Dataset Information

De novo assembly, transcriptome characterization, and simple sequence repeat marker development in duckweed Lemna gibba.

ABSTRACT: Lemna gibba is a species of duckweed showing great potential in bioenergy production and wastewater treatment. However, the relevant transcriptomic and genomic resources are very limited for this species, which dramatically hinders its genetic diversity and genome mapping researches. In this work, ~?233.5 million clean reads were generated from L. gibba by Illumina paired-end sequencing, and subsequently they were de novo assembled into 131,870 unigenes, of which 61,622 were annotated and 43,319 were expressed with Fragments Per Kilobase of transcript per Million fragments mapped (FPKM)?>?5. In total, 19,297 simple sequence repeats (SSRs) were identified from 15,261 SSR-containing unigenes. Dinucleotide (78.4%) were the most abundant SSRs, followed by tri- (14.9%), tetra- (4.1%), and penta-nucleotides (1.5%). The top three motifs were AG/CT (69.9%), AC/GT (6.5%), and ATC/ATG (4.9%). Further analysis revealed that the presence of SSR motif was independent of the expression level for a given gene. Based on the sequence of these SSR-containing unigenes, a total of 10,292 SSR markers were developed, of which only 2671 were further retained after removing those derived from unannotated or extra-low expressed (e.g., FPKM???5) unigenes. Finally, a subset of 70 SSR markers was randomly selected and examined in nine diverse L. gibba genotypes for the PCR amplification and polymorphism, as well as in other duckweed species for the inter-specifically amplifiability. This work is the first report on the transcriptome-based large-scale SSR markers development and analysis in L. gibba. The transcriptome generated and the SSR markers developed in this work will provide a valuable resource for genetic diversity assessment in L. gibba and also for species relationship investigation in Lemnaceae family.

SUBMITTER: Fu L

PROVIDER: S-EPMC7036385 | biostudies-literature | 2020 Jan

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

De novo assembly, transcriptome characterization, and simple sequence repeat marker development in duckweed Lemna gibba.

Fu Lili L Ding Zehong Z Kumpeangkeaw Anuwat A Tan Deguan D Han Bingying B Sun Xuepiao X Zhang Jiaming J

Physiology and molecular biology of plants : an international journal of functional plant biology 20191209 1

Lemna gibba is a species of duckweed showing great potential in bioenergy production and wastewater treatment. However, the relevant transcriptomic and genomic resources are very limited for this species, which dramatically hinders its genetic diversity and genome mapping researches. In this work, ~ 233.5 million clean reads were generated from L. gibba by Illumina paired-end sequencing, and subsequently they were de novo assembled into 131,870 unigenes, of which 61,622 were annota ...[more]

PMID: 32158126

Similar Datasets

Project description:BackgroundDuckweeds are small, rapidly growing aquatic flowering plants. Due to their ability for biomass production at high rates they represent promising candidates for biofuel feedstocks. Duckweeds are also excellent model organisms because they can be maintained in well-defined liquid media, usually reproduce asexually, and because genomic resources are becoming increasingly available. To demonstrate the utility of duckweed for integrated metabolic studies, we examined the metabolic adaptation of growing Lemna gibba cultures to different nutritional conditions.ResultsTo establish a framework for quantitative metabolic research in duckweeds we derived a central carbon metabolism network model of Lemna gibba based on its draft genome. Lemna gibba fronds were grown with nitrate or glutamine as nitrogen source. The two conditions were compared by quantification of growth kinetics, metabolite levels, transcript abundance, as well as by 13C-metabolic flux analysis. While growing with glutamine, the fronds grew 1.4 times faster and accumulated more protein and less cell wall components compared to plants grown on nitrate. Characterization of photomixotrophic growth by 13C-metabolic flux analysis showed that, under both metabolic growth conditions, the Calvin-Benson-Bassham cycle and the oxidative pentose-phosphate pathway are highly active, creating a futile cycle with net ATP consumption. Depending on the nitrogen source, substantial reorganization of fluxes around the tricarboxylic acid cycle took place, leading to differential formation of the biosynthetic precursors of the Asp and Gln families of proteinogenic amino acids. Despite the substantial reorganization of fluxes around the tricarboxylic acid cycle, flux changes could largely not be associated with changes in transcripts.ConclusionsThrough integrated analysis of growth rate, biomass composition, metabolite levels, and metabolic flux, we show that Lemna gibba is an excellent system for quantitative metabolic studies in plants. Our study showed that Lemna gibba adjusts to different nitrogen sources by reorganizing central metabolism. The observed disconnect between gene expression regulation and metabolism underscores the importance of metabolic flux analysis as a tool in such studies.

