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1.
Biochem Genet ; 61(3): 879-900, 2023 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-36214954

RESUMEN

Since the late 1980s, the oak decline has affected the Zagros oak forests in western Iran. Persian oak (Quercus brantii L.) the most important tree species of these forests has been damaged more than any other plant species. In the present study, the RNA sequencing technique was used for the first time to identify key genes and molecular mechanisms involved in Persian oak decline. The RNA was extracted from the leaves of healthy and declined oak trees, and sequenced using the Illumina HiSeq 2500 platform (2 × 150 bp paired-end reads). De novo transcriptome assembly of Persian oak revealed 56,743 unigenes and 6049 differentially expressed genes (DEGs) between declined and control samples. The results of gene ontology analysis showed that most of the DEGs involved in oak decline belong to the group of stress-responsive genes. In general, oak decline samples showed significant reductions in gene expression associated with "photosynthesis and storage of sugar" and "protein synthesis and related processes." Additionally, DEGs related to the starch degradation pathway were up-regulated, whereas DEGs associated with acetate-mevalonate (MVA), biosynthesis of lignin, and lignases pathways were down-regulated. The present study's findings can be an effective step in identifying the genes involved in oak decline and deciphering the relationship between this phenomenon and biotic and abiotic stresses.


Asunto(s)
Quercus , Quercus/genética , Quercus/metabolismo , RNA-Seq , Perfilación de la Expresión Génica , Transcriptoma , Análisis de Secuencia de ARN , Regulación de la Expresión Génica de las Plantas
2.
Biochem Genet ; 61(5): 1867-1879, 2023 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-36877417

RESUMEN

Target-site resistance (TSR) and non-target-site resistance (NTSR) to herbicides in arable weeds are increasing rapidly all over the world and threatening universal food safety. Resistance to herbicides that inhibit ACCase activity has been identified in wild oat. In this study, expression of ACC1, ACC2, CYP71R4 and CYP81B1 genes under herbicide stress conditions were studied in two TSR (resistant in the residue Ile1781-Leu and Ile2041-Asn of ACCase) biotypes, two NTSR biotypes and one susceptible biotype of A. ludoviciana for the first time. Treated and untreated biotypes with ACCase-inhibitor clodinafop propargyl herbicide were sampled from the stem and leaf tissues at 24 h after treatment. Our results showed an increase in gene expression levels in different tissues of both types of resistance biotypes that occurred under herbicide treatment compared with non-herbicide treatment. In all samples, the expression levels of leaf tissue in all studied genes were higher than in stem tissue. The results of ACC gene expression showed that the expression level of ACC1 was significantly higher than that of ACC2. Also, expression levels of TSR biotypes were higher than NTSR biotypes for the ACC1 gene. For both CYP71R4 and CYP81B1 genes, the expression ratio increased significantly in TSR and NTSR biotypes in different tissues after herbicide treatment. In contrast, the expression levels of CYP genes in NTSR biotypes were higher than in TSR biotypes. Our results support the hypothesis that the reaction of plants to herbicide is carried out through a different regulation of genes, which can be the result of the interaction of resistance type in the target or non-target-site.


Asunto(s)
Avena , Herbicidas , Avena/genética , Herbicidas/farmacología
3.
Genomics ; 113(2): 693-705, 2021 03.
Artículo en Inglés | MEDLINE | ID: mdl-33485953

RESUMEN

Lentil cultivation could be challenged by combined heat and drought stress in semi-arid regions. We used RNA-seq approach to profile transcriptome changes of Lens culinaris exposed to individual and combined heat and drought stresses. It was determined that most of the differentially expressed genes observed in response to combined stress, could not be identified by analysis of transcriptome exposed to corresponding individual stresses. Interestingly, this study results revealed that the expression of ribosome generation and protein biosynthesis and starch degradation pathways related genes were uniquely up-regulated under the combined stress. Although multiple genes related to antioxidant activity were up-regulated in response to all stresses, variation in types and expression levels of these genes under the combined stress were higher than that of individual stresses. Using this comparative approach, for the first time, we reported up-regulation of several TF, CDPK, CYP, and antioxidant genes in response to combined stress in plants.


