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1.
Sci Rep ; 13(1): 22478, 2023 12 18.
Artículo en Inglés | MEDLINE | ID: mdl-38110487

RESUMEN

Bio-based fertilizers (BBFs) recovered from animal manure are promising products to optimise resources recovery and generate high agricultural yields. However, their fertilization value may be limited and it is necessary to enrich BBFs with microbial consortia to enhance their fertilization value. Three specific microbial consortia were developed according to the characteristics of three different BBFs produced from manure (bio-dried solid fraction, solid fraction of digestate and biochar) to enhance plant growth and product quality. A greenhouse pot experiment was carried out with tomato plants grown with microbiologically activated BBFs applied either as N-organic fertilizers or as an organic amendment. A next generation sequencing analysis was used to characterise the development of each rhizospheric community. All the activated BBFs gave enhanced tomato yields (fresh and dry weight) compared with the non-activated treatments and similar to, or higher than, chemical fertilization. Concerning the tomato fruits' organoleptic quality, lycopene and carotenoids concentrations were improved by biological activation. Metagenomic analysis points at Trichoderma as the main driver of the positive effects, with the effects of added bacteria being negligible or limited at the early stages after fertilization. In the context of the circular economy, the activated BBFs could be used to replace synthetic fertilisers, reducing costs and environmental burdens and increasing production.


Asunto(s)
Solanum lycopersicum , Trichoderma , Animales , Fertilizantes , Estiércol , Agricultura , Suelo
2.
Bioresour Technol ; 374: 128781, 2023 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-36828223

RESUMEN

Thin-layer (TL) photobioreactors (PBRs) are characterised by high productivity. However, their use is limited to lab/pilot-scale, and a deeper level of characterisation is needed to reach industrial scale and test the resistance of multiple microalgae. Here, the performance and composition of eight microalgal communities cultivated in the two main TLs design (thin-layer cascade (TLC) and thin-layer raceway pond (RW)) were investigated through Illumina sequencing. Chlorella vulgaris showed robustness in both designs and often acted as an "invasive" species. Inoculum and reactor type brought variability. Eukaryotic microalgae inocula led to a more robust and stable community (higher similarity), however, RWs were characterised by a higher variability and did not favour the eukaryotic microalgae. The only cyanobacterial inoculum, Nostoc piscinale, was maintained, however the community was variable between designs. The reactor design had an effect on the N cycle with the TLC and RW configurations, enhancing nitrification and denitrification respectively.


Asunto(s)
Chlorella vulgaris , Microalgas , Aguas Residuales , Estanques , Fotobiorreactores , Biomasa , Bacterias/genética
3.
Bioresour Technol ; 363: 127979, 2022 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-36126844

RESUMEN

Centrate is a low-cost alternative to synthetic fertilizers for microalgal cultivation, reducing environmental burdens and remediation costs. Adapted microalgae need to be selected and characterised to maximise biomass production and depuration efficiency. Here, the performance and composition of six microalgal communities cultivated both on synthetic media and centrate within semi-open tubular photobioreactors were investigated through Illumina sequencing. Biomass grown on centrate, exposed to a high concentration of ammonium, showed a higher quantity of nitrogen (5.6% dry weight) than the biomass grown on the synthetic media nitrate (3.9% dry weight). Eukaryotic inocula were replaced by other microalgae while cyanobacterial inocula were maintained. Communities were generally similar for the same inoculum between media, however, inoculation with cyanobacteria led to variability within the eukaryotic community. Where communities differed, centrate resulted in a higher richness and diversity. The higher nitrogen of centrate possibly led to higher abundance of genes coding for N metabolism enzymes.


Asunto(s)
Compuestos de Amonio , Cianobacterias , Microalgas , Compuestos de Amonio/metabolismo , Biomasa , Fertilizantes , Microalgas/metabolismo , Nitratos/metabolismo , Nitrógeno/metabolismo , Fotobiorreactores/microbiología , Crecimiento Sostenible , Aguas Residuales
4.
Bioresour Technol ; 360: 127619, 2022 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-35842066

RESUMEN

Microalgae cultivation is proposed as an effective system for pathogens reduction and wastewater depuration, however, a full characterisation of the risks is still needed. Two raceways were inoculated with Scenedesmus, one using wastewater and the other using a fertilizer medium. Microbial community and pathogen presence were explored by next generation sequencing (NGS), commercial qPCR array and plate counts. These methods proved to be complementary for a full characterization of community structure and potential risks. Media and sampling locations contributed to shape communities and pathogenic loads. The main pathogenic genera detected were Arcobacter and Elizabethkingia (mainly in wastewater) with an important presence of Aeromonas (all samples). A lower presence of pathogens was detected in fertilizer samples, while wastewater showed a reduction from inlet to outlet. Raceways showed potential as an effective biotreatment, with most of the retained pathogens released in the outlet and only a minor part settled in the biomass.


