Identification and bioinformatics analysis of novel-miR16 target gene ZjTCP4 in Chinese jujube

doi:10.3969/j.issn.1004-1524.20230488

Acta Agriculturae Zhejiangensis ›› 2024, Vol. 36 ›› Issue (3): 534-543.DOI: 10.3969/j.issn.1004-1524.20230488

• Horticultural Science • Previous Articles Next Articles

Identification and bioinformatics analysis of novel-miR16 target gene ZjTCP4 in Chinese jujube

ZHANG Luhe¹^,²(), WANG Duofeng¹, ZHANG De¹, ZHANG Guangzhong¹, ZHAO Tong¹, LYU Binyan¹, ZHANG Yangjun¹, LI Yi²^,^*()

1. Gansu Academy of Forestry Sciences, Lanzhou 730020, China
2. College of Forestry, Gansu Agricultural University, Lanzhou 730070, China

Received:2023-04-12 Online:2024-03-25 Published:2024-04-09

Abstract

Abstract:

TCPs transcription factor regulates plant organogenesis and morphological configuration mainly through regulating cell proliferation and plant hormone pathways, so as to explore the sequence characteristics and expression characteristics of ZjTCP4 transcription factor of Chinese jujube, in order to provide reference for the mechanism study of TCP transcription factor response to drought stress, and lay a foundation for the subsequent molecular breeding of jujube tree. The novel-miR16 secondary structure has the typical hairpin structure of miRNA and is a member of the jujube miR319 family. Most of the degradation sites of target gene ZjTCP4 were located at the 10th and 11th nucleotides of the novel-miR16 binding sites. Bioinformatics analysis showed that the full-length cDNA sequence of ZjTCP4 gene was 1 386 bp, predicted to encode 452 amino acid polypeptide, isoelectric point (pI) of 6.51, molecular weight of 49 091.55 u. It should be an intramembranous or extracellular protein, a non-secretory protein, a hydrophilic protein, located in the nucleus. The amino acid sequence encoded by ZjTCP4 gene of Chinese jujube is most similar to AtTCP4 of Arabidopsis thaliana, and has high similarity to AtTCP17, AtTCP18 and AtTCP12 of Arabidopsis thaliana. Novel-miR16 was up-regulated in response to drought stress, while its target gene ZjTCP4 was down-regulated under drought stress. Novel-miR16 can regulate ZjTCP4 transcription factor in response to drought stress. Under drought stress, the up-regulated expression of novel-miR16 leads to down-regulated expression of target gene ZjTCP4 (CCG006181) to improve drought tolerance of Chinese jujube.

Key words: Chinese jujube, microRNA, TCP transcription factor, bioinformatics, subcellular localization

CLC Number:

S665.1

ZHANG Luhe, WANG Duofeng, ZHANG De, ZHANG Guangzhong, ZHAO Tong, LYU Binyan, ZHANG Yangjun, LI Yi. Identification and bioinformatics analysis of novel-miR16 target gene ZjTCP4 in Chinese jujube[J]. Acta Agriculturae Zhejiangensis, 2024, 36(3): 534-543.

Figures/Tables 8

Fig.1 The secondary hairpin structures of novel-miR16 in Chinese jujube Mature novel-miR16 sequences are in red.

Table 1 Predicted target genes of novel-miR16 in Chinese jujube

novel-miR16序列 Sequence of novel-miR16	靶基因 Target Acc	靶基因注释 Target gene annotation
UUGGACUGAAGGGAGCUCCC	CCG006181	transcription factor TCP4-like

Fig.2 Verification of novel-miR16 target mRNA in Chinese jujube

Fig.3 Result of electrophoresis of PCR products M, 2 000 bp ladder Marker; 1, PCR products.

Fig.4 Bioinformatics analysis of protein encoded by ZjTCP4 gene A, Tertiary structure prediction of protein encoded by ZjTCP4 gene;B, Transmembrane domain analysis of protein encoded by ZjTCP4 gene; C, Signal peptide prediction of protein encoded by ZjTCP4 gene;D, Post-translational modification prediction of protein encoded by ZjTCP4 gene; E, Hydrophobicity analysis of protein encoded by ZjTCP4 gene.

Fig.5 Phylogenetic tree of protein encoded by ZjTCP4

Fig.6 Subcellular localization of the ZjTCP4 protein From left to right, superimposed photos of GFP green fluorescent protein, CHI, bright field and three channels of CK/gene, respectively.

Fig.7 The expression level of novel-miR16 and ZjTCP4 under drought stress in Chinese jujube No identical lowercase letters between different time points of the same gene indicate significant differences (P<0.05).

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Identification and bioinformatics analysis of novel-miR16 target gene ZjTCP4 in Chinese jujube

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