Asunto(s)
Sequías , Respuesta al Choque Térmico , Lens (Planta)/genética , Transcriptoma , Regulación de la Expresión Génica de las Plantas , Lens (Planta)/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Regulación hacia Arriba
4.
Mol Biol Rep ; 47(9): 7009-7016, 2020 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-32886329

RESUMEN

Vinblastine and vincristine are two important anti-cancer drugs that are synthesized by the Terpenoid Indole Alkaloids (TIAs) pathway in periwinkle (Catharanthus roseus). The major challenge in the pharmaceutical industry is the low production rate of these Alkaloids. TIA pathway is affected by elicitors, such as salicylic acid (SA). This study aimed to investigate the expression pattern of some key genes in TIAs pathway under SA treatment. Foliar application of SA (0.01 and 0.1 mM) was used and leaves samples were taken at 0, 12, 18, 24 and 48 h after the treatment. qRT-PCR was used to investigate the expression pattern of Chorismate mutase (Cm), tryptophan decarboxylase (Tdc), Geraniol-10-hydroxylase (G10h), Secologanin synthase (Sls), Strictosidine synthase (Str), Desacetoxyvindoline-4-hydroxylase (D4h) and Deacetylvindoline-4-O-acetyltransferase (Dat) genes, following the SA treatment. The results of this experiment showed that transcript levels of Tdc, G10h, Sls, Str, D4h and Dat genes were significantly up-regulated in both SA concentration treatments. Furthermore, the highest transcript levels of Dat was observed after 48 h of the SA treatments. qRT-PCR results suggests that SA induces transcription of major genes involved in Alkaloids biosynthesis in Catharanthus roseus. It can be concluded that up-regulation of Tdc, G10h, Sls, Str, D4h and Dat genes can result in a higher production rate of Vinblastine and vincristine Alkaloids.


Asunto(s)
Catharanthus/metabolismo , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Proteínas de Plantas/biosíntesis , Ácido Salicílico/farmacología , Alcaloides de Triptamina Secologanina/metabolismo , Catharanthus/genética , Proteínas de Plantas/genética
5.
Mol Biol Rep ; 46(5): 5005-5017, 2019 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-31317454

RESUMEN

The receptor like kinases (RLKs) belong to the RLK/Pelle superfamily, one of the largest gene families in plants. RLKs play an important role in plant development, as well as in response to biotic and abiotic stresses. The lysine motif receptor like kinases (LysM-RLKs) are a subfamily of RLKs containing at least one lysine motif (LysM) that are involved in the perception of elicitors or pathogen-associated molecular patterns (PAMPs). In the present study, 77 putative RLKs genes and three receptor like proteins were identified in potato (Solanum tuberosum) genome, following a genome-wide search. The 77 potato RLK proteins are classified into two major phylogenetic groups based on their kinase domain amino acid sequence similarities. Out of 77 RLKs, 10 proteins had at least one LysM. Among them three RLP proteins were found in potato genome with either 2 or three tandem LysM but these lacked a cytoplasmic kinase domain. Expression analyses of a potato LysM-RLKs (StLysM-RLK05) was carried out by a Real time RT-PCR, following inoculation of potato leaves and immature tubers with late blight and common scab pathogens, respectively. The expression was significantly higher in resistant than in susceptible following S. scabies inoculation. The StLysM-RLK05 sequence was verified and it was polymorphic in scab susceptible cultivar. The present study provides an overview of the StLysM-RLKs gene family in potato genome. This information is helpful for future functional analysis of such an important protein family, in Solanaceae species.


Asunto(s)
Proteínas Serina-Treonina Quinasas/genética , Solanum tuberosum/genética , Secuencia de Aminoácidos/genética , Simulación por Computador , Evolución Molecular , Estudio de Asociación del Genoma Completo/métodos , Filogenia , Proteínas de Plantas/genética , Proteínas Quinasas/genética , Solanum tuberosum/metabolismo
6.
Mol Biol Rep ; 46(1): 415-424, 2019 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-30448893