Asunto(s)
Microalgas , Scenedesmus , Biomasa , Fertilizantes , Fotobiorreactores , Aguas Residuales/química
5.
Molecules ; 27(2)2022 Jan 10.
Artículo en Inglés | MEDLINE | ID: mdl-35056737

RESUMEN

This paper demonstrated the growth ability of twelve algae-microbial consortia (AC) isolated from organic wastes when a pig slurry-derived wastewater (NFP) was used as growth substrate in autotrophic cultivation. Nutrient recovery, biochemical composition, fatty acid and amino acid profiles of algae consortia were evaluated and compared. Three algae-microbial consortia, i.e., a Chlorella-dominated consortium (AC_1), a Tetradesmus and Synechocystis co-dominated consortium (AC_10), and a Chlorella and Tetradesmus co-dominated consortium (AC_12) were found to have the best growth rates (µ of 0.55 ± 0.04, 0.52 ± 0.06, and 0.58 ± 0.03 d-1, respectively), which made them good candidates for further applications. The ACs showed high carbohydrates and lipid contents but low contents of both proteins and essential amino acids, probably because of the low N concentration of NFP. AC_1 and AC_12 showed optimal ω6:ω3 ratios of 3.1 and 3.6, which make them interesting from a nutritional point of view.


Asunto(s)
Microalgas/química , Microalgas/crecimiento & desarrollo , Aminoácidos/análisis , Animales , Biomasa , Ácidos Grasos/análisis , Concentración de Iones de Hidrógeno , Consorcios Microbianos/fisiología , Nitrógeno , Porcinos , Aguas Residuales
6.
Bioresour Technol ; 347: 126416, 2022 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-34838970

RESUMEN

Pure microalgae cultivation in organic wastes may be hampered by their low adaptation to extreme growth conditions and by the risk of microbial contamination. This work aimed to isolate self-adapted microalgae-microbial consortia able to survive in organic wastes characterized by extreme conditions, to be then proposed for technological application in removing carbon and nutrients from wastes' streams. To do so, sixteen organic wastes with different origins and consistency were sampled. Twelve microbial consortia were isolated from wastes and their eukaryotic and prokaryotic compositions were analyzed by next generation sequencing. Eight eukaryotic communities were dominated by Chlorophyta, led by Chlorella, able to survive in different wastes regardless of chemical-biological properties. Tetradesmus, the second most represented genus, grew preferentially in substrates with less stressing chemical-physical parameters. Chlorella and Tetradesmus were mostly isolated from cow slurry and derived wastes which proved to be the best local residual organic source.


Asunto(s)
Chlorella , Microalgas , Biomasa , Carbono , Consorcios Microbianos
7.
Plant Physiol Biochem ; 168: 27-42, 2021 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-34619596

RESUMEN

The investigation of the adaptive strategies of wild plant species to extreme environments is a challenging issue, which favors the identification of new traits for plant resilience. We investigated different traits which characterize the root-soil interaction of Parietaria judaica, a wild plant species commonly known as "Pellitory-of-the-wall". P. judaica adopts the acidification-reduction strategy (Strategy I) for iron (Fe) acquisition from soil, and it can complete its life cycle in highly calcareous environments without any symptoms of chlorosis. In a field-to-lab approach, the microbiome associated with P. judaica roots was analyzed in spontaneous plants harvested from an urban environment consisting in an extremely calcareous habitat. Also, the phenolics and carboxylates content and root plasticity and exudation were analyzed in P. judaica plants grown under three different controlled conditions mimicking the effect of calcareous environments on Fe availability: results show that P. judaica differentially modulates root plasticity under different Fe availability-impaired conditions, and that it induces, to a high extent, the exudation of caffeoylquinic acid derivatives under calcareous conditions, positively impacting Fe solubility.


