Cloning and Expression Analysis of GmZAT12 Gene in Soybean

doi:10.7668/hbnxb.20192746

Abstract

Abstract:

Zinc finger proteins are transcription factors widely studied in eukaryotes,and play important roles in plant growth and development,and responses to stresses.In order to deeply understand the gene function of zinc finger protein in soybean,the full-length CDS sequence of GmZAT12 gene was cloned from Shangdou 1201,and the characteristics of coded protein by this gene was predicted by bioinformatics analysis.The subcellular localization of GmZAT12 protein was detected by the tobacco epidermal injection system, and the expression pattern of GmZAT12 gene in different tissues of soybean and abiotic stress was analyzed by Real-time PCR (qRT-PCR) technology. The results showed that GmZAT12 CDS contained 516 bp,encoding 171 amino acids and the molecular weight of the protein was 19.264 28 ku with a theoretical isoelectric point(pI)of 9.02.Its main components were random coils and α-helices and it contained 20 phosphorylation sites, mainly serine phosphorylation sites.Sequence analysis indicated that GmZAT12 possessed two conserved C2H2 zinc finger domains.The result of subcellular location indicated that GmZAT12 protein was localized in the nucleus.The results of qRT-PCR showed that CmZAT12 gene expressed mainly in roots,leaves and seeds of soybean,while low expression in flower and stem, and was induced by high temperature,low temperature,NaCl and ABA.The fact implied that this gene might be involved in abiotic stress signaling pathways.

Key words: Soybean, GmZAT12 gene, Cloning, Expression analysis, qRT-PCR

ZHANG Jinyu, XU Xinjuan, CHAO Maoni, ZHANG Xiaohong, WU Xiangyuan, GAO Jitao, HUANG Zhongwen. Cloning and Expression Analysis of GmZAT12 Gene in Soybean[J]. Acta Agriculturae Boreali-Sinica, 2022, 37(3): 1-7. doi: 10.7668/hbnxb.20192746.

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Figures/Tables 9

Fig.1 Detection of PCR product of GmZAT12 in soybean M.Marker DL2000;1.GmZAT12。

Fig.2 Prediction for phosphorylation sites of GmZAT12 protein

Fig.3 Secondary structure analysis of GmZAT12

Fig.4 Sequence alignment of GmZAT12 with other ZAT homologs from different plant species GmZAT12 homologous proteins used are listed as follows:GmZF1(Glyma.15G040700),GmZF2(Glyma.12G065800)and GmZAT8(Glyma.11G142500.1)from soybean(Glycine max);OsZAT12(XP_015638278.1)and OsZFP16(AAP74357.1)from rice(Oryza sativa);ZmZAT12(XP_008654280.1)and ZmZAT8(XP_008660288.1)form maize(Zea mays);AtZAT11(AT2G37430),AtZAT18(AT3G53600),AtZAT12(AT5G59820)and AtZF1(At2G28710)from Arabidopsis;HaZAT11(XP_021973789.1)from sunflower(Helianthus annuus);SlZAT11(XP_004239776.1)from tomato(Solanum lycopersicum),and MtZAT11(XP_003621801.1)from Medicago truncatula.The same as Fig.5.

Fig.5 Phylogenetic tree of GmZAT12 and other plant ZAT proteins

Fig.6 Tissue expression analysis of GmZAT12 Different lowercase letters indicate significant difference.The same as Fig.8-9.

Fig.7 Subcellular location of GmZAT12 1.Dark field;2.Bright field;3.Merge.

Fig.8 Express analysis of GmZAT12 under different temperature stress treatments

Fig.9 Expression analysis of GmZAT12 under NaCl and ABA treatments

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