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1.
Molecules ; 24(1)2019 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-30609669

RESUMO

Trigonella foenum-graecum L. (fenugreek) is a valuable resource of producing diosgenin which serves as a substrate for synthesizing more than two hundred kinds of steroidal drugs. Phytochemical analysis indicated that methyl jasmonate (MeJA) efficiently induced diosgenin biosynthesis in fenugreek seedlings. Though early steps up to cholesterol have recently been elucidated in plants, cytochrome P450 (CYP)- and glycosyltransferase (GT)-encoding genes involved in the late steps from cholesterol to diosgenin remain unknown. This study established comparative fenugreek transcriptome datasets from the MeJA-treated seedlings and the corresponding control lines. Differential gene expression analysis identified a number of MeJA-induced CYP and GT candidate genes. Further gene expression pattern analysis across a different MeJA-treating time points, together with a phylogenetic analysis, suggested specific family members of CYPs and GTs that may participate in the late steps during diosgenin biosynthesis. MeJA-induced transcription factors (TFs) that may play regulatory roles in diosgenin biosynthesis were also discussed. This study provided a valuable genetic resource to functionally characterize the genes involved in diosgenin biosynthesis, which will push forward the production of diosgenin in microbial organisms using a promising synthetic biology strategy.


Assuntos
Diosgenina/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Transcriptoma , Trigonella/genética , Trigonella/metabolismo , Biologia Computacional/métodos , Ontologia Genética , Redes e Vias Metabólicas , Anotação de Sequência Molecular , Filogenia , Fatores de Transcrição , Trigonella/classificação
2.
J Pharm Biomed Anal ; 149: 403-409, 2018 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-29154110

RESUMO

Siberian ginseng (Eleutherococcus senticosus, Araliaceae) and roseroot (Rhodiola rosea, Rosaceae) are popular herbal supplements which have been shown to improve resilience to conditions such as stress and exhaustion. Using DNA barcoding methods we tested 25 Siberian ginseng and 14 roseroot products which are widely available to UK customers to test whether the herbal ingredient stated on the label is also in the product. All Siberian ginseng supplements contained E. senticosus, however, 36% also contained an Eleutherococcus species other than E. senticosus. In three out of the 13 roseroot products which produced amplifiable DNA, we could only retrieve sequences matching alfalfa (declared on the product label) and fenugreek (not declared). In the other 10 supplements Rhodiola was detected but only five matched the target species R. rosea. As DNA can get severely degraded during the manufacturing process we did not take the absence of Rhodiola DNA as proof for a compromised product. Contamination could explain the presence of non-target species such as fenugreek but is unlikely to be account for the detection of congeneric Rhodiola species in roseroot preparations. Our results therefore suggest that the substitution or mixing of the target medicinal ingredient in these two popular supplements with other species is common.


Assuntos
Suplementos Nutricionais/análise , Eleutherococcus/química , Contaminação de Alimentos/análise , Rotulagem de Alimentos , Rhodiola/química , Código de Barras de DNA Taxonômico , Suplementos Nutricionais/normas , Eleutherococcus/genética , Medicago sativa/química , Medicago sativa/genética , Filogenia , Rhodiola/genética , Trigonella/química , Trigonella/genética , Reino Unido
3.
Planta ; 245(5): 977-991, 2017 May.
Artigo em Inglês | MEDLINE | ID: mdl-28161815

RESUMO

MAIN CONCLUSION: Representational difference analysis of cDNA was performed and differential products were sequenced and annotated. Candidate genes involved in biosynthesis of diosgenin in fenugreek were identified. Detailed mechanism of diosgenin synthesis was proposed. Fenugreek (Trigonella foenum-graecum L.) is a valuable medicinal and crop plant. It belongs to Fabaceae family and has a unique potential to synthesize valuable steroidal saponins, e.g., diosgenin. Elicitation (methyl jasmonate) and precursor feeding (cholesterol and squalene) were used to enhance the content of sterols and steroidal sapogenins in in vitro grown plants for representational difference analysis of cDNA (cDNA-RDA). To identify candidate genes involved in diosgenin biosynthesis, differential, factor-specific libraries were subject to the next-generation sequencing. Approximately 9.9 million reads were obtained, trimmed, and assembled into 31,491 unigenes with an average length of 291 bp. Then, functional annotation and gene ontogeny enrichment analysis was performed by aligning all-unigenes with public databases. Within the transcripts related to sterol and steroidal saponin biosynthesis, we discovered novel candidate genes of diosgenin biosynthesis and validated their expression using quantitative RT-PCR analysis. Based on these findings, we supported the idea that diosgenin is biosynthesized from cycloartenol via cholesterol. This is the first report on the next-generation sequencing of cDNA-RDA products. Analysis of the transcriptomes enriched in low copy sequences contributed substantially to our understanding of the biochemical pathways of steroid synthesis in fenugreek.