Asunto(s)
Microbiota , Parietaria , Hierro , Fenoles , Raíces de Plantas , Suelo
8.
Eur Surg Res ; 61(2-3): 72-82, 2020.
Artículo en Inglés | MEDLINE | ID: mdl-33080605

RESUMEN

BACKGROUND: Vascular endothelial growth factor (VEGF) is a subfamily of growth factors involved in angiogenesis; CD34+ cells are normally found in endothelial progenitor cells and endothelial cells of blood vessels. Colonic adenomatous polyps may not always be completely removable endoscopically, and a preoperative diagnosis might still be necessary. The aim of the study was to evaluate whether VEGF-A, VEGF-C and CD34 mRNA expression along colorectal carcinogenesis steps can implement NICE (Narrow-Band Imaging International Colorectal Endoscopic) classification in the diagnosis of malignancy in colorectal polypoid lesions. METHODS: Seventy-one subjects with colonic adenoma or cancer who underwent screening narrow-band imaging (NBI) colonoscopy were prospectively enrolled in the MICCE1 project (Treviso center). Polyps were classified according to the NICE classification. Real-time RT-PCR for VEGF-A, VEGF-C and CD34 mRNA expression was performed. Nonparametric statistics, receiver-operating characteristic curve analysis and logistic multiple regression analysis were used. RESULTS: VEGF-A and CD34 mRNA expression was significantly higher in sessile adenomas than in polypoid ones (p < 0.001 and p = 0.01, respectively). VEGF-A, VEGF-C and CD34 mRNA expression was significantly higher in adenocarcinoma than in adenoma (p = 0.01, p = 0.01 and p = 0.01, respectively). The accuracy of VEGF-A, VEGF-C and CD34 mRNA expression for prediction of malignancy was 0.79 (95% CI 0.65-0.90), 0.81 (95% CI 0.66-0.91) and 0.80 (95% CI 0.65-0.90), respectively, while the accuracy of the NICE classification was 0.85 (95% CI 0.72-0.94). The determination coefficient R2, which indicates the amount of the variability explained by a regression model, for NICE classification alone was 0.24 (p < 0.001). A regression model that included NICE classification and VEGF-C mRNA expression showed an R2 = 0.39 as well as a model including NICE classification and CD34 mRNA levels. CONCLUSIONS: This study demonstrated that VEGF-C and CD34 mRNA levels might be useful to stratify colorectal polyps in different risk of progression classes by implementing the accuracy of the NICE classification. Studies on in vivo detection of these markers are warranted.


Asunto(s)
Adenocarcinoma/metabolismo , Antígenos CD34/metabolismo , Neoplasias del Colon/metabolismo , Factor A de Crecimiento Endotelial Vascular/metabolismo , Factor C de Crecimiento Endotelial Vascular/metabolismo , Adenocarcinoma/diagnóstico por imagen , Adenoma/diagnóstico por imagen , Adenoma/metabolismo , Biomarcadores de Tumor/metabolismo , Neoplasias del Colon/diagnóstico por imagen , Colonoscopía , Femenino , Humanos , Masculino , Persona de Mediana Edad , Imagen de Banda Estrecha , Neovascularización Patológica , Estudios Prospectivos
9.
Eur Surg Res ; 60(5-6): 186-195, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31597147

RESUMEN

BACKGROUND: Interferon gamma (IFNγ) and tumor necrosis factor-related weak inducer of apoptosis (TWEAK) molecules seem to have a potential effect on angiogenic factors such as vascular endothelial growth factor (VEGF). The aim of this study was to assess a possible interplay between IFNγ and TWEAK cytokines and VEGF machinery in the different steps of colorectal carcinogenesis. METHODS: A total of 92 subjects with colonic adenoma or cancer who underwent screening colonoscopy or surgery were prospectively enrolled. Polypoid lesion tissue samples were collected and frozen. Real-time reverse transcription polymerase chain reaction for IFNγ, TWEAK, and VEGF-A mRNA expression was performed. Immunoassays for VEGF-A, VEGF-C, VEGFR-1, VEGFR-2, and VEGFR-3 were also performed. Nonparametric statistics, receiver operating characteristic curve analysis, and logistic multiple regression analysis were used. RESULTS: IFNγ and TWEAK mRNA expression was higher in patients with T2 or more advanced colorectal cancer than in those with adenomas or T1 cancer (p < 0.001 and p = 0.01, respectively). IFNγ and TWEAK mRNA expression levels directly correlated with VEGF-A mRNA expression levels (rho = 0.44, p < 0.001 and rho = 0.29, p = 0.004, respectively). On the contrary, IFNγ and TWEAK mRNA expression levels inversely correlated with VEGF-C protein levels (rho = -0.29, p = 0.04 and rho = -0.31, p = 0.03, respectively). Similarly, IFNγ and TWEAK mRNA expression levels inversely correlated with VEGFR2 protein levels (rho = -0.38, p = 0.033 and rho = -0.40, p = 0.025, respectively). CONCLUSION: This study showed that in colorectal polypoid lesions, IFNγ and TWEAK expressions are directly correlated to VEGF-A expression but inversely correlated with VEGFR2 levels, suggesting a possible feedback mechanism in the regulation of VEGF-A expression.


