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1.
Int J Environ Health Res ; 34(2): 1100-1112, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-37036296

RESUMO

Tetraclinis articulata essential oil proved to be effective in controlling tomato grey mould, so we would investigate its effect on some tomato defense mechanisms. The pretreatment of Botrytis cinerea infected tomato plants with TAEO emulsion enhanced the activity of antioxidant enzymes activity SOD, CAT, APX, and GPX, and total polyphenols content and it decreased IC50 of free radical-scavenging activity and H2O2 content. Results showed amelioration in antioxidant status in TAEO emulsion treated and B. cinerea infected plants indicating that treatment decreased infection in tomato plants. The qRT-PCR analysis of defense genes expression Chitinase SlChi, transcription factors SlWRKY and SlAP2/ERF, Lipoxygenase SlLOX, and Thioredoxin SlTRX showed that they were up-regulated as early as 12 hpi sustained with a second increase at 48 hpi in TAEO emulsion pretreated and infected plants. These results suggest the potential use of TAEO emulsion as natural product to induce tomato antioxidant status and activate defense genes.


Assuntos
Fungicidas Industriais , Óleos Voláteis , Solanum lycopersicum , Fungicidas Industriais/toxicidade , Óleos Voláteis/farmacologia , Peróxido de Hidrogênio , Emulsões , Antioxidantes/farmacologia , Fungos , Doenças das Plantas/prevenção & controle , Doenças das Plantas/genética
2.
Physiol Mol Biol Plants ; 27(1): 135-150, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33627968

RESUMO

Faba bean (Vicia faba L.) is the major food legume crop in Tunisia. However, its growth and yield is strongly affected by water-limited environments. In this study, osmotic stress exhibited a negative effect on Bachar and Badii cultivar. Nevertheless, the deteriorating effects of osmotic stress were relatively low on studied parameters of Bachar due to its better efficiency to reduce oxidative damage by increasing enzymatic activities such as catalase (CAT), superoxide dismutase (SOD) and ascorbate peroxidase (APX), accumulation of total chlorophyll (Chlt), soluble sugars and leaf relative water content (RWC). GC-MS analysis determined a total of 11 soluble carbohydrates induced by osmotic stress and differentially accumulated in the both cultivars. Bachar showed elevated levels of mannose, glucose, galactose, ribose, rhamnose and myo-inositol which might help to maintain osmotic adjustment, membranes and proteins protection from the damaging effect of reactive oxygen species. Sugar metabolism related genes (VfNINV3, VfPHS2, VfFRK4, VfHXK1, VfGPI1, VfSTP1.1, VfpGlcT1.1, VfSTP5.1, VfpGlcT1.2, VfSWEET2.1, VfVINV2, VfSUS1, VfPGM1, VfSUT1.1, VfGPT1, VfSPS1, VfSPP1, VfPHS1, VfSUT4.1 and VfTMT1.1) were differentially expressed in both cultivars demonstrating their important roles in sugar accumulation. Most of these genes were upregulated in the leaves of Bachar under moderate and severe stress, which could lead to increase glycolysis and tricarboxylic acid cycle in order to accelerate energy production, necessary to increase osmotic regulation and consequently enhancing the osmotic stress tolerance in that cultivar. Overall, sugars accumulation ability can be used as a useful indicator for the osmotic stress tolerant potential in faba bean breeding programs. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at (10.1007/s12298-021-00935-1).

3.
Physiol Mol Biol Plants ; 26(6): 1173-1186, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32549681

RESUMO

Drought stress is one of the most prevalent environmental factors limiting faba bean (Vicia faba L.) crop productivity. ß-aminobutyric acid (BABA) is a non-protein amino acid that may be involved in the regulation of plant adaptation to drought stress. The effect of exogenous BABA application on physiological, biochemical and molecular responses of faba bean plants grown under 18% PEG-induced drought stress were investigated. The results showed that the application of 1 mM of BABA improved the drought tolerance of faba bean. The application of BABA increased the leaf relative water content, leaf photosynthesis rate (A), transpiration rate (E), and stomatal conductance (gs), thereby decreased the water use efficiency. Furthermore, exogenous application of BABA decreased production of hydrogen peroxide (H2O2), malondialdehyde and electrolyte leakage levels, leading to less cell membrane damage due to oxidative stress. Regarding osmoprotectants, BABA application enhanced the accumulation of proline, and soluble sugars, which could improve the osmotic adjustment ability of faba bean under drought challenge. Interestingly, mended antioxidant enzyme activities like catalase, guaiacol peroxidase, ascorbate peroxidase and superoxide dismutase and their transcript levels may lead to counteract the damaging effects of oxidative stress and reducing the accumulation of harmful substances in BABA-treated faba bean plants. In addition, exogenous BABA significantly induced the accumulation of drought tolerance-related genes like VfMYB, VfDHN, VfLEA, VfERF, VfNCED, VfWRKY, VfHSP and VfNAC in leaves and roots, suggesting that BABA might act as a signal molecule to regulate the expression of drought tolerance-related genes.

