RESUMO
Diacylglycerol kinase (DGK) is an enzyme that plays a pivotal role in abiotic and biotic stress responses in plants by transforming the diacylglycerol into phosphatidic acid. However, there is no report on the characterization of soybean DGK genes in spite of the availability of the soybean genome sequence. In this study, we performed genome-wide analysis and expression profiling of the DGK gene family in the soybean genome. We identified 12 DGK genes (namely GmDGK1-12) which all contained conserved catalytic domains with protein lengths and molecular weights ranging from 436 to 727 amino acids (aa) and 48.62 to 80.93 kDa, respectively. Phylogenetic analyses grouped GmDGK genes into three clusters-cluster I, cluster II, and cluster III-which had three, four, and five genes, respectively. The qRT-PCR analysis revealed significant GmDGK gene expression levels in both leaves and roots coping with polyethylene glycol (PEG), salt, alkali, and salt/alkali treatments. This work provides the first characterization of the DGK gene family in soybean and suggests their importance in soybean response to abiotic stress. These results can serve as a guide for future studies on the understanding and functional characterization of this gene family.
Assuntos
Diacilglicerol Quinase/genética , Perfilação da Expressão Gênica , Genômica , Glycine max/enzimologia , Glycine max/genética , Família Multigênica , Estresse Fisiológico/genética , Motivos de Aminoácidos , Sequência de Aminoácidos , Cromossomos de Plantas/genética , Diacilglicerol Quinase/química , Diacilglicerol Quinase/metabolismo , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Especificidade de Órgãos/genética , Filogenia , Regiões Promotoras Genéticas/genética , Domínios Proteicos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Frações Subcelulares/metabolismoRESUMO
Human basic fibroblast growth factor (hFGF2) is widely recognized for accelerating skin wound healing in both animal models and randomized clinical trials. However, the low skin permeation and bioavailability of hFGF2 remain the most limiting factors in the pharmacological application. For the first time, Camelina Lipid Droplets (CLD) delivery system was displayed important virtue, by promoting the skin absorption of hFGF2, which is a key factor that accelerates the skin wound repair, and provide a new alternative for skin therapy. In this study, we used the CLD as a safer material to prepare the nanoparticles, which were characterized by size and morphology. Our data revealed that particle sizes of Camelina Lipid Droplets linked to hFGF2 (CLD-hFGF2) were around 133.5 nm; it also displayed that the complex of CLD-hFGF2 penetrates the skin barrier in deeper than an individual hFGF2. This suggests that once the hFGF2 is fixed onto the surface of CLD, it can cross the stratum corneum and play a therapeutic role into the dermis. Furthermore, we demonstrated that CLD-hFGF2 enhances fibroblast migration, and significantly improves skin regeneration for accelerating wound healing without any significant toxicity. This paper highlights the importance of CLD as an emerging delivery system; it is also providing a new and applicable therapeutic research direction through enhancing the skin permeation of hFGF2 to accelerate wound healing.
Assuntos
Gotículas Lipídicas , Cicatrização , Animais , Humanos , Tamanho da Partícula , Pele/metabolismo , Absorção CutâneaRESUMO
Calcineurin B-like protein-interacting protein kinases (CIPKs) are key elements of plant abiotic stress signaling pathways. CIPKs are SOS2 (Salt Overly Sensitive 2)-like proteins (protein kinase S [PKS] proteins) which all contain a putative FISL motif. It seems that the FISL motif is found only in the SOS2 subfamily of protein kinases. In this study, the full-length cDNA of a soybean CIPK gene (GmPKS4) was isolated and was revealed to have an important role in abiotic stress responses. A qRT-PCR analysis indicated that GmPKS4 expression is upregulated under saline conditions or when exposed to alkali, salt-alkali, drought, or abscisic acid (ABA). A subcellular localization assay revealed the presence of GmPKS4 in the nucleus and cytoplasm. Further studies on the GmPKS4 promoter suggested it affects soybean resistance to various stresses. Transgenic Arabidopsis thaliana and soybean hairy roots overexpressing GmPKS4 had increased proline content as well as high antioxidant enzyme activities but decreased malondialdehyde levels following salt and salt-alkali stress treatments. Additionally, GmPKS4 overexpression activated reactive oxygen species scavenging systems, thereby minimizing damages due to oxidative and osmotic stresses. Moreover, upregulated stress-related gene expression levels were detected in lines overexpressing GmPKS4 under stress conditions. In conclusion, GmPKS4 improves soybean tolerance to salt and salt-alkali stresses. The overexpression of GmPKS4 enhances the scavenging of reactive oxygen species, osmolyte synthesis, and the transcriptional regulation of stress-related genes.