Asunto(s)
Neoplasias Colorrectales/irrigación sanguínea , Citocina TWEAK/genética , Interferón gamma/genética , Neovascularización Patológica/etiología , Factor A de Crecimiento Endotelial Vascular/genética , Adulto , Anciano , Anciano de 80 o más Años , Neoplasias Colorrectales/etiología , Citocina TWEAK/análisis , Femenino , Humanos , Interferón gamma/análisis , Modelos Logísticos , Masculino , Persona de Mediana Edad , Estudios Prospectivos , ARN Mensajero/análisis , Factor A de Crecimiento Endotelial Vascular/análisis , Receptor 2 de Factores de Crecimiento Endotelial Vascular/análisis
10.
Environ Microbiol ; 2018 May 22.
Artículo en Inglés | MEDLINE | ID: mdl-29786171

RESUMEN

It has been previously shown that the transgenic overexpression of the plant root vacuolar proton pumps H+ -ATPase (V-ATPase) and H+ -PPase (V-PPase) confer tolerance to drought. Since plant-root endophytic bacteria can also promote drought tolerance, we hypothesize that such promotion can be associated to the enhancement of the host vacuolar proton pumps expression and activity. To test this hypothesis, we selected two endophytic bacteria endowed with an array of in vitro plant growth promoting traits. Their genome sequences confirmed the presence of traits previously shown to confer drought resistance to plants, such as the synthesis of nitric oxide and of organic volatile organic compounds. We used the two strains on pepper (Capsicuum annuum L.) because of its high sensitivity to drought. Under drought conditions, both strains stimulated a larger root system and enhanced the leaves' photosynthetic activity. By testing the expression and activity of the vacuolar proton pumps, H+ -ATPase (V-ATPase) and H+ -PPase (V-PPase), we found that bacterial colonization enhanced V-PPase only. We conclude that the enhanced expression and activity of V-PPase can be favoured by the colonization of drought-tolerance-inducing bacterial endophytes.

12.
Tumour Biol ; 39(3): 1010428317692263, 2017 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-28347226

RESUMEN

Colorectal cancer incidence in patients undergoing screening protocols is decreasing because of the higher rate of discovered preneoplastic colonic lesions; however, adenomatous polyps may not always be removable endoscopically and surgery may still be necessary. The aim of this study was to assess the vascular endothelial growth factor expression in the different steps of colorectal carcinogenesis to explore its potential role as a marker of malignancy in polypoid lesions. A total of 92 subjects with colonic adenoma or cancer who underwent screening colonoscopy or surgery were prospectively enrolled. Real-time reverse transcription polymerase chain reaction for VEGF-A messenger RNA expression and immunohistochemistry for VEGF-A were performed. Immunoassays for VEGF-A, VEGF-C, VEGFR-1, VEGFR-2, and VEGFR-3 were also performed. Non-parametric statistics, receiver operating characteristic curve analysis, and logistic multiple regression analysis were used. VEGF-A messenger RNA expression was higher in patients with high-grade dysplasia or colorectal cancer than in those with low-grade dysplasia adenomas (p = 0.01). At immunohistochemistry, VEGF-A expression was significantly higher in colorectal cancer patients compared to dysplastic adenomas (p < 0.001), and the accuracy of VEGF-A expression for prediction of malignancy was 91.7 (95% confidence interval = 78.7-97.9). VEGF-C protein expression was lower in colorectal cancer patients than in simple adenomas (p = 0.02). VEGF-A levels were directly correlated to polyp size (rho = 0.73, p = 0.0062). Multivariate analysis demonstrated that malignancy and polyp size were independent predictors of VEGF-A mucosal levels. This study demonstrated that the VEGF-A expression changes along the colorectal carcinogenesis pathway showing a neat step up at the passage from high-grade dysplasia to invasive cancer. This feature might potentially be useful to stratify colorectal polyps in different risks of progression classes. Moreover, the high level of VEGF-A expression predicted the presence of lymphovascular invasion with good accuracy.