Assuntos
Acetatos/metabolismo , Ciclopentanos/metabolismo , Diosgenina/metabolismo , Oxilipinas/metabolismo , Fitosteróis/metabolismo , Reguladores de Crescimento de Planta/metabolismo , Transcriptoma , Trigonella/genética , DNA Complementar/genética , Sequenciamento de Nucleotídeos em Larga Escala , Análise de Sequência de DNA , Trigonella/metabolismo
4.
Plant Cell Rep ; 35(5): 1189-203, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-26956134

RESUMO

KEY MESSAGE: Coexpression of two antifungal genes ( NPR1 and defensin ) in transgenic peanut results in the development of resistance to two major fungal pathogens, Aspergillus flavus and Cercospora arachidicola. Fungal diseases have been one of the principal causes of crop losses with no exception to peanut (Arachis hypogeae L.), a major oilseed crop in Asia and Africa. To address this problem, breeding for fungal disease resistance has been successful to some extent against specific pathogens. However, combating more than one fungal pathogen via breeding is a major limitation in peanut. In the present study, we demonstrated the potential use of co-overexpression of two genes, NPR1 and defensin isolated from Brassica juncea and Trigonella foenum-graecum respectively; that offered resistance towards Aspergillus flavus in peanut. The transgenic plants not only resisted the mycelial growth but also did not accumulate aflatoxin in the seeds. Resistance was also demonstrated against another pathogen, Cercospora arachidicola at varied levels; the transgenic plants showed both reduction in the number of spots and delay in the onset of disease. PCR, Southern and Western blot analysis confirmed stable integration and expression of the transgenes in the transgenic plants. The combinatorial use of the two pathogen resistance genes presents a novel approach to mitigate two important fungal pathogens of peanut.


Assuntos
Arachis/imunologia , Resistência à Doença/genética , Mostardeira/genética , Doenças das Plantas/imunologia , Proteínas de Plantas/metabolismo , Trigonella/genética , Arachis/genética , Arachis/microbiologia , Ascomicetos/fisiologia , Aspergillus flavus/fisiologia , Defensinas/genética , Defensinas/metabolismo , Expressão Gênica , Doenças das Plantas/microbiologia , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Plântula/genética , Plântula/imunologia , Plântula/microbiologia , Sementes/genética , Sementes/imunologia , Sementes/microbiologia , Transformação Genética , Transgenes
5.
Int J Mol Sci ; 16(12): 29889-99, 2015 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-26694357

RESUMO

The effects of methyl jasmonate (MeJA), an elicitor of plant defense mechanisms, on the biosynthesis of diosgenin, a steroidal saponin, were investigated in six fenugreek (Trigonella foenum-graecum) varieties (Gujarat Methi-2, Kasuri-1, Kasuri-2, Pusa Early Branching, Rajasthan Methi and Maharashtra Methi-5). Treatment with 0.01% MeJA increased diosgenin levels, in 12 days old seedlings, from 0.5%-0.9% to 1.1%-1.8%. In addition, MeJA upregulated the expression of two pivotal genes of the mevalonate pathway, the metabolic route leading to diosgenin: 3-hydroxy-3-methylglutaryl-CoA reductase (HMG) and sterol-3-ß-glucosyl transferase (STRL). In particular, MeJA increased the expression of HMG and STRL genes by 3.2- and 22.2-fold, respectively, in the Gujarat Methi-2 variety, and by 25.4- and 28.4-fold, respectively, in the Kasuri-2 variety. Therefore, MeJA may be considered a promising elicitor for diosgenin production by fenugreek plants.


