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1.
Nat Methods ; 20(7): 1010-1020, 2023 07.
Artigo em Inglês | MEDLINE | ID: mdl-37202537

RESUMO

The Cell Tracking Challenge is an ongoing benchmarking initiative that has become a reference in cell segmentation and tracking algorithm development. Here, we present a significant number of improvements introduced in the challenge since our 2017 report. These include the creation of a new segmentation-only benchmark, the enrichment of the dataset repository with new datasets that increase its diversity and complexity, and the creation of a silver standard reference corpus based on the most competitive results, which will be of particular interest for data-hungry deep learning-based strategies. Furthermore, we present the up-to-date cell segmentation and tracking leaderboards, an in-depth analysis of the relationship between the performance of the state-of-the-art methods and the properties of the datasets and annotations, and two novel, insightful studies about the generalizability and the reusability of top-performing methods. These studies provide critical practical conclusions for both developers and users of traditional and machine learning-based cell segmentation and tracking algorithms.


Assuntos
Benchmarking , Rastreamento de Células , Rastreamento de Células/métodos , Aprendizado de Máquina , Algoritmos
2.
Nano Lett ; 22(4): 1604-1608, 2022 Feb 23.
Artigo em Inglês | MEDLINE | ID: mdl-35129990

RESUMO

Generally speaking, for a semiconductor, the temperature dependence of excitonic emission corresponds to that of its band gap. However, an anomalous behavior is exhibited by the excitonic luminescence of diamond where as the temperature increases (from 10 to 300 K), its indirect exciton luminescence peak displays a spectral-distinguishable blue shift, whereas the indirect band-gap absorption shows a weak red shift. According to experimental high-resolution deep-ultraviolet spectra and theoretical analysis, the weak red shift of its indirect band gap is ascribed to its large Debye temperature (ΘD ≈ 2220 K), which makes the lattice constant change comparatively little in a large temperature range, so the change of its band gap is relatively small; in this case, as the temperature rises, the thermal population of valence-band holes that moves to a high-energy state far away from the Fermi surface contributes to the macroscopic blue shift of its excitonic emission.

3.
Plant J ; 105(3): 771-785, 2021 02.
Artigo em Inglês | MEDLINE | ID: mdl-33160290

RESUMO

Plants have evolved numerous receptor-like kinases (RLKs) that modulate environmental stress responses. However, little is known regarding soybean (Glycine max) RLKs. We have previously identified that Glycine soja Ca2+ /CAM-binding RLK (GsCBRLK) is involved in salt tolerance. Here, we report that soluble NSF attachment protein receptor proteins BET1s mediate subcellular localization of calmodulin-binding receptor-like cytoplasmic kinases CRCK1s to modulate salt stress responses. Direct interaction between GsCBRLK and GsBET11a was initially identified via yeast two-hybrid and bimolecular fluorescence complementation assays. Further analysis demonstrated conserved interaction between BET1s and CRCK1s. GsCBRLK interacted with all BET1 proteins in wild soybean (Glycine soja) and Arabidopsis, and GsBET11a strongly associated with GsCRCK1a-1d, but slightly with AtCRCK1. In addition, GsBET11a interacted with GsCBRLK via its C-terminal transmembrane domain (TMD), where the entire TMD, not the sequence, was critical for the interaction. Moreover, the N-terminal variable domain (VD) of GsCBRLK was responsible for interacting with GsBET11a, and the intensity of interaction between GsCBRLK/AtCRCK1 and GsBET11a was dependent on VD. Furthermore, GsBET11a was able to mediate the GsCBRLK subcellular localization via direct interaction with VD. Additionally, knockout of AtBET11 or AtBET12 individually did not alter GsCBRLK localization, while GsBET11a expression caused partial internalization of GsCBRLK from the plasma membrane (PM). We further suggest the necessity of GsCBRLK VD for its PM localization via N-terminal truncation assays. Finally, GsBET11a was shown to confer enhanced salt stress tolerance when overexpressed in Arabidopsis and soybean. These results revealed the conserved and direct interaction between BET1s and CRCK1s, and suggested their involvement in salt stress responses.


Assuntos
Glycine max/fisiologia , Proteínas de Plantas/metabolismo , Proteínas SNARE/metabolismo , Estresse Salino/fisiologia , Arabidopsis/genética , Proteínas Quinases Dependentes de Cálcio-Calmodulina/metabolismo , Membrana Celular/metabolismo , Secas , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Domínios e Motivos de Interação entre Proteínas , Proteínas SNARE/genética
4.
Bioinformatics ; 37(24): 4844-4850, 2021 12 11.
Artigo em Inglês | MEDLINE | ID: mdl-34329376

RESUMO

MOTIVATION: Live cell segmentation is a crucial step in biological image analysis and is also a challenging task because time-lapse microscopy cell sequences usually exhibit complex spatial structures and complicated temporal behaviors. In recent years, numerous deep learning-based methods have been proposed to tackle this task and obtained promising results. However, designing a network with excellent performance requires professional knowledge and expertise and is very time-consuming and labor-intensive. Recently emerged neural architecture search (NAS) methods hold great promise in eliminating these disadvantages, because they can automatically search an optimal network for the task. RESULTS: We propose a novel NAS-based solution for deep learning-based cell segmentation in time-lapse microscopy images. Different from current NAS methods, we propose (i) jointly searching non-repeatable micro architectures to construct the macro network for exploring greater NAS potential and better performance and (ii) defining a specific search space suitable for the live cell segmentation task, including the incorporation of a convolutional long short-term memory network for exploring the temporal information in time-lapse sequences. Comprehensive evaluations on the 2D datasets from the cell tracking challenge demonstrate the competitiveness of the proposed method compared to the state of the art. The experimental results show that the method is capable of achieving more consistent top performance across all ten datasets than the other challenge methods. AVAILABILITYAND IMPLEMENTATION: The executable files of the proposed method as well as configurations for each dataset used in the presented experiments will be available for non-commercial purposes from https://github.com/291498346/nas_cellseg. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.


Assuntos
Aprendizado Profundo , Redes Neurais de Computação , Microscopia , Imagem com Lapso de Tempo , Processamento de Imagem Assistida por Computador/métodos
5.
Brain ; 144(8): 2541-2553, 2021 09 04.
Artigo em Inglês | MEDLINE | ID: mdl-33792674

RESUMO

Direct electrical stimulation, the transient 'lesional' method probing brain function, has been utilized in identifying the language cortex and preserving language function during epilepsy and neuro-oncological surgeries for about a century. However, comparison of functional maps of the language cortex across languages/continents based on cortical stimulation remains unclear. We conducted a retrospective multicentre study including four cohorts of direct electrical stimulation mapping from four centres across three continents, where three indigenous languages (English, French and Mandarin) are spoken. All subjects performed the two most common language tasks: number counting and picture naming during stimulation. All language sites were recorded and normalized to the same brain template. Next, Spearman's correlation analysis was performed to explore the consistency of the distributions of the language cortex across centres, a kernel density estimation to localize the peak coordinates, and a hierarchical cluster analysis was performed to detect the crucial epicenters. A total of 598 subjects with 917 speech arrest sites (complete interruption of ongoing counting) and 423 anomia sites (inability to name or misnaming) were included. Different centres presented highly consistent distribution patterns for speech arrest (Spearman's coefficient r ranged from 0.60 to 0.85, all pair-wise correlations P < 0.05), and similar patterns for anomia (Spearman's coefficient r ranged from 0.37 to 0.80). The combinational speech arrest map was divided into four clusters: cluster 1 mainly located in the ventral precentral gyrus and pars opercularis, which contained the peak of speech arrest in the ventral precentral gyrus; cluster 2 in the ventral and dorsal precentral gyrus; cluster 3 in the supplementary motor area; cluster 4 in the posterior superior temporal gyrus and supramarginal gyrus. The anomia map revealed two clusters: one was in the posterior part of the superior and middle temporal gyri, which peaked at the posterior superior temporal gyrus; and the other within the inferior frontal gyrus, peaked at the pars triangularis. This study constitutes the largest series to date of language maps generated from direct electrical stimulation mapping. The consistency of data provides evidence for common language networks across languages, in the context of both speech and naming circuit. Our results not only clinically offer an atlas for language mapping and protection, but also scientifically provide better insight into the functional organization of language networks.