Asunto(s)
Neoplasias Colorrectales/metabolismo , Factor A de Crecimiento Endotelial Vascular/biosíntesis , Adulto , Anciano , Anciano de 80 o más Años , Carcinogénesis/genética , Carcinogénesis/metabolismo , Carcinogénesis/patología , Estudios de Casos y Controles , Neoplasias Colorrectales/genética , Neoplasias Colorrectales/patología , Neoplasias Colorrectales/cirugía , Femenino , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana Edad , Invasividad Neoplásica , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Factor A de Crecimiento Endotelial Vascular/genética
13.
Oncotarget ; 7(21): 30109-18, 2016 May 24.
Artículo en Inglés | MEDLINE | ID: mdl-27097112

RESUMEN

AIM: The BRAF mutation is a rare pathogenetic alternative to KIT/PDGFRA mutation in GIST and causes Imatinib resistance. A recent description of KIT and BRAF mutations co-occurring in an untreated GIST has challenged the concept of their being mutually exclusive and may account for ab initio resistance to Imatinib, even in the presence of Imatinib-sensitive KIT mutations. BRAF sequencing is generally limited to KIT/PDGFRA wild-type cases. Hence, the frequency of concomitant mutations may be underestimated. METHODS: We screened for KIT (exon 9, 11 ,13 ,17), PDGFRA (exon 12,14, 18) and BRAF (exon 15) mutations a series of 407 GIST. Additionally, we evaluated the BRAF V600E mutation-specific antibody, VE1, as a surrogate for V600E mutation, on a series of 313 GIST (24 on whole sections, 288 cases on tissue array), including 6 cases molecularly ascertained to carry the BRAF V600E mutation. RESULTS: No concomitant KIT/BRAF or PDGFRA/BRAF mutations were detected. BRAF mutation was detected only in one case, wild-type for KIT/PDGFRA. All the 6 BRAF-mutant cases stained positive with the VE1 antibody. A weak VE1 expression was observed in 14/287 (4.9%) BRAF wild-type cases, as observed also in 2/6 BRAF-mutant cases. Overall in our series, sensitivity and specificity of the VE1 antobody were 100% and 95.1%, respectively. CONCLUSIONS: The concomitance of BRAF mutation with either KIT or PDGFRA mutation is rare in GIST. In these tumors, moderate/strong VE1 immunoreactivity is a valuable surrogate for molecular analysis. Instead, genotyping is warranted in the presence of weak VE1 staining.


Asunto(s)
Neoplasias Gastrointestinales/genética , Tumores del Estroma Gastrointestinal/genética , Proteínas Proto-Oncogénicas B-raf/genética , Proteínas Proto-Oncogénicas c-kit/genética , Receptor alfa de Factor de Crecimiento Derivado de Plaquetas/genética , Adulto , Anciano , Anciano de 80 o más Años , Anticuerpos Monoclonales/inmunología , Antineoplásicos/farmacología , Antineoplásicos/uso terapéutico , Análisis Mutacional de ADN/métodos , Resistencia a Antineoplásicos , Exones , Femenino , Neoplasias Gastrointestinales/patología , Tumores del Estroma Gastrointestinal/tratamiento farmacológico , Tumores del Estroma Gastrointestinal/patología , Genotipo , Humanos , Mesilato de Imatinib/farmacología , Mesilato de Imatinib/uso terapéutico , Inmunohistoquímica , Masculino , Persona de Mediana Edad , Mutación , Proteínas Proto-Oncogénicas B-raf/inmunología , Análisis de Secuencia de ADN , Análisis de Matrices Tisulares
14.
Clin Sarcoma Res ; 5: 7, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-25699170

