RESUMEN
Amino acid permeases (AAPs) are proteins of the integral membrane that play important roles in plant growth, development, and responses to various stresses. The molecular functions of several AAPs were characterized in Arabidopsis and rice, but there is still limited information on wheat. Here, we identified 51 AAP genes (TaAAPs) in the wheat genome, classified into six groups based on phylogenetic and protein structures. The chromosome location and gene duplication analysis showed that gene duplication events played a crucial role in the expansion of the TaAAPs gene family. Collinearity relationship analysis revealed several orthologous AAPs between wheat and other species. Moreover, cis-element analysis of promoter regions and transcriptome data suggested that the TaAAPs can respond to salt stress. A TaAAP1 gene was selected and transformed in wheat. Overexpressing TaAAP1 enhanced salt tolerance by increasing the expression of ethylene synthesis genes (TaACS6/TaACS7/TaACS8) and accumulating more ethylene. The present study provides an overview of the AAP family in the wheat genome as well as information on systematics, phylogenetics, and gene duplication, and shows that overexpressing TaAAP1 enhances salt tolerance by regulating ethylene production. These results serve as a theoretical foundation for further functional studies on TaAAPs in the future.
Asunto(s)
Arabidopsis , Tolerancia a la Sal , Tolerancia a la Sal/genética , Triticum/genética , Filogenia , Etilenos , Sistemas de Transporte de Aminoácidos/genética , Arabidopsis/genéticaRESUMEN
Freezing stress is a major factor limiting production and geographical distribution of temperate crops. Elongator is a six subunit complex with histone acetyl-transferase activity and is involved in plant development and defense responses in Arabidopsis thaliana. However, it is unknown whether and how an elongator responds to freezing stress in plants. In this study, we found that wheat elongator subunit 4 (TaELP4) negatively regulates freezing tolerance through ethylene signaling. TaELP4 promoter contained cold response elements and was up-regulated in freezing stress. Subcellular localization showed that TaELP4 and AtELP4 localized in the cytoplasm and nucleus. Silencing of TaELP4 in wheat with BSMV-mediated VIGS approach significantly elevated tiller survival rate compared to control under freezing stress, but ectopic expression of TaELP4 in Arabidopsis increased leaf damage and survival rate compared with Col-0. Further results showed that TaELP4 positively regulated ACS2 and ACS6 transcripts, two main limiting enzymes in ethylene biosynthesis. The determination of ethylene content showed that TaELP4 overexpression resulted in more ethylene accumulated than Col-0 under freezing stress. Epigenetic research showed that histone H3K9/14ac levels significantly increased in coding/promoter regions of AtACS2 and AtACS6 in Arabidopsis. RT-qPCR assays showed that the EIN2/EIN3/EIL1-CBFs-COR pathway was regulated by TaELP4 under freezing stress. Taken together, our results suggest that TaELP4 negatively regulated plant responses to freezing stress via heightening histone acetylation levels of ACS2 and ACS6 and increasing their transcription and ethylene accumulation.
Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Etilenos/metabolismo , Congelación , Regulación de la Expresión Génica de las Plantas , Histonas/genética , Histonas/metabolismo , Plantas Modificadas Genéticamente/metabolismo , Triticum/genética , Triticum/metabolismoRESUMEN
BACKGROUND AND AIMS: The brittle rachis trait is a feature of many wild grasses, particularly within the tribe Triticeae. Wild Hordeum and Triticum species form a disarticulation layer above the rachis node, resulting in the production of wedge-type dispersal units. In Aegilops longissima, only one or two of the nodes in the central portion of its rachis are brittle. In Triticeae species, the formation of a disarticulation layer above the rachis node requires the co-transcription of the two dominant and complementary genes Btr1 and Btr2. This study aims to establish whether homologues of Btr1 and/or Btr2 underlie the unusual brittle rachis phenotype observed in Ae. longissima. METHODS: Scanning electron microscopy was used to examine the disarticulation surfaces. Quantitative RT-PCR and RNA in situ hybridization experiments were used to identify gene expression in the immature inflorescence. KEY RESULTS: Analysis based on scanning electron microscopy was able to demonstrate that the disarticulation surfaces formed in the Ae. longissima rachis are morphologically indistinguishable from those formed in the rachises of wild Hordeum and Triticum species. RNA in situ hybridization showed that in the immature Ae. longissima inflorescence, the intensity of Btr1 transcription varied from high at the rachis base to low at its apex, while that of Btr2 was limited to the nodes in the central to distal portion of the rachis. CONCLUSIONS: The disarticulation pattern shown by Ae. longissima results from the limitation of Btr1 and Btr2 co-expression to nodes lying in the centre of the rachis.