5.
BMC Plant Biol ; 18(1): 262, 2018 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-30382818

RESUMO

BACKGROUND: Celiac disease (CD) is an autoimmune disorder affecting genetically predisposed individuals whose dietary gluten proteins trigger an inflammatory reaction in the small intestine. Gluten is found in the seeds of cereals like bread wheat (Triticum aestivum ssp. aestivum) and spelt (Triticum aestivum ssp. spelta). The development of new varieties lacking immunogenic peptides is one of the strategies currently investigated to address the CD problem. Among gluten proteins, α-gliadins display the strongest immunogenicity with four main T-cell stimulatory epitopes. The objective of this work was to study the expression of α-gliadin epitopes related to CD in a wide collection of 121 spelt accessions (landraces and varieties, spring and winter accessions) from different provenances, and to analyze the correlation between the presence of epitope sequences in gDNA and their expression (cDNA). The effect of environmental factors (harvest year and N fertilization) on the epitope expression was also investigated. RESULTS: TaqMan probes targeting the canonical form of the epitopes were used to evaluate the epitope expression levels. Significant variations in the amount of epitope transcripts were identified between accessions and according to the provenances. Spring accessions showed a significantly higher immunogenicity than winter ones and no influence of spelt breeding on the epitope expression levels could be assessed when comparing landraces and varieties from Northwestern Europe. No correlation was observed between quantitative PCR results obtained from cDNA and gDNA for 45 accessions tested, stressing the need to use markers focusing on epitope transcripts rather than on genomic sequences. A relative stability of the amount of epitopes expressed by a same accession across four harvest years was detected. The fertilization strategy, evaluated through seven N fertilization modalities applied to two commercial spelt varieties, did not influence the epitope expression of the first variety, whereas it had a slight effect for the second one. CONCLUSIONS: The results obtained in this work showed that the CD-related epitope expression greatly fluctuated among the spelt accessions studied. This expression was not correlated to the epitope genomic occurrence and environmental factors had almost no influence on the amount of epitope transcripts.


Assuntos
Epitopos/genética , Gliadina/imunologia , Triticum/genética , Doença Celíaca/etiologia , Doença Celíaca/imunologia , Fertilizantes , Regulação da Expressão Gênica de Plantas , Gliadina/genética , Humanos , Nitrogênio/metabolismo , Reação em Cadeia da Polimerase/métodos , Triticum/imunologia
6.
Mol Plant Microbe Interact ; 30(11): 855-865, 2017 11.
Artigo em Inglês | MEDLINE | ID: mdl-28726589

RESUMO

Invasive plant pathogens have developed the ability to modify the metabolism of their host, promoting metabolic processes that facilitate the growth of the pathogen at the general expense of the host. The particular enzymatic process SUMOylation, which performs posttranslational modification of target proteins, leading to changes in many aspects of protein activity and, hence, metabolism, has been demonstrated to be active in many eukaryotic organisms, both animals and plants. Here, we provide experimental evidence that indicates that, in leaves of Solanum tuberosum that have been infected by Phytophthora infestans, the SUMO (small ubiquitin-like modifier) pathway enzymes of the host are partially under transcriptional control exerted by the oomycete. Using a recently developed approach that employs three-dimensional gels, we show that, during the infection process, the abundances of most of the known SUMO conjugates of S. tuberosum change significantly, some decreasing, but many increasing in abundance. The new proteomic approach has the potential to greatly facilitate investigation of the molecular events that take place during the invasion by a pathogen of its host plant.


