De novo Sequencing and Analysis of Salvia hispanica Tissue-Specific Transcriptome and Identification of Genes Involved in Terpenoid Biosynthesis.
ABSTRACT: Salvia hispanica (commonly known as chia) is gaining popularity worldwide as a healthy food supplement due to its low saturated fatty acid and high polyunsaturated fatty acid content, in addition to being rich in protein, fiber, and antioxidants. Chia leaves contain plethora of secondary metabolites with medicinal properties. In this study, we sequenced chia leaf and root transcriptomes using the Illumina platform. The short reads were assembled into contigs using the Trinity software and annotated against the Uniprot database. The reads were de novo assembled into 103,367 contigs, which represented 92.8% transcriptome completeness and a diverse set of Gene Ontology terms. Differential expression analysis identified 6151 and 8116 contigs significantly upregulated in the leaf and root tissues, respectively. In addition, we identified 30 contigs belonging to the Terpene synthase (TPS) family and demonstrated their evolutionary relationships to tomato TPS family members. Finally, we characterized the expression of S. hispanica TPS members in leaves subjected to abiotic stresses and hormone treatments. Abscisic acid had the most pronounced effect on the expression of the TPS genes tested in this study. Our work provides valuable community resources for future studies aimed at improving and utilizing the beneficial constituents of this emerging healthy food source.
Project description:Salvia hispanica L. (chia) is a member of the mint family that is cultivated for its seeds. The majority of seed content in chia is comprised of omega fatty acids. Furthermore, chia seeds are also rich in fiber and minerals. The human health potential of chia seeds have driven studies of dietary effects, however there is little genetic or genomic studies available. In this study we obtained RNA from seeds, shoots, cotyledons, leaf primordia, nodes, racemes, and flower tissues from different developmental stages to generate an expression atlas for chia. RNA was sequenced on an Illumina Hiseq 2500. Sequence reads were assembled de novo to produce transcripts. Sequence reads were aligned to the chia transcriptome assembly to generate counts for each tissue type. Differentially expressed transcripts were determined for each tissue type.
Project description:Chia (Salvia hispanica L.) is a small seed that comes from an annual herbaceous plant, Salvia hispanica L. In recent years, usage of Chia seeds has tremendously grown due to their high nutritional and medicinal values. Chia was cultivated by Mesopotamian cultures, but then disappeared for centuries until the middle of the 20th century, when it was rediscovered. Chia seeds contain healthy ?-3 fatty acids, polyunsaturated fatty acids, dietary fiber, proteins, vitamins, and some minerals. Besides this, the seeds are an excellent source of polyphenols and antioxidants, such as caffeic acid, rosmarinic acid, myricetin, quercetin, and others. Today, chia has been analyzed in different areas of research. Researches around the world have been investigating the benefits of chia seeds in the medicinal, pharmaceutical, and food industry. Chia oil is today one of the most valuable oils on the market. Different extraction methods have been used to produce the oil. In the present study, an extensive overview of the chemical composition, nutritional properties, and antioxidant and antimicrobial activities, along with extraction methods used to produce chia oil, will be discussed.
Project description:Chia (Salvia hispanica L.) is an annual short-day plant whose growth has not been studied extensively in low-altitudes and at temperatures outside of its optimal range. The objective of this study was to describe the growth dynamics of a chia crop from an experimental plantation in south-east Mexico, on three different sowing dates. The chia grew at temperatures (18-37°C) and an altitude (9 m a.s.l.) outside of the recommended conditions (20-30°C, 500-1000 m a.s.l.). Three individual-plant responses were measured weekly, before seed harvest: height, number of leaves and number of inflorescences. Three theoretical nonlinear growth models were fitted to the data, a different model for each response. Mixed-effect model parameters were estimated by maximum likelihood, and the goodness of fit for each model was evaluated using two criteria: Modeling Efficiency and Root Mean Square Error. Chia seed yield was also measured in each treatment. Estimated parameters for plant height confirmed that medium sowing time (MST) and late sowing time (LST) plants had smaller heights than the early sowing time (EST) plants. Moreover, at the end of their life cycle, EST plants had a greater number of leaves and inflorescences, and higher seed yield. All of these differences were associated to the extended time of vegetative growth of EST plants favored by optimal photoperiod and temperature. Growth dynamics of chia during its ontogenic phases was explored, in more detail, with relative growth parameters derived from fitted models: a decrease in photoperiod influences the beginning of the reproductive phase, with the consequent reduction in speed of vegetative growth. In addition, nonlinear mixed-effects models can be useful in understanding the relation between growth parameters, plant maturity, and the suitable time for chia seed harvest. Our results suggest chia crops are adaptable to non-conventional environmental conditions.