Asunto(s)
Aegilops , Hordeum , Desarticulación , Genes de Plantas , Hordeum/genética , Triticum/genéticaRESUMEN
KEY MESSAGE: This study provides a link between a de novo gene and novel phenotype in wheat-rye hybrids that can be used as a model for induced de novo genetic variation. Wide hybridization can produce de novo DNA variation that may cause novel phenotypes. However, there is still a lack of specific links between changed genes and novel phenotypes in wide hybrids. The well-studied high-molecular-weight glutenin subunit (HMW-GS) genes in tribe Triticeae provide a useful model for addressing this issue. In this study, we investigated the feasibility of a wheat-rye hybridization method for inducing de novo phenotypes using the Glu-1Dx2.2 subunit as an example. We developed three hexaploid wheat lines with normal fertility and a Glu-1Dx2.2 variant, named Glu-1Dx2.2 (v) , derived from three F1 hybrids. The wild-type Glu-1Dx2.2 has two direct repeats of 295 bp length separated by an intervening 101 bp in its central repetitive region. In the mutant Glu-1Dx2.2 (v) , one copy of the repeats and the intervening sequence were deleted, probably through homology-dependent illegitimate recombination (IR). This study provides a direct link between a de novo allele and novel phenotype. Our results indicate that the wheat-rye method may be a useful tool to induce de novo genetic variations that broaden the genetic diversity for wheat improvement.
Asunto(s)
Glútenes/genética , Hibridación Genética , Secale/genética , Triticum/genética , Alelos , Clonación Molecular , ADN de Plantas/genética , Fenotipo , Análisis de Secuencia de ADNRESUMEN
The mechanism of action of low-density lipoprotein receptor related protein 4 (LRP4) is mediated largely via the Agrin-LRP4-MuSK signalling pathway in the nervous system. LRP4 contributes to the development of synapses in the peripheral nervous system (PNS). It interacts with signalling molecules such as the amyloid beta-protein precursor (APP) and the wingless type protein (Wnt). Its mechanisms of action are complex and mediated via interaction between the pre-synaptic motor neuron and post-synaptic muscle cell in the PNS, which enhances the development of the neuromuscular junction (NMJ). LRP4 may function differently in the central nervous system (CNS) than in the PNS, where it regulates ATP and glutamate release via astrocytes. It mayaffect the growth and development of the CNS by controlling the energy metabolism. LRP4 interacts with Agrin to maintain dendrite growth and density in the CNS. The goal of this article is to review the current studies involving relevant LRP4 signaling pathways in the nervous system. The review also discusses the clinical and etiological roles of LRP4 in neurological illnesses, such as myasthenia gravis, Alzheimer's disease and epilepsy. In this review, we provide a theoretical foundation for the pathogenesis and therapeutic application of LRP4 in neurologic diseases.
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Agrina , Proteínas Relacionadas con Receptor de LDL , Proteínas Relacionadas con Receptor de LDL/metabolismo , Agrina/metabolismo , Péptidos beta-Amiloides/metabolismo , Proteínas Tirosina Quinasas Receptoras/metabolismo , Unión Neuromuscular/metabolismoRESUMEN
BACKGROUND: The association of BCAAs (isoleucine, leucine, and valine) with cardiovascular and cerebrovascular diseases has been widely recognized by researchers, but there is limited evidence to support the relationship between BCAAs and multiple chronic conditions (MCCs) in older adults. This study aimed to explore the correlation between BCAA levels in the diets of older adults and MCCs. METHODS: Based on a health management cohort project in Nanshan District of Shenzhen, 4278 individuals over 65 years old were selected as participants via multi-stage stratified sampling from May 2018 to December 2019. Data were collected using a validated semi-quantitative food frequency questionnaire, as well as anthropometric and chronic disease reports. MCC was defined as the coexistence of two or more chronic diseases, namely, hypertension, dyslipidemia, diabetes, CAD, stroke, CKD, and CLD. Multivariate unconditional logistic regression analysis was used to analyze the relationship between dietary BCAAs and MCCs in older adults, and then, gender stratification analysis was performed. A restricted cubic spline model (a fitted smooth curve) was used to determine the dose-response relationship of isoleucine with MCCs. RESULTS: A total of 4278 older adults aged 65 and above were included in this study, with an average age of 72.73 ± 5.49 years. The cohort included 1861 males (43.50%). Regardless of whether confounding factors were corrected, isoleucine was a risk factor for MCCs (OR = 3.388, 95%CI:1.415,8.109). After gender stratification, the relationships between dietary isoleucine and MCCs (OR = 6.902, 95%CI:1.875,25.402) and between leucine (OR = 0.506,95%CI:0.309,0.830) and MCCs were significant in women, but not in men. No significant association between valine and MCCs was observed. In addition, isoleucine was a risk factor for MCCs when its intake was greater than 4.297 g/d. CONCLUSION: Isoleucine may play an important role in regulating age-related diseases. BCAAs such as isoleucine can be used as risk markers for MCCs in older adults.