RESUMEN

Structural mutations providing herbicide resistance may cause a modification of the three dimensional structure of a protein which will lead to a decrease in the herbicide efficacy. Wild oat (Avena ludoviciana Durieu.) is an increasingly disruptive weed in areas of intensive cereal production, thus the aim of this research was to identify mutations conferring resistance to ACCase-inhibitor herbicides at greenhouse, laboratory and in silico scales. Among the selected biotypes, No. 3 in the position 1781 (Ile1781-Leu) and No. 14 in the position 2041 (Ile2041-Asn), showed resistance to ACCase-inhibitor. The above mutations were confirmed using the specific primers and PCR-based methods. Analysis of molecular docking indicated that residues of Trp1948 and Pro2001 are important in the binding site and showed remarkable variation in the mutation types. Using molecular dynamic simulation analysis, we demonstrated that mutation types changed the conformation of the enzyme. These changes resulted in compressed conformation in the active site, which limited the availability of binding herbicide-enzyme. In present, no crystallography molecular structure and modeling reported on the ACCase of plants and this study investigated interactions of clodinafop propargyl and ACCase CT domain in A. ludoviciana by modeling, docking and simulations for the first time. Totally, bioinformatics analysis as well as PCR-based method confirmed that herbicide resistance conferred by nucleotide mutations in the gene sequence.


Asunto(s)
Acetil-CoA Carboxilasa/genética , Avena/efectos de los fármacos , Propionatos/farmacología , Piridinas/farmacología , Acetil-CoA Carboxilasa/antagonistas & inhibidores , Avena/genética , Resistencia a los Herbicidas/genética , Herbicidas/farmacología , Simulación del Acoplamiento Molecular/métodos , Mutación , Proteínas de Plantas/genética , Propionatos/metabolismo , Piridinas/metabolismo
7.
Cell Mol Biol (Noisy-le-grand) ; 64(4): 113-118, 2018 Mar 31.
Artículo en Inglés | MEDLINE | ID: mdl-29631693

RESUMEN

Weeds are serious problem in crop production and wild oat is a grass weed of economic and agronomic significance. We need to extend our basic knowledge of weeds especially in molecular genetics and gene expression. For study of gene expression by semi-quantitative and quantitative PCR, it is recommended that normalization of reference genes be carried out in order to select the most stable reference gene for a precise gene expression study. The purpose of this research was evaluation of four reference genes in response to treated and untreated (control) by herbicide in two tissues (stem and leaf) of non-target site resistance wild oat (A. ludoviciana). Four candidate reference genes including Actin, Ef1α (elongation factor 1 alpha), GAPDH (glyceraldehyde 3-phosphate dehydrogenase) and TBP (TATA-box-binding protein) were used to determine stable reference gene exposed to the herbicide using the statistical methods of NormFinder, BestKeeper and delta-Ct. NormFinder indicated that TBP and Actin genes are the best combination of two genes for normalizing calculations (with a combined gene stability value of 0.012) for qPCR analysis under herbicide stress in different tissues of non-target site resistance wild oat. Based on the statistical results, the Ef1α gene was identified as the unstable reference gene. Totally, according to results of this study, TBP gene is the most stable reference gene and therefore, this gene can be used as a reference gene for future studies of quantitative PCR analysis of herbicide stress-responsive gene expression in wild oat and potentially in other grass weed species.


Asunto(s)
Avena/efectos de los fármacos , Regulación de la Expresión Génica de las Plantas , Genes Esenciales , Proteínas de Plantas/genética , Reacción en Cadena de la Polimerasa/normas , Proteína de Unión a TATA-Box/genética , Actinas/genética , Actinas/metabolismo , Avena/genética , Avena/crecimiento & desarrollo , Avena/metabolismo , Perfilación de la Expresión Génica , Gliceraldehído 3-Fosfato Deshidrogenasa (NADP+)/genética , Gliceraldehído 3-Fosfato Deshidrogenasa (NADP+)/metabolismo , Herbicidas/toxicidad , Factor 1 de Elongación Peptídica/genética , Factor 1 de Elongación Peptídica/metabolismo , Hojas de la Planta/efectos de los fármacos , Hojas de la Planta/genética , Hojas de la Planta/crecimiento & desarrollo , Hojas de la Planta/metabolismo , Proteínas de Plantas/metabolismo , Tallos de la Planta/efectos de los fármacos , Tallos de la Planta/genética , Tallos de la Planta/crecimiento & desarrollo , Tallos de la Planta/metabolismo , Estrés Fisiológico , Proteína de Unión a TATA-Box/metabolismo
8.
J Sci Food Agric ; 96(4): 1333-40, 2016 Mar 15.
Artículo en Inglés | MEDLINE | ID: mdl-25903322

