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Identification of ERF genes in peanuts and functional analysis of AhERF008 and AhERF019 in abiotic stress response

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Abstract

Ethylene-responsive factor (ERF) play an important role in regulating gene expression in plant development and response to stresses. In peanuts (Arachis hypogaea L.), which produce flowers aerially and pods underground, only a few ERF genes have been identified so far. This study identifies 63 ERF unigenes from 247,313 peanut EST sequences available in the NCBI database. The phylogeny, gene structures, and putative conserved motifs in the peanut ERF proteins were analysed. Comparative analysis revealed the absence of two subgroups (A1 and A3) of the ERF family in peanuts; only 10 subgroups were identified in peanuts compared to 12 subgroups in Arabidopsis and soybeans. AP2/ERF domains were found to be conserved among peanuts, Arabidopsis, and soybeans. Outside the AP2/ERF domain, many soybean-specific conserved motifs were also detected in peanuts. The expression analysis of ERF family genes representing each clade revealed differential expression patterns in response to biotic and abiotic stresses. Overexpression of AhERF008 influenced the root gravity of Arabidopsis, whereas overexpression of AhERF019 enhanced tolerance to drought, heat, and salt stresses in Arabidopsis. The information generated in this study will be helpful to further investigate the function of ERFs in plant development and stress response.

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Acknowledgments

This work was funded by the National Natural Science Foundation of China (No. 31071456), the National Basic Research Program of China (2013CB127803), the China Agriculture Research System (No. CARS-14), and the China Postdoctoral Science Foundation (No. 2013M530782).

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Authors and Affiliations

  1. Key Laboratory of Biology and Genetic Improvement of Oil Crops, Ministry of Agriculture, Oil Crops Research Institute of Chinese Academy of Agricultural Sciences, Wuhan, China

    Liyun Wan, Yanshan Wu, Jiaquan Huang, Yong Lei, Liying Yan, Huifang Jiang & Boshou Liao

  2. Institute of Agro-products Processing Science and Technology, Chinese Academy of Agricultural Sciences, Beijing, 100193, China

    Xiaofeng Dai

  3. The State Key Laboratory of Plant Cell and Chromosome Engineering, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing, China

    Juncheng Zhang

  4. International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad, India

    Rajeev K Varshney

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  1. Liyun Wan
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ESM Table 1

Primer sequences of the peanut ERF genes referenced in this article. (XLSX 18 kb)

ESM Table 2

Characteristics of the peanut ERF genes. (XLSX 11 kb)

ESM Table 3

ESTs coding the 63 ERF factors identified in peanuts. (XLSX 2592 kb)

ESM Table 4

Summary of conserved motifs (CMs) within the AhERF family by comparative analysis with soybeans. (XLSX 37 kb)

ESM Table 5

Expression patterns of the peanut stress responsive ERF genes. (XLSX 12 kb)

ESM Table 6

Annotated information for the peanut ERFs from the NCBI database. (XLSX 10 kb)

ESM Fig. 1

Phylogenetic tree of peanut and soybean ERF proteins. (GIF 198 kb)

High resolution image (TIFF 763 kb)

ESM Fig. 2

Amino acid sequence alignment of the AP2/ERF DNA-binding domains from the 63 peanut and 98 soybean ERF proteins described by Nakano et al. (2006) using ClustalW. (GIF 6292 kb)

High resolution image (TIFF 6292 kb)

ESM Fig. 3

Unrooted phylogenetic tree of peanut ERF proteins. (GIF 30 kb)

High resolution image (TIFF 926 kb)

ESM Fig. 4

Phylogenetic relationships among peanut CBF/DREB subfamily (group A) unigenes. (GIF 24 kb)

High resolution image (TIFF 1846 kb)

ESM Fig. 5

Phylogenetic relationships among the peanut ERF subfamily (group B) unigenes. (GIF 54 kb)

High resolution image (TIFF 1802 kb)

ESM Fig. 6

EAR motif-like sequences conserved in the C-terminal region of subgroups A-5 and B-1 in peanuts and soybeans. (A) Amino acid sequence alignment of the C-terminal region of proteins from subgroup A-5. (B) Amino acid sequence alignment of the C-terminal region of proteins from subgroup B-1 proteins. (GIF 164 kb)

High resolution image (TIFF 2029 kb)

ESM Fig. 7

Soybean-specific sequence motifs conserved in subgroups A-6, B-1, and B-2 of the ERF family are also exist in peanuts. (A) Amino acid sequence alignment of proteins from subgroup A-6. (B) Amino acid sequence alignment of proteins from subgroup B-1. (C) Amino acid sequence alignment of proteins from subgroup B-2. (GIF 128 kb)

High resolution image (TIFF 3518 kb)

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Wan, L., Wu, Y., Huang, J. et al. Identification of ERF genes in peanuts and functional analysis of AhERF008 and AhERF019 in abiotic stress response. Funct Integr Genomics 14, 467–477 (2014). https://doi.org/10.1007/s10142-014-0381-4

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