Dataset Information

Comparative Transcriptome and Metabolic Profiling Analysis of Buckwheat (Fagopyrum Tataricum (L.) Gaertn.) under Salinity Stress.

ABSTRACT: Tartary buckwheat (Fagopyrum tataricum (L.) Gaertn.) is a nutritional crop, which has high flavonoid content. However, buckwheat is a salt sensitive glycophyte cereal crop and the growth and grain yield of buckwheat are significantly affected by soil salinity. In this study, we performed a comprehensive analysis of the transcriptome and metabolome of salt treated-buckwheat to understand the effects of salinity on buckwheat. A total of 50,681,938 clean reads were acquired from all samples. We acquired 94,950 unigenes with a mean length of 1133 bp and N50 length of 1900 bp assembly. Of these, 63,305 unigenes (66.7%) were matched in public databases. Comparison of the transcriptome expression patterns between control and salt treated groups showed that 4098 unigenes were up-regulated and 3292 unigenes were down-regulated significantly. Further, we found that genes involved with amino acid, lipid and nucleotide metabolism were most responsive to salt stress. Additionally, many genes involved in secondary metabolite biosynthesis changed significantly following treatment. Those affected included phenylpropanoid biosynthesis and flavonoid biosynthesis. Chromatographic analysis was used to examine the differences in concentration of flavonoids, carotenoids, amino acids and organic acids in the samples following treatment. There was a significant increase in rutin (12.115 mg/g dry weight), following salt stress; whereas, six carotenoids (lutein, zeaxanthin, 13Z-?-carotene, ?-carotene, E-?-carotene and 9Z-?-carotene) did not significantly respond to salt stress. Ultimately, our data acts as a valuable resource for future research on buckwheat and can be used as the basis for future analysis focused on gene-to-metabolite networks in buckwheat.

SUBMITTER: Ma W

PROVIDER: S-EPMC6835380 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature

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Comparative Transcriptome and Metabolic Profiling Analysis of Buckwheat (<i>Fagopyrum Tataricum</i> (L.) Gaertn.) under Salinity Stress.

Ma Weibo W Kim Jae Kwang JK Jia Caihua C Yin Feifan F Kim Hyo Jin HJ Akram Waheed W Hu Xuebo X Li Xiaohua X

Metabolites 20191014 10

Tartary buckwheat (<i>Fagopyrum tataricum</i> (L.) Gaertn.) is a nutritional crop, which has high flavonoid content. However, buckwheat is a salt sensitive glycophyte cereal crop and the growth and grain yield of buckwheat are significantly affected by soil salinity. In this study, we performed a comprehensive analysis of the transcriptome and metabolome of salt treated-buckwheat to understand the effects of salinity on buckwheat. A total of 50,681,938 clean reads were acquired from all samples. ...[more]

PMID: 31614965

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Project description:BackgroundTartary buckwheat is an important minor crop species with high nutritional and medicinal value and is widely planted worldwide. Cultivated Tartary buckwheat plants are tall and have hollow stems that lodge easily, which severely affects their yield and hinders the development of the Tartary buckwheat industry.MethodsHeifeng No. 1 seeds were treated with ethylmethanesulfonate (EMS) to generate a mutant library. The dwarf mutant ftdm was selected from the mutagenized population, and the agronomic characteristics giving rise to the dwarf phenotype were evaluated. Ultra-fast liquid chromatography-electrospray ionization tandem mass spectrometry (UFLC-ESI-MS/MS) was performed to determine the factors underlying the different phenotypes between the wild-type (WT) and ftdm plants. In addition, RNA sequencing (RNA-seq) was performed via the HiSeq 2000 platform, and the resulting transcriptomic data were analysed to identify differentially expressed genes (DEGs). Single-nucleotide polymorphism (SNP) variant analysis revealed possible sites associated with dwarfism. The expression levels of the potential DEGs between the WT and ftdm mutant were then measured via qRT-PCR and fragments per kilobase of transcript per million mapped reads (FPKM).ResultThe plant height (PH) of the ftdm mutant decreased to 42% of that of the WT, and compared with the WT, the mutant and had a higher breaking force (BF) and lower lodging index (LI). Lower GA4 and GA7 contents and higher contents of jasmonic acid (JA), salicylic acid (SA) and brassinolactone (BR) were detected in the stems of the ftdm mutant compared with the WT. Exogenous application of GAs could not revert the dwarfism of the ftdm mutant. On the basis of the transcriptomic analysis, 146 homozygous SNP loci were identified. In total, 12 DEGs with nonsynonymous mutations were ultimately identified, which were considered potential candidate genes related to the dwarf trait. When the sequences of eight genes whose expression was downregulated and four genes whose expression was upregulated were compared, SKIP14, an F-box protein whose sequence is 85% homologous to that of SLY1 in Arabidopsis, presented an amino acid change (from Ser to Asn) and was expressed at a lower level in the stems of the ftdm mutant compared with the WT. Hence, we speculated that this amino acid change in SKIP14 resulted in a disruption in GA signal transduction, indirectly decreasing the GA content and downregulating the expression of genes involved in GA biosynthesis or the GA response. Further studies are needed to determine the molecular basis underlying the dwarf phenotype of the ftdm mutant.ConclusionWe report a Tartary buckwheat EMS dwarf mutant, ftdm, suitable for high-density planting and commercial farming. A significant decrease in GA4 and GA7 levels was detected in the ftdm mutant, and 12 DEGs expressed in the stems of the ftdm mutant were selected as candidates of the dwarfing gene. One nonsynonymous mutation was detected in the SKIP14 gene in the ftdm mutant, and this gene had a lower transcript level compared with that in the WT.