RESUMEN

BACKGROUND: Contamination of food and agricultural crops by Fusarium species is a major concern of food spoilage and a potential public health hazard. In the present study, natural contamination of maize and wheat samples from main cultivation areas of Iran by Fusarium species belonging to the Giberella fujikuroi species complex was evaluated, with special attention to the ability of the isolates to produce fumonisin B1 (FB1 ). RESULTS: A total of 55 Fusarium isolates were obtained from 27/32 maize samples (84.4%) and 11/15 wheat samples (73.3%). They were identified as F. verticillioides (47.3%), F. proliferatum (47.3%), F. fujikuroi (1.8%), F. nygamai (1.8%) and F. redolens (1.8%) by sequence analysis of translation elongation factor 1-α (TEF1-α). Twenty-two of 55 Fusarium isolates belonging to F. proliferatum (23.6%), F. verticillioides (14.5%) and F. fujikuroi (1.8%) produced FB1 in the concentration range 230.4-9565.0 µg mL(-1) . The dendrogram resulting from the TEF1-α profile showed that the genotypes were divided into clusters I, II and III, of which cluster III contained only F. redolens, its first report from Iran. CONCLUSION: On the basis of in vitro FB1 biosynthesis of the analyzed strains, the high degree of contamination of maize and wheat with Fusarium strains reported here should be considered as a potential public health threat, because a meaningful number of the isolates were found to produce hazardous levels of carcinogenic FB1 .


Asunto(s)
Carcinógenos Ambientales , Fumonisinas/metabolismo , Gibberella/clasificación , Gibberella/aislamiento & purificación , Triticum/microbiología , Zea mays/microbiología , ADN de Hongos/análisis , ADN de Hongos/química , Microbiología de Alimentos , Fumonisinas/análisis , Fusarium/clasificación , Fusarium/aislamiento & purificación , Irán , Factor 1 de Elongación Peptídica/genética , Filogenia
9.
Planta ; 240(2): 409-21, 2014 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-24893853

RESUMEN

MAIN CONCLUSION: Expression of amylosucrase in potato resulted in larger starch granules with rough surfaces and novel physico-chemical properties, including improved freeze-thaw stability, higher end viscosity, and better enzymatic digestibility. Starch is a very important carbohydrate in many food and non-food applications. In planta modification of starch by genetic engineering has significant economic and environmental benefits as it makes the chemical or physical post-harvest modification obsolete. An amylosucrase from Neisseria polysaccharea fused to a starch-binding domain (SBD) was introduced in two potato genetic backgrounds to synthesize starch granules with altered composition, and thereby to broaden starch applications. Expression of SBD-amylosucrase fusion protein in the amylose-containing potato resulted in starch granules with a rough surface, a twofold increase in median granule size, and altered physico-chemical properties including improved freeze-thaw stability, higher end viscosity, and better enzymatic digestibility. These effects are possibly a result of the physical interaction between amylosucrase and starch granules. The modified larger starches not only have great benefit to the potato starch industry by reducing losses during starch isolation, but also have an advantage in many food applications such as frozen food due to its extremely high freeze-thaw stability.


Asunto(s)
Glucosiltransferasas/metabolismo , Solanum tuberosum/metabolismo , Almidón/metabolismo , Glucosiltransferasas/genética , Solanum tuberosum/genética
10.
Biotechnol Adv ; 71: 108296, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38042311

RESUMEN

Classical plant breeding methods are limited in their ability to confer disease resistance on plants. However, in recent years, advancements in molecular breeding and biotechnological have provided new approaches to overcome these limitations and protect plants from disease. Antimicrobial peptides (AMPs) constitute promising agents that may be able to protect against infectious agents. Recently, peptides have been recombinantly produced in plants at scale and low cost. Because AMPs are less likely than conventional antimicrobials to elicit resistance of pathogenic bacteria, they open up exciting new avenues for agricultural applications. Here, we review recent advances in the design and production of bioactive recombinant AMPs that can effectively protect crop plants from diseases.


