Taiwan as an Austronesian ancestral homeland with a more distant origin in South China, whereas alternative models favor South China to North Vietnam or a Southeast Asian origin. We test these propositions by studying phylogeography of paper mulberry, a common East Asian tree species introduced and clonally propagated since prehistoric times across the Pacific for making barkcloth, a practical and symbolic component of Austronesian cultures. Using the hypervariable chloroplast ndhF-rpl32 sequences of 604 samples collected from East Asia, Southeast Asia, and Oceanic islands (including 19 historical herbarium specimens from Near and Remote Oceania), 48 haplotypes are detected and haplotype cp-17 is predominant in both Near and Remote Oceania. Because cp-17 has an unambiguous Taiwanese origin and cp-17-carrying Oceanic paper mulberries are clonally propagated, our data concur with expectations of Taiwan as the Austronesian homeland, providing circumstantial support for the "out of Taiwan" hypothesis. Our data also provide insights into the dispersal of paper mulberry from South China "into North Taiwan," the "out of South China-Indochina" expansion to New Guinea, and the geographic origins of post-European introductions of paper mulberry into Oceania.

Definition of Eight Mulberry Species in the Genus Morus by Internal Transcribed Spacer-Based Phylogeny.

Mulberry, belonging to the order Rosales, family Moraceae, and genus Morus, has received attention because of both its economic and medicinal value, as well as for its important ecological function. The genus Morus has a worldwide distribution, however, its taxonomy remains complex and disputed. Many studies have attempted to classify Morus species, resulting in varied numbers of designated Morus spp. To address this issue, we used information from internal transcribed spacer (ITS) genetic sequences to study the taxonomy of all the members of generally accepted genus Morus. We found that intraspecific 5.8S rRNA sequences were identical but that interspecific 5.8S sequences were diverse. M. alba and M. notabilis showed the shortest (215 bp) and the longest (233 bp) ITS1 sequence length, respectively. With the completion of the mulberry genome, we could identify single nucleotide polymorphisms within the ITS locus in the M. notabilis genome. From reconstruction of a phylogenetic tree based on the complete ITS data, we propose that the Morus genus should be classified into eight species, including M. alba, M. nigra, M. notabilis, M. serrata, M. celtidifolia, M. insignis, M. rubra, and M. mesozygia. Furthermore, the classification of the ITS sequences of known interspecific hybrid clones into both paternal and maternal clades indicated that ITS variation was sufficient to distinguish interspecific hybrids in the genus Morus.

A novel sucrose hydrolase from the bombycoid silkworms Bombyx mori, Trilocha varians, and Samia cynthia ricini with a substrate specificity for sucrose.

Although membrane-associated sucrase activity has been detected in the midgut of various lepidopteran species, it has not yet been identified and characterized at the molecular level. In the present study, we identified a novel sucrose hydrolase (SUH) gene from the following three bombycoid silkworms: Bombyx mori, Trilocha varians, and Samia cynthia ricini and named them BmSuh, TvSuh, and ScSuh, respectively. The EST dataset showed that BmSuh is one of the major glycoside hydrolase genes in the larval midgut of B. mori. These genes were almost exclusively expressed in the larval midgut in all three species, mainly at the feeding stage. SUHs are classified into the glycoside hydrolase family 13 and show significant homology to insect maltases. Enzymatic assays revealed that recombinant SUHs were distinct from conventional maltases and exhibited substrate specificity for sucrose. The recombinant BmSUH was less sensitive to sugar-mimic alkaloids than TvSUH and ScSUH, which may explain the reason why the sucrase activity in the B. mori midgut was less affected by the sugar-mimic alkaloids derived from mulberry.

Large-scale intersubspecific recombination in the plant-pathogenic bacterium Xylella fastidiosa is associated with the host shift to mulberry.

