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"Rubio, María Belén"
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Trichoderma harzianum Volatile Organic Compounds Regulated by the THCTF1 Transcription Factor Are Involved in Antifungal Activity and Beneficial Plant Responses
by
Gualtieri, Liberata
,
Monte, Enrique
,
Rubio, María Belén
in
Antifungal activity
,
Arabidopsis
,
Benzaldehyde
2023
The transcription factor THCTF1 from Trichoderma harzianum, previously linked to the production of 6-pentyl-2H-pyran-2-one (6-PP) derivatives and antifungal activity against Fusarium oxysporum, has been related in this study to conidiation, production of an array of volatile organic compounds (VOCs) and expression of methyltransferase genes. VOCs emitted by three T. harzianum strains (wild type T34, transformant ΔD1-38 that is disrupted in the Thctf1 gene encoding the transcription factor THCTF1, and ectopic integration transformant ΔJ3-16) were characterized by Proton Transfer Reaction-Quadrupole interface-Time-Of-Flight-Mass Spectrometry (PTR-Qi-TOF-MS). Thctf1 disruption affected the production of numerous VOCs such as the antifungal volatiles 2-pentyl furan and benzaldehyde which were under-emitted, and acetoine, a plant systemic defense inductor, which was over-emitted. Biological assays show that VOCs regulated by THCTF1 are involved in the T. harzianum antifungal activity against Botrytis cinerea and in the beneficial effects leading to Arabidopsis plant development. The VOC blend from the disruptant ΔD1-38: (i) inhibited Arabidopsis seed germination for at least 26 days and (ii) when applied to Arabidopsis seedlings resulted in increased jasmonic acid- and salicylic acid-dependent defenses.
Journal Article
Microbiological Evaluation of the Disinfecting Potential of UV-C and UV-C Plus Ozone Generating Robots
by
Monte, Enrique
,
Rubio, María Belén
,
Morán-Diez, María Eugenia
in
Antiinfectives and antibacterials
,
Bacillus subtilis
,
Bacteria
2021
This study examined the microbicidal activity of ultraviolet (UV)-C185–256-nm irradiance (robot 1) and ozone generated at UV-C185-nm by low-pressure mercury vapor lamps (robot 2) adapted to mobile robotic devices for surface decontamination, which was achieved in less than 1 h. Depending on their wall structure and outer envelopes, many microorganisms display different levels of resistance to decontaminating agents. Thus, the need for novel disinfection approaches is further exacerbated by the increased prevalence of multidrug-resistant bacteria, as well as the potential of novel microorganisms, with the ability to cause disease outbreaks. To set up a rapid and effective approach for microorganisms propagation prevention, we focused on the effects of UV-C and ozone on a distinct microorganism survival ratio. A set of microorganisms, including Escherichia coli, Micrococcus luteus, Saccharomyces cerevisiae, Trichoderma harzianum, and Bacillus subtilis, were used to evaluate the disinfection power of UV-C and UV-C plus ozone generating robots. UV-C disinfection can be suited to ad hoc tasks, is easy to operate, requires low maintenance, does not have the need for the storage of dangerous chemicals, and does not produce by-products that may affect human health and the environment. The robotic cumulative irradiation technology developed (fluence accumulated values of 2.28 and 3.62 mJ cm−2, for robot 1 and 2, respectively), together with the production of ozone (with a maximum peak of 0.43 ppm) capable of reaching UV-C shaded surfaces, and analyzed in the current study, despite being designed for the need to reduce the risk of epidemic outbreaks in real-life scenarios, represents a versatile tool that could be employed for air and surface disinfection within many circumstances that are faced daily.
Journal Article
Effectiveness of composts and Trichoderma strains for control of Fusarium wilt of tomato
by
NICOLÁS, Carlos
,
HERMOSA, Rosa
,
RUBIO, María Belén
in
antagonism
,
Biological control
,
Botrytis cinerea
2015
Fusarium wilt caused by Fusarium oxysporum f. sp. lycopersici (FOL) is a major limiting disease in tomato production in Morocco. Commercial and locally produced Moroccan composts and peat were found to reduce Fusarium wilt in tomato plants. We explored the presence of Trichoderma strains in these materials, and in six soils sampled in the North West of Morocco, where a low incidence of Fusarium wilt had been previously observed. The most abundant Trichoderma-like fungus was selected from each soil, compost or peat sample. Twelve Trichoderma strains were isolated and identified molecularly. Trichoderma asperellum CT9 and Trichoderma virens ST11 showed the greatest overall antagonistic activity against FOL, Rhizoctonia solani, Botrytis cinerea and Pythium ultimum. The three strains evaluated in in planta tests, CT9, ST11 and T. virens ST10, reduced tomato Fusarium wilt, and strain ST11 also promoted growth of tomato plants.