Assuntos
Acetatos/farmacologia , Ciclopentanos/farmacologia , Diosgenina/metabolismo , Oxilipinas/farmacologia , Plântula/metabolismo , Trigonella/metabolismo , Biomassa , Vias Biossintéticas/efeitos dos fármacos , Vias Biossintéticas/genética , Eletroforese em Gel de Ágar , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Genes Essenciais , Genes de Plantas , Plântula/anatomia & histologia , Plântula/efeitos dos fármacos , Trigonella/efeitos dos fármacos , Trigonella/genética
6.
Genet Mol Res ; 13(4): 10464-81, 2014 Dec 12.
Artigo em Inglês | MEDLINE | ID: mdl-25511030

RESUMO

In this study, we analyzed the correlation between genetic variation based on random amplified polymorphic DNA (RAPD), acid phosphatase, and glutamate-oxaloacetate transaminase isozymes, and amino acid composition with the antioxidant potential status of 7 wild Trigonella foenum-graecum L. accessions collected from diverse ecogeographical regions. RAPD revealed that 90 DNA products had highly polymorphism value (94.12%) based on band numbers, with sizes ranging from 50-2100 base pairs, and band intensity. Of 49 DNA polymorphic bands, 31 unique and 3 monomorphic bands were scored. Acid phosphatase and glutamate-oxaloacetate transaminase showed total polymorphism values of 90.00 and 93.75%, respectively, based on zymogram number, relative front (Rf), and optical intensity. Because isozymes are composed of amino acids, they were analyzed using high-performance liquid chromatography, which revealed the presences of 16 amino acids of variable content ranging from 13.21-15.35%, 9 of which are essential amino acids in humans. RAPD and isozymes showed similarly high estimates of genetic variability. Genetic relationships revealed by unweighted pair group method with arithmetic mean clustering analysis based on data obtained from all primers of RAPD and each isozyme were very similar. The antioxidant potential based on free radical scavenging, 2, 2-diphenyl-1-picrylhydrazyl, b-carotene-linoleate, total phenolic, and flavonoid contents values were variable among accessions. We found that fenugreek is a valuable genetic resource with high antioxidant activity. Their genotypes, based on data and clustering of RAPD, isozymes, and variable amino acid contents, combined with their antioxidant potential statues are important in fenugreek breeding and improvement programs.


Assuntos
Fosfatase Ácida/genética , Aspartato Aminotransferase Citoplasmática/genética , Polimorfismo Genético , Trigonella/genética , Fosfatase Ácida/metabolismo , Antioxidantes/metabolismo , Aspartato Aminotransferase Citoplasmática/metabolismo , Depuradores de Radicais Livres/metabolismo , Genótipo , Oxirredução , Filogenia , Técnica de Amplificação ao Acaso de DNA Polimórfico , Trigonella/metabolismo
7.
Plant Physiol Biochem ; 83: 217-24, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25179433

RESUMO

The amino acid sequence of Fenugreek ß-amylase is not available in protein data bank. Therefore, an attempt has been made to identify the catalytic amino acid residues of enzyme by employing studies of pH dependence of enzyme catalysis, chemical modification and bioinformatics. Treatment of purified Fenugreek ß-amylase with EDAC in presence of glycine methyl ester and sulfhydryl group specific reagents (IAA, NEM and p-CMB), followed a pseudo first-order kinetics and resulted in effective inactivation of enzyme. The reaction with EDAC in presence of NTEE (3-nitro-l-tyrosine ethylester) resulted into modification of two carboxyl groups per molecule of enzyme and presence of one accessible sulfhydryl group at the active site, per molecule of enzyme was ascertained by titration with DTNB. The above results were supported by the prevention of inactivation of enzyme in presence of substrate. Based on MALDI-TOF analysis of purified Fenugreek ß-amylase and MASCOT search, ß-amylase of Medicago sativa was found to be the best match. To further confirm the amino acid involved in catalysis, homology modelling of ß-amylase of M. sativa was performed. The sequence alignment, superimposition of template and target models, along with study of interactions involved in docking of sucrose and maltose at the active site, led to identification of Glu187, Glu381 and Cys344 as active site residues.


Assuntos
Simulação de Acoplamento Molecular , Proteínas de Plantas , Trigonella , beta-Amilase , Domínio Catalítico , Proteínas de Plantas/química , Proteínas de Plantas/genética , Trigonella/enzimologia , Trigonella/genética , beta-Amilase/química , beta-Amilase/genética
8.
Food Chem ; 148: 188-95, 2014 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-24262545

RESUMO

This study was conducted to evaluate the influence of developmental stages (vegetative, flowering and fruiting) of mixoploid fenugreek aerial parts on their chemical composition and allelopathic potential, assessed on lettuce germination and seedling growth. Aqueous and organic extracts significantly delayed germination, reduced its rate and affected seedling growth. Ethyl acetate and methanol extracts of aerial parts harvested at vegetative stage were the most toxic for lettuce germination and seedling growth, respectively. LC-MS/MS analysis of T. foenum-graecum aerial parts methanolic extract showed nine different flavonol glycosides (quercetin and kaempferol glucosides). Chemical composition of aerial parts differed with the developmental stage; indeed, at the vegetative and fruiting stages, analysis revealed the presence of 9 compounds as compared to only 6 compounds at the flowering stage. Thus, it is necessary to follow the qualitative changes of allelochemicals production at different developmental stages to identify the most productive one.