Homologous recombination plays an important role in the structuring of genetic variation of many bacteria; however, its importance in adaptive evolution is not well established. We investigated the association of intersubspecific homologous recombination (IHR) with the shift to a novel host (mulberry) by the plant-pathogenic bacterium Xylella fastidiosa. Mulberry leaf scorch was identified about 25 years ago in native red mulberry in the eastern United States and has spread to introduced white mulberry in California. Comparing a sequence of 8 genes (4,706 bp) from 21 mulberry-type isolates to published data (352 isolates representing all subspecies), we confirmed previous indications that the mulberry isolates define a group distinct from the 4 subspecies, and we propose naming the taxon X. fastidiosa subsp. morus. The ancestry of its gene sequences was mixed, with 4 derived from X. fastidiosa subsp. fastidiosa (introduced from Central America), 3 from X. fastidiosa subsp. multiplex (considered native to the United States), and 1 chimeric, demonstrating that this group originated by large-scale IHR. The very low within-type genetic variation (0.08% site polymorphism), plus the apparent inability of native X. fastidiosa subsp. multiplex to infect mulberry, suggests that this host shift was achieved after strong selection acted on genetic variants created by IHR. Sequence data indicate that a single ancestral IHR event gave rise not only to X. fastidiosa subsp. morus but also to the X. fastidiosa subsp. multiplex recombinant group which infects several hosts but is the only type naturally infecting blueberry, thus implicating this IHR in the invasion of at least two novel native hosts, mulberry and blueberry.

Protective effect of mulberry flavonoids on sciatic nerve in alloxan-induced diabetic rats

Mulberry leaves (Morus alba L.) are a traditional Chinese medicine for blood serum glucose reduction. This study evaluated the protective effects of mulberry flavonoids on sciatic nerve in alloxan-induced diabetic rats. In this study, 80 Sprague-Dawley rats were divided into five groups: A (control), B (diabetic treated with saline), C-D (diabetic treated with 0.3, 0.1 g/kg mulberry flavonoids once a day for 8 weeks) and E (diabetic treated with 0.3 mg/kg methycobal). The diabetic condition was induced by intraperitoneal injection of 200 mg/kg alloxan dissolved in saline. At the end of the experimental period, blood, and tissue samples were obtained for biochemical and histopathological investigation. Treatment with 0.3 g/kg mulberry flavonoids significantly inhibited the elevated serum glucose (P< 0.01). The increased myelin sheath area (P< 0.01), myelinated fiber cross-sectional area and extramedullary fiber number (P< 0.05) were also reduced in alloxan-induced rats treated with 0.3 g/kg mulberry flavonoids. 0.3 g/kg mulberry flavonoids also markedly decreased onion-bulb type myelin destruction and degenerative changes of mitochondria and Schwann cells. These findings demonstrate that mulberry flavonoids may improve the recovery of a severe peripheral nerve injury in alloxan-induced diabetic rats and is likely to be useful as a potential treatment on peripheral neuropathy (PN) in diabetic rats.

Folhas de amoreira (Morus alba L.) é um medicamento tradicional chinês para a redução da glicose no soro sanguíneo. Avaliaram-se, neste trabalho, os efeitos protetores dos flavonóides de amora no nervo ciático em ratos diabéticos aloxano-induzidos. Dividiram-se 80 ratos Sprague-Dawley em cinco grupos: A (controle), B (diabétidos tratados com solução salina), C-D (diabéticos tratados com 0,3, 0,1 g/kg) e E (diabéticos tratados com 0,3 mg de metilcobal).A diabetes foi induzida por injeção intraperitoneal de 200 mg/kg de aloxana dissolvida em solução salina. No final do período experimental, obtiveram-se amostras de sangue e de tecido para investigação bioquímica e histopatológica. O tratamento com 0,3 g/kg de flavonóides da amoreira inibiu, significativamente, a elevação de glicose no soro (p <0,01). O aumento da área da bainha de mielina (p <0,01), da área de fibra da seção transversal e do número de fibras mielinizadas extramedulares (p <0,05) foi também reduzido em ratos aloxânicos, tratados com 0,3 g/kg flavonóides de amora. Flavonóides da amoreira na dose de 0,3 g/kg também diminuiram, acentuadamente, a destruição da mielina do tipo bulbo de cebola e as alterações degenerativas das células mitocôndrias e das células de Schwann. Estes resultados demonstram que os flavonóides da amoreira podem melhorar a recuperação de uma lesão nervosa periférica grave em ratos com diabetes, induzida por aloxana, e parece ser útil como tratamento potencial para a neuropatia periférica (PN) em ratos diabéticos.

Development and characterization of microsatellite markers for Morus spp. and assessment of their transferability to other closely related species.

BACKGROUND: Adoption of genomics based breeding has emerged as a promising approach for achieving comprehensive crop improvement. Such an approach is more relevant in the case of perennial species like mulberry. However, unavailability of genomic resources of co-dominant marker systems has been the major constraint for adopting molecular breeding to achieve genetic enhancement of Mulberry. The goal of this study was to develop and characterize a large number of locus specific genic and genomic SSR markers which can be effectively used for molecular characterization of mulberry species/genotypes. RESULT: We analyzed a total of 3485 DNA sequences including genomic and expressed sequences (ESTs) of mulberry (Morus alba L.) genome. We identified 358 sequences to develop appropriate microsatellite primer pairs representing 222 genomic and 136 EST regions. Primers amplifying locus specific regions of Dudia white (a genotype of Morus alba L), were identified and 137 genomic and 51 genic SSR markers were standardized. A two pronged strategy was adopted to assess the applicability of these SSR markers using mulberry species and genotypes along with a few closely related species belonging to the family Moraceae viz., Ficus, Fig and Jackfruit. While 100% of these markers amplified specific loci on the mulberry genome, 79% were transferable to other related species indicating the robustness of these markers and the potential they hold in analyzing the molecular and genetic diversity among mulberry germplasm as well as other related species. The inherent ability of these markers in detecting heterozygosity combined with a high average polymorphic information content (PIC) of 0.559 ranging between 0.076 and 0.943 clearly demonstrates their potential as genomic resources in diversity analysis. The dissimilarity coefficient determined based on Neighbor joining method, revealed that the markers were successful in segregating the mulberry species, genotypes and other related species into distinct clusters. CONCLUSION: We report a total of 188 genomic and genic SSR markers in Morus alba L. A large proportion of these markers (164) were polymorphic both among mulberry species and genotypes. A substantial number of these markers (149) were also transferable to other related species like Ficus, Fig and Jackfruit. The extent of polymorphism revealed and the ability to detect heterozygosity among the cross pollinated mulberry species and genotypes render these markers an invaluable genomic resource that can be utilized in assessing molecular diversity as well as in QTL mapping and subsequently mulberry crop improvement through MAS.

Comparison of inhibitory activities and mechanisms of five mulberry plant bioactive components against α-glucosidase.

The α-glucosidase inhibitory effects of five bioactive components, namely 1-deoxynojirimycin, cyanidin-3-glucoside, cyanidin-3-rutinoside, resveratrol and oxyresveratrol contained in mulberry (Morus, Moraceae) plants have been compared. Spectroscopy methods were employed to compare their α-glucosidase inhibitory mechanisms. The results revealed that 1-deoxynojirimycin (competitive), resveratrol and oxyresveratrol (noncompetitive) were stronger inhibitors than acarbose, while cyanidin-3-glucoside and cyanidin-3-rutinoside (mix competitive and noncompetitive) showed modest activities. 1-Deoxynojirimycin, resveratrol and oxyresveratrol could quench the fluorescence spectra statically by forming stable complexes, while the quenching of cyanidin-3-rutinoside and cyanidin-3-glucoside belonged to dynamic quenching by the collision of molecules. The interactions between ligands and α-glucosidase were mainly driven by hydrophobic force, or hydrogen bonding consequently induced conformational changes and reduced surface hydrophobicity. Docking results suggested that they could bind to α-glucosidase at different sites. This work provides useful information for the understanding of the ligands-α-glucosidase interactions and identifies oxyresveratrol as a potent α-glucosidase inhibitor.