Assuntos
Interações Hospedeiro-Patógeno , Phytophthora infestans/fisiologia , Proteômica/métodos , Solanum tuberosum/metabolismo , Solanum tuberosum/microbiologia , Sumoilação , Evolução Molecular , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Genótipo , Interações Hospedeiro-Patógeno/genética , Doenças das Plantas/microbiologia , Folhas de Planta/metabolismo , Folhas de Planta/microbiologia , Proteínas de Plantas/metabolismo , Proteoma/metabolismo , Solanum tuberosum/genética , Fatores de Tempo
7.
Physiol Mol Biol Plants ; 23(2): 397-409, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-28461727

RESUMO

The genetic diversity of 21 faba bean populations was examined using morphological and molecular markers. DNA was extracted from 189 individuals and 8 microsatellite markers were genotyped individually in these 21 populations. A total of 53 alleles were obtained in all populations, with an average of 6.62 alleles per locus. The expected and observed heterozygosity was 0.38 and 0.62 respectively. The average polymorphism index content of SSR markers was 0.61, ranging from 0.31 to 0.81. The unweighted pair group method with arithmetic mean dendrogram clustered all the populations into two groups, each for them subdivided into 3 sub-groups according to geographical origin. Morphological variation showed that the populations were not grouped according to their geographical origin. Therefore, patterns of differentiation of morphological traits did not coincide with molecular differentiation, indicating that morphological variation does not reflect genetic subdivision in studied faba bean populations. Analysis of molecular variance revealed high levels of genetic variation (83%) within population and provides a good base for designing genetic improvement programs. The result of Principal Component Analysis (PCA) revealed that three dimensional principal components (PC1, PC2 and PC3) contributed 40.56% of the total variability and accounted with values of 20.64, 11.22 and 8.70%, respectively. Cluster analysis based on PCA indicated three separate groups of populations. The genetic relationships found between the 21 populations samples were the same in both the PCA and STRUCTURE analysis which support the results observed. These data may serve as a foundation for the development of faba bean breeding programs.

8.
Plant Cell Environ ; 39(1): 62-79, 2016 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-26082265

RESUMO

To counter environmental cues, cultivated tomato (Solanum lycopersicum L.) has evolved adaptive mechanisms requiring regulation of downstream genes. The dehydration-responsive element-binding protein 2 (DREB2) transcription factors regulate abiotic stresses responses in plants. Herein, we isolated a novel DREB2-type regulator involved in salinity response, named SlDREB2. Spatio-temporal expression profile together with investigation of its promoter activity indicated that SlDREB2 is expressed during early stages of seedling establishment and in various vegetative and reproductive organs of adult plants. SlDREB2 is up-regulated in roots and young leaves following exposure to NaCl, but is also induced by KCl and drought. Its overexpression in WT Arabidopsis and atdreb2a mutants improved seed germination and plant growth in presence of different osmotica. In tomato, SlDREB2 affected vegetative and reproductive organs development and the intronic sequence present in the 5' UTR drives its expression. Physiological, biochemical and transcriptomic analyses showed that SlDREB2 enhanced plant tolerance to salinity by improvement of K(+) /Na(+) ratio, and proline and polyamines biosynthesis. Exogenous hormonal treatments (abscisic acid, auxin and cytokinins) and analysis of WT and 35S::SlDREB2 tomatoes hormonal contents highlighted SlDREB2 involvement in abscisic acid biosynthesis/signalling. Altogether, our results provide an overview of SlDREB2 mode of action during early salt stress response.


Assuntos
Regulação da Expressão Gênica de Plantas , Solanum lycopersicum/genética , Fatores de Transcrição/metabolismo , Transcriptoma , Ácido Abscísico/farmacologia , Arabidopsis/genética , Arabidopsis/fisiologia , Sequência de Bases , Desidratação , Secas , Perfilação da Expressão Gênica , Solanum lycopersicum/fisiologia , Dados de Sequência Molecular , Mutação , Análise de Sequência com Séries de Oligonucleotídeos , Reguladores de Crescimento de Plantas/farmacologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raízes de Plantas/genética , Raízes de Plantas/fisiologia , Plantas Geneticamente Modificadas , Tolerância ao Sal , Plântula/genética , Plântula/fisiologia , Análise de Sequência de DNA , Cloreto de Sódio/farmacologia , Estresse Fisiológico , Fatores de Transcrição/genética
9.
BMC Plant Biol ; 15: 85, 2015 Mar 12.
Artigo em Inglês | MEDLINE | ID: mdl-25888402

