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Molecular characterization of a thioredoxin h gene (HbTRX1) from Hevea brasiliensis showing differential expression in latex between self-rooting juvenile clones and donor clones

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Abstract

The cDNA code of thioredoxin h, designated as HbTRX1, was isolated from Hevea brasiliensis by rapid amplification of cDNA ends. HbTRX1 contained a 542-bp open reading frame encoding 123 amino acids. The deduced HbTRX1 protein showing high identity to thioredoxin h of other plant species was predicted to possess the conserved catalytic site WCXPC. Semiquantitative reverse transcription-polymerase chain reaction analysis revealed that HbTRX1 was constitutively expressed in all tested tissues. HbTRX1 transcripts accumulated at relatively low levels in the flower, somatic embryo, and leaves, while HbTRX1 transcripts accumulated at relatively high levels in the callus and latex. The HbTRX1 transcript was expressed at different levels, with higher levels in self-rooting juvenile clones than in their donor clones. HbTRX1 was expressed in Escherichia coli, and its activity was demonstrated using the dithiothreitol-dependent insulin assay. This work provides a basis for studying the biological function of thioredoxin h in rubber tree.

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Abbreviations

ORF:

Open reading frame

RACE:

Rapid amplification of cDNA ends

RT-PCR:

Reverse transcription-polymerase chain reaction

UTR:

Untranslated region

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Acknowledgments

This work was supported by the National Nonprofit Institute Research Grant of CATAS-ITBB (ITBBZD0712) and the National Natural Science Foundation of China (30960307). We deeply thank Professor Xiong-Ting Chen (Institute of Tropical Biosciences and Biotechnology, Chinese Academy of Tropical Agricultural Sciences) for his collaboration in the treatment of plant materials.

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Authors and Affiliations

  1. Key Laboratory of Tropical Crop Biotechnology, Ministry of Agriculture, Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou, 571101, People’s Republic of China

    Hui-Liang Li, Hui-Zhong Lu, Dong Guo & Shi-Qing Peng

  2. Key Laboratory of Rubber Biology, Ministry of Agriculture, Institute of Rubber, Chinese Academy of Tropical Agricultural Sciences, Danzhou, 571737, People’s Republic of China

    Wei-Min Tian

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  1. Hui-Liang Li
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  2. Hui-Zhong Lu
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  3. Dong Guo
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Corresponding author

Correspondence to Shi-Qing Peng.

Additional information

Hui-Liang Li and Hui-Zhong Lu contributed equally to this work.

Electronic supplementary material

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11033_2010_321_MOESM1_ESM.tif

Supplementary 1 Sequence alignment of the deduced HbTRX1 and other plant thioredoxins h. VvTRX (Vitis vinifera, ABX79345), ClTRX (Codonopsis lanceolata, BAE16559), GmTRX (Glycine max, ABV71991), HbTRX (H. brasiliensis, AF133127.1), PtTRX (Populus tremula × Populus, AF483266), ATTRX1 (A. thaliana, P29448), ATTRX2 (A. thaliana, NM_123358), ATTRX3 (A. thaliana, NM_123664), ATTRX4 (A. thaliana, NM_101829), ATTRX5 (A. thaliana, NM_103588), ATTRX6 (A. thaliana, AAD39316), ATTRX7 (A. thaliana, AAG52561), and ATTRX8 (A. thaliana, AAG51342). Amino acid residues that are identical in all six sequences are shaded in black, and well-conserved residues are shaded in gray. (TIFF 1143 kb)

11033_2010_321_MOESM2_ESM.tif

Supplementary 2 Phylogenetic analysis of HbTRX1 and other species by MEGA version 3.1 from CLUSTAL W alignments. The thioredoxins h used in the evolutionary analysis are retrieved from the GenBank, including VvTRX (V. vinifera, ABX79345), ClTRX (C. lanceolata, BAE16559), GmTRX (G. max, ABV71991), HbTRX (H. brasiliensis, AF133127.1), PtTRX (P. tremula × Populus, AF483266), ATTRX1 (A. thaliana, P29448), ATTRX2 (A. thaliana, NM_123358), ATTRX3 (A. thaliana, NM_123664), ATTRX4 (A. thaliana, NM_101829), ATTRX5 (A. thaliana, NM_103588), ATTRX6 (A. thaliana, AAD39316), ATTRX7 (A. thaliana, AAG52561), and ATTRX8 (A. thaliana, AAG51342). (TIFF 372 kb)

11033_2010_321_MOESM3_ESM.tif

Supplementary 3 Nucleotide sequence of the promoter region of HbTRX1. The start code is boxed. The cis-acting regulatory elements are underlined. (TIFF 406 kb)

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Li, HL., Lu, HZ., Guo, D. et al. Molecular characterization of a thioredoxin h gene (HbTRX1) from Hevea brasiliensis showing differential expression in latex between self-rooting juvenile clones and donor clones. Mol Biol Rep 38, 1989–1994 (2011). https://doi.org/10.1007/s11033-010-0321-x

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