Assuntos
Orobanche/crescimento & desenvolvimento , Exsudatos de Plantas/química , Plantas Daninhas/crescimento & desenvolvimento , Trigonella/química , Controle de Plantas Daninhas/métodos , Agricultura , Alelopatia , Germinação/efeitos dos fármacos , Orobanche/efeitos dos fármacos , Exsudatos de Plantas/metabolismo , Exsudatos de Plantas/farmacologia , Plantas Daninhas/efeitos dos fármacos , Ploidias , Sementes/efeitos dos fármacos , Sementes/crescimento & desenvolvimento , Espectrometria de Massas em Tandem , Trigonella/genética , Trigonella/metabolismo
9.
Genet Mol Res ; 12(4): 6284-98, 2013 Dec 04.
Artigo em Inglês | MEDLINE | ID: mdl-24338424

RESUMO

Protein and DNA polymorphismswere surveyed among seven accessions of wild fenugreek (Trigonellafoenum-graecum L.) to estimate their genetic diversity and relationships. Samples were obtained from diverse ecogeographical areas in Saudi Arabia and Yemen. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis of seed storage protein showed genetic variations among fenugreek germplasms, both quantitatively and qualitatively, generating a total of 168 polypeptide bands with different molecular weights ranging from 4.5 to 300 kDa. Twenty-six of these bands were polymorphic, with a considerable polymorphism value (80.00%). Furthermore, restriction fragment length polymorphism (RFLP) analysis was also employed, which was based on the ability of four restriction enzymes (EagI, EcoRI, FspI, and HindIII) to cleave genomic DNA of the plant materials at specific target nucleotide sequences into different numbers of DNA fragments. RFLP analysis revealed 166 fragments with known sequences and variable lengths ranging from 80 to 4000 bp with a highly degree of polymorphism (88.71%). Data derived from SDS-PAGE or RFLP analyses were used to produce dendrograms, which clustered the studied fenugreek accessions into different groups based on the unweighted pair group method with arithmetic mean (UPGMA). The resulting relationships indicated that these two marker techniques were nearly equivalent, but not identical, with respect to phylogenetic information. In conclusion, SDS-PAGE analysis of seed proteins should be augmented with RFLP analysis of DNA for reliable estimates of genetic diversity among fenugreek germplasms.


Assuntos
Proteínas de Armazenamento de Sementes/metabolismo , Sementes/genética , Trigonella/genética , Análise por Conglomerados , DNA de Plantas/genética , DNA de Plantas/isolamento & purificação , Eletroforese em Gel de Poliacrilamida , Expressão Gênica , Filogenia , Polimorfismo de Fragmento de Restrição , Proteínas de Armazenamento de Sementes/genética , Sementes/metabolismo , Trigonella/metabolismo
10.
Funct Integr Genomics ; 13(4): 435-43, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-24022215

RESUMO

Plant defensins are small (45 to 54 amino acids) positively charged antimicrobial peptides produced by the plant species, which can inhibit the growth of a broad range of fungi at micro-molar concentrations. These basic peptides share a common characteristic three-dimensional folding pattern with one α-helix and three ß-sheets that are stabilized by eight disulfide-linked cysteine residues. Instead of using two single-gene constructs, it is beneficial when two effective genes are made into a single fusion gene with one promoter and terminator. In this approach, we have linked two plant defensins namely Trigonella foenum-graecum defensin 2 (Tfgd2) and Raphanus sativus antifungal protein 2 (RsAFP2) genes by a linker peptide sequence (occurring in the seeds of Impatiens balsamina) and made into a single-fusion gene construct. We used pET-32a+ vector system to express Tfgd2-RsAFP2 fusion gene with hexahistidine tag in Escherichia coli BL21 (DE3) pLysS cells. Induction of these cells with 1 mM IPTG achieved expression of the fusion protein. The solubilized His6-tagged recombinant fusion protein was purified by immobilized-metal (Ni2+) affinity column chromatography. The final yield of the fusion protein was 500 ng/µL. This method produced biologically active recombinant His6-tagged fusion protein, which exhibited potent antifungal action towards the plant pathogenic fungi (Botrytis cinerea, Fusarium moniliforme, Fusarium oxysporum, Phaeoisariopsis personata and Rhizoctonia solani along with an oomycete pathogen Phytophthora parasitica var nicotianae) at lower concentrations under in vitro conditions. This strategy of combining activity of two defensin genes into a single-fusion gene will definitely be a promising utility for biotechnological applications.