RESUMO

BACKGROUND: Cytokinins (CKs) are involved in response to various environmental cues, including salinity. It has been previously reported that enhancing CK contents improved salt stress tolerance in tomato. However, the underlying mechanisms of CK metabolism and signaling under salt stress conditions remain to be deciphered. RESULTS: Two tomato isopentenyltransferases, SlIPT3 and SlIPT4, were characterized in tomato and Arabidopsis. Both proteins displayed isopentenyltransferase (IPT) activity in vitro, while their encoding genes exhibited different spatio-temporal expression patterns during tomato plant development. SlIPT3 and SlIPT4 were affected by the endogenous CK status, tightly connected with CKs feedback regulation, as revealed by hormonal treatements. In response to salt stress, SlIPT3 and SlIPT4 were strongly repressed in tomato roots, and differently affected in young and old leaves. SlIPT3 overexpression in tomato resulted in high accumulation of different CK metabolites, following modifications of CK biosynthesis-, signaling- and degradation-gene expression. In addition, 35S::SlIPT3 tomato plants displayed improved tolerance to salinity consecutive to photosynthetic pigments and K(+)/Na(+) ratio retention. Involvement of SlIPT3 and SlIPT4 in salt stress response was also observed in Arabidopsis ipt3 knock-out complemented plants, through maintenance of CK homeostasis. CONCLUSIONS: SlIPT3 and SlIPT4 are functional IPTs encoded by differently expressed genes, distinctively taking part in the salinity response. The substantial participation of SlIPT3 in CK metabolism during salt stress has been determined in 35S::SlIPT3 tomato transformants, where enhancement of CKs accumulation significantly improved plant tolerance to salinity, underlining the importance of this phytohormone in stress response.


Assuntos
Alquil e Aril Transferases/fisiologia , Arabidopsis/fisiologia , Citocininas/metabolismo , Regulação da Expressão Gênica de Plantas , Tolerância ao Sal , Solanum lycopersicum/enzimologia , Solanum lycopersicum/fisiologia , Alquil e Aril Transferases/genética , Arabidopsis/genética , Solanum lycopersicum/embriologia , Solanum lycopersicum/genética , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/fisiologia
10.
Plant Physiol ; 164(4): 1967-90, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24567191

RESUMO

The zinc finger superfamily includes transcription factors that regulate multiple aspects of plant development and were recently shown to regulate abiotic stress tolerance. Cultivated tomato (Solanum lycopersicum Zinc Finger2 [SIZF2]) is a cysteine-2/histidine-2-type zinc finger transcription factor bearing an ERF-associated amphiphilic repression domain and binding to the ACGTCAGTG sequence containing two AGT core motifs. SlZF2 is ubiquitously expressed during plant development, and is rapidly induced by sodium chloride, drought, and potassium chloride treatments. Its ectopic expression in Arabidopsis (Arabidopsis thaliana) and tomato impaired development and influenced leaf and flower shape, while causing a general stress visible by anthocyanin and malonyldialdehyde accumulation. SlZF2 enhanced salt sensitivity in Arabidopsis, whereas SlZF2 delayed senescence and improved tomato salt tolerance, particularly by maintaining photosynthesis and increasing polyamine biosynthesis, in salt-treated hydroponic cultures (125 mm sodium chloride, 20 d). SlZF2 may be involved in abscisic acid (ABA) biosynthesis/signaling, because SlZF2 is rapidly induced by ABA treatment and 35S::SlZF2 tomatoes accumulate more ABA than wild-type plants. Transcriptome analysis of 35S::SlZF2 revealed that SlZF2 both increased and reduced expression of a comparable number of genes involved in various physiological processes such as photosynthesis, polyamine biosynthesis, and hormone (notably ABA) biosynthesis/signaling. Involvement of these different metabolic pathways in salt stress tolerance is discussed.


