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BACKGROUND: Salt stress significantly reduces soybean yield. To improve salt tolerance in soybean, it is important to mine the genes associated with salt tolerance traits. RESULTS: Salt tolerance traits of 286 soybean accessions were measured four times between 2009 and 2015. The results were associated with 740,754 single nucleotide polymorphisms (SNPs) to identify quantitative trait nucleotides (QTNs) and QTN-by-environment interactions (QEIs) using three-variance-component multi-locus random-SNP-effect mixed linear model (3VmrMLM). As a result, eight salt tolerance genes (GmCHX1, GsPRX9, Gm5PTase8, GmWRKY, GmCHX20a, GmNHX1, GmSK1, and GmLEA2-1) near 179 significant and 79 suggested QTNs and two salt tolerance genes (GmWRKY49 and GmSK1) near 45 significant and 14 suggested QEIs were associated with salt tolerance index traits in previous studies. Six candidate genes and three gene-by-environment interactions (GEIs) were predicted to be associated with these index traits. Analysis of four salt tolerance related traits under control and salt treatments revealed six genes associated with salt tolerance (GmHDA13, GmPHO1, GmERF5, GmNAC06, GmbZIP132, and GmHsp90s) around 166 QEIs were verified in previous studies. Five candidate GEIs were confirmed to be associated with salt stress by at least one haplotype analysis. The elite molecular modules of seven candidate genes with selection signs were extracted from wild soybean, and these genes could be applied to soybean molecular breeding. Two of these genes, Glyma06g04840 and Glyma07g18150, were confirmed by qRT-PCR and are expected to be key players in responding to salt stress. CONCLUSIONS: Around the QTNs and QEIs identified in this study, 16 known genes, 6 candidate genes, and 8 candidate GEIs were found to be associated with soybean salt tolerance, of which Glyma07g18150 was further confirmed by qRT-PCR.
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Interação Gene-Ambiente , Genes de Plantas , Glycine max , Polimorfismo de Nucleotídeo Único , Locos de Características Quantitativas , Tolerância ao Sal , Glycine max/genética , Glycine max/fisiologia , Tolerância ao Sal/genética , Locos de Características Quantitativas/genética , FenótipoRESUMO
UNLABELLED: Autographa californica multiple nucleopolyhedrovirus orf132 (named ac132) has homologs in all genome-sequenced group I nucleopolyhedroviruses. Its role in the viral replication cycle is unknown. In this study, ac132 was shown to express a protein of around 28 kDa, which was determined to be associated with the nucleocapsids of both occlusion-derived virus and budded virus. Confocal microscopy showed that AC132 protein appeared in central region of the nucleus as early as 12 h postinfection with the virus. It formed a ring zone at the periphery of the nucleus by 24 h postinfection. To investigate its role in virus replication, ac132 was deleted from the viral genome by using a bacmid system. In the Sf9 cell culture transfected by the ac132 knockout bacmid, infection was restricted to single cells, and the titer of infectious budded virus was reduced to an undetectable level. However, viral DNA replication and the expression of late genes vp39 and odv-e25 and a reporter gene under the control of the very late gene p10 promoter were unaffected. Electron microscopy showed that nucleocapsids, virions, and occlusion bodies were synthesized in the cells transfected by an ac132 knockout bacmid, but the formation of the virogenic stroma and occlusion bodies was delayed, the numbers of enveloped nucleocapsids were reduced, and the occlusion bodies contained mainly singly enveloped nucleocapsids. AC132 was found to interact with envelope protein ODV-E18 and the viral DNA-binding protein P6.9. The data from this study suggest that ac132 possibly plays an important role in the assembly and envelopment of nucleocapsids. IMPORTANCE: To our knowledge, this is the first report on a functional analysis of ac132. The data presented here demonstrate that ac132 is required for production of the budded virus and multiply enveloped occlusion-derived virus of Autographa californica multiple nucleopolyhedrovirus. This article reveals unique phenotypic changes induced by ac132 deletion on the virus and multiple new findings on ac132.
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Nucleocapsídeo/metabolismo , Nucleopoliedrovírus/fisiologia , Proteínas Estruturais Virais/metabolismo , Montagem de Vírus , Animais , Deleção de Genes , Microscopia Eletrônica de Transmissão , Peso Molecular , Nucleocapsídeo/genética , Nucleocapsídeo/ultraestrutura , Nucleopoliedrovírus/genética , Células Sf9 , Spodoptera , Proteínas Estruturais Virais/química , Proteínas Estruturais Virais/genética , Vírion/ultraestrutura , Liberação de VírusRESUMO
The lack of biosafety and insufficient delivery efficiency of gene-carriers are still obstacles to human gene therapy. This paper reported highly biocompatible chitosan (CS) functionalized Prussian blue (PB) nanoparticles (designated as CS/PB NPs) for photocontrollable gene delivery. The ultra-small size (â¼3 nm), positive charge and high physiological stability of CS/PB NPs make it suitable to be a nonviral vector. In addition, CS/PB NPs could effectively convert the near infrared (NIR) light into heat due to its strong absorption in the NIR region, assisting the uptake of NPs by cells. Upon NIR light irradiation, CS/PB NPs showed superior gene transfection efficiency, much higher than that of free polyethylenimine (PEI). Both in vitro and in vivo experiments demonstrated that CS/PB NPs had excellent biocompatiblity. This work also encourages further exploration of the CS/PB NPs as a photocontrollable nanovector for combined photothermal and gene therapy.
