title: Molecular characterization and functional analysis by heterologous expression in E. coli under diverse abiotic stresses for OsLEA5, the atypical hydrophobic LEA protein from Oryza sativa L.
abstract: In this study, we report the molecular characterization and functional analysis of OsLEA5 gene, which belongs to the atypical late embryogenesis abundant (LEA) group 5C from Oryza sativa L. The cDNA of OsLEA5 contains a 456 bp ORF encoding a polypeptide of 151 amino acids with a calculated molecular mass of 16.5 kDa and a theoretical pI of 5.07. The OsLEA5 polypeptide is rich in Leu (10%), Ser (8.6%), and Asp (8.6%), while Cys, Trp, and Gln residue contents are very low, which are 2, 1.3, and 1.3%, respectively. Bioinformatic analysis revealed that group 5C LEA protein subfamily contains a Pfam:LEA_2 domain architecture and is highly hydrophobic, intrinsically ordered with largely β-sheet and specific amino acid composition and distribution. Real-time PCR analysis showed that OsLEA5 was expressed in different tissue organs during different development stages of rice. The expression levels of OsLEA5 in the roots and panicles of full ripe stage were dramatically increased. The results of stress tolerance and cell viability assay demonstrated that recombinant E. coli cells producing OsLEA5 fusion protein exhibited improved resistance against diverse abiotic stresses: high salinity, osmotic, freezing, heat, and UV radiation. The OsLEA5 protein confers stabilization of the LDH under different abiotic stresses, such as heating, freeze-thawing, and drying in vitro. The combined results indicated that OsLEA5 protein was a hydrophobic atypical LEA and closely associated with resistance to multiple abiotic stresses. This research offered the valuable information for the development of crops with enhanced resistance to diverse stresses.
authors: Key Laboratory of Biorheological Science and Technology, Ministry of Education, College of Bioengineering, Chongqing University, Chongqing, China.
authors: Shuai
authors: S
authors: He
authors: Lili
authors: L
authors: Tan
authors: Zongli
authors: Z
authors: Hu
authors: Guoping
authors: G
authors: Chen
authors: Guixue
authors: G
authors: Wang
authors: Tingzhang
authors: T
authors: Hu
mesh_terms: D000383:Agriculture / Q000379:methods; D000596:Amino Acids / Q000235:genetics; D019295:Computational Biology; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli; D035264:Flowers / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental / Q000235:genetics / Q000502:physiology*; D057927:Hydrophobic and Hydrophilic Interactions; D016366:Open Reading Frames / Q000235:genetics; D012275:Oryza / Q000235:genetics*; D009997:Osmotic Pressure; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism*; D018517:Plant Roots / Q000378:metabolism; D011487:Protein Conformation; D060888:Real-Time Polymerase Chain Reaction; D054712:Salinity; D013312:Stress, Physiological / Q000235:genetics / Q000502:physiology*; D013696:Temperature
chemical_list: D000596:Amino Acids; D018076:DNA, Complementary; D010940:Plant Proteins; C066246:late embryogenesis abundant protein, plant