Assuntos
Defensinas/toxicidade , Proteínas de Plantas/toxicidade , Raphanus/genética , Proteínas Recombinantes de Fusão/toxicidade , Trigonella/genética , Sequência de Aminoácidos , Sequência de Bases , Defensinas/genética , Fungicidas Industriais/toxicidade , Fusarium/efeitos dos fármacos , Dados de Sequência Molecular , Phytophthora/efeitos dos fármacos , Proteínas de Plantas/genética , Proteínas Recombinantes de Fusão/genética
11.
Plant Mol Biol ; 79(3): 243-58, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-22527750

RESUMO

Galactomannans are hemicellulosic polysaccharides composed of a (1 â†’ 4)-linked ß-D-mannan backbone substituted with single-unit (1 â†’ 6)-α-linked D-galactosyl residues. Developing fenugreek (Trigonella foenum-graecum) seeds are known to accumulate large quantities of galactomannans in the endosperm, and were thus used here as a model system to better understand galactomannan biosynthesis and its regulation. We first verified the specific deposition of galactomannans in developing endosperms and determined that active accumulation occurred from 25 to 38 days post anthesis (DPA) under our growth conditions. We then examined the expression levels during seed development of ManS and GMGT, two genes encoding backbone and side chain synthetic enzymes. Based on transcript accumulation dynamics for ManS and GMGT, cDNA libraries were constructed using RNA isolated from endosperms at four ages corresponding to before, at the beginning of, and during active galactomannan deposition. DNA from these libraries was sequenced using the 454 sequencing technology to yield a total of 1.5 million expressed sequence tags (ESTs). Through analysis of the EST profiling data, we identified genes known to be involved in galactomannan biosynthesis, as well as new genes that may be involved in this process, and proposed a model for the flow of carbon from sucrose to galactomannans. Measurement of in vitro ManS and GMGT activities and analysis of sugar phosphate and nucleotide sugar levels in the endosperms of developing fenugreek seeds provided data consistent with this model. In vitro enzymatic assays also revealed that the ManS enzyme from fenugreek endosperm preferentially used GDP-mannose as the substrate for the backbone synthesis.


Assuntos
Endosperma/metabolismo , Etiquetas de Sequências Expressas , Mananas/biossíntese , Trigonella/embriologia , Trigonella/metabolismo , Northern Blotting , Reação em Cadeia da Polimerase em Tempo Real , Sementes/metabolismo , Trigonella/genética
12.
Bull Environ Contam Toxicol ; 88(5): 659-65, 2012 May.
Artigo em Inglês | MEDLINE | ID: mdl-22392005

RESUMO

In the present study effects of herbicides glyphosate (GP), alachlor (AL) and maleic hydrazide (MH) is studied on mitotic cells of Trigonella foenum-graecum L. Seeds of T. foenum-graecum L. treated with a series of concentrations ranging from 0.1%, 0.2%, 0.3%, 0.4% and 0.5% for 1, 2 and 6 h and their effect on mitotic index and chromosomal aberrations was studied. The results indicate that these herbicides reduced mitotic index in dose-dependent manner. In addition, increase in the percentage of abnormal mitotic plates was observed in herbicide treated groups which was both concentration and time dependent. Commonly observed abnormalities were c-mitosis, laggards, bridges, stickiness, c-anaphase, precocious separation, un-equal distribution and fragments. The result of the present investigation indicates that commonly used herbicides GP, AL and MH have significant genotoxic effect on T. foenum-graecum plant.


Assuntos
Acetamidas/toxicidade , Glicina/análogos & derivados , Herbicidas/toxicidade , Hidrazida Maleica/toxicidade , Mutagênicos/toxicidade , Trigonella/efeitos dos fármacos , Aberrações Cromossômicas/induzido quimicamente , Glicina/toxicidade , Mitose/efeitos dos fármacos , Trigonella/genética , Trigonella/crescimento & desenvolvimento
13.
Ann Bot ; 109(4): 773-82, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22186276

