RESUMO
Soybean ß-conglycinin is a major allergen that adversely affects the nutritional properties of soybean. Soybean deficient in ß-conglycinin is associated with low allergenicity and high nutritional value. Long intergenic noncoding RNAs (lincRNAs) regulate gene expression and are considered important regulators of essential biological processes. Despite increasing knowledge of the functions of lincRNAs, relatively little is known about the effects of lincRNAs on the accumulation of soybean ß-conglycinin. The current study presents the identification of a lincRNA lincCG1 that was mapped to the intergenic noncoding region of the ß-conglycinin α-subunit locus. The full-length lincCG1 sequence was cloned and found to regulate the expression of soybean seed storage protein (SSP) genes via both cis- and trans-acting regulatory mechanisms. Loss-of-function lincCG1 mutations generated using the clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9) system led to the deficiency of the allergenic α'-, α-, and ß-subunits of soybean ß-conglycinin as well as higher content of proteins, sulfur-containing amino acids, and free arginine. The dominant null allele LincCG1, and consequently, the ß-conglycinin-deficient phenotype associated with the lincCG1-gene-edited line was stably inherited by the progenies in a Mendelian fashion. The dominant null allele LincCG1 may therefore be exploited for engineering/developing novel hypoallergenic soybean varieties. Furthermore, Cas9-free and ß-conglycinin-deficient homozygous mutant lines were obtained in the T1 generation. This study is the first to employ the CRISPR/Cas9 technology for editing a lincRNA gene associated with the soybean allergenic protein ß-conglycinin. Moreover, this study reveals that lincCG1 plays a crucial role in regulating the expression of the ß-conglycinin subunit gene cluster, besides highlighting the efficiency of employing the CRISPR/Cas9 system for modulating lincRNAs, and thereby regulating soybean seed components.
Assuntos
Antígenos de Plantas , Sistemas CRISPR-Cas , Edição de Genes , Globulinas , Glycine max , RNA Longo não Codificante , Proteínas de Armazenamento de Sementes , Proteínas de Soja , Proteínas de Armazenamento de Sementes/genética , Proteínas de Armazenamento de Sementes/química , Globulinas/genética , Globulinas/metabolismo , Globulinas/química , Glycine max/genética , Glycine max/metabolismo , Antígenos de Plantas/genética , Antígenos de Plantas/química , Proteínas de Soja/genética , Proteínas de Soja/metabolismo , Proteínas de Soja/química , RNA Longo não Codificante/genética , Regulação da Expressão Gênica de Plantas , Sementes/genética , Sementes/metabolismo , Sementes/químicaRESUMO
The growth and development of soybean plants can be affected by both abiotic and biotic stressors, such as saline-alkali stress and Phytophthora root rot. In this study, we identified a stress-related gene-GmARM-whose promoter contained several hormone-response and stress-regulatory elements, including ABRE, TCA element, STRE, and MBS. qRT-PCR analysis showed that the expression of GmARM was the highest in seeds at 55 days after flowering. Furthermore, this gene was upregulated after exposure to saline-alkali stress and Phytophthora root rot infection at the seedling stage. Thus, we generated GmARM mutants using the CRISPR-Cas9 system to understand the role of this gene in stress response. T3 plants showed significantly improved salt tolerance, alkali resistance, and disease resistance, with a significantly higher survival rate than the wildtype plants. Moreover, mutations in GmARM affected the expression of related stress-resistance genes, indicating that GmARM mutants achieved multiple stress tolerance. Therefore, this study provides a foundation for further exploration of the genes involved in resistance to multiple stresses in soybean that can be used for breeding multiple stress-resistance soybean varieties.
Assuntos
Sistemas CRISPR-Cas , Edição de Genes , Glycine max , Estresse Fisiológico , Glycine max/genética , Glycine max/fisiologia , Glycine max/microbiologia , Edição de Genes/métodos , Estresse Fisiológico/genética , Resistência à Doença/genética , Doenças das Plantas/microbiologia , Doenças das Plantas/genética , Plantas Geneticamente Modificadas/genética , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Phytophthora/fisiologia , Genes de PlantasRESUMO
Pericytes, a specific type of mesenchymal cell that surround the basement membrane of pulmonary venules and capillaries. They are crucial pathological features observed in individuals with the severe lung disease of pulmonary fibrosis (PF). The presence of pericytes leads to inflammation and fibrosis in the lung interstitium and alveolar space due to the release of various cytokines and chemokines. Pericytes also stimulate the proliferation and activation of fibroblasts, thereby promoting the progression of PF. Previous studies examining the mechanism of action of pericytes have primarily focused on cell signal transduction pathways, cell growth and death processes, and the synthesis and breakdown of extracellular matrix (ECM). Notably, the transforming growth factor-ß (TGF-ß) and Wnt signaling pathways have been associated with the action of pericytes in driving the progression of PF. It is therefore clear that pericytes play an essential role in the development of PF, while also offering possible avenues for targeted therapeutic intervention against this condition. The current article provides a comprehensive review on how pericytes contribute to inflammatory responses, as well as their importance for understanding the mechanism of PF. In addition, this review discusses the potential use of pericyte-targeted approaches for the treatment of patients affected by this debilitating lung disease.
Assuntos
Pericitos , Fibrose Pulmonar , Pericitos/patologia , Pericitos/metabolismo , Humanos , Fibrose Pulmonar/patologia , Fibrose Pulmonar/metabolismo , Animais , Fator de Crescimento Transformador beta/metabolismo , Transdução de Sinais , Matriz Extracelular/metabolismo , Via de Sinalização WntRESUMO
High-performance devices of quartz glass demand an atomic surface, which induces a challenge for chemical mechanical polishing (CMP) with a high material removal rate (MRR). Moreover, traditional CMP usually employs toxic and corrosive slurries, leading to the pollution of the environment. To overcome these challenges, a novel green photocatalytic CMP is proposed. In the CMP, SiO2@TiO2 core-shell abrasives were developed, and the CMP slurry included the developed abrasives, sodium carbonate, hydrogen peroxide and sorbitol. After photocatalytic CMP, the surface roughness Sa of quartz glass is 0.185 nm, with a scanning area of 50 × 50 µm2, and the MRR is 8.64 µm h-1. To the best of our knowledge, the MRR is the highest on such a big area of atomic surface for quartz glass. X-ray photoelectron spectroscopy reveals that SiO2@TiO2 core-shell abrasives were used as photocatalysts motivated by simulated solar light, generating electrons and holes and producing hydroxyl radicals through hydrogen peroxide. As a result, OH- could combine with Si atoms on the surface of quartz glass, forming Si-OH-Si bonds. Then the formed bonds were removed based on the balance between chemical and mechanical functions. The proposed CMP, developed SiO2@TiO2 abrasives and slurry provide new insights to achieve an atomic surface of quartz glass with a high MRR.