Assuntos
Anomia/fisiopatologia , Mapeamento Encefálico/métodos , Encéfalo/fisiopatologia , Distúrbios da Fala/fisiopatologia , Fala/fisiologia , Neoplasias Encefálicas/fisiopatologia , Neoplasias Encefálicas/cirurgia , Estimulação Elétrica , Epilepsias Parciais/fisiopatologia , Epilepsias Parciais/cirurgia , Humanos , Idioma , Estudos Retrospectivos
6.
Sensors (Basel) ; 22(14)2022 Jul 07.
Artigo em Inglês | MEDLINE | ID: mdl-35890799

RESUMO

Cognitive biometrics is an emerging branch of biometric technology. Recent research has demonstrated great potential for using cognitive biometrics in versatile applications, including biometric recognition and cognitive and emotional state recognition. There is a major need to summarize the latest developments in this field. Existing surveys have mainly focused on a small subset of cognitive biometric modalities, such as EEG and ECG. This article provides a comprehensive review of cognitive biometrics, covering all the major biosignal modalities and applications. A taxonomy is designed to structure the corresponding knowledge and guide the survey from signal acquisition and pre-processing to representation learning and pattern recognition. We provide a unified view of the methodological advances in these four aspects across various biosignals and applications, facilitating interdisciplinary research and knowledge transfer across fields. Furthermore, this article discusses open research directions in cognitive biometrics and proposes future prospects for developing reliable and secure cognitive biometric systems.


Assuntos
Biometria , Cognição
7.
Sensors (Basel) ; 23(1)2022 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-36616657

RESUMO

Deep learning has become a predominant method for solving data analysis problems in virtually all fields of science and engineering. The increasing complexity and the large volume of data collected by diverse sensor systems have spurred the development of deep learning methods and have fundamentally transformed the way the data are acquired, processed, analyzed, and interpreted. With the rapid development of deep learning technology and its ever-increasing range of successful applications across diverse sensor systems, there is an urgent need to provide a comprehensive investigation of deep learning in this domain from a holistic view. This survey paper aims to contribute to this by systematically investigating deep learning models/methods and their applications across diverse sensor systems. It also provides a comprehensive summary of deep learning implementation tips and links to tutorials, open-source codes, and pretrained models, which can serve as an excellent self-contained reference for deep learning practitioners and those seeking to innovate deep learning in this space. In addition, this paper provides insights into research topics in diverse sensor systems where deep learning has not yet been well-developed, and highlights challenges and future opportunities. This survey serves as a catalyst to accelerate the application and transformation of deep learning in diverse sensor systems.


Assuntos
Aprendizado Profundo , Software , Engenharia
8.
Small ; 17(2): e2005368, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33319918

RESUMO

2D materials have a great potential for wide-range applications due to their adjustable bandgap characteristics and special crystal structures. ß-HgI2 is a new 2D van der Waals inorganic molecular crystal material with a wide bandgap of 4.03 eV, on whose preparation and properties there are few relevant reports due to the feature of instability of molecular crystals. Here, an economical method to control the synthesis of large-size 2D ß-HgI2 single crystal by using a mineralizer-assisted solution is reported. According to angle-resolved polarization Raman spectroscopy and first-principles optical absorption calculation, 2D ß-HgI2 flake has a strong in-plane anisotropic light scattering characteristic and high optical absorption dichroism (az /ay  = 3.4), which is due to a low in-plane symmetry of the orthorhombic structure of ß-HgI2 . More importantly, due to the molecular crystal structure of ß-HgI2 , its sensitivity to temperature is less than that of 2D materials such as MoS2 , which has been confirmed by temperature-dependent Raman spectroscopy. In the work, more 2D inorganic molecular crystals are studied in the aspect of growth, which provides a theoretical basis for 2D molecular crystal optoelectronic devices' potential applications.

9.
Opt Lett ; 45(7): 1719-1721, 2020 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-32235982

RESUMO

In this Letter, we describe an aperiodic-oscillation emission phenomenon originating from the Fabry-Perot effect in the deep-ultraviolet backscattering fluorescence spectrum of the $c$c-plane AlGaN film, which is related to the dispersion of its ordinary refractive index near the band edge. Based on this fluorescence spectrum, the ordinary refractive index of the AlGaN film near the band edge could be directly obtained. Certainly, by means of variable angle spectroscopic ellipsometry, the ordinary refractive index of the AlGaN film could be also achieved. Comparing the results obtained by both methods, we discovered that the refractive indices are quite similar, which suggests that the aperiodic-oscillation fluorescence spectrum is also a reliable approach to capture the refractive index of anisotropic optical films.

