Crops ›› 2024, Vol. 40 ›› Issue (2): 23-29.doi: 10.16035/j.issn.1001-7283.2024.02.004

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Cloning, Bioinformatics and Expression Analysis of FtERF Gene in Fagopyrum tataricum

Zhang Jun1(), Cai Suyun1, Xu Zihao1, Hou Lei1, He Runli1(), Yin Guifang2, Wang Lihua2(), Wang Yanqing2, Lu Wenjie2, Sun Daowang2   

  1. 1College of Traditional Chinese Medicine and Food Engineering, Shanxi University of Traditional Chinese Medicine, Taiyuan 030619, Shanxi, China
    2Biotechnology and Genetic Germplasm Resources Research Institute, Yunnan Academy of Agricultural Sciences / Yunnan Provincial Key Laboratory of Agricultural Biotechnology / Key Laboratory of Southwestern Crop Gene Resources and Germplasm Innovation, Ministry of Agriculture and Rural Affairs, Kunming 650201, Yunnan, China
  • Received:2022-12-30 Revised:2023-04-26 Online:2024-04-15 Published:2024-04-15

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Abstract:

Ethylene response factor plays an important role in plant development and cell metabolism. ERF gene was cloned from Tartary buckwheat by RT-PCR, and its bioinformatics and differential expression were analyzed. The results showed that the length of open reading frame of FtERF gene was 729 bp, encoding 243 amino acids, molecular weight of 26.08 kD, isoelectric point of 9.22, belonging to the hydrophilic protein. FtERF didn't contain signal peptides and transmembrane helical structure, and was located in the nucleus of the cell. The secondary and tertiary structures of the proteins were highly similar. FtERF contained the inhibitory motif DLNxxP, and phylogenetic evolution suggested that FtERF and ERF4 were closely related. The results showed that the expression of Tartary buckwheat in different parts of thick-shell were higher than those in thin-shell Tartary buckwheat, and the expression of Tartary buckwheat in mature period was higher than that in immature period.

Key words: Tartary buckwheat, Ethylene response factor, Bioinformatics, Expression analysis

Table 1

Sequence information of primers used in the experiment"

引物名称
Primer name		 引物序列
Primer sequence		 用途
Usage
FtERF-F ATGGCACCCAAGCATAAGG 基因克隆
FtERF-R GGCGAGATCTGAAGGCGG
FtERF-F GGTTTCACGGCAGGGATT 实时荧光定量PCR
FtERF-R CACGATAGACGATGAGGCAGA
H3-F AAGAAGTCCCACAGATACCGC
H3-R AGCCTCCTGAAGAGCTAGCAC

Fig.1

PCR electrophoresis of FtERF gene M: SM0331 DNA Marker; 1: ERF gene fragment of Yunqiao 1; 2: ERF gene fragment of Xiaomiqiao."

Fig.2

Conservative domain analysis of FtERF"

Fig.3

Hydrophobicity analysis of FtERF"

Fig.4

Transmembrane domain analysis of FtERF"

Fig.5

Peptide signal prediction of FtERF"

Fig.6

Phosphorylation site prediction of FtERF"

Fig.7

Secondary structure prediction of FtERF protein"

Fig.8

Tertiary structure prediction of FtERF protein"

Fig.9

Multiple sequence alignment of FtERF protein The EAR domain suppression motif is in the blue box."

Fig.10

Phylogenetic tree of FtERF and ERF proteins of other species"

Fig.11

Relative expression levels of FtERF gene in different parts and different development periods of Tartary buckwheat “*”and“**”indicate significant difference at 0.05 and 0.01 levels, respectively."

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