trifolium pratense Dotaz Zobrazit nápovědu
The red cloverl seems to be a prospective source of flavonoids and isoflavonoids with phytoestrogenic activity. Young seedlings of four different varieties of Trifolium pratense L. were employed to derive callus and suspension cultures. The optimal medium for cultivation is Gamborg's cultivating medium with an addition of 2 mg x l(-1) of 2,4-dichlorphenoxyacetic acid and 2 mg x l(-1) of 6-benzylaminopurine. On the basis of growth and production characteristics, subcultivating intervals of 29 to 43 days in callus culture and 16 to 23 days in suspension culture were determined. TLC and HPLC revealed that the explant culture of Trifolium pratense L. contains the isoflavonoid formononetin.
- MeSH
- flavonoidy biosyntéza MeSH
- kultivační techniky MeSH
- Trifolium metabolismus MeSH
- Publikační typ
- anglický abstrakt MeSH
- časopisecké články MeSH
- Názvy látek
- flavonoidy MeSH
Important substances of secondary metabolism include flavonoids and isoflavonoids. The Trifolium pratense L. (Fabacae) suspension culture is characterized by low production of these metabolites and therefore we tried to influence the production output with elicitation. From their origin point of view, the elicitors are divided into two groups - biotic and abiotic. The latter group includes, for instance, the salts of heavy metals. Our work was aimed at observing the effect of the copper sulphate abiotic elicitor on the production of the Trifolium pratense L. suspension culture (variety DO-8 and variety DO-9) that was cultivated in Gamborg media supplemented with 2 mg l(-1) of 2,4-dichlorophenoxyacetic acid and 2 mg l(-1) of 6-benzylaminopurine. The maximum increase in the flavonoid production took place, when compared with the test check, during the 168-hour application of the 100 micromol concentration. The DO-8 variety isoflavonoids production was stimulated namely during the 48-hour application of the I micromol concentration; the best elicitation effect of the DO-9 variety was achieved with the 168-hour application of the 10 micromol concentration.
- MeSH
- flavonoidy biosyntéza MeSH
- síran měďnatý farmakologie MeSH
- techniky tkáňových kultur MeSH
- Trifolium metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- flavonoidy MeSH
- síran měďnatý MeSH
PREMISE OF THE STUDY: Red clover (Trifolium pratense) is an important forage plant from the legume family with great importance in agronomy and livestock nourishment. Nevertheless, assembling its medium-sized genome presents a challenge, given current hardware and software possibilities. Next-generation sequencing technologies enable us to generate large amounts of sequence data at low cost. In this study, the genome assembly and red clover genome features are presented. METHODS: First, assembly software was assessed using data sets from a closely related species to find the best possible combination of assembler plus error correction program to assemble the red clover genome. The newly sequenced genome was characterized by repetitive content, number of protein-coding and nonprotein-coding genes, and gene families and functions. Genome features were also compared with those of other sequenced plant species. KEY RESULTS: Abyss with Echo correction was used for de novo assembly of the red clover genome. The presented assembly comprises ∼314.6 Mbp. In contrast to leguminous species with comparable genome sizes, the genome of T. pratense contains a larger repetitive portion and more abundant retrotransposons and DNA transposons. Overall, 47 398 protein-coding genes were annotated from 64 761 predicted genes. Comparative analysis revealed several gene families that are characteristic for T. pratense. Resistance genes, leghemoglobins, and nodule-specific cystein-rich peptides were identified and compared with other sequenced species. CONCLUSIONS: The presented red clover genomic data constitute a resource for improvement through molecular breeding and for comparison to other sequenced plant species.
- Klíčová slova
- Fabaceae, Trifolium pratense, assessment of assembly software, de novo assembly, genome annotation, red clover,
- MeSH
- DNA rostlinná analýza MeSH
- genom rostlinný * MeSH
- mapování chromozomů MeSH
- rostlinné geny * MeSH
- rostlinné proteiny genetika MeSH
- sekvence nukleotidů MeSH
- sekvenční analýza DNA MeSH
- Trifolium genetika MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- DNA rostlinná MeSH
- rostlinné proteiny MeSH
The important substances of secondary metabolism include flavonoids and isoflavonoids. The Trifolium pratense L. (Fabaceae) suspension culture's yield of these metabolites is low, thus an attempt was made to increase the production by elicitation. The endogenous signal substances of the plants' defensive responses include jasmonic acid that also functions as a biotic elicitor in the case of exogenous application. In the experiment the authors monitored the impact of 4 different concentrations of jasmonic acid on the Trifolium pratense L. (variety DO-8 and variety DO-9) suspension culture's yield. The culture was cultivated in Gamborg medium to which 2 mg.l(-1) of 2,4-dichlorophenoxyacetic acid and 2 mg.l(-1) of 6-benzylaminopurine were added. The maximum increase in the production of flavonoids was achieved, when compared with the control samples, with both varieties of the 500 micromol concentration (DO-9, 6-hour application by 140%; DO-8, 24-hour application by 65%). The production of isoflavonoids (genistin, daidzein, genistein, and formononetine) was best stimulated in both varieties by the 50 micromol concentration (in the case of DO-9 variety, the 48-hour application increased the content of genistin by up to 845%).