10.
Opt Lett ; 45(6): 1313-1316, 2020 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-32163952

RESUMO

Raman tensors, one of the basic physical properties of ${{\rm MoS}_2}$MoS2, are rarely reported. Here, angle-resolved polarized Raman scatterings on basal and cross planes of layered ${{\rm MoS}_2}$MoS2 were carried out using the geometry configuration of parallel polarization, and the Raman tensors of three optical vibration modes were systematically studied. As a polar vibration mode, the differential polarizability of the ${{\rm A}_{1{\rm g}}}$A1g mode corresponding to the Raman tensor along the $c$c direction is larger than that along the $a$a direction. In addition, it is also larger than that formed by ${{\rm E}_{2{\rm g}}}$E2g and ${{\rm E}_{1{\rm g}}}$E1g modes. All the experimental results above are beneficial to the understanding of inelastic light-scattering process of ${{\rm MoS}_2}$MoS2.

11.
Bioorg Med Chem ; 28(2): 115254, 2020 01 15.
Artigo em Inglês | MEDLINE | ID: mdl-31866272

RESUMO

Idiopathic pulmonary fibrosis (IPF) is a serious and fatal lung disease, with a median survival of only 3-5 years from diagnosis. Janus kinase 3 (JAK3) has a well-established role in the pathogenesis of various autoimmune diseases, including rheumatoid arthritis (RA) and autoimmune-related pulmonary fibrosis. In this study, through the use of a conformationally-constrained design strategy, a series of thieno[3,2-d]pyrimidines were synthesized as potent JAK3 inhibitors for the treatment of IPF. Among them, the most potent JAK3 inhibitor, namely 8e (IC50 = 1.38 nM), significantly reduced the degree of airsacculitis and fibrosis according to hematoxylin-eosin (HE) staining assay for the lung tissue in the bleomycin (BLM)-induced pulmonary fibrosis mouse model. The clear reduction of the lung collagen deposition by the determination of Masson and hydroxyproline (HYP) content also demonstrated its efficacy in the treatment of fibrosis. In addition, 8e also reduced the expression of the inflammatory markers IL-6, IL-17A, TNF-α and malondialdehyde (MDA) in lung tissue, which indicated its higher anti-inflammatory activity compared with that of the reference agents (nintedanib and gefitinib). Furthermore, it possessed low cytotoxicity against normal human bronchial epithelia (HBE) cells (IC50 > 39.0 µM) and C57BL mice. All these evaluated biological properties suggest that 8e may be a potential JAK3 inhibitor for the treatment of IPF.


Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Fibrose Pulmonar Idiopática/tratamento farmacológico , Janus Quinase 3/antagonistas & inibidores , Inibidores de Proteínas Quinases/farmacologia , Pirimidinas/farmacologia , Animais , Anti-Inflamatórios não Esteroides/síntese química , Anti-Inflamatórios não Esteroides/química , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Células Epiteliais/efeitos dos fármacos , Feminino , Humanos , Fibrose Pulmonar Idiopática/metabolismo , Fibrose Pulmonar Idiopática/patologia , Janus Quinase 3/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Simulação de Acoplamento Molecular , Estrutura Molecular , Inibidores de Proteínas Quinases/síntese química , Inibidores de Proteínas Quinases/química , Pirimidinas/síntese química , Pirimidinas/química , Relação Estrutura-Atividade
12.
BMC Plant Biol ; 19(1): 25, 2019 01 14.
Artigo em Inglês | MEDLINE | ID: mdl-30642256

RESUMO

Following publication of the original article [1], the author reported that their given name was misspelled.