Assuntos
Arabidopsis/fisiologia , Proteínas de Plantas/metabolismo , Proteínas Repressoras/metabolismo , Tolerância ao Sal , Solanum lycopersicum/fisiologia , Ácido Abscísico/metabolismo , Ácido Abscísico/farmacologia , Sequência de Aminoácidos , Arabidopsis/efeitos dos fármacos , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Hidroponia , Solanum lycopersicum/efeitos dos fármacos , Solanum lycopersicum/genética , Solanum lycopersicum/crescimento & desenvolvimento , Dados de Sequência Molecular , Análise de Sequência com Séries de Oligonucleotídeos , Pressão Osmótica , Fotossíntese/efeitos dos fármacos , Fotossíntese/genética , Reguladores de Crescimento de Plantas/farmacologia , Proteínas de Plantas/química , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Poliaminas/metabolismo , Proteínas Repressoras/química , Proteínas Repressoras/genética , Salinidade , Tolerância ao Sal/efeitos dos fármacos , Tolerância ao Sal/genética , Transdução de Sinais , Cloreto de Sódio/farmacologia , Transcrição Gênica/efeitos dos fármacos , Regulação para Cima/efeitos dos fármacos , Regulação para Cima/genética
11.
BMC Genomics ; 15: 1043, 2014 Nov 29.
Artigo em Inglês | MEDLINE | ID: mdl-25433532

RESUMO

BACKGROUND: Scab, caused by the fungus Venturia inaequalis, is one of the most important diseases of cultivated apple. While a few scab resistance genes (R genes) governing qualitative resistance have been isolated and characterized, the biological roles of genes governing quantitative resistance, supposed to be more durable, are still unknown. This study aims to investigate the molecular mechanisms involved in the partial resistance of the old Belgian apple cultivar 'Président Roulin' against V. inaequalis. RESULTS: A global gene expression analysis was conducted in 'Président Roulin' (partially resistant) and in 'Gala' (susceptible) challenged by V. inaequalis by using the cDNA-AFLP method (cDNA-Amplified Fragment Length Polymorphism). Transcriptome analysis revealed significant modulation (up- or down-regulation) of 281 out of approximately 20,500 transcript derived fragments (TDFs) in 'Président Roulin' 48 hours after inoculation. Sequence annotation revealed similarities to several genes encoding for proteins belonging to the NBS-LRR and LRR-RLK classes of plant R genes and to other defense-related proteins. Differentially expressed genes were sorted into functional categories according to their gene ontology annotation and this expression signature was compared to published apple cDNA libraries by Gene Enrichment Analysis. The first comparison was made with two cDNA libraries from Malus x domestica uninfected leaves, and revealed in both libraries a signature of enhanced expression in 'Président Roulin' of genes involved in response to stress and photosynthesis. In the second comparison, the pathogen-responsive TDFs from the partially resistant cultivar were compared to the cDNA library from inoculated leaves of Rvi6 (HcrVf2)-transformed 'Gala' lines (complete disease resistance) and revealed both common physiological events, and notably differences in the regulation of defense response, the regulation of hydrolase activity, and response to DNA damage. TDFs were in silico mapped on the 'Golden Delicious' apple reference genome and significant co-localizations with major scab R genes, but not with quantitative trait loci (QTLs) for scab resistance nor resistance gene analogues (RGAs) were found. CONCLUSIONS: This study highlights possible candidate genes that may play a role in the partial scab resistance mechanisms of 'Président Roulin' and increase our understanding of the molecular mechanisms involved in the partial resistance against apple scab.


Assuntos
Resistência à Doença/genética , Perfilação da Expressão Gênica , Interações Hospedeiro-Patógeno/genética , Malus/genética , Doenças das Plantas/genética , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados , Ascomicetos , Impressões Digitais de DNA , DNA Complementar , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Malus/metabolismo , Malus/microbiologia , Anotação de Sequência Molecular , Dados de Sequência Molecular , Estresse Oxidativo , Doenças das Plantas/microbiologia , Locos de Características Quantitativas , Análise de Sequência de DNA
12.
Mol Biol Rep ; 41(9): 6181-94, 2014 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24973883

RESUMO

Fusarium head blight (FHB), predominantly caused by Fusarium graminearum, is a devastating disease that poses a serious threat to wheat (Triticum aestivum L.) production worldwide. A suppression subtractive hybridization cDNA library was constructed from F. graminearum infected spikes of a resistant Belgian winter wheat, Centenaire, exhibiting Type II resistance to FHB in order to identify differentially expressed members of full-size ABCG family. Members of the ABCG family are pleiotropic drug transporters allowing the movement of structurally unrelated metabolites, including pathogens-derived virulent compounds, across biological membranes and could be potentially involved in resistance to plant pathogens. In this study, five new full-size ABCG transporter expressed sequence tags TaABCG2, TaABCG3, TaABCG4, TaABCG5 and TaABCG6 have been identified. Time-course gene expression profiling between the FHB resistant Centenaire and the susceptible Robigus genotype showed that the newly isolated transcripts were differentially expressed up to 72 h-post inoculation. The respective genes encoding these transcripts were mapped to corresponding wheat chromosomes or chromosomal arms known to harbor quantitative trait loci for FHB resistance. Interestingly, these ABCG transcripts were also induced by deoxynivalenol (DON) treatment of germinating wheat seeds and the toxin treatment inhibited root and hypocotyl growth. However, the hypocotyl of the FHB resistant cultivar Centenaire was less affected than that of the susceptible cultivar Robigus, reflecting more likely the genotype-dependent differential expression pattern of the identified ABCG genes. This work emphasizes the potential involvement of ABCG transporters in wheat resistance to FHB, at least in part through the detoxification of the pathogen-produced DON.