RESUMEN
A new strategy for the construction of a sensitive and stable electrochemiluminescent platform based on titanate nanotubes (TNTs) and Nafion composite modified electrode for luminol is described, TNTs contained composite modified electrodes that showed some photocatalytic activity toward luminol electrochemiluminescence emission, and thus could dramatically enhance luminol light emission. This extremely sensitive and stable platform allowed a decrease of the experiment electrochemiluminescence luminol reagent. In addition, in luminol solution at low concentrations, we compared the capabilities of a bare glassy carbon electrode with the TNT composite modified electrode for hydrogen peroxide detection. The results indicated that compared with glassy carbon electrode this platform was extraordinarily sensitive to hydrogen peroxide. Therefore, by combining with an appropriate enzymatic reaction, this platform would be a sensitive matrix for many biomolecules.
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Carbono/química , Técnicas Electroquímicas , Luminiscencia , Luminol/química , Nanotubos/química , Titanio/química , Electrodos , Vidrio/químicaRESUMEN
The ankle-link complex (ALC) consists of USH2A, WHRN, PDZD7, and ADGRV1 and plays an important role in hair cell development. At present, its architectural organization and signaling role remain unclear. By establishing Adgrv1 Y6236fsX1 mutant mice as a model of the deafness-associated human Y6244fsX1 mutation, the authors show here that the Y6236fsX1 mutation disrupts the interaction between adhesion G protein-coupled receptor V subfamily member 1 (ADGRV1) and other ALC components, resulting in stereocilia disorganization and mechanoelectrical transduction (MET) deficits. Importantly, ADGRV1 inhibits WHRN phosphorylation through regional cAMP-PKA signaling, which in turn regulates the ubiquitination and stability of USH2A via local signaling compartmentalization, whereas ADGRV1 Y6236fsX1 does not. Yeast two-hybrid screening identified the E3 ligase WDSUB1 that binds to WHRN and regulates the ubiquitination of USH2A in a WHRN phosphorylation-dependent manner. Further FlAsH-BRET assay, NMR spectrometry, and mutagenesis analysis provided insights into the architectural organization of ALC and interaction motifs at single-residue resolution. In conclusion, the present data suggest that ALC organization and accompanying local signal transduction play important roles in regulating the stability of the ALC.
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Sordera , Animales , Humanos , Ratones , Proteínas Portadoras/genética , Sordera/genética , Sordera/metabolismo , Proteínas de la Matriz Extracelular/química , Proteínas de la Matriz Extracelular/genética , Proteínas de la Matriz Extracelular/metabolismo , Mutación/genética , FosforilaciónRESUMEN
Seed dispersal among wild species belonging to the tribe Triticeae is typically achieved by the formation of a brittle rachis. The trait relies on the development of a disarticulation layer, most frequently above the rachis node (resulting in wedge type dispersal units), but in some species below the rachis node (resulting in barrel type dispersal units). The genes responsible for the former type are the complementary pair Btr1 and Btr2, while the genetic basis of the latter type has yet to be determined. Aegilops tauschii forms barrel type dispersal units and previous study showed this species lacked an intact copy of Btr1. Here it has been demonstrated that Ae. tauschii carries two of Btr2; and that Btr2 transcript is present in a region below the rachis node where the abscission zone forms. The implication is that in this species, the Btr2 product is involved in the formation of barrel type dispersal units.
RESUMEN
In many non-cultivated angiosperm species, seed dispersal is facilitated by the shattering of the seed head at maturity; in the Triticeae tribe, to which several of the world's most important cereals belong, shattering takes the form of a disarticulation of the rachis. The products of the genes Btr1 and Btr2 are both required for disarticulation to occur above the rachis nodes within the genera Hordeum (barley) and Triticum/Aegilops (wheat). Here, it has been shown that both Btr1 and Btr2 are specific to the Triticeae tribe, although likely paralogs (Btr1-like and Btr2-like) are carried by the family Poaceae including Triticeae. Aegilops tauschii (the donor of the bread wheat D genome) lacks a copy of Btr1 and disarticulation in this species occurs below, rather than above the rachis node; thus, the product of Btr1 appears to be required for disarticulation to occur above the rachis node.