Project description:BackgroundTartary buckwheat (Fagopyrum tataricum Gaertn.) is a widely cultivated medicinal and edible crop with excellent economic and nutritional value. The development of tartary buckwheat seeds is a very complex process involving many expression-dependent physiological changes and regulation of a large number of genes and phytohormones. In recent years, the gene regulatory network governing the physiological changes occurring during seed development have received little attention.ResultsHere, we characterized the seed development of tartary buckwheat using light and electron microscopy and measured phytohormone and nutrient accumulation by using high performance liquid chromatography (HPLC) and by profiling the expression of key genes using RNA sequencing with the support of the tartary buckwheat genome. We first divided the development of tartary buckwheat seed into five stages that include complex changes in development, morphology, physiology and phytohormone levels. At the same time, the contents of phytohormones (gibberellin, indole-3-acetic acid, abscisic acid, and zeatin) and nutrients (rutin, starch, total proteins and soluble sugars) at five stages were determined, and their accumulation patterns in the development of tartary buckwheat seeds were analyzed. Second, gene expression patterns of tartary buckwheat samples were compared during three seed developmental stages (13, 19, and 25 days postanthesis, DPA), and 9 765 differentially expressed genes (DEGs) were identified. We analyzed the overlapping DEGs in different sample combinations and measured 665 DEGs in the three samples. Furthermore, expression patterns of DEGs related to phytohormones, flavonoids, starch, and storage proteins were analyzed. Third, we noted the correlation between the trait (physiological changes, nutrient changes) and metabolites during seed development, and discussed the key genes that might be involved in the synthesis and degradation of each of them.ConclusionWe provided abundant genomic resources for tartary buckwheat and Polygonaceae communities and revealed novel molecular insights into the correlations between the physiological changes and seed development of tartary buckwheat.

Project description:BackgroundTartary buckwheat seed development is an extremely complex process involving many gene regulatory pathways. MicroRNAs (miRNAs) have been identified as the important negative regulators of gene expression and performed crucial regulatory roles in various plant biological processes. However, whether miRNAs participate in Tartary buckwheat seed development remains unexplored.ResultsIn this study, we first identified 26 miRNA biosynthesis genes in the Tartary buckwheat genome and described their phylogeny and expression profiling. Then we performed small RNA (sRNA) sequencing for Tartary buckwheat seeds at three developmental stages to identify the miRNAs associated with seed development. In total, 230 miRNAs, including 101 conserved and 129 novel miRNAs, were first identified in Tartary buckwheat, and 3268 target genes were successfully predicted. Among these miRNAs, 76 exhibited differential expression during seed development, and 1534 target genes which correspond to 74 differentially expressed miRNAs (DEMs) were identified. Based on integrated analysis of DEMs and their targets expression, 65 miRNA-mRNA interaction pairs (25 DEMs corresponding to 65 target genes) were identified that exhibited significantly opposite expression during Tartary buckwheat seed development, and 6 of the miRNA-mRNA pairs were further verified by quantitative real-time polymerase chain reaction (qRT-PCR) and ligase-mediated rapid amplification of 5' cDNA ends (5'-RLM-RACE). Functional annotation of the 65 target mRNAs showed that 56 miRNA-mRNA interaction pairs major involved in cell differentiation and proliferation, cell elongation, hormones response, organogenesis, embryo and endosperm development, seed size, mineral elements transport, and flavonoid biosynthesis, which indicated that they are the key miRNA-mRNA pairs for Tartary buckwheat seed development.ConclusionsOur findings provided insights for the first time into miRNA-mediated regulatory pathways in Tartary buckwheat seed development and suggested that miRNAs play important role in Tartary buckwheat seed development. These findings will be help to study the roles and regulatory mechanism of miRNAs in Tartary buckwheat seed development.

Dataset Information

Comparative Transcriptome and Metabolic Profiling Analysis of Buckwheat (Fagopyrum Tataricum (L.) Gaertn.) under Salinity Stress.

Publications

Comparative Transcriptome and Metabolic Profiling Analysis of Buckwheat (<i>Fagopyrum Tataricum</i> (L.) Gaertn.) under Salinity Stress.

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