Journal Article
Tomato progeny inherit resistance to the nematode Meloidogyne javanica linked to plant growth induced by the biocontrol fungus Trichoderma atroviride
by
Araújo Filho, Jerônimo Vieira de
,
Castillo, Pablo
,
Medeiros, Hugo Agripino de
in
38/39
,
38/77
,
631/449/2169/2674
2017
Root-knot nematodes (RKN) are major crop pathogens worldwide.
Trichoderma
genus fungi are recognized biocontrol agents and a direct activity of
Trichoderma atroviride
(Ta) against the RKN
Meloidogyne javanica
(Mj), in terms of 42% reduction of number of galls (NG), 60% of number of egg masses and 90% of number of adult nematodes inside the roots, has been observed in tomato grown under greenhouse conditions. An
in vivo
split-root designed experiment served to demonstrate that Ta induces systemic resistance towards Mj, without the need for the organisms to be in direct contact, and significantly reduces NG (20%) and adult nematodes inside tomato roots (87%). The first generation (F1) of Ta-primed tomato plants inherited resistance to RKN; although, the induction of defenses occurred through different mechanisms, and in varying degrees, depending on the Ta-Mj interaction. Plant growth promotion induced by Ta was inherited without compromising the level of resistance to Mj, as the progeny of Ta-primed plants displayed increased size and resistance to Mj without fitness costs. Gene expression results from the defense inductions in the offspring of Ta-primed plants, suggested that an auxin-induced reactive oxygen species production promoted by Ta may act as a major defense strategy during plant growth.
Journal Article
Accuracy of high-risk HPV DNA PCR, p16(INK4a) immunohistochemistry or the combination of both to diagnose HPV-driven oropharyngeal cancer
2022
Background
The incidence of high-risk human papillomavirus (hrHPV)-driven head and neck squamous cell carcinoma, in particular oropharyngeal cancers (OPC), is increasing in high-resource countries. Patients with HPV-induced cancer respond better to treatment and consequently have lower case-fatality rates than patients with HPV-unrelated OPC. These considerations highlight the importance of reliable and accurate markers to diagnose truly HPV-induced OPC.
Methods
The accuracy of three possible test strategies, i.e. (a) hrHPV DNA PCR (DNA), (b) p16
(INK4a)
immunohistochemistry (IHC) (p16), and (c) the combination of both tests (considering joint DNA and p16 positivity as positivity criterion), was analysed in tissue samples from 99 Belgian OPC patients enrolled in the HPV-AHEAD study. Presence of HPV E6*I mRNA (mRNA) was considered as the reference, indicating HPV etiology.
Results
Ninety-nine OPC patients were included, for which the positivity rates were 36.4%, 34.0% and 28.9% for DNA, p16 and mRNA, respectively. Ninety-five OPC patients had valid test results for all three tests (DNA, p16 and mRNA). Using mRNA status as the reference, DNA testing showed 100% (28/28) sensitivity, and 92.5% (62/67) specificity for the detection of HPV-driven cancer. p16 was 96.4% (27/28) sensitive and equally specific (92.5%; 62/67). The sensitivity and specificity of combined p16 + DNA testing was 96.4% (27/28) and 97.0% (65/67), respectively. In this series, p16 alone and combined p16 + DNA missed 1 in 28 HPV driven cancers, but p16 alone misclassified 5 in 67 non-HPV driven as positive, whereas combined testing would misclassify only 2 in 67.
Conclusions
Single hrHPV DNA PCR and p16
(INK4a)
IHC are highly sensitive but less specific than using combined testing to diagnose HPV-driven OPC patients. Disease prognostication can be encouraged based on this combined test result.