13.
Plant Cell Environ ; 42(1): 145-157, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-29664126

RESUMO

The plant sucrose nonfermenting kinase 1 (SnRK1) kinases play the central roles in the processes of energy balance, hormone perception, stress resistance, metabolism, growth, and development. However, the functions of these kinases are still elusive. In this study, we used GsSnRK1 of wild soybean as bait to perform library-scale screens by the means of yeast two-hybrid to identify its interacting proteins. The putative interactions were verified by yeast retransformation and ß-galactosidase assays, and the selected interactions were further confirmed in planta by bimolecular fluorescence complementation and biochemical Co-IP assays. Protein phosphorylation analyses were carried out by phos-tag assay and anti-phospho-(Ser/Thr) substrate antibodies. Finally, we obtained 24 GsSnRK1 interactors and several putative substrates that can be categorized into SnRK1 regulatory ß subunit, protein modification, biotic and abiotic stress-related, hormone perception and signalling, gene expression regulation, water and nitrogen transport, metabolism, and unknown proteins. Intriguingly, we first discovered that GsSnRK1 interacted with and phosphorylated the components of soybean nodulation and symbiotic nitrogen fixation. The interactions and potential functions of GsSnRK1 and its associated proteins were extensively discussed and analysed. This work provides plausible clues to elucidate the novel functions of SnRK1 in response to variable environmental, metabolic, and physiological requirements.


Assuntos
Glycine max/enzimologia , Proteínas de Plantas/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Imunoprecipitação , Fosforilação , Glycine max/metabolismo , Especificidade por Substrato , Técnicas do Sistema de Duplo-Híbrido
14.
BMC Plant Biol ; 18(1): 234, 2018 Oct 13.
Artigo em Inglês | MEDLINE | ID: mdl-30316294

RESUMO

BACKGROUND: Even though bicarbonate alkaline stress is a serious threat to crop growth and yields, it attracts much fewer researches than high salinity stress. The basic leucine zipper (bZIP) transcription factors have been well demonstrated to function in diverse abiotic stresses; however, their biological role in alkaline tolerance still remains elusive. In this study, we functionally characterized a bZIP gene from Glycine soja GsbZIP67 in bicarbonate alkaline stress responses. RESULTS: GsbZIP67 was initially identified as a putative bicarbonate responsive gene, on the basis of previous RNA-seq data of 50 mM NaHCO3-treated Glycine soja roots. GsbZIP67 protein possessed a conserved bZIP domain, and belonged to the group S2 bZIP, which is yet less well-studied. Our studies showed that GsbZIP67 targeted to nucleus in Arabidopsis protoplasts, and displayed transcriptional activation activity in yeast cells. The quantitative real-time PCR analyses unraveled the bicarbonate stress responsive expression and tissue specific expression of GsbZIP67 in wild soybean. Further phenotypic analysis illustrated that GsbZIP67 overexpression in alfalfa promoted plant growth under bicarbonate alkaline stress, as evidenced by longer roots and shoots. Furthermore, GsbZIP67 overexpression also modified the physiological indices of transgenic alfalfa under bicarbonate alkaline stress. In addition, the expression levels of several stress responsive genes were also augmented by GsbZIP67 overexpression. CONCLUSIONS: Collectively, in this study, we demonstrated that GsbZIP67 acted as a positive regulator of plant tolerance to bicarbonate alkaline stress. These results provide direct genetic evidence of group S2 bZIPs in bicarbonate alkaline stress, and will facilitate further studies concerning the cis-elements and/or downstream genes targeted by GsbZIP67 in stress responses.


Assuntos
Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Bicarbonatos/toxicidade , Regulação da Expressão Gênica de Plantas , Glycine max/genética , Medicago sativa/fisiologia , Álcalis/toxicidade , Sequência de Aminoácidos , Fatores de Transcrição de Zíper de Leucina Básica/genética , Núcleo Celular/metabolismo , Genes Reporter , Medicago sativa/genética , Fenótipo , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raízes de Plantas/genética , Raízes de Plantas/fisiologia , Brotos de Planta/genética , Brotos de Planta/fisiologia , Plantas Geneticamente Modificadas , Transporte Proteico , Alinhamento de Sequência , Estresse Fisiológico
15.
Physiol Plant ; 164(2): 145-162, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-29243826

