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Characterization of Rice Group 3 LEA Genes in Developmental Stages and Under Abiotic Stress

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Abstract

We performed a BLAST search of the rice database and screened four group 3 late embryogenesis abundant genes (OsG3LEA-47.3, OsG3LEA-41.9, OsG3LEA-20.5, and OsG3LEA-24.5) that shared characteristics of canonical G3LEAs such as multiple copies of consensus motif, hydrophilic, structural intrinsic disorder, thermostability, abscisic acid (ABA)-responsiveness, and high G and C content in gene sequence. Under nonstress conditions, OsG3LEA-20.5 and OsG3LEA-24.5 were constitutively expressed, whereas OsG3LEA-47.3 and OsG3LEA-41.9 were expressed in a stage- or tissue-specific manner. Transcripts of OsG3LEA-20.5 and OsG3LEA-24.5 accumulated under salt, ABA, and cold stress treatment. By contrast, OsG3LEA-47.3 and OsG3LEA-41.9 showed less responsiveness to stress in tillering and heading stages, respectively. All genes showed enhanced transcript levels after pollination and embryo development. To investigate the functions, we overexpressed OsG3LEA-47.3, OsG3LEA-41.9, OsG3LEA-20.5, and OsG3LEA-24.5 in Arabidopsis (47.3-ox, 41.9-ox, 20.5-ox, and 24.5-ox). Analysis of tolerance to drought stress revealed higher recovery after 14 days of dehydration treatment for 47.3-ox or 41.9-ox than the wild type (WT), 20.5-ox, or 24.5-ox. In addition, 47.3-ox and 24.5-ox plants showed higher survival than WT, 41.9-ox, and 20.5-ox plants under heat treatment, which induced similar expression patterns of heat shock protein genes in WT, 47.3-ox, 41.9-ox, 20.5-ox, and 24.5-ox plants. In vitro chaperone activity with the model substrate citrate synthase was comparable between OsG3LEA-47.3 and OsHSP16.9A, a rice molecular chaperone providing thermoprotection in vivo and in vitro. These results suggest that rice G3LEAs share many physical and biological features but function in contrasting ways.

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Abbreviations

ABA:

Abscisic acid

CS:

Citrate synthase

GFP:

Green fluorescence protein

HS:

Heat shock

HSPs:

Heat shock proteins

IPTG:

Isopropyl-β-d-thiogalactopyranoside

LEAs:

Late embryogenesis abundant proteins

RCBF2 :

C-repeated binding factor 2 gene

WT:

Wild type

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Acknowledgments

This work was supported by the National Science Council, Taiwan, ROC (no. NSC97-2313-B-008-001-MY3) to CH Yeh.

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Authors and Affiliations

  1. Graduate Institute of Systems Biology and Bioinformatics, National Central University, Taoyuan, 32001, Taiwan

    Yi-Ting Ke & Ching-Hui Yeh

  2. Department of Life Sciences, National Central University, Taoyuan, 32001, Taiwan

    Yi-Ting Ke, Chung-An Lu, Shaw-Jye Wu & Ching-Hui Yeh

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  1. Yi-Ting Ke
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  2. Chung-An Lu
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Correspondence to Ching-Hui Yeh.

Electronic supplementary material

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Supplementary Table S1

Oligonucleotides used in RT-PCR and coding region (CD) cloning. (DOCX 15 kb)

Supplementary Fig. S1

Hydropathy profiles for the deduced rice group 3 LEA proteins OsG3LEA-47.3, OsG3LEA-41.9, OsG3LEA-20.5, and OsG3LEA-24.5. (PPTX 296 kb)

Supplementary Fig. S2

Schematic representation of the structural features of the deduced rice group 3 LEA proteins OsG3LEA-47.3, OsG3LEA-41.9, OsG3LEA-20.5, and OsG3LEA-24.5. The disorder prediction agrees with the standard 0.5 threshold. (PPTX 120 kb)

Supplementary Fig. S3

The growth stage and sampling points in TNG67. (PPTX 151 kb)

Supplementary Fig. S4

Overall appearance of Col-0 WT and OsG3LEA-47.3 Arabidopsis transgenic plants grown on soil for 24 days at 23oC. (PPTX 557 kb)

Supplementary Fig. S5

Leaf number at flowering of OsG3LEA-41.9 transgenic Arabidopsis plants. (PPTX 53 kb)

Supplementary Fig. S6

Concentrations of OsG3LEA-47.3 with citrate synthase (CS)under 43oC. (PPTX 118 kb)

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Ke, YT., Lu, CA., Wu, SJ. et al. Characterization of Rice Group 3 LEA Genes in Developmental Stages and Under Abiotic Stress. Plant Mol Biol Rep 34, 1003–1015 (2016). https://doi.org/10.1007/s11105-016-0983-1

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