RESUMEN

BACKGROUND: Recently a few cases of synovial sarcoma (SS) of the abdominal viscera have been reported, raising awareness about the potential for confusion between this entity and KIT-negative gastrointestinal stromal tumors (GIST). We report the clinicopathological, immunophenotypical and molecular features of fifteen more SS occurring in the stomach (8 cases), epigastric region (one case), small intestine (one case), large intestine (three cases), involving both the terminal ileum and the caecum (one case) and liver (one case). METHODS: Immunostains for SMA, DESMIN, CD34, CD117, S100, EMA, CK AE1/3, TLE1, CD56, CD99, BCL2, DOG1 were performed. Rearrangement of SS18 gene region was screened in all cases: by conventional karyotype in one case, the remaining cases were screened either by interphase FISH or Q-PCR or both. RESULTS: Ten patients were male and five female, with an age range of 17-61 years (median 44). Tumor size ranged from 2 to 15 cm (median 8). Mitoses per 10 HPF ranged from 4 to 27 (median 9.5). Eleven tumors were monophasic fibrous SS, one biphasic SS and three poorly differentiated SS. SMA, Desmin, CD34, CD117 and S100 were negative in all cases, whereas EMA and/or CK AE1/AE3 were positive in all cases. TLE1, BCL2 and CD56 were positive in all tested cases. DOG1 was positive in one case. SS18 gene region rearrangement was demonstrated in all cases. A fusion transcript was amplified in eight cases: either SS18-SSX2 or SS18-SSX1 respectively in four cases each. CONCLUSIONS: SS is increasingly recognized at visceral sites. Molecular analyses play a key role when dealing with usual histotypes in unusual sites. Correct diagnosis is crucial for appropriate therapy.

15.
Front Plant Sci ; 4: 326, 2013.
Artículo en Inglés | MEDLINE | ID: mdl-23986768

RESUMEN

Iron uptake in dicots depends on their ability to induce a set of responses in root cells including rhizosphere acidification through H(+) extrusion and apoplastic Fe(III) reduction by Fe(III)-chelate reductase. These responses must be sustained by metabolic rearrangements aimed at providing the required NAD(P)H, ATP and H(+). Previous results in Fe-deficient cucumber roots showed that high H(+) extrusion is accompanied by increased phosphoenolpyruvate carboxylase (PEPC) activity, involved in the cytosol pH-stat; moreover (31)P-NMR analysis revealed increased vacuolar pH and decreased vacuolar [inorganic phosphate (Pi)]. The opposite was found in soybean: low rhizosphere acidification, decreased PEPC activity, vacuole acidification, and increased vacuolar [Pi]. These findings, highlighting a different impact of the Fe deficiency responses on cytosolic pH in the two species, lead to hypothesize different roles for H(+) and Pi movements across the tonoplast in pH homeostasis. The role of vacuole in cytosolic pH-stat involves the vacuolar H(+)-ATPase (V-ATPase) and vacuolar H(+)-pyrophosphatase (V-PPase) activities, which generating the ΔpH and ΔΨ, mediate the transport of solutes, among which Pi, across the tonoplast. Fluxes of Pi itself in its two ionic forms, H2PO4 (-) predominating in the vacuole and HPO4 (2-) in the cytosol, may be involved in pH homeostasis owing to its pH-dependent protonation/deprotonation reactions. Tonoplast enriched fractions were obtained from cucumber and soybean roots grown with or without Fe. Both V-ATPase and V-PPase activities were analyzed and the enrichment and localization of the corresponding proteins in root tissues were determined by Western blot and immunolocalization. V-ATPase did not change its activity and expression level in response to Fe starvation in both species. V-PPase showed a different behavior: in cucumber roots its activity and abundance were decreased, while in Fe-deficient soybean roots they were increased. The distinct role of the two H(+) pumps in Pi fluxes between cytoplasm and vacuole in Fe-deficient cucumber and soybean root cells is discussed.

16.
Plant Physiol Biochem ; 57: 168-74, 2012 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-22705592

RESUMEN

The regulation exerted by the Fe status in the plant on Fe deficiency responses was investigated in Cucumis sativus L. roots at both biochemical and molecular levels. Besides the two activities strictly correlated with Fe deficiency response, those of the Fe(III)-chelate reductase and the high affinity Fe transporter, we considered also H(+)-ATPase (EC 3.6.3.6) and phosphoenolpyruvate carboxylase (EC 4.1.1.31), that have been shown to be involved in this response. Both enzymatic activities and gene expression were monitored using a split root system. Absence of Fe induced the expression of the four transcripts, accompanied by an increase in the corresponding enzymatic activities. The application of the split root technique gave some information about the regulation of Fe uptake. In fact, 24 h after split root application, transcripts were still high and comparable to those of the -Fe control in the Fe-supplied half side, while in the -Fe side there was a drop in the expression and the relative enzymatic activities. Major changes occurred after 48 and 72 h. The coordinated regulation of these responses is discussed.


