RESUMO
BACKGROUND: We previously identified six drought-inducible CC-type glutaredoxins in cassava cultivars, however, less is known about their potential role in the molecular mechanism by which cassava adapted to abiotic stress. RESULTS: Herein, we investigate one of cassava drought-responsive CC-type glutaredoxins, namely MeGRXC3, that involved in regulation of mannitol-induced inhibition on seed germination and seedling growth in transgenic Arabidopsis. MeGRXC3 overexpression up-regulates several stress-related transcription factor genes, such as PDF1.2, ERF6, ORA59, DREB2A, WRKY40, and WRKY53 in Arabidopsis. Protein interaction assays show that MeGRXC3 interacts with Arabidopsis TGA2 and TGA5 in the nucleus. Eliminated nuclear localization of MeGRXC3 failed to result mannitol-induced inhibition of seed germination and seedling growth in transgenic Arabidopsis. Mutation analysis of MeGRXC3 indicates the importance of conserved motifs for its transactivation activity in yeast. Additionally, these motifs are also indispensable for its functionality in regulating mannitol-induced inhibition of seed germination and enhancement of the stress-related transcription factors in transgenic Arabidopsis. CONCLUSIONS: MeGRXC3 overexpression confers mannitol sensitivity in transgenic Arabidopsis possibly through interaction with TGA2/5 in the nucleus, and nuclear activity of MeGRXC3 is required for its function.
Assuntos
Glutarredoxinas/genética , Manihot/genética , Pressão Osmótica/fisiologia , Proteínas de Plantas/genética , Motivos de Aminoácidos , Arabidopsis , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Fatores de Transcrição de Zíper de Leucina Básica/genética , Fatores de Transcrição de Zíper de Leucina Básica/metabolismo , Núcleo Celular/genética , Núcleo Celular/metabolismo , Secas , Regulação da Expressão Gênica de Plantas , Germinação/efeitos dos fármacos , Glutarredoxinas/metabolismo , Manitol/farmacologia , Pressão Osmótica/efeitos dos fármacos , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Plântula/genética , Plântula/crescimento & desenvolvimento , Sementes/efeitos dos fármacos , Sementes/genética , Sementes/crescimento & desenvolvimento , Técnicas do Sistema de Duplo-HíbridoRESUMO
Quercetin is one of the main flavonoids in the human diet and mainly found in different plant tissues, including seeds, flowers, leaves, stems, and roots. However, its biological function in plant tissues, especially in seeds, is unknown. In this study, the seed germination and subsequent seedling growth of Apocynum pictum and A. venetum under osmotic stress (400 mmol L-1 mannitol) supplemented with 5 µmol L-1 quercetin were evaluated after 7, 14, and 21 days of germination. Results showed that quercetin improved the germination percentage and seed vigor, as indicated by the higher germination energy, shoot length, root length, dry weight, fresh weight, and chlorophyll content in A. pictum and A. venetum seedlings under the mannitol compared with those under the mannitol alone. Quercetin decreased H2O2 and O2- production and cell membrane damage, and mostly increased the gene expression of superoxide dismutase, peroxidase, catalase, chalcone synthase and flavonol synthase in A. pictum and A. venetum seedlings under the mannitol compared with those under the mannitol alone. In addition, the germination energy of A. pictum was 21.57% higher than that of A. venetum, and the gene expression of key enzymes in quercetin biosynthesis in A. pictum was mostly higher than that in A. venetum after 1 and 7 days of germination. These results indicated that quercetin was an effective anti-osmotic agent that alleviated the adverse effect of mannitol-induced osmotic stress on seed germination and seed vigor, and A. pictum seeds were more osmotic resistant than A. venetum seeds.