- MeSH
- cyklopentany farmakologie MeSH
- flavonoidy biosyntéza MeSH
- isoflavony biosyntéza MeSH
- kultivované buňky MeSH
- oxylipiny farmakologie MeSH
- regulátory růstu rostlin farmakologie MeSH
- Trifolium metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- cyklopentany MeSH
- flavonoidy MeSH
- isoflavony MeSH
- jasmonic acid MeSH Prohlížeč
- oxylipiny MeSH
- regulátory růstu rostlin MeSH
Vanadium compounds increased the content and release of distinct isoflavones in a Trifolium pratense suspension culture. Regarding transport-mechanism inhibitors, the process was mostly facilitated by ABC proteins and vesicular transport. The transport of isoflavones and other secondary metabolites is an important part of metabolism within plants and cultures in vitro regarding their role in defence against various abiotic and biotic stressors. This research focuses on the way how to increase production and exudation of isoflavones by application of chemical elicitor and the basic identification of their transport mechanisms across cell membranes. The release of five isoflavones (genistin, genistein, biochanin A, daidzein, and formononetin) into a nutrient medium was determined in a Trifolium pratense var. DO-8 suspension culture after two vanadium compound treatments and cultivation for 24 and 48 h. The NH4VO3 solution caused a higher concentration of isoflavones in the medium after 24 h. This increased content of secondary metabolites was subsequently suppressed by distinct transport-mechanism inhibitors. The transport of isoflavones in T. pratense was mostly affected by ABC inhibitors from the multidrug-resistance-associated protein subfamily, but the genistein concentration in the medium was lower after treatment with multidrug-resistance protein subfamily inhibitors. Brefeldin A, which blocks vesicular transport, also decreased the concentration of some isoflavones in the nutrient medium.
- Klíčová slova
- Abiotic elicitation, Plasma-membrane transport, Red clover, Secondary metabolites,
- MeSH
- biologický transport účinky léků MeSH
- buněčná membrána účinky léků metabolismus MeSH
- isoflavony metabolismus MeSH
- Trifolium účinky léků metabolismus MeSH
- vanad farmakologie MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- isoflavony MeSH
- vanad MeSH
The production of secondary metabolites in Trifolium pratense L. suspension culture of the family of legume plants (Fabaceae) is low, and therefore there was an attempt to increase it by elicitation. New synthetic substance, 2-(2-fluoro-6-nitrobenzylsulfanyl)pyridine-4-carbothioamide, was tested as elicitor--a substance that showed the best elicitation effect after 48-hour application of 1 μmol L⁻¹ concentration. Maximum contents of genistin (11.60 mg g⁻¹ DW), daidzein (8.31 mg g⁻¹ DW), and genistein (1.50 mg g⁻¹ DW) were recorded, and the production of these isoflavonoids thus significantly increased, when compared with the control, by 152%, 151%, and 400%. The maximum content of flavonoids (5.78 mg g⁻¹ DW) and the increase in the production by 142%, when compared with the control, were induced by 6-hour application of 100 μmol L⁻¹ concentration. The tested substance showed to be an effective elicitor of phenylpropane metabolism.
- Klíčová slova
- CHROMOSOMES *, EXPERIMENTAL LAB STUDY *,
- MeSH
- chromozomy * MeSH
- Trifolium * MeSH
- výzkum * MeSH
- Publikační typ
- časopisecké články MeSH
Plant-rhizobia symbiosis can activate key genes involved in regulating nodulation associated with biological nitrogen fixation (BNF). Although the general molecular basis of the BNF process is frequently studied, little is known about its intraspecific variability and the characteristics of its allelic variants. This study's main goals were to describe phenotypic and genotypic variation in the context of nitrogen fixation in red clover (Trifolium pretense L.) and identify variants in BNF candidate genes associated with BNF efficiency. Acetylene reduction assay validation was the criterion for selecting individual plants with particular BNF rates. Sequences in 86 key candidate genes were obtained by hybridization-based sequence capture target enrichment of plants with alternative phenotypes for nitrogen fixation. Two genes associated with BNF were identified: ethylene response factor required for nodule differentiation (EFD) and molybdate transporter 1 (MOT1). In addition, whole-genome population genotyping by double-digest restriction-site-associated sequencing (ddRADseq) was performed, and BNF was evaluated by the natural 15N abundance method. Polymorphisms associated with BNF and reflecting phenotype variability were identified. The genetic structure of plant accessions was not linked to BNF rate of measured plants. Knowledge of the genetic variation within BNF candidate genes and the characteristics of genetic variants will be beneficial in molecular diagnostics and breeding of red clover.
