Bioinformatics and Expression Pattern Analysis of Senescence-associated Gene TaSAG39 in Wheat

doi:10.7668/hbnxb.20192703

Abstract

Abstract:

Cysteine protease,a kind of hydrolase in the process of protein degradation,participates in the senescence and maturation of plants and plays an important role in the growth and development of plants.In this study,a senesence-specific cysteine protease(SAG39)was screened from the previously obtained transcriptome data of wheat at senescence stage.In order to study the role of TaSAG39 gene in wheat at senescence,the gene structure and expression pattern of TaSAG39 were analyzed by bioinformatics analysis and Real-time fluorescence quantitative(qRT-PCR)technology.The results showed that the amino acid sequence encoded by the cysteine protease gene SAG39 had the closest relationship with the ancestral species of wheat,such as Aegilops tauschii,Triticum dicoccoides and Triticum durum,and it included the unique active site Cys-His-Asn and EFNIN structures of the papain sub-family.The full-length genomes of TaSAG39-5A,TaSAG39-5B and TaSAG39-5D were 1 455,1 435,1 439 bp,respectively,and the length of the coding sequence was 1 041,1 050,1 038 bp,respectively,with each containing two exons and one intron.The TaSAG39s proteins consisted of 346,349,345 amino acids.With relative molecular weight of about 37 ku and isoelectric point of 5.53—5.67,these proteins were stable negatively charged hydrophilic proteins.Its main components were random coils and α-helix,and there were signal peptides in the N segment of the protein.It had the conserved Inhibittor_I29 and Petidase_C1 domains,and contained 35—37 phosphorylation sites,mainly serine and threonine phosphorylation sites.TaSAG39 protein may interact with cysteine protease inhibitors and 14-3-3 protein.The promoter contained cis-acting elements relative to abscisic acid,methyl jasmonate,auxin,light,drought,low temperature and stress response.The results of qRT-PCR confirmed that gene TaSAG39 was induced to express by natural aging,darkness,drought,methyl jasmonate,high temperature and auxin,among which natural aging was the most significant.

Key words: Wheat, TaSAG39, Cysteine protease, Bioinformatics analysis, Expression pattern

SUN Lili, JIN Xiujuan, ZHAO Kai, Md Ashraful Islam, LU Juan, WANG Shuguang, SUN Daizhen. Bioinformatics and Expression Pattern Analysis of Senescence-associated Gene TaSAG39 in Wheat[J]. Acta Agriculturae Boreali-Sinica, 2022, 37(2): 18-27. doi: 10.7668/hbnxb.20192703.

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Figures/Tables 14

Fig.1 Phylogenetic tree analysis of TaSAG39 protein

Fig.2 Comparative analysis of TaSAG39 amino acid sequences

^*.The disulfide bond site;The yellow box.The structure of EFNIN;The blue box.The cysteine protease activity(Cys-His-Asn); The red line.The conserved domain Inhibitor_I29 and Peptidase_C1.

Fig.3 Amino acid composition of TaSAG39

Tab.1 Physicochemical properties of TaSAG39 protein

项目Item	TaSAG39-5A	TaSAG39-5B	TaSAG39-5D
氨基酸数量/个 Number of amino acids	346	349	345
分子质量/ku Molecular weight	36.67	36.90	36.53
不稳定系数Instability index	35.99	38.07	34.59
脂肪指数Aliphatic index	72.63	71.43	71.13
平均亲水性Grand average of hydropathicity	-0.183	-0.122	-0.174
理论等电点Theoretical Pi	5.53	5.67	5.62
带正电残基(Arg + Lys) Positively charged residues	36	35	35
带负电残基(Asp + Glu) Negatively charged residues	42	41	40
分子式Formula	C₁₅₇₈H₂₄₇₃N₄₇₃O₅₀₃S₁₇	C₁₅₀₅H₂₄₈₈N₄₇₄O₄₉₉S₁₉	C₁₅₇₆H₂₄₅₇N₄₇₁O₄₉₉S₁₇

Fig.4 Prediction of secondary structure of TaSAG39 protein

Fig.5 Prediction of tertiary structure of TaSAG39 protein

Fig.6 Prediction of signal peptide of TaSAG39 protein

Fig.7 Prediction of phosphorylation site of TaSAG39 protein

Fig.8 Conserved motif analysis of TaSAG39 protein

Fig.9 Conserved domains of TaSAG39 protein

Fig.10 Analysis of TaSAG39 gene's structure

Fig.11 The analysis of TaSAG39 protein interaction networks

Fig.12 Cis-acting elements in the promoter region of TaSAG39

Fig.13 Expression pattern of TaSAG39 under different treatments
Different lowercase letters indicate significant difference in the expression of TaSAG39 gene in different samples at 0.05 level.

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