Assuntos
Transportadores de Cassetes de Ligação de ATP/genética , Fusarium/metabolismo , Regulação da Expressão Gênica de Plantas , Tricotecenos/metabolismo , Triticum/genética , Sequência de Aminoácidos , Mapeamento Cromossômico , Cromossomos de Plantas/genética , Etiquetas de Sequências Expressas , Perfilação da Expressão Gênica , Biblioteca Gênica , Interações Hospedeiro-Patógeno/genética , Dados de Sequência Molecular , Filogenia , Doenças das Plantas/microbiologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Locos de Características Quantitativas , RNA de Plantas/genética , Triticum/microbiologia , Regulação para Cima
13.
J Microbiol Methods ; 217-218: 106890, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38272400

RESUMO

Research into phytase production is useful for improving the efficiency of animal production, reducing environmental impact, and contributing to the development of sustainable and efficient animal production systems. This study aims to investigate the potential of yeast strains for phytase biosynthesis in nutrient media. Phytase is a phosphomonoesterase (E.C 3.1.3.8) catalyzing in a ladder-like manner the dephosphorylation of phytic acid and its salts, with various resulting myo-inositol phosphates and phosphoric acid. Yeasts of the genera Saccharomyces, Zygosaccharomyces, Candida, and Pichia were evaluated in a two-step screening procedure for phytase production. One hundred and eighteen strains were screened in the first stage, which was conducted on four types of solid culture media containing calcium phytate as the selected background. On PSM medium, many strains were found to form halos as early as the 24th hour of development. Several strains with significant potential for enzyme production were evaluated in the second step of the screening. It was conducted in a liquid culture medium. In conclusion, the strain C. melibiosica 2491 was selected for further studies when cultured in a YPglu culture medium. Further research will focus on finding suitable conditions that increase the biosynthesis of the enzyme, which is of significant technological and practical interest for animal nutrition.


Assuntos
6-Fitase , Saccharomyces cerevisiae , Animais , Pichia , Candida , Fosfatos de Inositol , Ácido Fítico
14.
Mol Biol Rep ; 40(10): 5883-906, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-24078158

RESUMO

The physiological, biochemical and molecular mechanisms regulating the initiation of a regenerative pathway remain partially unknown. Efforts to identify the biological features that confer transformation ability, or the tendency of some cells to induce transgene silencing, would help to improve plant genetic engineering. The objective of our study was to monitor the evolution of plant cell competencies in relation to both in vitro tissue culture regeneration and the genetic transformation properties. We used a simple wheat regeneration procedure as an experimental model for studying the regenerative capacity of plant cells and their receptivity to direct gene transfer over the successive steps of the regenerative pathway. Target gene profiling studies and biochemical assays were conducted to follow some of the mechanisms triggered during the somatic-to-embryogenic transition (i.e. dedifferentiation, cell division activation, redifferentiation) and affecting the accessibility of plant cells to receive and stably express the exogenous DNA introduced by bombardment. Our results seem to indicate that the control of cell-cycle (S-phase) and host defense strategies can be crucial determinants of genetic transformation efficiency. The results from studies conducted at macro-, micro- and molecular scales are then integrated into a holistic approach that addresses the question of tissue culture and transgenesis competencies more broadly. Through this multilevel analysis we try to establish functional links between both regenerative capacity and transformation receptiveness, and thereby to provide a more global and integrated vision of both processes, at the core of defense/adaptive mechanisms and survival, between undifferentiated cell proliferation and organization.