RESUMO

Alkaline stress is a major form of abiotic stress that severely inhibits plant growth and development, thus restricting crop productivity. However, little is known about how plants respond to alkali. In this study, a slow-type anion channel homolog 3 gene, GsSLAH3, was isolated and functionally characterized. Bioinformatics analysis showed that the GsSLAH3 protein contains 10 transmembrane helices. Consistently, GsSLAH3 was found to locate on plasma membrane by transient expression in onion epidermal cells. In wild soybeans, GsSLAH3 expression was induced by NaHCO3 treatment, suggesting its involvement in plant response to alkaline stress. Ectopic expression of GsSLAH3 in yeast increased sensitivity to alkali treatment. Dramatically, overexpression of GsSLAH3 in Arabidopsis thaliana enhanced alkaline tolerance during the germination, seedling and adult stages. More interestingly, we found that transgenic lines also improved plant tolerance to KHCO3 rather than high pH treatment. A nitrate content analysis of Arabidopsis shoots showed that GsSLAH3 overexpressing lines accumulated more NO3- than wild-type. In summary, our data suggest that GsSLAH3 is a positive alkali responsive gene that increases bicarbonate resistance specifically.


Assuntos
Bicarbonatos/farmacologia , Proteínas de Plantas/metabolismo , Compostos de Potássio/farmacologia , Arabidopsis/efeitos dos fármacos , Arabidopsis/genética , Arabidopsis/metabolismo , Membrana Celular/efeitos dos fármacos , Membrana Celular/metabolismo , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/efeitos dos fármacos , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo
16.
Physiol Plant ; 164(3): 268-278, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-29578245

RESUMO

Ubiquitin is a highly conserved protein with multiple essential regulatory functions through the ubiquitin-proteasome system. Even though its functions in the ubiquitin-mediated protein degradation pathway are very well characterized, the function of ubiquitin genes in the regulation of the alkaline stress response is not fully established. In this study, we identified 12 potential UBQ genes in the Glycine soja genome, and analyzed their evolutionary relationship, conserved domains and promoter cis-elements. We also explored the expression profiles of G. soja UBQ genes under alkaline stress, based on the transcriptome sequencing. We found that the expression of GsUBQ10 was significantly induced by alkaline stress, and the function of GsUBQ10 was characterized by overexpression in transgenic alfalfa (Medicago sativa). Our results suggested that GsUBQ10 transgenic lines significantly improved the alkaline tolerance in alfalfa. The GsUBQ10 transgenic lines showed lower relative membrane permeability, lower malon dialdehyde content and higher catalase activity than in the wild-type plants. This indicates that GsUBQ10 is involved in regulating the reactive oxygen species accumulation under alkaline stress. Taken together, we identified an ubiquitin gene GsUBQ10 from G. soja, which plays a positive role in responses to alkaline stress in alfalfa.


Assuntos
Glycine max/metabolismo , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Ubiquitina/metabolismo , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Medicago sativa/genética , Medicago sativa/metabolismo , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/genética , Glycine max/genética , Estresse Fisiológico/genética , Estresse Fisiológico/fisiologia
17.
Plant J ; 86(6): 514-29, 2016 06.
Artigo em Inglês | MEDLINE | ID: mdl-27121031

RESUMO

Although research has extensively illustrated the molecular basis of plant responses to salt and high-pH stresses, knowledge on carbonate alkaline stress is poor and the specific responsive mechanism remains elusive. We have previously characterized a Glycine soja Ca(2+) /CAM-dependent kinase GsCBRLK that could increase salt tolerance. Here, we characterize a methionine sulfoxide reductase (MSR) B protein GsMSRB5a as a GsCBRLK interactor by using Y2H and BiFc assays. Further analyses showed that the N-terminal variable domain of GsCBRLK contributed to the GsMSRB5a interaction. Y2H assays also revealed the interaction specificity of GsCBRLK with the wild soybean MSRB subfamily proteins, and determined that the BoxI/BoxII-containing regions within GsMSRBs were responsible for their interaction. Furthermore, we also illustrated that the N-terminal basic regions in GsMSRBs functioned as transit peptides, which targeted themselves into chloroplasts and thereby prevented their interaction with GsCBRLK. Nevertheless, deletion of these regions allowed them to localize on the plasma membrane (PM) and interact with GsCBRLK. In addition, we also showed that GsMSRB5a and GsCBRLK displayed overlapping tissue expression specificity and coincident expression patterns under carbonate alkaline stress. Phenotypic experiments demonstrated that GsMSRB5a and GsCBRLK overexpression in Arabidopsis enhanced carbonate alkaline stress tolerance. Further investigations elucidated that GsMSRB5a and GsCBRLK inhibited reactive oxygen species (ROS) accumulation by modifying the expression of ROS signaling, biosynthesis and scavenging genes. Summarily, our results demonstrated that GsCBRLK and GsMSRB5a interacted with each other, and activated ROS signaling under carbonate alkaline stress.