- Klíčová slova
- associated genes, associated polymorphisms, biological nitrogen fixation, genome-wide association, red clover,
- MeSH
- alely MeSH
- fenotyp MeSH
- fixace dusíku genetika MeSH
- genotyp MeSH
- interakce mikroorganismu a hostitele MeSH
- kořeny rostlin genetika mikrobiologie MeSH
- polymorfismus genetický * MeSH
- Rhizobium fyziologie MeSH
- rostlinné geny genetika MeSH
- sekvenční analýza DNA metody MeSH
- symbióza genetika MeSH
- Trifolium genetika mikrobiologie MeSH
- Publikační typ
- časopisecké články MeSH
Subterranean clover is an important annual forage legume, whose diploidy and inbreeding nature make it an ideal model for genomic analysis in Trifolium. We reported a draft genome assembly of the subterranean clover TSUd_r1.1. Here we evaluate genome mapping on nanochannel arrays and generation of a transcriptome atlas across tissues to advance the assembly and gene annotation. Using a BioNano-based assembly spanning 512 Mb (93% genome coverage), we validated the draft assembly, anchored unplaced contigs and resolved misassemblies. Multiple contigs (264) from the draft assembly coalesced into 97 super-scaffolds (43% of genome). Sequences longer than >1 Mb increased from 40 to 189 Mb giving 1.4-fold increase in N50 with total genome in pseudomolecules improved from 73 to 80%. The advanced assembly was re-annotated using transcriptome atlas data to contain 31 272 protein-coding genes capturing >96% of the gene content. Functional characterization and GO enrichment confirmed gene expression for response to water deprivation, flavonoid biosynthesis and embryo development ending in seed dormancy, reflecting adaptation to the harsh Mediterranean environment. Comparative analyses across Papilionoideae identified 24 893 Trifolium-specific and 6325 subterranean-clover-specific genes that could be mined further for traits such as geocarpy and grazing tolerance. Eight key traits, including persistence, improved livestock health by isoflavonoid production in addition to important agro-morphological traits, were fine-mapped on the high-density SNP linkage map anchored to the assembly. This new genomic information is crucial to identify loci governing traits allowing marker-assisted breeding, comparative mapping and identification of tissue-specific gene promoters for biotechnological improvement of forage legumes.
- Klíčová slova
- BioNano, Legume comparative genomics, advanced reference assembly, forage legumes, gene expression, transcriptome,
- MeSH
- genom rostlinný genetika MeSH
- genomika metody MeSH
- sekvenční analýza DNA metody MeSH
- Trifolium genetika MeSH
- Publikační typ
- časopisecké články MeSH
PREMISE OF THE STUDY: The phenotype of an individual can be modified by the environment experienced by its predecessors, a phenomenon called transgenerational or maternal effects. These effects are studied mostly across sexual generations and are thought to be mediated also by epigenetic variation. However, we do not know how important transgenerational effects are across asexual generations of clonal plants. METHODS: We investigated the role of different drought intensities and durations experienced by parental plants of Trifolium repens on the growth of offspring ramets after transplantation of clonal cuttings to control conditions. We also treated half of the plants with 5-azacytidine, which is a demethylating agent, to test the potential role of DNA methylation on transgenerational effects. KEY RESULTS: Transgenerational effects were manifested as increased biomass of offspring ramets if parental plants experienced medium drought applied for a short period and decreased biomass of offspring ramets if parental plants experienced intense drought for a short period. These transgenerational effects were not observed for offspring of parents from the same treatments if these were treated with 5-azacytidine, whose application significantly decreased the amount of 5-methyl-2'-deoxycytidine in plants. CONCLUSIONS: Transgenerational effects might play an important role in the clonal plant Trifolium repens and are probably mediated by epigenetic variation. The growth and behavior of clonal plants might be affected not only by the ambient environment but also by environments that are no longer present at the time of clonal reproduction. This phenomenon can have yet unacknowledged ecological and evolutionary implications for clonal plants.
- Klíčová slova
- 5-azacytidine, DNA methylation, HPLC, asexual reproduction, drought, epigenetic variation, plant behavior,
- MeSH
- biomasa MeSH
- fenotyp MeSH
- fyziologický stres * MeSH
- nepohlavní rozmnožování * MeSH
- období sucha * MeSH
- Trifolium růst a vývoj fyziologie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