Assuntos
Perfilação da Expressão Gênica , Genes de Plantas/genética , Regeneração/genética , Sementes/genética , Transformação Genética , Triticum/embriologia , Triticum/genética , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Triticum/enzimologia
15.
Mol Biol Rep ; 40(2): 1569-77, 2013 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-23086274

RESUMO

Somatic embryogenesis is a useful tool of plant breeding. In this context, a procedure for inducing somatic embryogenesis in Prunus incisa leaf explants had been previously developed. The original in vitro protocol relies on picloram treatments and exposure to darkness as inductive conditions, the best frequency of embryogenesis being obtained on the second leaf (F(2)) exposed to 4 µM picloram during 30 days. The morphological and biochemical changes observed during somatic embryogenesis occur in response to alterations in gene expression regulation patterns. A molecular study was conducted in order to provide deeper insight into the fundamental biological factors involved in the induction of this process using a gene candidate strategy and semi-quantitative reverse transcription polymerase chain reaction analysis. So far, no sequence data related to somatic embryogenesis has been available in cherry. In the present study, we cloned and sequenced cDNA fragments of putative genes encoding auxin-binding protein, cell cycle regulator and somatic embryogenesis receptor kinase. Time-course differential transcript accumulations were observed for all investigated genes in leaves or derived callus tissues during the observation period (first month of culture). Their possible involvement in the sequential steps of the embryogenic pathway (dedifferentiation, cell proliferation, differentiation through somatic embryogenesis) is presented and discussed.


Assuntos
Folhas de Planta/metabolismo , Proteínas de Plantas/genética , Prunus/metabolismo , Sequência de Aminoácidos , Sequência de Bases , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/metabolismo , Clonagem Molecular , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Proteína Quinase 6 Ativada por Mitógeno/genética , Proteína Quinase 6 Ativada por Mitógeno/metabolismo , Dados de Sequência Molecular , Folhas de Planta/embriologia , Folhas de Planta/genética , Proteínas de Plantas/metabolismo , Técnicas de Embriogênese Somática de Plantas , Prunus/embriologia , Prunus/genética , Análise de Sequência de DNA , Homologia de Sequência
16.
Methods Mol Biol ; 2967: 75-83, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37608104

RESUMO

Cocoa (Theobroma cacao L.) is an international commodity used as an ingredient in the manufacturing of chocolate making its authentication a key issue in the cocoa chain. Various molecular techniques have been increasingly applied for quality requirements. These issues highlight the need for techniques that allow the extraction and detection of cocoa DNA from highly processed cocoa products and chocolate. The applicability of real-time PCR to highly processed cocoa-derived products for authentication purposes depends on the possibility of extracting high-quality and amplifiable DNA and further developing efficient PCR tests. This methodology herein describes the use of a classical CTAB method providing DNA suitable for TaqMan real-time PCR amplification. Real-time PCR is a simple and fast method, with a high potential application in a wide range of food products. The main features of this technique are focused on two DNA targets, one located in the nuclear genome (vicilin-li PCR test) and a second one based on chloroplast DNA (lipids PCR test), which successfully passed the performance criteria considering the specificity, sensitivity, efficiency of amplification, robustness, and applicability in processed cocoa-derived products and chocolate.


Assuntos
Cacau , Chocolate , Cacau/genética , Reação em Cadeia da Polimerase em Tempo Real , Alimentos , Comércio
17.
Foods ; 12(11)2023 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-37297478

RESUMO

The present work is focused on the physicochemical characteristics, chemical composition, and some biological activities of Koelreuteria paniculata seed oil. The glyceride oil, obtained with a Soxhlet apparatus by extraction with hexane, was characterized by a relatively high oil content (over 20%), and it is defined as a non-drying oil (iodine value-44 gI2/100 g) with good oxidative stability (over 50 h). There were identified 11 fatty acids, 6 sterols, 3 tocopherols, and 6 phospholipids, as the last group was reported for the first time. The major components among them were-monounsaturated eicosenoic and oleic acids, ß-sitosterol, ß-tocopherol, and phosphatidylcholine. The in vitro tests demonstrated DNA protective activity and a lack of cytotoxicity of the oil, data that has been reported for the first time. The in vitro MTT test of the oil on HT-29 and PC3 cell lines did not indicate antitumor activity. The seed oil studied contains valuable bio-components, which have proven benefits for human health, and that is why it could be used in food, cosmetic, and pharmaceutical products.

