Cloning and Expression of AP2/ERF Transcription Factor Gene ShERF3 in Sugarcane and Subcellular Localization of Its Encoded Protein

doi:10.13560/j.cnki.biotech.bull.1985.2022-1400

Abstract

Abstract:

AP2/ERF transcription factors play an important role on regulating plant growth and development and involving in biotic and abiotic stresses resistances in plants. Cloning and gene function analysis of ShERF3 may provide gene resource for sugarcane resistance genetic improvement. The ShERF3 gene was cloned from‘ROC22’sugarcane plants based on the transcriptome data. Then the structure, subcellular location and gene expression pattern of ShERF3 were analyzed with real time PCR, bioinformation and rice protoplast subcellular location methods. The results showed that 1 142 bp cDNA sequence of ShERF3 was obtained. It contained a 1 053 bp integral opening reading frame encoding 350 amino acids. The predicted encoding protein of ShERF3 containing a conserved AP2 domain belonged to the ERF subgroup of AP2/ERF family. The homology analysis indicated that ShERF3 was highly homologous with ERF3 from Sorghum bicolor and Panicum virgatum and ERF118 from Panicum hallii and Setaria italica. It predicted that ShERF3 is a unstable hydrophobic protein. The subcellular location analysis showed that ShERF3 located in the nulear of rice protoplast cells. ShERF3 mainly expressed in sugarcane mature internodes and relatively lower expressed in the leaves and roots. In addition, the expression of ShERF3 was firstly down-regulated and then up-regulated under PEG treatment. And the expression of ShERF3 decreased with the extension of stress time under NaCl treatment. These results indicates that ShERF3 actively responds to draught and salt stresses. It may play a key role on regulating sugarcane stem development, draught and salt stresses responses.

Key words: sugarcane, ShERF3, abiotic stress, expression analysis

ZHAO Xue-ting, GAO Li-yan, WANG Jun-gang, SHEN Qing-qing, ZHANG Shu-zhen, LI Fu-sheng. Cloning and Expression of AP2/ERF Transcription Factor Gene ShERF3 in Sugarcane and Subcellular Localization of Its Encoded Protein[J]. Biotechnology Bulletin, 2023, 39(6): 208-216.

Figures/Tables 8

Fig. 1 PCR amplification electrophoresis of sugarcane ShERF3 cDNA M：DSMT marker 2000；1：ShERF3

Fig. 2 Conserved domains of protein ShERF3 in sugarcane

Fig. 3 Amino acid sequence alignment of ShERF3

Fig. 4 Phylogenetic tree of ShERF3 and other AP2/ERF family members

Fig. 5 Prediction of secondary structure of ShERF3 in sugarcane The blue is α-helix; red is extended- strand; green is beta turn; purple is random coil

Fig. 6 Subcellular localization results of ShERF3

Fig. 7 Expression analysis of ShERF3 gene in different parts of ‘ROC22’ IL: Immature leaves; ML: mature leaves; S1: stem node 1-2; S2: stem node 4-5; S3: stem node 7-8; S4: stem node 10-11; S5: stem node 12-13; S6: stem node 14-15; Rs: root system. Different lowercases indicate significant differences at the 0.05 level. The same below

Fig. 8 Expressions of SheRF3 gene under drought and salt stress

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