Asunto(s)
Antiinfecciosos , Péptidos Catiónicos Antimicrobianos , Péptidos Catiónicos Antimicrobianos/genética , Péptidos Catiónicos Antimicrobianos/química , Péptidos Antimicrobianos , Plantas/genética , Antiinfecciosos/química , Biotecnología
11.
PLoS One ; 19(4): e0297334, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38574179

RESUMEN

Potato tubers are rich sources of various nutrients and unique sources of starch. Many genes play major roles in different pathways, including carbohydrate metabolism during the potato tuber's life cycle. Despite substantial scientific evidence about the physiological and morphological development of potato tubers, the molecular genetic aspects of mechanisms underlying tuber formation have not yet been fully understood. In this study, for the first time, RNA-seq analysis was performed to shed light on the expression of genes involved in starch biosynthesis during potato tuber development. To this end, samples were collected at the hook-like stolon (Stage I), swollen tips stolon (Stage II), and tuber initiation (Stage III) stages of tuber formation. Overall, 23 GB of raw data were generated and assembled. There were more than 20000 differentially expressed genes (DEGs); the expression of 73 genes involved in starch metabolism was further studied. Moreover, qRT-PCR analysis revealed that the expression profile of the starch biosynthesis DEGs was consistent with that of the RNA-seq data, which further supported the role of the DEGs in starch biosynthesis. This study provides substantial resources on potato tuber development and several starch synthesis isoforms associated with starch biosynthesis.


Asunto(s)
Solanum tuberosum , Solanum tuberosum/metabolismo , Perfilación de la Expresión Génica , Tubérculos de la Planta/metabolismo , Metabolismo de los Hidratos de Carbono/genética , Almidón/metabolismo , Regulación de la Expresión Génica de las Plantas
12.
Plant Biotechnol J ; 11(4): 470-9, 2013 May.
Artículo en Inglés | MEDLINE | ID: mdl-23231535

RESUMEN

The Escherichia coli glycogen branching enzyme (GLGB) was fused to either the C- or N-terminus of a starch-binding domain (SBD) and expressed in two potato genetic backgrounds: the amylose-free mutant (amf) and an amylose-containing line (Kardal). Regardless of background or construct used, a large amount of GLGB/SBD fusion protein was accumulated inside the starch granules, however, without an increase in branching. The presence of GLGB/SBD fusion proteins resulted in altered morphology of the starch granules in both genetic backgrounds. In the amf genetic background, the starch granules showed both amalgamated granules and porous starch granules, whereas in Kardal background, the starch granules showed an irregular rough surface. The altered starch granules in both amf and Kardal backgrounds were visible from the initial stage of potato tuber development. High-throughput transcriptomic analysis showed that expression of GLGB/SBD fusion protein in potato tubers did not affect the expression level of most genes directly involved in the starch biosynthesis except for the up-regulation of a beta-amylase gene in Kardal background. The beta-amylase protein could be responsible for the degradation of the extra branches potentially introduced by GLGB.


Asunto(s)
Enzima Ramificadora de 1,4-alfa-Glucano/metabolismo , Plantas Modificadas Genéticamente/metabolismo , Solanum tuberosum/metabolismo , Almidón/metabolismo , Enzima Ramificadora de 1,4-alfa-Glucano/genética , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Plantas Modificadas Genéticamente/genética , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismo , Solanum tuberosum/genética
13.
3 Biotech ; 12(2): 52, 2022 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-35127307

RESUMEN

Selection of transgenic plants by using genes encoding screenable markers of plant origin with health benefit properties, such as anthocyanin is an important aim in plant genetic engineering. In this study, Malus domestica MYB10 (MdMYB10) gene, was used for Agrobacterium tumefaciens-mediated transformation of two SBS-02 and SBS-04 sugar beet lines. The impact of different light regimes on plant tissue culture from a combination of light, dark/light and dark was investigated. The results of this study showed that the MdMYB10 gene was successfully integrated into the selected purple transgenic lines, suggesting that the expression of MdMYB10 gene in sugar beet shoots can be used as a screenable markers for transformation, possibly replacing antibiotic resistant genes. Furthermore, the results of the antibacterial activity of transgenic plants extracts showed that the total extract obtained from transgenic lines significantly (P < 0.01) inhibited the growth and development of Enterococcus faecium and Enterococcus faecalis bacteria compared to the non-transgenic plants. The results of this study showed that the combination of betalain with vancomycin demonstrated a synergistic antimicrobial effect, also, suggesting that the expression of MdMYB10 may play a dual role by accumulating betalain and exhibiting a screenable markers function. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-022-03120-7.