RESUMEN
Saxitoxin (STX) has high toxicity, and is water soluble, acid stable and thermostable. Therefore, STX in seawater can be accumulated by marine organism to form bioaccumulation. To ensure the safety of seafood for consumption, it is crucial to accurately determine trace STX in seawater and seafood. We herein developed a novel magnetic electrochemical immunosensor for ultra-sensitive detection of STX in seawater and seafood by using non-competitive strategy. The immunosensor employs STX-specific antibody-functionalized magnetic beads (MBs) for STX recognition, palladium-doped graphitic carbon nitride (g-C3N4-PdNPs) peroxidase mimetic for catalyzing H2O2-mediated oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) to generate signal. The immunosensor combines the merits of g-C3N4-PdNPs peroxidase mimetic, non-competitive strategy, MBs-based antibody recognition and magnetic gold electrode, and thus has excellent stability, lower cost, no risk of false positive result, high sensitivity and strong ability resist to matrix interference. The proposed immunosensor has been successfully used to detect trace STX in seawater and shellfish samples with a detection limit of 1.2â¯pg/mL (4.0â¯×â¯10-12 M), a recovery of 93-107% and a relative standard deviation (RSD, nâ¯=â¯5) <â¯5%. The success of this study provided a promising approach for the rapid and on-site detection of trace STX in seawater and seafood.
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Técnicas Biosensibles , Técnicas Electroquímicas , Saxitoxina/aislamiento & purificación , Alimentos Marinos/análisis , Oro/química , Humanos , Límite de Detección , Saxitoxina/química , MariscosRESUMEN
We herein developed a novel biosensor for the visual detection of trace uranyl ion (UO2(2+)) in aqueous environment with high sensitivity and specificity by using DNAzyme-functionalized magnetic beads (MBs) for UO2(2+) recognition and gold nano-particles (AuNPs)-based enzymatic catalysis oxidation of TMB (3,3',5,5'-tetramethylbenzidine sulfate) for signal generation. The utilization of MBs facilitates the magnetic separation and collection of sensing system from complex sample solution, which leads to more convenient experimental operation and more strong resistibility of the biosensor to the matrix of sample, and the utilization of AuNPs-based enzymatic catalysis amplification greatly improved the sensitivity of the biosensor. Compared with the previous DNAzyme-based UO2(2+) sensors, the proposed biosensor has outstanding advantages such as relative high sensitivity and specificity, operation convenience, low cost and more strong resistibility to the matrix of sample. It can be used to detect as low as 0.02 ppb (74 pM) of UO2(2+) in aqueous environment by only naked-eye observation and 1.89 ppt (7.0 pM) of UO2(2+) by UV-visible spectrophotometer with a recovery of 93-99% and a RSD ≤ 5.0% (n=6) within 3h. Especially, the visual detection limit of 0.02 ppb (74 pM) is much lower than the maximum allowable level of UO2(2+) (130 nM) in the drinking water defined by the U.S. Environmental Protection Agency (EPA), indicating that our method meets the requirement of rapid and on-site detection of UO2(2+) in the aqueous environment by only naked-eye observation.
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Técnicas Biosensibles/métodos , ADN Catalítico/aislamiento & purificación , Compuestos de Uranio/aislamiento & purificación , Catálisis , Colorimetría , Agua Potable/análisis , Oro/química , Límite de Detección , Campos Magnéticos , Nanopartículas del Metal/química , Estados Unidos , Compuestos de Uranio/toxicidadRESUMEN
A novel fluorescent biosensor for detecting uranyl ion (UO2(2+)) in aqueous environment has been developed based on the specific recognition of DNAzyme and the fluorescence quenching ability of molybdenum disulfide (MoS2) nanosheets. The DNAzyme contains a DNA enzyme strand and a 6-carboxylfluorescein (FAM)-labeled DNA substrate strand. We demonstrated that MoS2 nanosheets have low affinity to the substrate-enzyme complex DNAzyme. Whereas, in the presence of UO2(2+), UO2(2+) can specifically cleave DNAzyme to release FAM-labeled single-strand DNA and the released FAM-labeled single-strand DNA can be firmly adsorbed on the surface of MoS2 nanosheets, which resulted in an obvious decrease of fluorescence intensity. This provided a sensing platform for the rapid, simple and sensitive fluorescent detection of UO2(2+). By using the sensing platform, a sensitive and selective fluorescent method for the rapid detection of UO2(2+) has been developed. In comparison with previous biosensor, the proposed method has obvious analytical advantage such as relatively high sensitivity and good stability, short analytical time and low cost. It can be used to detect as low as 2.14 nM of UO2(2+) in aqueous environment with a recovery of 96-102% and a RSD<5% (n=6). The success of this study provides a promising alternative for the rapid and on-site detection of UO2(2+) in environmental monitoring.