A germin-like protein gene of rice increased superoxide dismutase activity in transformed tobacco

Yasmin, Tayyaba; Mumtaz, Abdul Samad; Mahmood, Tariq; Naqvi, Syed Muhammad Saqlan

doi:10.1007/s10535-015-0524-z

Cited by 21 publications

(12 citation statements)

References 30 publications

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“…The influence of plant defence by GLPs is likely related to their SOD activity [23,27,52]. Superoxide produced by NADPH oxidase or peroxidases in response to pathogen attack is predicted to be dis-mutated to H 2 O 2 by the GLPs, accounting for the accumulation of H 2 O 2 [41].…”

Section: Resultsmentioning

confidence: 99%

Genome-Wide Characterization and Expression Analysis of the Germin-Like Protein Family in Rice and Arabidopsis

Chen

et al. 2016

IJMS

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Previous studies have shown that germin-like proteins (GLPs) are present ubiquitously in rice and Arabidopsis. However, the understanding regarding their role in development and abiotic/biotic stress resistance remains limited. In the present study, we report genome-wide identification, characterisation, subcellular localization, enzyme activity, and expression analysis of the GLP gene family in rice and Arabidopsis to study their functions. In total, 43 and 32 GLPs in the rice and Arabidopsis genome were identified based on a systematic analysis, respectively. The GLP genes were clustered into six clades based on phylogenetic analysis, and many stress and developmental-related cis-elements were detected in promoters of GLP genes. In addition, subcellular location and superoxide dismutase (SOD) analysis demonstrated that the random selected OsGLP genes on chromosomes 8 and 4 of rice were expressed in the cell wall with SOD activity. Overall, our results showed that tandem duplication events, especially the clusters of tandem duplication genes on chromosome 8 in rice, play a major role in expansion of the GLP family and thus increase our understanding of the role of the GLP family in abiotic/biotic stress and development.

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Section: Resultsmentioning

confidence: 99%

Genome-Wide Characterization and Expression Analysis of the Germin-Like Protein Family in Rice and Arabidopsis

Chen

et al. 2016

IJMS

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“…Germin-like proteins (GLPs) compose a diverse family of plant glycoproteins belonging to the cupin superfamily. GLPs in cereals and other plant species may have oxalate oxidase (OXO) or superoxide dismutase (SOD) activity, producing H 2 O 2 (Yasmin et al, 2015), however GLPs from different species were reported to be involved in biotic as well as abiotic stresses and also in the growth and development (Barman and Banerjee, 2015). For example, transgenic plants with increased levels of germin like protein subfamily 2 member 1 (AT1G09560) exhibited reduced primary root and enhanced lateral root growth suggesting a role as regulatory components of root architecture (Ham et al, 2012) in line with reported responses (Figure 1).…”

Section: Discussionmentioning

confidence: 99%

Protein Profiling of Arabidopsis Roots Treated With Humic Substances: Insights Into the Metabolic and Interactome Networks

et al. 2018

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Background and Aim: Humic substances (HSs) influence the chemical and physical properties of the soil, and are also known to affect plant physiology and nutrient uptake. This study aimed to elucidate plant metabolic pathways and physiological processes influenced by HS activity.Methods: Arabidopsis roots were treated with HS for 8 h. Quantitative mass spectrometry-based proteomics analysis of root proteins was performed using the iTRAQ (Isobaric Tag for Relative and Absolute Quantification) technique. Out of 902 protein families identified and quantified for HS treated vs. untreated roots, 92 proteins had different relative content. Bioinformatic tools such as STRING, KEGG, IIS and Cytoscape were used to interpret the biological function, pathway analysis and visualization of network amongst the identified proteins.Results: From this analysis it was possible to evaluate that all of the identified proteins were functionally classified into several categories, mainly redox homeostasis, response to inorganic substances, energy metabolism, protein synthesis, cell trafficking, and division.Conclusion: In the present study an overview of the metabolic pathways most modified by HS biological activity is provided. Activation of enzymes of the glycolytic pathway and up regulation of ribosomal protein indicated a stimulation in energy metabolism and protein synthesis. Regulation of the enzymes involved in redox homeostasis suggest a pivotal role of reactive oxygen species in the signaling and modulation of HS-induced responses.

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“…This assay showed that Os RGLP1 had equivalent activity to E. coli SODA (Figure 3D-F), confirming that Os RGLP1 purified from K12 shuffle E. coli was properly folded and active. Os RGLP1 expressed in plants is reported to confer high-temperature resistant SOD activity [15, 16, 26]. To test the thermal stability of Os RGLP1 expressed from E. coli , purified protein was incubated at different temperatures and then tested for residual SOD activity (Figure 3G).…”

Section: Resultsmentioning

confidence: 99%

“…Os RGLP1 has been successfully transformed into Nicotiana tabacum (tobacco), Solanum tuberosum (potato) and Medicago truncatula (barrelclover) [13, 14]. Os RGLP1-transformed transgenic plants have higher SOD activity than wild type plants, and enhanced stress tolerance due to this heat resistant SOD activity [15]. Heterologous expression of Os RGLP1 in potato also induces resistance against infection by Fusarium oxysporum f.sb.…”

Section: Introductionmentioning

confidence: 99%

N-glycosylation onOryza SativaRoot Germin-like Protein 1 is conserved but not required for stability or activity

Rubbab

Pegg

Phung

et al. 2021

Preprint

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Germin and germin-like proteins (GLPs) are a broad family of extracellular glycoproteins ubiquitously distributed in plants. Overexpression of Oryza sativa root germin like protein 1 (OsRGLP1) enhances superoxide dismutase (SOD) activity in transgenic plants. Here, we report bioinformatic analysis and heterologous expression of OsRGLP1 to study the role of glycosylation on OsRGLP1 protein stability and activity. Sequence analysis of OsRGLP1 homologs identified diverse N-glycosylation sequons, one of which was highly conserved. We therefore expressed OsRGLP1 in glycosylation-competent Saccharomyces cerevisiae as a Maltose Binding Protein (MBP) fusion. Mass spectrometry analysis of purified OsRGLP1 showed it was expressed by S. cerevisiae in both N-glycosylated and unmodified forms. Glycoprotein thermal profiling showed little difference in the thermal stability of the glycosylated and unmodified protein forms. Circular Dichroism spectroscopy of MBP-OsRGLP1 and a N-Q glycosylation-deficient variant showed that both glycosylated and unmodified MBP-OsRGLP1 had similar secondary structure, and both forms had equivalent SOD activity. Together, we concluded that glycosylation was not critical for OsRGLP1 protein stability or activity, and it could therefore likely be produced in Escherichia coli without glycosylation. Indeed, we found that OsRGLP1 could be efficiently expressed and purified from K12 shuffle E. coli with a specific activity of 1251 Units/mg. In conclusion, we find that some highly conserved N-glycosylation sites are not necessarily required for protein stability or activity, and describe a suitable method for production of OsRGLP1 which paves the way for further characterization and use of this protein.

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A germin-like protein gene of rice increased superoxide dismutase activity in transformed tobacco

Cited by 21 publications

References 30 publications

Genome-Wide Characterization and Expression Analysis of the Germin-Like Protein Family in Rice and Arabidopsis

Genome-Wide Characterization and Expression Analysis of the Germin-Like Protein Family in Rice and Arabidopsis

Protein Profiling of Arabidopsis Roots Treated With Humic Substances: Insights Into the Metabolic and Interactome Networks

N-glycosylation onOryza SativaRoot Germin-like Protein 1 is conserved but not required for stability or activity

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