Project description:Salvia hispanica (chia) constituted an important crop for pre-Columbian civilizations and is considered a superfood for its rich content of essential fatty acids and proteins. In this study, we performed the first comprehensive comparative transcriptome analysis between seeds from cultivated varieties and from accessions collected from native wild populations in Mexico. From the 69,873 annotated transcripts assembled de novo, enriched functional categories and pathways revealed that the lipid metabolism was one of the most activated processes. Expression changes were detected among wild and cultivated groups and among growth conditions in transcripts responsible for triacylglycerol and fatty acid synthesis and degradation. We also quantified storage protein fractions that revealed variation concerning nutraceutical proteins such as albumin and glutelin. Genetic diversity estimated with 23,641 single nucleotide polymorphisms (SNPs) revealed that most of the variation remains in the wild populations, and that a wild-type cultivated variety is genetically related to wild accessions. Additionally, we reported 202 simple sequence repeat (SSRs) markers useful for population genetic studies. Overall, we provided transcript variation that can be used for breeding programs to further develop chia varieties with enhanced nutraceutical traits and tools to explore the genetic diversity and history of this rediscovered plant.
Project description:Chia (Salvia hispanica L.), a member of the mint family (Lamiaceae), is a rediscovered crop with great importance in health and nutrition and is also the highest known terrestrial plant source of heart-healthy omega-3 fatty acid, alpha linolenic acid (ALA). At present, there is no public genomic information or database available for this crop, hindering research on its genetic improvement through genomics-assisted breeding programs. The first comprehensive analysis of the global transcriptome profile of developing Salvia hispanica L. seeds, with special reference to lipid biosynthesis is presented in this study. RNA from five different stages of seed development was extracted and sequenced separately using the Illumina GAIIx platform. De novo assembly of processed reads in the pooled transcriptome using Trinity yielded 76,014 transcripts. The total transcript length was 66,944,462 bases (66.9 Mb), with an average length of approximately 880 bases. In the molecular functions category of Gene Ontology (GO) terms, ATP binding and nucleotide binding were found to be the most abundant and in the biological processes category, the metabolic process and the regulation of transcription-DNA-dependent and oxidation-reduction process were abundant. From the EuKaryotic Orthologous Groups of proteins (KOG) classification, the major category was "Metabolism" (31.97%), of which the most prominent class was 'carbohydrate metabolism and transport' (5.81% of total KOG classifications) followed by 'secondary metabolite biosynthesis transport and catabolism' (5.34%) and 'lipid metabolism' (4.57%). A majority of the candidate genes involved in lipid biosynthesis and oil accumulation were identified. Furthermore, 5596 simple sequence repeats (SSRs) were identified. The transcriptome data was further validated through confirmative PCR and qRT-PCR for select lipid genes. Our study provides insight into the complex transcriptome and will contribute to further genome-wide research and understanding of chia. The identified novel UniGenes will facilitate gene discovery and creation of genomic resource for this crop.
Project description:Chia (Salvia hispanica), an herbaceous plant native to Latin America, has become important in the last 20 years due to its beneficial effects on health. Here, we present the first record and identification of two viruses in chia plants. The comparison of the complete nucleotide sequences showed the presence of two viral species with the typical genome organization of bipartite New World begomovirus, identified as Sida mosaic Bolivia virus 2 and Tomato yellow spot virus, according to the ICTV taxonomic criteria for begomovirus classification. DNA-A from Sida mosaic Bolivia virus 2 exhibited 96.1% nucleotide identity with a Bolivian isolate of Sida micrantha, and Tomato yellow spot virus showed 95.3% nucleotide identity with an Argentine bean isolate. This is the first report of begomoviruses infecting chia as well as of the occurrence of Sida mosaic Bolivia virus 2 in Argentina.
Project description:: p-Coumaric acid (p-CA) belongs to a family of natural esters of hydroxycinnamic acid compounds that have been shown to modulate plant growth and metabolism. In this study, we investigated the effect of exogenous p-CA on plant growth, reactive oxygen species (ROS)-induced oxidative damage, photosynthetic metabolism, osmolyte content and changes in superoxide dismutase (SOD) enzymatic activity. Exogenous p-CA improved Salvia hispanica (chia) growth by significantly enhancing shoot length, fresh and dry weights coupled with augmented levels of total chlorophyll and carotenoid contents. Furthermore, p-CA also triggered an induction in proline, glycine betaine (GB) and superoxide (O2?-) levels while no changes were observed for hydrogen peroxide (H2O2) and downstream malondialdehyde (MDA) content. Also, no change in SOD activity was observed in the p-CA treatment relative to the control. Therefore, the results suggest that exogenous p-CA improves chia seedling growth possibly via activation of a ROS-signalling pathway involving O2?- under the control of proline accumulation.