RESUMO

BACKGROUND AND AIMS: Satellite DNA is a genomic component present in virtually all eukaryotic organisms. The turnover of highly repetitive satellite DNA is an important element in genome organization and evolution in plants. Here we assess the presence and physical distribution of the repetitive DNA E180 family in Medicago and allied genera. Our goals were to gain insight into the karyotype evolution of Medicago using satellite DNA markers, and to evaluate the taxonomic and phylogenetic signal of a satellite DNA family in a genus hypothesized to have a complex evolutionary history. METHODS: Seventy accessions from Medicago, Trigonella, Melilotus and Trifolium were analysed by PCR to assess the presence of the repetitive E180 family, and fluorescence in situ hybridization (FISH) was used for physical mapping in somatic chromosomes. KEY RESULTS: The E180 repeat unit was PCR-amplified in 37 of 40 taxa in Medicago, eight of 12 species of Trigonella, six of seven species of Melilotus and in two of 11 Trifolium species. Examination of the mitotic chromosomes revealed that only 13 Medicago and two Trigonella species showed FISH signals using the E180 probe. Stronger hybridization signals were observed in subtelomeric and interstitial loci than in the pericentromeric loci, suggesting this satellite family has a preferential genomic location. Not all 13 Medicago species that showed FISH localization of the E180 repeat were phylogenetically related. However, nine of these species belong to the phylogenetically derived clade including the M. sativa and M. arborea complexes. CONCLUSIONS: The use of the E180 family as a phylogenetic marker in Medicago should be viewed with caution. Its amplification appears to have been produced through recurrent and independent evolutionary episodes in both annual and perennial Medicago species as well as in basal and derived clades.


Assuntos
DNA de Plantas/genética , DNA Satélite/genética , Evolução Molecular , Medicago/genética , Fluxo Gênico , Marcadores Genéticos , Melilotus/genética , Dados de Sequência Molecular , Técnicas de Amplificação de Ácido Nucleico , Filogenia , Sequências Repetitivas de Ácido Nucleico , Especificidade da Espécie , Trifolium/genética , Trigonella/genética
14.
Pak J Biol Sci ; 15(9): 426-36, 2012 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-24163952

RESUMO

Systematic breeding efforts on fenugreek have so far been neglected in Ethiopia. For this, 143 random samples of fenugreek accessions along with a commercial variety were used in this study to evaluate the potential of the land races. The field experiment was conducted at Haramaya University research station during 2011 main cropping season. Treatments were arranged in a 12x12 simple lattice design. The highest biomass and seed yielding accessions were generally concentrated more in the categories of yellow and green seed colors. When compared with the commercial variety, above 27% of the tested accessions performed significantly better in terms of seed yield indicating that significant yield gains could be secured by simple selection. However, further evaluation over wider environments is necessary to arrive at conclusive points for such quantitative traits. Green and yellow seeded accessions are widely distributed over all the country and over half of the accessions (63%) had green seed color. High seed yield bearing accessions were those collected from northwest and central part of Ethiopia, while accessions collected from eastern and northwestern Ethiopia were strikingly bold seed size. This variability would provide a basis for improving the crop in breeding program.


Assuntos
Produtos Agrícolas/crescimento & desenvolvimento , Sementes/crescimento & desenvolvimento , Trigonella/crescimento & desenvolvimento , Biomassa , Cruzamento , Cor , Etiópia , Fitoterapia , Plantas Medicinais , Seleção Genética , Fatores de Tempo , Trigonella/classificação , Trigonella/genética
15.
Planta Med ; 77(7): 765-70, 2011 May.
Artigo em Inglês | MEDLINE | ID: mdl-21104608

RESUMO

Flavonoids belong to a group of plant natural products with variable phenolic structures and play important roles in protection against biotic and abiotic stress. Fenugreek (Trigonella foenum-graecum L.) seeds and stems contain flavonol glycosides and isoflavone derivatives. Up to now, the molecular features of fenugreek flavonoid biosynthesis have not been characterized. Here we present cloning of a cDNA encoding a chalcone isomerase (namely TFGCHI-1) from the leaves of T. foenum-graecum which convert chalcones to flavanones in vitro. Transformation of Arabidopsis loss-of-function TT5 (CHI) mutant with a TFGCHI-1 cDNA complemented TT5 and produced higher levels of flavonol glycosides than wild-type Col-0.


Assuntos
Liases Intramoleculares/genética , Proteínas de Plantas/genética , Trigonella/enzimologia , Sequência de Aminoácidos , Arabidopsis/genética , Sequência de Bases , Clonagem Molecular , DNA Complementar/genética , Escherichia coli/genética , Escherichia coli/metabolismo , Flavonoides/biossíntese , Vetores Genéticos/genética , Glicosídeos/biossíntese , Liases Intramoleculares/metabolismo , Modelos Moleculares , Dados de Sequência Molecular , Filogenia , Proteínas de Plantas/metabolismo , Sementes/química , Trigonella/genética
16.
N Biotechnol ; 28(2): 110-7, 2011 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-20851781

RESUMO

Many plants used as functional foods or for medicinal purposes have been criticized for their inconsistent physiological effects. Variation in genotype and environmental conditions under which plants are produced can contribute to this inconsistency in biochemical composition. Fenugreek (Trigonella foenum-graecum L.) is a medicinal plant that not only can lower blood glucose and cholesterol levels in animals, but also can be used as a forage crop for livestock feed. Seed content for the bioactive compounds diosgenin, galactomannan and 4-hydroxyisoleucine was characterized for ten fenugreek genotypes under rainfed and irrigated conditions. High and low seed yielding genotype/environment combinations were identified that possessed distinct biochemical and seed production traits. In general high seed yielding genotype/environment combinations exhibited a more stable biochemical composition and consisted largely of irrigated fenugreek. This research indicates that comprehensive biochemical analysis of plant products would facilitate the development of more reliable produce for use by the functional food/medicinal plant industry.


Assuntos
Alimento Funcional , Plantas Medicinais/química , Trigonella/química , Ração Animal , Animais , Produtos Agrícolas , Diosgenina/análise , Humanos , Isoleucina/análogos & derivados , Isoleucina/análise , Mananas/análise , Fenótipo , Trigonella/genética
17.
J Biochem Mol Biol ; 39(3): 278-83, 2006 May 31.
Artigo em Inglês | MEDLINE | ID: mdl-16756756

RESUMO

Defensins are small cysteine rich peptides with a molecular mass of 5-10 kDa and some of them exhibit potent antifungal activity. We have cloned the coding region of a cDNA of 225 bp cysteine rich defensin, named as Tfgd1, from the legume Trigonella foenum-graecum. The amino acid sequence deduced from the coding region comprised 74 amino acids, of which the N-terminal 27 amino acids constituted the signal peptide and the mature peptide comprised 47 amino acids. The protein is characterized by the presence of eight cysteine resisdues, conserved in the various plant defensins forming four disulphide bridges, which stabilize the mature peptide. The recombinant protein expressed in E coli exhibited antifungal activity against the broad host range fungus, Rhizoctonia solani and the peanut leaf spot fungus, Phaeoisariopsis personata.


Assuntos
Ascomicetos/efeitos dos fármacos , Defensinas/genética , Defensinas/farmacologia , Rhizoctonia/efeitos dos fármacos , Trigonella , Clonagem Molecular , Defensinas/química , Testes de Sensibilidade Microbiana , Proteínas de Plantas/química , Proteínas de Plantas/genética , Proteínas de Plantas/farmacologia , Proteínas Recombinantes de Fusão/farmacologia , Análise de Sequência de DNA , Trigonella/química , Trigonella/genética , Trigonella/fisiologia
18.
BMC Plant Biol ; 4: 13, 2004 Jul 30.
Artigo em Inglês | MEDLINE | ID: mdl-15285785

RESUMO

BACKGROUND: Various species of genus Trigonella are important from medical and culinary aspect. Among these, Trigonella foenum-graecum is commonly grown as a vegetable. This anti-diabetic herb can lower blood glucose and cholesterol levels. Another species, Trigonella caerulea is used as food in the form of young seedlings. This herb is also used in cheese making. However, little is known about the genetic variation present in these species. In this report we describe the use of ISSR and RAPD markers to study genetic diversity in both, Trigonella foenum-graecum and Trigonella caerulea. RESULTS: Seventeen accessions of Trigonella foenum-graecum and nine accessions of Trigonella caerulea representing various countries were analyzed using ISSR and RAPD markers. Genetic diversity parameters (average number of alleles per polymorphic locus, percent polymorphism, average heterozygosity and marker index) were calculated for ISSR, RAPD and ISSR+RAPD approaches in both the species. Dendrograms were constructed using UPGMA algorithm based on the similarity index values for both Trigonella foenum-graecum and Trigonella caerulea. The UPGMA analysis showed that plants from different geographical regions were distributed in different groups in both the species. In Trigonella foenum-graecum accessions from Pakistan and Afghanistan were grouped together in one cluster but accessions from India and Nepal were grouped together in another cluster. However, in both the species accessions from Turkey did not group together and fell in different clusters. CONCLUSIONS: Based on genetic similarity indices, higher diversity was observed in Trigonella caerulea as compared to Trigonella foenum-graecum. The genetic similarity matrices generated by ISSR and RAPD markers in both species were highly correlated (r = 0.78 at p = 0.001 for Trigonella foenum-graecum and r = 0.98 at p = 0.001 for Trigonella caerulea) indicating congruence between these two systems. Implications of these observations in the analysis of genetic diversity and in supporting the possible Center of Origin and/or Diversity for Trigonella are discussed.


