Characterization of OsLEA1a and its inhibitory effect on the resistance of  E. coli  to diverse abiotic stresses

Hu, Tingzhang; Zhang, Nong; Fu, Meiling; Qin, Jining; Huang, Xiaoyun

doi:10.1016/j.ijbiomac.2016.06.056

Cited by 16 publications

(16 citation statements)

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“…Sorbitol (0.6, 0.8, and 1.0 M) and PEG 4000 (5, 10, and 20% w/v) were used to decrease osmotic potential and mimic dehydration, and H 2 O 2 (0.1, 0.5, and 1.0 M) was tested to promote oxidant conditions. In all experiments, absorbance at 600 nm (OD 600 ) was used to measure the bacterial growth in liquid media (Liu and Zheng, 2005; Wu et al, 2014; Hu et al, 2016). All experiments were carried out in three biological replicates each replicate was done at least three times.…”

Section: Methodsmentioning

confidence: 99%

Insights on Structure and Function of a Late Embryogenesis Abundant Protein from Amaranthus cruentus: An Intrinsically Disordered Protein Involved in Protection against Desiccation, Oxidant Conditions, and Osmotic Stress

et al. 2017

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Late embryogenesis abundant (LEA) proteins are part of a large protein family that protect other proteins from aggregation due to desiccation or osmotic stresses. Recently, the Amaranthus cruentus seed proteome was characterized by 2D-PAGE and one highly accumulated protein spot was identified as a LEA protein and was named AcLEA. In this work, AcLEA cDNA was cloned into an expression vector and the recombinant protein was purified and characterized. AcLEA encodes a 172 amino acid polypeptide with a predicted molecular mass of 18.34 kDa and estimated pI of 8.58. Phylogenetic analysis revealed that AcLEA is evolutionarily close to the LEA3 group. Structural characteristics were revealed by nuclear magnetic resonance and circular dichroism methods. We have shown that recombinant AcLEA is an intrinsically disordered protein in solution even at high salinity and osmotic pressures, but it has a strong tendency to take a secondary structure, mainly folded as α-helix, when an inductive additive is present. Recombinant AcLEA function was evaluated using Escherichia coli as in vivo model showing the important protection role against desiccation, oxidant conditions, and osmotic stress. AcLEA recombinant protein was localized in cytoplasm of Nicotiana benthamiana protoplasts and orthologs were detected in seeds of wild and domesticated amaranth species. Interestingly AcLEA was detected in leaves, stems, and roots but only in plants subjected to salt stress. This fact could indicate the important role of AcLEA protection during plant stress in all amaranth species studied.

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Section: Methodsmentioning

confidence: 99%

Insights on Structure and Function of a Late Embryogenesis Abundant Protein from Amaranthus cruentus: An Intrinsically Disordered Protein Involved in Protection against Desiccation, Oxidant Conditions, and Osmotic Stress

et al. 2017

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“…The transformed E. coli (pET28a- DofLEA ) cell were grown in our lab as described previously103037. Briefly, DofLEA genes were inserted into the pET28a vector and then transformed into the E. coli host strain BL21 (DE3).…”

Section: Methodsmentioning

confidence: 99%

“…Heat and salt tolerance assays were performed as described previously103037 (shown in Figure 6S). The transformed E. coli (pET28a- DofLEA ) colonies grown on Luria-Bertani (LB)+ kanamycin plates were used to inoculate starting cultures, which were incubated at 37 °C for 12–16 h. The cultures were diluted 50-fold using fresh LB medium, and incubation continued at 37 °C until the mid-log phase (2–3 h, OD 600 = 0.6).…”

Section: Methodsmentioning

confidence: 99%

Functional insights into the late embryogenesis abundant (LEA) protein family from Dendrobium officinale (Orchidaceae) using an Escherichia coli system