Journal Article
HPV DNA genotyping, HPV E6I mRNA detection, and p16 INK4a /Ki-67 staining in Belgian head and neck cancer patient specimens, collected within the HPV-AHEAD study
2021
The main risk factors for head and neck cancer (HNC) are tobacco and alcohol use. However, an important fraction of oropharyngeal cancer (OPC) is caused by human papillomaviruses (HPV), a subgroup with increasing incidence in several western countries.
As part of the HPV-AHEAD study, we assessed the role of HPV infection in 772 archived tissue specimens of Belgian HNC patients: 455 laryngeal (LC), 106 oral cavity (OCC), 99 OPC, 76 hypopharyngeal (HC), and 36 unspecified parts of the head and neck. All specimens were tested for HPV DNA (21 genotypes); whereof all HPV DNA-positives, all HPV DNA-negative OPCs and a random subset of HPV DNA-negatives of the other HNC-sites were tested for the presence of type-specific HPV RNA and p16
over-expression.
The highest HPV DNA prevalence was observed in OPC (36.4 %), and was significantly lower (p < 0.001) in the other HNCs (OCC:7.5 %, LC:6.6 %). HPV16 was the most common HPV-genotype in all HNCs. Approximately 83.0 % of the HPV DNA-positive OPCs tested HPV RNA or p16-positive, compared to about 37.5 % and 44.0 % in OCC and LC, respectively. Estimation of the attributable fraction of an HPV infection in HNC was very similar for HPV RNA or p16 in addition to DNA-positivity; with 30 % for OPC, and 3 % for OCC and LC.
Our study confirms the heterogeneity of HPV DNA prevalence across anatomical sites in HNC, with a predominance of HPV16 in all sites. The estimated proportion of HPV-driven HNC in Belgium, during the period 1980-2014, was 10 times higher in OPC compared to OCC and LC.
Journal Article
Effect of inorganic N top dressing and Trichoderma harzianum seed-inoculation on crop yield and the shaping of root microbial communities of wheat plants cultivated under high basal N fertilization
by
Martínez de Alba, Ángel Emilio
,
Rubio Pérez, María Belén
,
Morán Díez, María Eugenia
in
Bacteria
,
bacterial composition
,
bulk soil
2020
This work was supported by the Spanish Government (Project RTI2018-099986-B-I00) and the European Regional Development Fund (FEDER) under the Regional Government of Castile and Leon support (Projects SA270P18 and Escalera de Excelencia CLU-2018-04). MI was granted by a contract by Diputación de Salamanca (Projects-2018), MM-D was granted by the Program II of Postdoctoral Fellows of the University of Salamanca, and AEMA was granted with a postdoctoral contract of the Regional Government of Castile and Leon (Project SA270P18).
Journal Article
ITrichoderma harzianum/I Volatile Organic Compounds Regulated by the THCTF1 Transcription Factor Are Involved in Antifungal Activity and Beneficial Plant Responses
by
Gualtieri, Liberata
,
Monte, Enrique
,
Rubio, María Belén
in
Antifungal agents
,
Arabidopsis thaliana
,
Genetic aspects
2023
The transcription factor THCTF1 from Trichoderma harzianum, previously linked to the production of 6-pentyl-2H-pyran-2-one (6-PP) derivatives and antifungal activity against Fusarium oxysporum, has been related in this study to conidiation, production of an array of volatile organic compounds (VOCs) and expression of methyltransferase genes. VOCs emitted by three T. harzianum strains (wild type T34, transformant ΔD1-38 that is disrupted in the Thctf1 gene encoding the transcription factor THCTF1, and ectopic integration transformant ΔJ3-16) were characterized by Proton Transfer Reaction-Quadrupole interface-Time-Of-Flight-Mass Spectrometry (PTR-Qi-TOF-MS). Thctf1 disruption affected the production of numerous VOCs such as the antifungal volatiles 2-pentyl furan and benzaldehyde which were under-emitted, and acetoine, a plant systemic defense inductor, which was over-emitted. Biological assays show that VOCs regulated by THCTF1 are involved in the T. harzianum antifungal activity against Botrytis cinerea and in the beneficial effects leading to Arabidopsis plant development. The VOC blend from the disruptant ΔD1-38: (i) inhibited Arabidopsis seed germination for at least 26 days and (ii) when applied to Arabidopsis seedlings resulted in increased jasmonic acid- and salicylic acid-dependent defenses.