Project description:In this study, we exposed a commonly used duckweed species-Lemna gibba L.-to twelve environmentally relevant metals and metalloids under laboratory conditions. The phytotoxic effects were evaluated in a multi-well-plate-based experimental setup by means of the chlorophyll fluorescence imaging method. This technique allowed the simultaneous measuring of the growth and photosynthetic parameters in the same samples. The inhibition of relative growth rates (based on frond number and area) and photochemical efficiency (Fv/Fo and Y(II)) were both calculated from the obtained chlorophyll fluorescence images. In the applied test system, growth-inhibition-based phytotoxicity endpoints proved to be more sensitive than chlorophyll-fluorescence-based ones. Frond area growth inhibition was the most responsive parameter with a median EC50 of 1.75 mg L-1, while Fv/Fo, the more responsive chlorophyll-fluorescence-based endpoint, resulted in a 5.34 mg L-1 median EC50 for the tested metals. Ag (EC50 0.005-1.27 mg L-1), Hg (EC50 0.24-4.87 mg L-1) and Cu (EC50 0.37-1.86 mg L-1) were the most toxic elements among the tested ones, while As(V) (EC50 47.15-132.18 mg L-1), Cr(III) (EC50 6.22-19.92 mg L-1), Se(VI) (EC50 1.73-10.39 mg L-1) and Zn (EC50 3.88-350.56 mg L-1) were the least toxic ones. The results highlighted that multi-well-plate-based duckweed phytotoxicity assays may reduce space, time and sample volume requirements compared to the standard duckweed growth inhibition tests. These benefits, however, come with lowered test sensitivity. Our multi-well-plate-based test setup resulted in considerably higher median EC50 (3.21 mg L-1) for frond-number-based growth inhibition than the 0.683 mg L-1 median EC50 derived from corresponding data from the literature with standardized Lemna-tests. Under strong acute phytotoxicity, frond parts with impaired photochemical functionality may become undetectable by chlorophyll fluorometers. Consequently, the plant parts that are still detectable display a virtually higher average photosynthetic performance, leading to an underestimation of phytotoxicity. Nevertheless, multi-well-plate-based duckweed phytotoxicity assays, combined with chlorophyll fluorescence imaging, offer definite advantages in the rapid screening of large sample series or multiple species/clones. As chlorophyll fluorescence images provide information both on the photochemical performance of the test plants and their morphology, a joint analysis of the two endpoint groups is recommended in multi-well-plate-based duckweed phytotoxicity assays to maximize the information gained from the tests.

Project description:Sainfoin (Onobrychis viciifolia) is a highly nutritious tannin-containing forage legume. In the diet of ruminants sainfoin can have anti-parasitic effects and reduce methane emissions under in vitro conditions. Many of these benefits have been attributed to condensed tannins or proanthocyanidins in sainfoin. A combination of increased use of industrially produced nitrogen fertilizer, issues with establishment and productivity in the first year and more reliable alternatives, such as red clover led to a decline in the use of sainfoin since the middle of the last century. In recent years there has been a resurgence of interest in sainfoin due to its potential beneficial nutraceutical and environmental attributes. However, genomic resources are scarce, thus hampering progress in genetic analysis and improvement. To address this we have used next generation RNA sequencing technology to obtain the first transcriptome of sainfoin. We used the library to identify gene-based simple sequence repeats (SSRs) and potential single nucleotide polymorphisms (SNPs).One genotype from each of five sainfoin accessions was sequenced. Paired-end (PE) sequences were generated from cDNA libraries of RNA extracted from 7 day old seedlings. A combined assembly of 92,772 transcripts was produced de novo using the Trinity programme. About 18,000 transcripts were annotated with at least one GO (gene ontology) term. A total of 63 transcripts were annotated as involved in the tannin biosynthesis pathway. We identified 3786 potential SSRs. SNPs were identified by mapping the reads of the individual assemblies against the combined assembly. After stringent filtering a total of 77,000 putative SNPs were identified. A phylogenetic analysis of single copy number genes showed that sainfoin was most closely related to red clover and Medicago truncatula, while Lotus japonicus, bean and soybean are more distant relatives.This work describes the first transcriptome assembly in sainfoin. The 92 K transcripts provide a rich source of SNP and SSR polymorphisms for future use in genetic studies of this crop. Annotation of genes involved in the condensed tannin biosynthesis pathway has provided the basis for further studies of the genetic control of this important trait in sainfoin.

Dataset Information

De novo assembly, transcriptome characterization, and simple sequence repeat marker development in duckweed Lemna gibba.

Publications

De novo assembly, transcriptome characterization, and simple sequence repeat marker development in duckweed <i>Lemna gibba</i>.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets