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Acta Agronomica Sinica ›› 2025, Vol. 51 ›› Issue (7): 1969-1978.doi: 10.3724/SP.J.1006.2025.44210

• RESEARCH NOTES • Previous Articles    

Promoter characterization and expression pattern analysis of the m6A methyltransferase gene SiMTA1 in foxtail millet

SHEN Ao1,2(), LIU Min2, NI Di-An1,*(), LIU Wei2,*()   

  1. 1School of Ecological Technology & Engineering, Shanghai Institute of Technology, Shanghai 201418, China
    2Institute of Crop Germplasm Resources, Shandong Academy of Agricultural Sciences, Jinan 250100, Shandong, China
  • Received:2024-12-17 Accepted:2025-04-30 Online:2025-07-12 Published:2025-05-12
  • Contact: *E-mail: wheiliu@163.com; E-mail: dani@sit.edu.cn
  • Supported by:
    Shandong Provincial Key Research and Development Program(2021LZGC025);National Natural Science Foundation of China(32171955);National Natural Science Foundation of China(32201736);Agricultural Science and Technology Innovation Project of the Shandong Academy of Agricultural Sciences(CXGC2023F13)

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Abstract:

MTA, a key RNA methyltransferase responsible for m6A methylation, also influences embryonic development and plays crucial roles in plant growth by interacting with other enzymes. In this study, the SiMTA1 gene (accession no. PQ801843) from foxtail millet was identified through homology alignment with Arabidopsis methyltransferase sequences. The gene is 4,239 bp in length, with a 2121 bp coding sequence (CDS) encoding a protein of 706 amino acids. Bioinformatic analyses were conducted on both the nucleotide and protein sequences, and cis-acting elements in the promoter region were characterized. The spatiotemporal expression patterns of SiMTA1 under various abiotic stresses and hormone treatments were examined using qRT-PCR. The results showed that SiMTA1 contains the MT-A70 domain, a subunit of N6-adenosine methyltransferase (MTase) that binds S-adenosylmethionine (SAM), and its secondary structure is mainly composed of random coils and α-helices. The SiMTA1 promoter harbors multiple stress- and hormone-responsive cis-elements. SiMTA1 is highly expressed in stem internodes during the heading stage of foxtail millet, and its expression is upregulated by salt, drought, auxin, cytokinin, and other treatments. These findings suggest that SiMTA1 may participate in developmental processes and responses to environmental and hormonal signals in foxtail millet. This study provides a theoretical foundation and a potential candidate gene for the genetic improvement of stress-resistant foxtail millet varieties.

Key words: foxtail millet, methyltransferase SiMTA1, promoter characteristics, stress and plant hormone response, spatiotemporal expression pattern

Fig. S1

SiMTA1-P promoter sequence"

Table 1

Primers used in this study"

引物名称Primer name 引物序列Primer sequence (5'-3')
Q-SiMTA1-F1 TGGCTGACGATGAAATGAGG
Q-SiMTA1-R1 GCCAGTTCGGATGATACGTT

Fig. 1

Multiple sequence alignment of MTA proteins from different plant species The accession numbers in NCBI are: AtMTA of Arabidopsis thaliana: NP_192814; OsMTA of rice: NP_001403747; TaMTA of wheat: XP_044403789; ZmMTA of maize: NP_001147883. The red box highlights the highly conserved region."

Fig. 2

Prediction of the SiMTA1 protein domain Numbers represent the amino acid positions of the corresponding sequences. The purple region represents a low-complexity sequence, while the gray box denotes the MT-A70 domain."

Fig. 3

Prediction of the secondary structure of the SiMTA1 Numbers represent the amino acid positions of the corresponding sequences. Blue: random coil; light purple: α-helix; dark purple: extended chain."

Fig. 4

Prediction of the tertiary structure of the SiMTA1 protein Different colors represent confidence levels (score range: 0-1).Dark blue: score > 0.9; blue: 0.9 > score > 0.7; yellow: 0.7 > score > 0.5; orange: score < 0.5."

