Project description:To investigate effects of intake of mulberry leaf extracts on hypercholesterolemia, we performed gene expression profiling on rat liver by microarray analysis. Microarray analysis revealed that mulberry leaf extracts up-regulated the gene expression involved in suppression of cholesterol synthesis and stimulation of innate-adaptive Immunity. Mice were fed a high-cholesterol diet without/with orally administration of mulberry leaf extracts for 4 weeks. Livers were taken for RNA extraction and hybridization on Agilent microarrays.
Project description:MicroRNAs (miRNAs) are endogenous non-coding small RNAs containing 21-24 nucleotides, and play an important function in the course of stand up to adversity. Identification of miRNA target mRNAs is essential for their functional analysis. In order to anatomize miRNA guided gene regulation under drought stress, we performed a transcriptome-wide experimental method using high throughput degradome sequencing to directly detect drought stress responsive miRNA targets in mulberry. In this study, drought library (DL) and contrast library (CL) which captured the cleaved mRNA were constructed from mulberry for sequencing by Illumina sequencer, and 409, 373 target genes for 30 conserved miRNA families and 990, 950 target genes for 199, 195 novel miRNAs were identified in CL and DL, respectively. Among those conserved miRNA families in DL, mno-miR156, mno-miR172 and mno-miR396 refer to the highest number of targets, indicating that mno-miR156, mno-miR172 and mno-miR396 families and their target genes might play a key important function in corresponding to drought stress in mulberry. By construction and analysis of miRNA-target network, we found that the DL independent networks consist of 838 miRNA-mRNA pairs (63.34%). Besides, the expression patterns of 11 target genes and 12 correspondent miRNAs were compared and analyzed by qRT-PCR. In addition, 5 of 6 miRNA targets were further verified by RNA ligase-mediated 5’ rapid amplification of cDNA ends (RLM-5’ RACE). Gene ontology (GO) annotations and Kyoto Encyclopaedia of Genes and Genomes (KEGG) pathway analysis showed that these target transcripts were implicated in a broad range of biological processes and various metabolic pathways. The characterization of target genes and the associated miRNAs in response to drought exposure provides a framework for understanding the molecular mechanism of resistance to drought in plants.
Project description:To investigate effects of intake of mulberry leaf extracts on hypercholesterolemia, we performed gene expression profiling on rat liver by microarray analysis. Microarray analysis revealed that mulberry leaf extracts up-regulated the gene expression involved in suppression of cholesterol synthesis and stimulation of innate-adaptive Immunity.
Project description:Mulberry (Morus alba L.) is considered a millenary medicinal plant and a food source for silkworms. Different M. alba extracts offer a variety of biological and pharmacological properties that are largely attributed to stilbenoids, a small group of phenylpropanoids, including resveratrol and oxyresveratrol. These are naturally present in non-renewable parts of mulberry trees, impeding their efficient extraction. As a way to bypass this spatiotemporal restriction, we generated cell suspensions from mulberry leaf petioles and demonstrated that the combined use of methyl jasmonate and two different types of cyclodextrins were able to elicit a high production of resveratrol and oxyresveratrol. As oxyresveratrol-producing enzymes are still unknown, we improved the structural and functional annotation of the mulberry genome by integrating short and long-read sequencing data and combined it with time series transcriptome, metabolite and proteome data in response to cell elicitation to identify a complete set of phenylpropanoid and stilbenoid related genes. These included 23 stilbene synthase (STS) genes and a group of six p-coumaroyl-CoA 2'-hydroxylases (C2’Hs), all being highly co-expressed with resveratrol and oxyresveratrol production. We transiently transformed grapevine (Vitis vinifera L.) calli and Nicotiana benthamiana plants to functionally validate the role of C2’Hs in the first committed step of oxyresveratrol synthesis, which provides an alternative substrate for STSs by hydroxylating p-coumaroyl-coA into 2’4’-dihydroxycinnamoyl-CoA. Finally, we offer tools for genomic and transcriptomic exploration also in the context of jasmonate elicitation aiding in the characterization of novel stilbenoid-modifying and regulatory genes in the Morus genus.
Project description:Autotetraploid carries several phenotypic changes with larger leaves and fruit compared to diploid. To analysis of phenotypic changes in mulberry autotetraploids on the transcriptome, we performed RNA-Seq analyses on mulberry leaf samples of diploid and autotetraploids using Illumina HiSEq 2000.