Cloning and expression analysis of Fagopyrum tataricum FtDELLA gene

doi:10.3969/j.issn.1004-1524.20230913

Acta Agriculturae Zhejiangensis ›› 2024, Vol. 36 ›› Issue (8): 1709-1718.DOI: 10.3969/j.issn.1004-1524.20230913

• Crop Science • Previous Articles Next Articles

Cloning and expression analysis of Fagopyrum tataricum FtDELLA gene

SUN Peiyuan(), RAN Bin, WANG Jiarui, LI Hongyou()

Research Center of Buckwheat Industry Technology, College of Life Sciences, Guizhou Normal University, Guiyang 550001, China

Received:2023-07-26 Online:2024-08-25 Published:2024-09-06
Contact: LI Hongyou

Abstract

Abstract:

DELLA protein is a negative regulatory protein in the gibberellin (GA) transduction pathway, which plays an important role in plant growth and development and stress response. In this experiment, a DELLA gene, named FtDELLA, was cloned from buckwheat using RT-PCR. The gene is 1 452 bp in CDS length and encodes 483 amino acids. Phylogenetic analysis showed that the FtDELLA protein contained the conserved DELLA conserved structural domains of the DELLA family. The qRT-PCR results showed that FtDELLA was highly expressed in stems and pre-grouting seeds, and its expression showed a decreasing trend during seed development. Under drought stress, the expression of FtDELLA showed a decreasing trend, indicating that it may be a negative regulatory gene for drought stress response in buckwheat. The results suggest that FtDELLA may play an important regulatory role in the growth and development of buckwheat and drought stress response. The results provide a theoretical basis for further research on the function and mechanism of action of FtDELLA in buckwheat.

Key words: Fagopyrum tataricum, FtDELLA, gene cloning, gibberellin, expression analysis

CLC Number:

S519

SUN Peiyuan, RAN Bin, WANG Jiarui, LI Hongyou. Cloning and expression analysis of Fagopyrum tataricum FtDELLA gene[J]. Acta Agriculturae Zhejiangensis, 2024, 36(8): 1709-1718.

Figures/Tables 10

Table 1 Primer sequences used in this study

引物名称 Primer name	引物序列 Primer sequence(5'-3')
FtDELLA-kF	ATGCCTGCAGGTCCGACGAT
FtDELLA-kR	TCAAGTCTCAGCCGAGTTTCG
FtDELLA-qF	TTCCATCACAGAGCTTCACC
FtDELLA-qR	GTTGTGGTTCGCCTCTTGTT
FtHLK-qF	TTCACGGGCACCATTCATGG
FtHLK-qR	AGGTGGAAGCTGAAGGAAGC
FtUPL7-qF	ATGTTGTGGGACGTGTGGTT
FtUPL7-qR	AAGCGATCCTCTGGTGATAG

Fig.1 PCR amplification result M, DL2 000 marker; 1, PCR amplification product.

Fig.2 Coding region and amino acid sequence of FtDELLA

Fig.3 Analysis of physical and chemical properties of FtDELLA A, Amino acid content distribution; B, FtDELLA phosphorylation modifications; C, FtDELLA amino acid pro- and hydrophobic predictions; D, FtDELLA signal peptide prediction map.

Fig.4 FtDELLA structure prediction A, Conserved structural domain; B, Prediction of secondary structure; the blue represents α-helix; the red represents β-sheet; the green represents β-turn; the purple represents random coil. C-a, Schematic representation of the N-terminal tertiary structure of the FtDELLA protein; C-b, Schematic representation of the C-terminal tertiary structure of the FtDELLA protein.

Fig.5 FtDELLA conserved motif analysis

Fig.6 Multiple sequence comparison between FtDELLA and DELLA proteins of other species

Fig.7 Phylogenetic tree of FtDELLA

Fig.8 FtDELLA expression in different tissue parts of buckwheat Seed 1, Seeds at 3 days after flowering; Seed 2, Seeds at 10 days after flowering; Seed 3, Seeds at 13 days after flowering.

Fig.9 Level of FtDELLA response under drought stress

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Cloning and expression analysis of Fagopyrum tataricum FtDELLA gene

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