18.
Plants (Basel) ; 12(4)2023 Feb 17.
Artigo em Inglês | MEDLINE | ID: mdl-36840268

RESUMO

Invasive species as sources of natural components are of increasing interest for scientific research. This is the case of Ailanthus altissima, which belongs to the top 100 of the most dangerous invasive plant species in Europe, and which is the subject of the present study. The purpose of the research was to analyze the main phenolic compounds in the flowers, leaves, and stem bark of A. altissima and determine the DNA-protective and antioxidant potential of their ethanolic extracts. HPLC profiling revealed the presence of 6 flavonoids and 10 phenolic acids, of which 15 were found in flowers, 14 in leaves, and 11 in the stem bark. Rutin (5.68 mg/g dw in flowers), hesperidin (2.67 mg/g dw in leaves) and (+)-catechin (2.15 mg/g dw in stem bark) were the best-represented flavonoids. Rosmarinic (10.32 mg/g dw in leaves) and salicylic (6.19 mg/g dw in leaves) acids were predominant among phenolic acids. All plant extracts tested showed in vitro antioxidant activity (determined by DPPH, ABTS, FRAP, and CUPRAC assays) and DNA-protection capacity (assay with supercoiled plasmid DNA-pUC19). The highest antioxidant activity was recorded in the flower parts (in the range from 661 to 893 mmol TE/g dw), followed by the leaves. A DNA protective potential for A. altissima leaf and flower extracts has not been established to date. In addition, the main microscopic diagnostic features of studied plant substances were described, with data for the flower parts being reported for the first time. The present study proves that A. altissima could be a natural source of DNA protection and antioxidants.

19.
Biology (Basel) ; 12(4)2023 Apr 03.
Artigo em Inglês | MEDLINE | ID: mdl-37106747

RESUMO

Application of date palm waste compost is quite beneficial in improving soil properties and crop growth. However, the effect of its application on soil microbial communities is less understood. High-throughput sequencing and quantitative real-time PCR (qPCR) were used to evaluate the effect of compost application on the soil microbial composition in a barley field during the tillering, booting and ripening stages. The results showed that compost treatment had the highest bacterial and fungal abundance, and its application significantly altered the richness (Chao1 index) and α-diversity (Shannon index) of fungal and bacterial communities. The dominant bacterial phyla found in the samples were Proteobacteria and Actinobacteria while the dominant fungal orders were Ascomycota and Mortierellomycota. Interestingly, compost enriched the relative abundance of beneficial microorganisms such as Chaetomium, Actinobacteriota, Talaromyces and Mortierella and reduced those of harmful microorganisms such as Alternaria, Aspergillus and Neocosmospora. Functional prediction based on Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) showed that amplicon sequence variant (ASV) sequences related to energy metabolism, amino acid metabolism and carbohydrate metabolism were associated with compost-treated soil. Based on Fungi Functional Guild (FUNGuild), identified fungi community metabolic functions such as wood saprotroph, pathotroph, symbiotroph and endophyte were associated with compost-treated soil. Overall, compost addition could be considered as a sustainable practice for establishing a healthy soil microbiome and subsequently improving the soil quality and barley crop production.

20.
Plants (Basel) ; 12(3)2023 Jan 23.
Artigo em Inglês | MEDLINE | ID: mdl-36771612

RESUMO

Proteomic analysis was performed to investigate the differentially abundant proteins (DAPs) in barley roots during the tillering stage. Bioinformatic tools were used to interpret the biological function, the pathway analysis and the visualisation of the network amongst the identified proteins. A total of 72 DAPs (33 upregulated and 39 downregulated) among a total of 2580 proteins were identified in response to compost treatment, suggesting multiple pathways of primary and secondary metabolism, such as carbohydrates and energy metabolism, phenylpropanoid pathway, glycolysis pathway, protein synthesis and degradation, redox homeostasis, RNA processing, stress response, cytoskeleton organisation, and phytohormone metabolic pathways. The expression of DAPs was further validated by qRT-PCR. The effects on barley plant development, such as the promotion of root growth and biomass increase, were associated with a change in energy metabolism and protein synthesis. The activation of enzymes involved in redox homeostasis and the regulation of stress response proteins suggest a protective effect of compost, consequently improving barley growth and stress acclimation through the reduction of the environmental impact of productive agriculture. Overall, these results may facilitate a better understanding of the molecular mechanism of compost-promoted plant growth and provide valuable information for the identification of critical genes/proteins in barley as potential targets of compost.

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