Asunto(s)
Cucumis sativus/metabolismo , Hierro/metabolismo , Raíces de Plantas/metabolismo , Cucumis sativus/enzimología , Regulación de la Expresión Génica de las Plantas , Fosfoenolpiruvato Carboxilasa/metabolismo , Raíces de Plantas/enzimología , ATPasas de Translocación de Protón/metabolismo
17.
J Plant Physiol ; 169(3): 242-8, 2012 Feb 15.
Artículo en Inglés | MEDLINE | ID: mdl-22078996

RESUMEN

The demand for iron in leguminous plants increases during symbiosis, as the metal is utilised for the synthesis of various Fe-containing proteins in both plant and bacteroids. However, the acquisition of this micronutrient is problematic due to its low bioavailability at physiological pH under aerobic conditions. Induction of root Fe(III)-reductase activity is necessary for Fe uptake and can be coupled to the rhizosphere acidification capacity linked to the H(+)-ATPase activity. Fe uptake is related to the expression of a Fe(2+) transporter (IRT1). In order to verify the possible role of nodules in the acquisition of Fe directly from the soil solution, the localization of H(+)-ATPase and IRT1 was carried out in common bean nodules by immuno-histochemical analysis. The results showed that these proteins were particularly abundant in the central nitrogen-fixing zone of nodules, around the periphery of infected and uninfected cells as well as in the vascular bundle of control nodules. Under Fe deficiency an over-accumulation of H(+)-ATPase and IRT1 proteins was observed especially around the cortex cells of nodules. The results obtained in this study suggest that the increase in these proteins is differentially localized in nodules of Fe-deficient plants when compared to the Fe-sufficient condition and cast new light on the possible involvement of nodules in the direct acquisition of Fe from the nutrient solution.


Asunto(s)
Deficiencias de Hierro , Phaseolus/enzimología , Proteínas de Plantas/metabolismo , ATPasas de Translocación de Protón/metabolismo , Transporte Biológico , Proteínas de Transporte de Catión/metabolismo , FMN Reductasa/metabolismo , Regulación de la Expresión Génica de las Plantas , Variación Genética , Genotipo , Inmunohistoquímica , Hierro/metabolismo , Fijación del Nitrógeno , Phaseolus/genética , Phaseolus/metabolismo , Phaseolus/microbiología , Reguladores del Crecimiento de las Plantas/metabolismo , Nodulación de la Raíz de la Planta , Raíces de Plantas/metabolismo , Rhizobium tropici/fisiología , Rizosfera , Nódulos de las Raíces de las Plantas/metabolismo , Simbiosis
18.
Plant Sci ; 181(2): 151-8, 2011 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-21683880

RESUMEN

Iron is an important nutrient in N(2)-fixing legume nodules. The demand for this micronutrient increases during the symbiosis establishment, where the metal is utilized for the synthesis of various iron-containing proteins in both the plant and the bacteroid. Unfortunately, in spite of its importance, iron is poorly available to plant uptake since its solubility is very low when in its oxidized form Fe(III). In the present study, the effect of iron deficiency on the activity of some proteins involved in Strategy I response, such as Fe-chelate reductase (FC-R), H(+)-ATPase, and phosphoenolpyruvate carboxylase (PEPC) and the protein level of iron regulated transporter (IRT1) and H(+)-ATPase proteins has been investigated in both roots and nodules of a tolerant (Flamingo) and a susceptible (Coco blanc) cultivar of common bean plants. The main results of this study show that the symbiotic tolerance of Flamingo can be ascribed to a greater increase in the FC-R and H(+)-ATPase activities in both roots and nodules, leading to a more efficient Fe supply to nodulating tissues. The strong increase in PEPC activity and organic acid content, in the Flamingo root nodules, suggests that under iron deficiency nodules can modify their metabolism in order to sustain those activities necessary to acquire Fe directly from the soil solution.


Asunto(s)
FMN Reductasa/metabolismo , Deficiencias de Hierro , Proteínas Reguladoras del Hierro/metabolismo , Phaseolus/metabolismo , Fosfoenolpiruvato Carboxilasa/metabolismo , Proteínas de Plantas/metabolismo , Transporte Biológico , Ácidos Carboxílicos/análisis , Compuestos Férricos/metabolismo , Regulación de la Expresión Génica de las Plantas , Genotipo , Hierro/análisis , Hierro/metabolismo , Nitrógeno/metabolismo , Fijación del Nitrógeno , Phaseolus/enzimología , Phaseolus/genética , Fosfoenolpiruvato Carboxilasa/genética , Enfermedades de las Plantas , ATPasas de Translocación de Protón/metabolismo , Nódulos de las Raíces de las Plantas/enzimología , Nódulos de las Raíces de las Plantas/genética , Nódulos de las Raíces de las Plantas/metabolismo , Simbiosis
19.
Tree Physiol ; 31(1): 102-13, 2011 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-21389006

