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Acta Agronomica Sinica ›› 2024, Vol. 50 ›› Issue (12): 3155-3164.doi: 10.3724/SP.J.1006.2024.44054

• RESEARCH NOTES • Previous Articles    

Cloning and expression analysis of transcription factor AhWRI1s in peanut

YIN Xiang-Zhen1(), ZHAO Jian-Xin2, HAO Cui-Cui2, PAN Li-Juan1, CHEN Na1, XU Jing1, JIANG Xiao1, ZHAO Xu-Hong1, WANG En-Qi2, CAO Huan2, YU Shan-Lin1, CHI Xiao-Yuan1,*()   

  1. 1Shandong Peanut Research Institute, Qingdao 266100, Shandong, China
    2College of Marine Science and Biological Engineering, Qingdao University of Science and Technology, Qingdao 266042, Shandong, China
  • Received:2024-03-29 Accepted:2024-08-15 Online:2024-12-12 Published:2024-09-03
  • Contact: *E-mail: chi000@126.com
  • Supported by:
    China Agriculture Research System of MOF and MARA(CARS-13);Agricultural Science and Technology Innovation Project of Shandong Academy of Agricultural Sciences(CXGC2023F20);Agricultural Science and Technology Innovation Project of Shandong Academy of Agricultural Sciences(CXGC2024F20);Key-Area Research and Development Program of Guangdong Province(2020B020219003);Major Scientific and Technological Project in Xinjiang(2022A02008-3);Taishan Scholar Project Funding, the Research and Development Program of Shandong Province (the Improved Variety Engineering Project)(2022LZGC007);Natural Science Foundation of Shandong Province(ZR2021QC172);Natural Science Foundation of Shandong Province(ZR2023QC146);Key Research and Development Plan of Shandong Province (Action Plan to Boost Scientific and Technological Innovation in Rural Revitalization)(2022TZXD0031)

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Abstract:

Peanut is one of the widely cultivated oil and economic crops worldwide and has become a major source of oil and protein for humans due to its high oil and protein content. With the increasing global demand for vegetable oil, improving the fatty acid composition and increasing the lipid content of peanut seeds has become a top priority in peanut breeding. Transcriptional regulators can modulate the expression of a series of genes in metabolic pathways related to lipid synthesis, significantly affecting lipid synthesis and metabolism. In this study, two transcription factors, AhWRI1-1 and AhWRI1-2, were cloned from the leaves of Huayu 33. The ORF of AhWRI1-1 was 1101 bp, encoding 366 amino acids, and the ORF of AhWRI1-2 was 1128 bp, encoding 375 amino acids. Bioinformatics analysis revealed that both AhWRI1-1 and AhWRI1-2 contained two AP2/EREBP conserved domains. The expression patterns of AhWRI1-1 and AhWRI1-2 in different tissues were detected by qRT-PCR. The results showed that AhWRI1-1 had the highest expression in seeds, suggesting its involvement in the regulation of fatty acid synthesis and oil accumulation, while AhWRI1-2 had the highest expression in hypocotyls, indicating its role in hypocotyl development. Additionally, the differences in the responses of AhWRI1-1 and AhWRI1-2 to abiotic stresses suggested that these transcription factors may play different roles under such conditions. Transcriptional activation experiments in yeast showed that both AhWRI1-1 and AhWRI1-2 possess transcriptional activation activities. This study lays the foundation for future in-depth functional studies of AhWRI1-1 and AhWRI1-2.

Key words: peanut, AP2/EREBP transcription factor, abiotic stress, gene expression analysis, transcriptional activation

Table 1

PCR primers"

引物名称
Primer name		 序列
Sequence (5′-3′)
AhWRI1-1-F ATGAAGAGGTCTCCTAATTCCTCT
AhWRI1-1-R TTAATAATTGGCATAGACATGCATATC
AhWRI1-2-F ATGGAAATGATGACAAAGAAA
AhWRI1-2-R TCACAGATAAGGGAGATTGC
qAhWRI1-1-F TGCCGAATTATCCGAGTT
qAhWRI1-1-R CAGCCATACACATAAGATTGAT
qAhWRI1-2-F TGAAGAGGAGTTGGATGTC
qAhWRI1-2-R TGTAGTGTTGGTAGTGGAAT

Fig. 1

Nucleotide sequence and deduced amino acid sequence of AhWRI1-1 (a) and AhWRI1-2 (b)"

Fig. 2

Alignment of WRI1 amino acid sequences from peanut and other species The two AP2/EREBP domains of WRI1 protein were marked by horizontal lines."

Fig. 3

Gene structure of AhWRI1-1 and AhWRI1-2"

Fig. 4

Phylogenetic analysis of plant WRI1s"

Fig. 5

Expression analysis of AhWRI1-1 and AhWRI1-2 in different tissues and at different seed developmental stages in peanut a: expression analysis of AhWRI1-1 in different tissues; b: expression analysis of AhWRI1-2 in different tissues; c: expression analysis of AhWRI1-1 in seeds at different developmental periods; d: expression analysis of AhWRI1-1 in seeds at different developmental periods. Actin11 was used as the internal reference gene. In a, b, c, and d, set the expression levels of AhWRI1-1 in seeds, AhWRI1-2 in hypocotyls, AhWRI1-1 in seeds at 48 d, and AhWRI1-2 in seeds at 12 d to 1, respectively, as reference points."

Fig. 6

Expression analysis of AhWRI1-1 (a) and AhWRI1-2 (b) under abiotic stresses and hormone treatment conditions Using Actin11 as the internal reference gene, set the expression levels of AhWRI1-1 and AhWRI1-2 at 0 h to 1, respectively, as reference points. ABA: abscisic acid; GA: gibberellic acid; ET: ethephon; JA: jasmonic acid; SA: salicylic acid."

Fig. 7

Analysis of transcriptional activation activity of AhWRI1-1 and AhWRI1-2"

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