Ling

Guo

2016

Sci Rep

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Late embryogenesis abundant (LEA) proteins, a diverse family, accumulate during seed desiccation in the later stages of embryogenesis. LEA proteins are associated with tolerance to abiotic stresses, such as drought, salinity and high or cold temperature. Here, we report the first comprehensive survey of the LEA gene family in Dendrobium officinale, an important and widely grown medicinal orchid in China. Based on phylogenetic relationships with the complete set of Arabidopsis and Oryza LEA proteins, 17 genes encoding D. officinale LEAs (DofLEAs) were identified and their deduced proteins were classified into seven groups. The motif composition of these deduced proteins was correlated with the gene structure found in each LEA group. Our results reveal the DofLEA genes are widely distributed and expressed in tissues. Additionally, 11 genes from different groups were introduced into Escherichia coli to assess the functions of DofLEAs. Expression of 6 and 7 DofLEAs in E. coli improved growth performance compared with the control under salt and heat stress, respectively. Based on qPCR data, all of these genes were up-regulated in various tissues following exposure to salt and heat stresses. Our results suggest that DofLEAs play an important role in responses to abiotic stress.

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“…Bacterial and yeast cells have been widely used to determine the effects of LEA proteins on abiotic stress tolerance (e.g., Hu et al 2016, Saucedo et al 2017. In this study, we investigated the survival and growth of E. coli cells overexpressing MtLEA proteins under abiotic stresses.…”

Section: Resultsmentioning

confidence: 99%

Three tandemly aligned LEA genes from Medicago truncatula confer differential protection to Escherichia coli against abiotic stresses

Zhang¹,

Wang²,

Liu³

et al. 2020

Biol. plant.

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Late embryogenesis abundant (LEA) proteins are important for abiotic stress tolerance in diverse organisms. Within the LEA protein superfamily, group 4 members are characterized by a conserved N-terminal region and a structurally disordered C-terminal region that varies regarding length and amino acid content. Previous in vitro assays have suggested that the conserved N-terminal region shared by group 4 LEA proteins is critical for forming an amphipathic α-helix and protecting enzymatic activities from the adverse effects of desiccation or freezing. However, the cellular roles of the varying C-terminal region remain largely to be characterized. Medicago truncatula contains five subgroup LEA4B proteins encoding loci of which three are tandemly arranged on chromosome 7 due to local gene duplication events. In this study, abiotic stresses and addition of abscisic acid (ABA) induced the transcription of the four LEA4B genes. Escherichia coli cells overexpressing the three tandemly aligned LEA genes indicated significantly increased tolerance to salt, osmotic, heat, and freezing stresses. However, the extent of the protective effects on the survival and growth of bacterial cells differed among the LEA proteins, potentially because of variations in the C-terminal region. This possibility was further supported by the observation that the protective effects of the native truncated MtLEA3140, which only contains a conserved N-terminal region, were inferior to the effects of the full-length mutant version. The results suggest that the structurally flexible C-terminal region of group 4 LEA proteins plays roles in protecting cells from damages caused by various abiotic stresses, and provide clues for elucidating the mechanisms underlying the intracellular functions of these proteins.

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Characterization of OsLEA1a and its inhibitory effect on the resistance of E. coli to diverse abiotic stresses

Cited by 16 publications

References 50 publications

Insights on Structure and Function of a Late Embryogenesis Abundant Protein from Amaranthus cruentus: An Intrinsically Disordered Protein Involved in Protection against Desiccation, Oxidant Conditions, and Osmotic Stress

Insights on Structure and Function of a Late Embryogenesis Abundant Protein from Amaranthus cruentus: An Intrinsically Disordered Protein Involved in Protection against Desiccation, Oxidant Conditions, and Osmotic Stress

Functional insights into the late embryogenesis abundant (LEA) protein family from Dendrobium officinale (Orchidaceae) using an Escherichia coli system

Three tandemly aligned LEA genes from Medicago truncatula confer differential protection to Escherichia coli against abiotic stresses

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