RESUMEN

We analysed Pyrus communis cv. Conference and Cydonia oblonga BA29, differently tolerant to lime-induced chlorosis, to identify the key mechanisms involved in their different performance under Fe deficiency induced by the absence of Fe (-Fe) or by the presence of bicarbonate (+FeBic). Under our experimental conditions, a decrease in root elongation was observed in BA29 under bicarbonate supply. Superoxide dismutase (SOD) and peroxidase (POD) activities were analysed and the relative isoforms were detected by native electrophoresis. The data obtained for both genotypes under -Fe and for BA29 +FeBic suggest the occurrence of overproduction of reactive oxygen species (ROS) and, at the same time, of a scarce capacity to detoxify them. The detection of ROS (O(2)(-) and H(2)O(2)) through histochemical localization supports these results and suggests that they could account for the modifications of mechanical properties of the cell wall during stress adaptation. On the other hand, in the cv. Conference +FeBic, an increase in non-specific POD activity was detected, confirming its higher level of protection in particular against H(2)O(2) accumulation. Peroxidases involved in lignification were assayed and histochemical analysis was performed. The results suggest that only in BA29 under bicarbonate supply can the presence of ROS in root apoplast be correlated with lignin deposits in external layers and in endodermis as a consequence of the shift of PODs towards a lignification role. We suggest that in BA29 the decrease in root growth could impair mineral nutrition, generating susceptibility to calcareous soils. In the cv. Conference, the allocation of new biomass to the root system could improve soil exploration and consequently Fe uptake.


Asunto(s)
Bicarbonatos/farmacología , Hierro/farmacología , Lignina/biosíntesis , Estrés Oxidativo/efectos de los fármacos , Pyrus/fisiología , Rosaceae/fisiología , Pared Celular/metabolismo , Clorofila/metabolismo , Genotipo , Peróxido de Hidrógeno/metabolismo , Hierro/metabolismo , Peroxidasas/metabolismo , Hojas de la Planta/efectos de los fármacos , Hojas de la Planta/genética , Hojas de la Planta/crecimiento & desarrollo , Hojas de la Planta/fisiología , Raíces de Plantas/efectos de los fármacos , Raíces de Plantas/genética , Raíces de Plantas/crecimiento & desarrollo , Raíces de Plantas/fisiología , Pyrus/efectos de los fármacos , Pyrus/genética , Pyrus/crecimiento & desarrollo , Especies Reactivas de Oxígeno/metabolismo , Rosaceae/efectos de los fármacos , Rosaceae/genética , Rosaceae/crecimiento & desarrollo , Suelo , Estrés Fisiológico , Superóxido Dismutasa/metabolismo
20.
BMC Med Genomics ; 2: 36, 2009 Jun 23.
Artículo en Inglés | MEDLINE | ID: mdl-19549311

RESUMEN

BACKGROUND: The presence of MLL rearrangements in acute leukemia results in a complex number of biological modifications that still remain largely unexplained. Armstrong et al. proposed MLL rearrangement positive ALL as a distinct subgroup, separated from acute lymphoblastic (ALL) and myeloblastic leukemia (AML), with a specific gene expression profile. Here we show that MLL, from both ALL and AML origin, share a signature identified by a small set of genes suggesting a common genetic disregulation that could be at the basis of mixed lineage leukemia in both phenotypes. METHODS: Using Affymetrix(R) HG-U133 Plus 2.0 platform, gene expression data from 140 (training set) + 78 (test set) ALL and AML patients with (24+13) and without (116+65) MLL rearrangements have been investigated performing class comparison (SAM) and class prediction (PAM) analyses. RESULTS: We identified a MLL translocation-specific (379 probes) signature and a phenotype-specific (622 probes) signature which have been tested using unsupervised methods. A final subset of 14 genes grants the characterization of acute leukemia patients with and without MLL rearrangements. CONCLUSION: Our study demonstrated that a small subset of genes identifies MLL-specific rearrangements and clearly separates acute leukemia samples according to lineage origin. The subset included well-known genes and newly discovered markers that identified ALL and AML subgroups, with and without MLL rearrangements.

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