Fig. 5

Schematic diagram of the distribution of cis-acting elements in the sense-strand of promoter SiMTA1-P Numbers represent the length of sequences corresponding to the positions of promoters, with different shapes representing different cis-acting elements."

Table 2

Partial cis-acting elements in the promoter SiMTA1-P"

顺式作用元件
Cis-acting element		 位置
Position		 序列
Sequence (5′-3′)		 生物学功能
Biological function
ABRE 17+
17-, 24-
843+
1156+
1585-
1898-		 CACGTG
ACGTG
AACCCGG
ACGTG
TACGGTC
GCCGCGTGGC		 含有ABRE基序的基因通常在ABA的调控下表达, 从而参与植物的逆境适应过程。
Genes containing the ABRE motif are typically expressed under the regulation of ABA, thereby participating in the plant’s stress adaptation process.
AS-1 1701- TGACG 与植物对生长素的响应有关, 参与调节生长素应答基因的表达, 影响植物的发育过程。
Associated with the plant’s response to auxin, it is involved in the regulation of auxin-responsive gene expression, thereby influencing the developmental processes of plants.
AuxRR-core 409+ GGTCCAT 生长素相关核心。
Auxin-related core.
CCGTCC-box 32+ CCGTCC 与植物的光信号传导和逆境胁迫响应有关。
Associated with plant light signaling and stress response.
CGTCA-motif 1701+ CGTCA 与植物对逆境(如干旱、盐胁迫等)的响应相关。
Related to the plant’s response to abiotic stresses, such as drought and salinity.
DRE 1737+ GCCGAC 与植物的干旱、低温和高盐胁迫响应有关。
Related to the plant’s response to drought, low-temperature, and high-salinity stresses.
G-box 24+, 1155- CACGTT 通常与逆境胁迫响应和植物激素信号传导有关。
Typically associated with stress responses and plant hormone signaling.
LTR 643-, 1116+ CCGAAA 与基因的低温响应有关。
Associated with the low-temperature response of genes.
MYB 817-, 876-
882-
1200-
1601+		 CAACCA
CAACAG
TAACTG
CAACCA		 参与多种生物学过程, 包括植物激素信号传导、生长发育、次生代谢以及逆境响应等。
Involved in a variety of biological processes, including plant hormone signaling, growth and development, secondary metabolism, and stress responses.
MYC 471+ CATTTG 参与多种生物学过程, 包括植物激素信号传导、生物钟调控以及逆境响应等。
Involved in a variety of biological processes, including plant hormone signaling, circadian clock regulation, and stress responses.
STRE 155-, 425-, 1238-, 1841+ AGGGG 参与植物对环境应激如冷、热、盐胁迫等的响应。
Involved in the plant’s response to environmental stresses such as cold, heat, and salt stress.

Fig. 6

Spatiotemporal expression patterns of SiMTA1 A: relative expression levels of SiMTA1 at different development stages; B: relative expression levels of SiMTA1 in different tissues during the booting stage. Different lowercase letters indicate significant differences at P < 0.05."

Fig. 7

Relative expression levels of SiMTA1 under different abiotic stress treatments A: relative expression levels of SiMTA1 under salt stress; B: relative expression levels of SiMTA1 under drought stress; C: relative expression levels of SiMTA1 under 4℃ stress. Different lowercase letters indicate significant differences at P < 0.05."

Fig. 8

Relative expression levels of SiMTA1 under different hormone treatments A: relative expression levels of SiMTA1 under IAA treatment; B: relative expression levels of SiMTA1 under 6-BA treatment; C: relative expression levels of SiMTA1 under GA3 treatment; D: relative expression levels of SiMTA1 under ACC treatment; E: relative expression levels of SiMTA1 under NAA treatment. Different lowercase letters indicate significant differences at P < 0.05."

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