Assuntos
Variação Genética , Técnica de Amplificação ao Acaso de DNA Polimórfico , Sequências Repetitivas de Ácido Nucleico , Trigonella/genética , DNA de Plantas/genética , DNA de Plantas/isolamento & purificação , Eletroforese em Gel de Ágar , Marcadores Genéticos/genética , Filogenia , Análise de Regressão , Especificidade da Espécie , Trigonella/classificação
19.
Plant Physiol ; 131(3): 1487-95, 2003 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-12644698

RESUMO

Galactomannans [(1-->6)-alpha-D-galactose (Gal)-substituted (1-->4)-beta-D-mannans] are major cell wall storage polysaccharides in the endosperms of some seeds, notably the legumes. Their biosynthesis in developing legume seeds involves the functional interaction of two membrane-bound glycosyltransferases, mannan synthase (MS) and galactomannan galactosyltransferase (GMGT). MS catalyzes the elongation of the mannan backbone, whereas GMGT action determines the distribution and amount of Gal substitution. Fenugreek (Trigonella foenum-graecum) forms a galactomannan with a very high degree of Gal substitution (Man/Gal = 1.1), and its GMGT has been characterized. We now report that the endosperm cell walls of the tobacco (Nicotiana tabacum) seed are rich in a galactomannan with a very low degree of Gal substitution (Man/Gal about 20) and that its depositional time course is closely correlated with membrane-bound MS and GMGT activities. Furthermore, we demonstrate that seeds from transgenic tobacco lines that express fenugreek GMGT constitutively in membrane-bound form have endosperm galactomannans with increased average degrees of Gal substitution (Man/Gal about 10 in T(1) generation seeds and about 7.5 in T(2) generation seeds). Membrane-bound enzyme systems from transgenic seed endosperms form galactomannans in vitro that are more highly Gal substituted than those formed by controls under identical conditions. To our knowledge, this is the first report of structural manipulation of a plant cell wall polysaccharide in transgenic plants via a biosynthetic membrane-bound glycosyltransferase.


Assuntos
Galactosiltransferases/metabolismo , Mananas/biossíntese , Sementes/genética , Tabaco/enzimologia , Trigonella/genética , Parede Celular/genética , Parede Celular/metabolismo , Galactose/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Regulação Enzimológica da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Teste de Complementação Genética , Mananas/metabolismo , Mutação , Plantas Geneticamente Modificadas , Sementes/enzimologia , Sementes/crescimento & desenvolvimento , Tabaco/genética , Tabaco/crescimento & desenvolvimento , Trigonella/enzimologia
20.
J Agric Food Chem ; 50(21): 5994-7, 2002 Oct 09.
Artigo em Inglês | MEDLINE | ID: mdl-12358471

RESUMO

A collection of 10 accessions of fenugreek (Trigonella foenum-graecum L.), an annual legume, was grown during two summers at three plot locations in western Canada to assess whether genetic (accession) and environmental factors (site and year of production) influenced levels of diosgenin, a steroidal sapogenin. The 60 harvested seed samples, each analyzed by single determinations on three subsamples of defatted and dried seed material, were hydrolyzed by a microscale procedure in water containing 2-propanol (70%) and sulfuric acid (1 M). The extracts were analyzed by capillary gas chromatography with 6-methyldiosgenin as internal standard. Diosgenin levels from mature seeds ranged from 0.28 to 0.92% (28-92 microg/10 mg). Analysis of variance on combined diosgenin levels from the three sites and two years revealed that accession, accession x year, and site x year effects were significant for diosgenin content, whereas site, year, and site x accession effects were not. Four accessions, CN 19062, CN 19067, CN 19070, and CN 19071, were identified with high levels of diosgenin on the basis of the 2-year data set. In these accessions, mean levels of diosgenin plus yamogenin from seven site years were estimated at 0.70, 0.98, 0.84, and 0.87%, respectively.


Assuntos
Diosgenina/análise , Sementes/química , Trigonella/química , 2-Propanol , Canadá , Cromatografia Gasosa , Meio Ambiente , Manipulação de Alimentos , Hidrólise , Especificidade da Espécie , Ácidos Sulfúricos , Trigonella/genética
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