Assuntos
Fabaceae/enzimologia , Fabaceae/metabolismo , Metionina Sulfóxido Redutases/metabolismo , Proteínas de Plantas/metabolismo , Arabidopsis/metabolismo , Proteínas Quinases Dependentes de Cálcio-Calmodulina/genética , Proteínas Quinases Dependentes de Cálcio-Calmodulina/metabolismo , Calmodulina/genética , Calmodulina/metabolismo , Proteínas de Ligação a Calmodulina/genética , Proteínas de Ligação a Calmodulina/metabolismo , Fabaceae/genética , Regulação da Expressão Gênica de Plantas , Metionina Sulfóxido Redutases/genética , Proteínas de Plantas/genética , Ligação Proteica , Espécies Reativas de Oxigênio/metabolismo
18.
Plant Mol Biol ; 95(3): 253-268, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28884328

RESUMO

KEY MESSAGE: Overexpression of Gshdz4 or GsNAC019 enhanced alkaline tolerance in transgenic Arabidopsis. We proved that Gshdz4 up-regulated both GsNAC019 and GsRD29B but GsNAC019 may repress the GsRD29B expression under alkaline stress. Wild soybean (Glycine soja) has a high tolerance to environmental challenges. It is a model species for dissecting the molecular mechanisms of salt-alkaline stresses. Although many NAC transcription factors play important roles in response to multiple abiotic stresses, such as salt, osmotic and cold, their mode of action in alkaline stress resistance is largely unknown. In our study, we identified a G. soja NAC gene, GsNAC019, which is a homolog of the Arabidopsis AtNAC019 gene. GsNAC019 was highly up-regulated by 50 mM NaHCO3 treatment in the roots of wild soybean. Further investigation showed that a well-characterized transcription factor, Gshdz4 protein, bound the cis-acting element sequences (CAATA/TA), which are located in the promoter of the AtNAC019/GsNAC019 genes. Overexpression of Gshdz4 positively regulated AtNAC019 expression in transgenic Arabidopsis, implying that AtNAC019/GsNAC019 may be the target genes of Gshdz4. GsNAC019 was demonstrated to be a nuclear-localized protein in onion epidermal cells and possessed transactivation activity in yeast cells. Moreover, overexpression of GsNAC019 in Arabidopsis resulted in enhanced tolerance to alkaline stress at the seedling and mature stages, but reduced ABA sensitivity. The closest Arabidopsis homolog mutant plants of Gshdz4, GsNAC019 and GsRD29B containing athb40, atnac019 and atrd29b were sensitive to alkaline stress. Overexpression or the closest Arabidopsis homolog mutant plants of the GsNAC019 gene in Arabidopsis positively or negatively regulated the expression of stress-related genes, such as AHA2, RD29A/B and KIN1. Moreover, this mutation could phenotypically promoted or compromised plant growth under alkaline stress, implying that GsNAC019 may contribute to alkaline stress tolerance via the ABA signal transduction pathway and regulate expression of the downstream stress-related genes.


Assuntos
Ácido Abscísico/farmacologia , Álcalis/farmacologia , Fabaceae/genética , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Proteínas de Plantas/genética , Fatores de Transcrição/genética , Adaptação Fisiológica/efeitos dos fármacos , Adaptação Fisiológica/genética , Arabidopsis/genética , Arabidopsis/metabolismo , Sequência de Bases , Bicarbonatos/farmacologia , Fabaceae/metabolismo , Reguladores de Crescimento de Plantas/farmacologia , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Regiões Promotoras Genéticas/genética , Ligação Proteica , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Homologia de Sequência do Ácido Nucleico , Cloreto de Sódio/farmacologia , Estresse Fisiológico , Fatores de Transcrição/metabolismo , Técnicas do Sistema de Duplo-Híbrido
19.
Plant Mol Biol ; 94(4-5): 509-530, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28681139