14.
Gene ; 807: 145952, 2022 Jan 10.
Artículo en Inglés | MEDLINE | ID: mdl-34500049

RESUMEN

Extreme temperature is one of the serious threats to crop production in present and future scenarios of global climate changes. Lentil (Lens culinaris) is an important crop, and there is a serious lack of genetic information regarding environmental and temperature stresses responses. This study is the first report of evaluation of key genes and molecular mechanisms related to temperature stresses in lentil using the RNA sequencing technique. De novo transcriptome assembly created 44,673 contigs and differential gene expression analysis revealed 7494 differentially expressed genes between the temperature stresses and control group. Basic annotation of generated transcriptome assembly in our study led to the identification of 2765 novel transcripts that have not been identified yet in lentil genome draft v1.2. In addition, several unigenes involved in mechanisms of temperature sensing, calcium and hormone signaling and DNA-binding transcription factor activity were identified. Also, common mechanisms in response to temperature stresses, including the proline biosynthesis, the photosynthetic light reactions balancing, chaperone activity and circadian rhythms, are determined by the hub genes through the protein-protein interaction networks analysis. Deciphering the mechanisms of extreme temperature tolerance would be a new way for developing crops with enhanced plasticity against climate change. In general, this study has identified set of mechanisms and various genes related to cold and heat stresses which will be useful in better understanding of the lentil's reaction to temperature stresses.


Asunto(s)
Lens (Planta)/crecimiento & desarrollo , Lens (Planta)/genética , Estrés Fisiológico/genética , Cambio Climático , Frío/efectos adversos , Respuesta al Choque por Frío/genética , Productos Agrícolas/genética , Perfilación de la Expresión Génica/métodos , Regulación de la Expresión Génica de las Plantas/genética , Respuesta al Choque Térmico/genética , Respuesta al Choque Térmico/fisiología , Calor/efectos adversos , Anotación de Secuencia Molecular/métodos , Fotosíntesis , Mapas de Interacción de Proteínas/genética , Temperatura , Transcriptoma/genética
15.
Plant Sci ; 304: 110820, 2021 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-33568310

RESUMEN

Fusarium head blight (FHB) is a destructive disease affecting cereal crops globally due to mycotoxin contamination of grains that reduce yield and quality. Among hundreds of QTLs identified for resistance, the QTL-Fhb1 is of significant interest even today, for its major contribution to FHB resistance. Previously, QTL-Fhb1 dissection based on a combined metabolo-genomics approach, identified a few potential resistance genes, including a NAC like transcription factor for FHB resistance. Sequencing and phylogenetic analysis confirmed NAC to be the wheat TaNAC032. Also, the quantitative RT-PCR studies revealed a greater induced expression of TaNAC032 in resistant NIL in comparison to susceptible NIL upon Fusarium graminearum (Fg) infection. The virus-induced gene silencing (VIGS) based functional validation of TaNAC032 in resistant NIL confirmed increased disease severity and fungal biomass. Metabolic profiling revealed low abundances of resistance-related (RR) metabolites in TaNAC032 silenced NIL-R compared to non-silenced. Silenced plants showed decreased transcript abundances of RR metabolite biosynthetic genes associated with a reduction in total lignin content in rachis, confirming the regulatory role of TaNAC032 in wheat in response to Fg infection. If TaNA032 is mutated in an FHB susceptible cultivar, it can be edited to enhance FHB resistance.


Asunto(s)
Fusarium , Genes de Plantas , Lignina/biosíntesis , Enfermedades de las Plantas/microbiología , Proteínas de Plantas/fisiología , Factores de Transcripción/fisiología , Triticum/microbiología , Regulación de la Expresión Génica de las Plantas/fisiología , Silenciador del Gen , Genes de Plantas/fisiología , Enfermedades de las Plantas/inmunología , Proteínas de Plantas/genética , Polimorfismo Genético/genética , Sitios de Carácter Cuantitativo , Reacción en Cadena en Tiempo Real de la Polimerasa , Alineación de Secuencia , Análisis de Secuencia de ADN , Factores de Transcripción/genética , Triticum/genética , Triticum/inmunología , Triticum/metabolismo
16.
Front Plant Sci ; 11: 1236, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-32903611