Journal Article
SARS-CoV-2 RNA Detection in Wastewater and Its Effective Correlation with Clinical Data during the Outbreak of COVID-19 in Salamanca
by
Martínez de Alba, Ángel Emilio
,
Muro, Antonio
,
García-Prieto, Juan Carlos
in
COVID-19
,
COVID-19 - diagnosis
,
COVID-19 - epidemiology
2024
Wastewater treatment plants (WWTPs) are the final stage of the anthropogenic water cycle where a wide range of chemical and biological markers of human activity can be found. In COVID-19 disease contexts, wastewater surveillance has been used to infer community trends based on viral abundance and SARS-CoV-2 RNA variant composition, which has served to anticipate and establish appropriate protocols to prevent potential viral outbreaks. Numerous studies worldwide have provided reliable and robust tools to detect and quantify SARS-CoV-2 RNA in wastewater, although due to the high dilution and degradation rate of the viral RNA in such samples, the detection limit of the pathogen has been a bottleneck for the proposed protocols so far. The current work provides a comprehensive and systematic study of the different parameters that may affect the detection of SARS-CoV-2 RNA in wastewater and hinder its quantification. The results obtained using synthetic viral RNA as a template allow us to consider that 10 genome copies per µL is the minimum RNA concentration that provides reliable and consistent values for the quantification of SARS-CoV-2 RNA. RT-qPCR analysis of wastewater samples collected at the WWTP in Salamanca (western Spain) and at six pumping stations in the city showed that below this threshold, positive results must be confirmed by sequencing to identify the specific viral sequence. This allowed us to find correlations between the SARS-CoV-2 RNA levels found in wastewater and the COVID-19 clinical data reported by health authorities. The close match between environmental and clinical data from the Salamanca case study has been confirmed by similar experimental approaches in four other cities in the same region. The present methodological approach reinforces the usefulness of wastewater-based epidemiology (WBE) studies in the face of future pandemic outbreaks.
Journal Article
HPV DNA genotyping, HPV E6I mRNA detection, and p16INK4a/Ki-67 staining in Belgian head and neck cancer patient specimens, collected within the HPV-AHEAD study
by
Vermeulen, Peter
,
Pawlita, Michael
,
Lucas, Eric
in
Head and neck neoplasms
,
Human papillomavirus
,
Human papillomavirus DNA tests
2021
•Heterogeneity of HPV DNA prevalence across anatomical sites in Belgian HNC patients.•HPV16 is the predominant genotype in all HNC sites.•Estimated proportion of HPV-driven HNC (1980–2014): 30 % for OPC, 3 % for OC and LC.
The main risk factors for head and neck cancer (HNC) are tobacco and alcohol use. However, an important fraction of oropharyngeal cancer (OPC) is caused by human papillomaviruses (HPV), a subgroup with increasing incidence in several western countries.
As part of the HPV-AHEAD study, we assessed the role of HPV infection in 772 archived tissue specimens of Belgian HNC patients: 455 laryngeal (LC), 106 oral cavity (OCC), 99 OPC, 76 hypopharyngeal (HC), and 36 unspecified parts of the head and neck. All specimens were tested for HPV DNA (21 genotypes); whereof all HPV DNA-positives, all HPV DNA-negative OPCs and a random subset of HPV DNA-negatives of the other HNC-sites were tested for the presence of type-specific HPV RNA and p16INK4a over-expression.
The highest HPV DNA prevalence was observed in OPC (36.4 %), and was significantly lower (p < 0.001) in the other HNCs (OCC:7.5 %, LC:6.6 %). HPV16 was the most common HPV-genotype in all HNCs. Approximately 83.0 % of the HPV DNA-positive OPCs tested HPV RNA or p16-positive, compared to about 37.5 % and 44.0 % in OCC and LC, respectively. Estimation of the attributable fraction of an HPV infection in HNC was very similar for HPV RNA or p16 in addition to DNA-positivity; with 30 % for OPC, and 3 % for OCC and LC.
Our study confirms the heterogeneity of HPV DNA prevalence across anatomical sites in HNC, with a predominance of HPV16 in all sites. The estimated proportion of HPV-driven HNC in Belgium, during the period 1980–2014, was 10 times higher in OPC compared to OCC and LC.
Journal Article