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Quitosana/química , Nanopartículas/química , Vetores Genéticos , Células HeLa , HumanosRESUMO
A novel multifunctional theranostic agent has been successfully fabricated by loading iron oxide nanoparticles into poly(lactic acid) (PLA) microcapsules followed by surface functionalization with graphene oxide. Both in vitro and in vivo experiments proved that the resulting microcapsules could serve as contrast agents to simultaneously enhance ultrasound, magnetic resonance and photoacoustic imaging. The composite microcapsules show good biocompatibility and rapid response to magnetic fields. Due to the strong absorption of the near-infrared light, the composite microcapsules could efficiently kill cancer cells upon NIR laser irradiation. In addition, it was found that such a photothermal effect could be obviously enhanced by applying an external magnetic field. In a nutshell, this multifunctional microcapsule can be developed as a promising platform that integrates multimodality imaging and therapy capabilities for effective cancer theranostics.
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This study was performed to investigate the effects of different regions of the Autographa califor nica multiple nucleopolyhedrovirus envelope protein E25 on its trafficking into nucleus and nuclear localization in host cells and on virus replication. Fourteen recombinant bacmids, each containing an e25 mutant with substitution or insertion of egfp, in the absence or presence of the native e25, were constructed and used to transfect Sf9 cells. The E25-EGFP fusion proteins and native E25 expressed in the cells transfect ed with individual recombinant bacmid were traced by autofluorescence from EGFP or by immuno-fluorescence assays. Confocal microscopy revealed that the E25-EGFP fusion protein with the N-domain (2-45aa) of E25 substituted by EGFP only distributed in the cytoplasm in transfected cells; and the fusion protein with EGFP inserted at the laa/2aa site of E25 completely remained outside of the nucleus and resided along the nuclear membrane. The E25-EGFPs with 46-118aa of E25 substituted by EGFP or with EGFP inserted at the 118aa/119aa site were present outside, across from the nuclear membrane or in nuclear plasm in dot-like shapes. The fusion proteins with the C-domain substituted by EGFP or with EGFP inserted at the site of 45/46aa or at the C-terminal formed a condensed ring or spread throughout the nucleus, in a similar manner to the E25 distributed in the cells transfected by the e25-knockout repair bacmid. These results prove that the N-terminal domain is critical for nuclear transportation of E25 and possibly to its position on the cytoplasm membrane as well; and the sequence downstream of the N-terminal domain also affects trafficking and nuclear localization of the protein. In cells transfected with bacmids containing both the native e25 and individual e25-egfp mutants, the E25-EGFP fusion proteins co-localized with E25 individually, showing similar patterns of subcellular localization as E25 mutants in the absence of native E25 in most cases, suggesting that the E25 likely exists and functions as dimmers or polymers. Production of infectious BV was dramatically reduced and even completely eliminated in most cases, either in the absence or presence of the native e25.
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Núcleo Celular/virologia , Mutação , Nucleopoliedrovírus/fisiologia , Proteínas Virais/genética , Proteínas Virais/metabolismo , Liberação de Vírus , Motivos de Aminoácidos , Animais , Núcleo Celular/metabolismo , Nucleopoliedrovírus/química , Nucleopoliedrovírus/genética , Transporte Proteico , Spodoptera/virologia , Proteínas Virais/química , Replicação ViralRESUMO
@#A lot of researches indicate the relationship between cognitive impairment and functional connectivity of default mode network,salience network and central executive network.The changes of networks are various in different cognitive dys-function.It is important to apply resting state functional magnetic resonance in the old adults with cognitive dysfunction.
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Objective To investigate the relationship between cholinergic pathway damage and the executive dysfunction of patients with different degrees of cognitive impairment caused by cerebral white matter lesions(WML). Methods From March,2016 to December,2017,115 patients were recruited,whose characteristics,such as age,gender, education,and history of hypertension,diabetes and stroke were recorded.According to the T2-weighted MRI,80 patients were defined as WML.WML patients were divided into cognitively normal(CN)group(n=41),vascular cognitive impairment of none dementia(VCIND)group(n=21)and vascular dementia(VaD)group(n=18)ac-cording to the result of Montreal Cognitive Assessment(MoCA)and Clinical Dementia Rating(CDR).Other 35 cases without WML and cognitive impairment were as control group.WML under MRI were evaluated with Cho-linergic Pathways Hyperintensities Scale(CHIPS).All the WML patients were assessed with Stroop Color-Word Test,Trail Making Test, Symbol Digital Modalities Test, and Verbal Fluence Test.The correlation between the scores of CHIPS and the executive tests were analysed. Results There was no significant difference in age, gender, level of education, and cardiovascular disease risk factors among four groups(P>0.05),but there were significant differences in scores of MoCA and CHIPS(F>25.781,P<0.001),while the score of MoCA was the least(P<0.01)and the scores of CHIPS were the most in VaD group (P<0.001).The CHIPS scores of left and bilateral hemisphere negatively correlated with all the scores of execu-tive tests(P<0.05),while that of the right hemisphere just correlated with the scores of some executive tests(P<0.05). Conclusion For cognitive impairment after WML,cholinergic pathway damage may relate with the executive function impairment,especially the damage in left cerebral hemisphere.
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@#Objective To explore the diversity of resting-state network of brain between the patients with leukoaraiosis and the healthy people. Methods 31 patients with leukoaraiosis (patients) and 27 healthy persons (controls) were checked with resting-state functional magnetic resonance imaging (rs-fMRI), and analyzed with the independent component analysis (ICA) to explore the resting-state functional brain network. Results The resting-state brain network was found in both the patients and the controls, which was coincident with the previous studies. The active areas were the same in both groups, and the activation was weaken in the patients than in the controls, especially in quadrate gyri, posterior cingulate cortex, superior temporal gyrus, superior parietal gyrus, anterior central gyrus, post central gyrus, insula and prefrontal cortex. Conclusion There is a significant diversity of resting-state network of brain between the patients with leukoaraiosis and healthy people in the activation of active areas.