RESUMEN

Antimicrobial peptides have been long known to confer resistance to plant pathogens. In this study, new recombinant peptides constructed from a dermaseptin B1 (DrsB1) peptide fused to a chitin-binding domain (CBD) from Avr4 protein, were used for Agrobacterium tumefaciens-mediated transformation of tobacco plants. Polymerase chain reaction (PCR), semi-quantitative RT-PCR, and western blotting analysis demonstrated the incorporation and expression of transgenes in tobacco genome and transgenic plants, respectively. In vitro experiments with recombinant peptides extracted from transgenic plants demonstrated a significant (P<0.01) inhibitory effect on the growth and development of plant pathogens. The DrsB1-CBD recombinant peptide had the highest antifungal activity against fungal pathogens. The expression of the recombinant peptides greatly protected transgenic plants from Alternaria alternata, Alternaria solani, Fusarium oxysporum, and Fusarium solani fungi, in comparison to Pythium sp. and Pythium aphanidermatum. Expression of new recombinant peptides resulted in a delay in the colonization of fungi and appearance of fungal disease symptoms from 6 days to more than 7 weeks. Scanning electron microscopy images revealed that the structure of the fungal mycelia appeared segmented, cling together, and crushed following the antimicrobial activity of the recombinant peptides. Greenhouse bioassay analysis showed that transgenic plants were more resistant to Fusarium and Pythium infections as compared with the control plants. Due to the high antimicrobial activity of the recombinant peptides against plant pathogens and novelty of recombinant peptides, this report shows the feasibility of this approach to generate disease resistance transgenic plants.

17.
Microbiologyopen ; 8(11): e837, 2019 11.
Artículo en Inglés | MEDLINE | ID: mdl-30912302

RESUMEN

Dermaseptin B1 (DrsB1), an antimicrobial cationic 31 amino acid peptide, is produced by Phyllomedusa bicolor. In an attempt to enhance the antimicrobial efficacy of DrsB1, the DrsB1 encoding 93 bp sequence was either fused to the N or C terminus of sequence encoding chitin-binding domain (CBD) of Avr4 gene from Cladosporium fulvum. Tobacco leaf disk explants were inoculated with Agrobacterium rhizogenes harboring pGSA/CBD-DrsB1 and pGSA/DrsB1-CBD expression vectors to produce hairy roots (HRs). Polymerase chain reaction (PCR) was employed to screen putative transgenic tobacco lines. Semi-quantitative RT-PCR and western blotting analysis indicated that the expression of recombinant genes were significantly higher, and recombinant proteins were produced in transgenic HRs. The recombinant proteins were extracted from the tobacco HRs and used against Pectobacterium carotovorum, Agrobacterium tumefaciens, Ralstonia solanacearum, and Xanthomonas campestris pathogenic bacteria and Alternaria alternata and Pythium sp. fungi. Two recombinant proteins had a statistically significant (p < 0.01) inhibitory effect on the growth and development of plant pathogens. The CBD-DrsB1 recombinant protein demonstrated a higher antibacterial effect, whereas the DrsB1-CBD recombinant protein demonstrated greater antifungal activity. Scanning electron microscopy images revealed that the structure of the fungal mycelia appeared segmented, adhered to each other, and crushed following the antimicrobial activity of the recombinant proteins. Due to the high antimicrobial activity of the recombinant proteins against plant pathogens, this strategy can be used to generate stable transgenic crop plants resistant to devastating plant pathogens.


Asunto(s)
Proteínas Anfibias/metabolismo , Antiinfecciosos/metabolismo , Péptidos Catiónicos Antimicrobianos/metabolismo , Expresión Génica , Nicotiana/metabolismo , Plantas Modificadas Genéticamente/metabolismo , Proteínas Recombinantes de Fusión/metabolismo , Alternaria/efectos de los fármacos , Alternaria/crecimiento & desarrollo , Proteínas Anfibias/genética , Péptidos Catiónicos Antimicrobianos/genética , Bacterias/efectos de los fármacos , Bacterias/crecimiento & desarrollo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Pruebas de Sensibilidad Microbiana , Plantas Modificadas Genéticamente/genética , Proteínas Recombinantes de Fusión/genética , Nicotiana/genética
18.
Mol Biotechnol ; 61(4): 253, 2019 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-30747381

RESUMEN

The original version of this article unfortunately contained a mistake in the unit "µg/l". The unit "µg/l" should be corrected to "µg/ml" throughout the paper.