RESUMO

KEY MESSAGE: Here we first found that GsERF71, an ERF factor from wild soybean could increase plant alkaline stress tolerance by up-regulating H+-ATPase and by modifing the accumulation of Auxin. Alkaline soils are widely distributed all over the world and greatly limit plant growth and development. In our previous transcriptome analyses, we have identified several ERF (ethylene-responsive factor) genes that responded strongly to bicarbonate stress in the roots of wild soybean G07256 (Glycine soja). In this study, we cloned and functionally characterized one of the genes, GsERF71. When expressed in epidermal cells of onion, GsERF71 localized to the nucleus. It can activate the reporters in yeast cells, and the C-terminus of 170 amino acids is essential for its transactivation activity. Yeast one-hybrid and EMSA assays indicated that GsERF71 specifically binds to the cis-acting elements of the GCC-box, suggesting that GsERF71 may participate in the regulation of transcription of the relevant biotic and abiotic stress-related genes. Furthermore, transgenic Arabidopsis plants overexpressing GsERF71 showed significantly higher tolerance to bicarbonate stress generated by NaHCO3 or KHCO3 than the wild type (WT) plants, i.e., the transgenic plants had greener leaves, longer roots, higher total chlorophyll contents and lower MDA contents. qRT-PCR and rhizosphere acidification assays indicated that the expression level and activity of H+-ATPase (AHA2) were enhanced in the transgenic plants under alkaline stress. Further analysis indicated that the expression of auxin biosynthetic genes and IAA contents were altered to a lower extent in the roots of transgenic plants than WT plants under alkaline stress in a short-term. Together, our data suggest that GsERF71 enhances the tolerance to alkaline stress by up-regulating the expression levels of H+-ATPase and by modifying auxin accumulation in transgenic plants.


Assuntos
Arabidopsis/metabolismo , Proteínas de Ligação a DNA/metabolismo , Glycine max/metabolismo , Proteínas de Plantas/metabolismo , Estresse Fisiológico/fisiologia , Fatores de Transcrição/metabolismo , Sequência de Aminoácidos , DNA de Plantas/metabolismo , Proteínas de Ligação a DNA/genética , Regulação Enzimológica da Expressão Gênica/fisiologia , Regulação da Expressão Gênica de Plantas/fisiologia , Concentração de Íons de Hidrogênio , Filogenia , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , ATPases Translocadoras de Prótons/genética , ATPases Translocadoras de Prótons/metabolismo , Glycine max/genética , Fatores de Transcrição/genética
20.
Int J Mol Sci ; 18(9)2017 Sep 19.
Artigo em Inglês | MEDLINE | ID: mdl-32961651

RESUMO

The PHD-finger family has been demonstrated to be involved in regulating plant growth and development. However, little information is given for its role in environmental stress responses. Here, we identified a total of 59 PHD family genes in the rice genome. These OsPHDs genes were located on eleven chromosomes and synteny analysis only revealed nine duplicated pairs within the rice PHD family. Phylogenetic analysis of all OsPHDs and PHDs from other species revealed that they could be grouped into two major clusters. Furthermore, OsPHDs were clustered into eight groups and members from different groups displayed a great divergence in terms of gene structure, functional domains and conserved motifs. We also found that with the exception of OsPHD6, all OsPHDs were expressed in at least one of the ten tested tissues and OsPHDs from certain groups were expressed in specific tissues. Moreover, our results also uncovered differential responses of OsPHDs expression to environmental stresses, including ABA (abscisic acid), water deficit, cold and high Cd. By using quantitative real-time PCR, we further confirmed the differential expression of OsPHDs under these stresses. OsPHD1/7/8/13/33 were differentially expressed under water deficit and Cd stresses, while OsPHD5/17 showed altered expression under water deficit and cold stresses. Moreover, OsPHD3/44/28 displayed differential expression under ABA and Cd stresses. In conclusion, our results provide valuable information on the rice PHD family in plant responses to environmental stress, which will be helpful for further characterizing their biological roles in responding to environmental stresses.

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