Project description:Omega-3 fatty acid desaturase (?-3 FAD, D15D) is a key enzyme for ?-linolenic acid (ALA) biosynthesis. Both chia (Salvia hispanica) and perilla (Perilla frutescens) contain high levels of ALA in seeds. In this study, the ?-3 FAD gene family was systematically and comparatively cloned from chia and perilla. Perilla FAD3, FAD7, FAD8 and chia FAD7 are encoded by single-copy (but heterozygous) genes, while chia FAD3 is encoded by 2 distinct genes. Only 1 chia FAD8 sequence was isolated. In these genes, there are 1 to 6 transcription start sites, 1 to 8 poly(A) tailing sites, and 7 introns. The 5'UTRs of PfFAD8a/b contain 1 to 2 purine-stretches and 2 pyrimidine-stretches. An alternative splice variant of ShFAD7a/b comprises a 5'UTR intron. Their encoded proteins harbor an FA_desaturase conserved domain together with 4 trans-membrane helices and 3 histidine boxes. Phylogenetic analysis validated their identity of dicot microsomal or plastidial ?-3 FAD proteins, and revealed some important evolutionary features of plant ?-3 FAD genes such as convergent evolution across different phylums, single-copy status in algae, and duplication events in certain taxa. The qRT-PCR assay showed that the ?-3 FAD genes of two species were expressed at different levels in various organs, and they also responded to multiple stress treatments. The functionality of the ShFAD3 and PfFAD3 enzymes was confirmed by yeast expression. The systemic molecular and functional features of the ?-3 FAD gene family from chia and perilla revealed in this study will facilitate their use in future studies on genetic improvement of ALA traits in oilseed crops.
Project description:The present study aimed to determine the role of Salvia hispanica L., (chia seed) fatty acid content in adipocyte lipid accumulation and human macrophage immunoregulatory potential. Chia seed fatty acid was extracted using hexane by the cold percolation method. A gas chromatography-mass spectrometry (GC-MS) analysis showed a 3:1 ratio of omega 3 and omega 6 fatty acid composition and it was more beneficial for human health. We treated it with increasing concentrations (0-6.4 ?g/mL) of chia seed fatty acid extract to determine the cytotoxicity on the preadipocytes and macrophage; no significant cytotoxicity was observed. Chia seed, in 0.2 and 0.4 ?g/mL doses, significantly arrested adipocyte hypertrophy and macrophage foam cell development. The gene expression levels of adipocyte confirmed the increased expression of adipocyte mitochondrial thermogenesis related genes, such as uncoupling protein-1 (UCP-1), peroxisome proliferator activated receptor gamma coactivator 1 alpha (PPAR?C1?) and PR domain containing 16 (PRDM16); and the down regulated expression of the lipid synthesis related gene sterol regulatory element binding of protein-1c (SREBP-1c). In addition, adipogenesis related genes, such as the proliferator activated receptor ? (PPAR?) and CCAAT/enhancer binding protein (C/EBP?) expressions, have been down regulated by chia seed treatment. Macrophage treated with chia seed-treated adipocyte condition media significantly inhibited the obesity associated inflammatory genes and protein expression levels, such as monocyte chemo attractant protein-1 (MCP-1), prostaglandins E2, interleukin-6, plasminogen activator inhibitor-1 (PAI-1) and tumor necrosis factor-? (TNF-?). In conclusion, a 3:1 ratio of omega 3 and omega 6 fatty acid composition of chia seed fatty acid content potentially inhibits lipid accumulation, and enhanced fatty acid oxidation, via UCP-1 and PRDM16 expression. Macrophage recruitment to adipocyte and the development of obesity associated inflammation was suppressed by chia seeds.
Project description:BACKGROUND:Recent studies show that galling Hymenoptera and Diptera are able to synthesize the plant hormone indole-3-acetic acid (auxin) from tryptophan and that plant response to insect-produced auxin is implicated in gall formation. We examined the leaf transcriptome of galled and ungalled leaves of individuals of the Hawaiian endemic plant Metrosideros polymorpha (Myrtaceae) subject to infestation by psyllid (Hemiptera) gall-makers in the genus Trioza (Triozidae). RESULTS:Transcript libraries were sequenced using Illumina technology and the reads assembled de novo into contigs. Functional identification of contigs followed a two-step procedure, first identifying contigs by comparison to the completely sequenced genome of the related Eucalyptus, followed by identifying the equivalent Arabidopsis gene using a pre-computed mapping between Eucalyptus and Arabidopsis genes. This allowed us to use the rich functional annotation of the Arabidopsis genome to assess the transcriptional landscape of galling in Metrosideros. Comparing galled and ungalled leaves, we find a highly significant enrichment of expressed genes with a gene ontology (GO) annotation to auxin response in the former. One gene consistently expressed in all galled trees examined but not detected in any libraries from ungalled leaves was the Metrosideros version of SMALL AUXIN UPREGULATED (SAUR) 67 which appears to be a marker for leaf-galling in Metrosideros. CONCLUSIONS:We conclude that an auxin response is involved in galling by Metrosideros psyllids. The possibility should therefore be considered that psyllids (like other insects examined) are able to synthesize auxin.