19.
Mol Biotechnol ; 61(4): 241-252, 2019 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-30649664

RESUMEN

Expression of strong antimicrobial peptides in plants is of great interest to combat a wide range of plant pathogens. To bring the Dermaseptin B1 (DrsB1) peptide to the intimate contact of the plant pathogens cell wall surface, the DrsB1 encoding sequence was fused to the C-terminal part of the two copies of the chitin-binding domain (CBD) of the Avr4 effector protein and used for Agrobacterium rhizogenes-mediated transformation. The expression of the recombinant protein in the tobacco hairy roots (HRs) was confirmed by molecular analysis. Antimicrobial activity analysis of the recombinant protein purified from the transgenic HRs showed that the (CBD)2-DrsB1 recombinant protein had a significant (p < 0.01) antimicrobial effect on the growth of different fungal and bacterial pathogens. The results of this study indicated that the recombinant protein had a higher antifungal activity against chitin-producing Alternaria alternata than Pythium spp. Scanning electron microscopy images demonstrated that the recombinant protein led to fungal hypha deformation, fragmentation, and agglutination of growing hypha, possibly by dissociating fungal cell wall components. In vitro evidences suggest that the expression of the (CBD)2-DrsB1 recombinant protein in plants by generating transgenic lines is a promising approach to produce disease-resistant plants, resistance to chitin-producing pathogenic fungi.


Asunto(s)
Proteínas Anfibias/genética , Péptidos Catiónicos Antimicrobianos/genética , Resistencia a la Enfermedad , Proteínas Fúngicas/química , Nicotiana/genética , Alternaria/efectos de los fármacos , Proteínas Anfibias/metabolismo , Proteínas Anfibias/farmacología , Péptidos Catiónicos Antimicrobianos/metabolismo , Péptidos Catiónicos Antimicrobianos/farmacología , Sitios de Unión , Quitina/metabolismo , Cladosporium/metabolismo , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Enfermedades de las Plantas/microbiología , Enfermedades de las Plantas/prevención & control , Raíces de Plantas/genética , Raíces de Plantas/crecimiento & desarrollo , Plantas Modificadas Genéticamente/crecimiento & desarrollo , Proteínas Recombinantes/farmacología , Nicotiana/crecimiento & desarrollo
20.
PLoS One ; 14(3): e0213305, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-30875373

RESUMEN

Salt stress is one of the major adverse environmental factors limiting crop productivity. Considering Iran as one of the bread wheat origins, we sequenced root transcriptome of an Iranian salt tolerant cultivar, Arg, under salt stress to extend our knowledge of the molecular basis of salinity tolerance in Triticum aestivum. RNA sequencing resulted in more than 113 million reads and about 104013 genes were obtained, among which 26171 novel transcripts were identified. A comparison of abundances showed that 5128 genes were differentially expressed due to salt stress. The differentially expressed genes (DEGs) were annotated with Gene Ontology terms, and the key pathways were identified using Kyoto Encyclopedia of Gene and Genomes (KEGG) pathway mapping. The DEGs could be classified into 227 KEGG pathways among which transporters, phenylpropanoid biosynthesis, transcription factors, glycosyltransferases, glutathione metabolism and plant hormone signal transduction represented the most significant pathways. Furthermore, the expression pattern of nine genes involved in salt stress response was compared between the salt tolerant (Arg) and susceptible (Moghan3) cultivars. A panel of novel genes and transcripts is found in this research to be differentially expressed under salinity in Arg cultivar and a model is proposed for salt stress response in this salt tolerant cultivar of wheat employing the DEGs. The achieved results can be beneficial for better understanding and improvement of salt tolerance in wheat.


Asunto(s)
Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Proteínas de Plantas/genética , Salinidad , Tolerancia a la Sal , Cloruro de Sodio/farmacología , Transcriptoma , Triticum/genética , Perfilación de la Expresión Génica , Secuenciación de Nucleótidos de Alto Rendimiento , Raíces de Plantas/efectos de los fármacos , Raíces de Plantas/genética , Raíces de Plantas/crecimiento & desarrollo , Triticum/efectos de los fármacos , Triticum/crecimiento & desarrollo
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