Cloning, transcription activity and expression analysis of actindepolymerizing factor 4, ZmADF4 in maize(Zea mays L.)

【Objective】To provide reference for studying the function and regulation pattern of maize ADF family gene and provide potential gene resources for stress resistance in maize, the actin-depolymerizing factor 4(ZmADF4) gene was cloned and its transcription activity and expression pattern were analyzed.【Method】The ZmADF4 gene was cloned from maize leaves by homologous cloning technology. The physicochemical properties, conserved domains, secondary and tertiary structures of the encoded protein were predicted by bioinformatics analysis. Subcellular localization was carried out by transient transformation of protoplasts, and its transcriptional activity was analyzed by yeast self activation. The expressions of ZmADF4 gene in different tissues were analyzed by RNA-Seq data downloaded and its expressions under different stress treatments were detected by real-time fluorescence quantitative PCR.【Result】The results showed that the ZmADF4 gene was successfully cloned from maize leaf. The full length coding region(CDS) of ZmADF4 was 420 bp, which encoded a 139 amino acids residues. The molecular weight of the protein was 15.855 kD and its theoretical isoelectric point(pI) was 7.66. The protein encoded by ZmADF4 gene belonged to hydrophilic protein, which had conserved domain ADF_gelsolin of typical ADF protein family and mainly located in cytoplasm. ZmADF4 gene had no self activating activity by transcriptional activity analysis. Further studies showed that the gene was differentially expressed in maize roots, stems, leaves, silks, anthers, embryo, endosperm and pericarps, with expressed highly in stems, pericarps and leaves. Moreover, the expressions of ZmADF4 gene changed under high temperature, low temperature, high salt stress, drought stress and abscisic acid(ABA) stress. The expression of ZmADF4 gene was up-regulated under high temperature, while down-regulated under low temperature and ABA stress. Under drought and high salt stress, the expression was first upregulated and then down-regulated.【Conclusion】ZmADF4 gene, belonging to ADF gene family, not only participates in the growth and development regulation of maize stalks, peel and leaves, but also in the adverse response regulation of low temperature, high temperature, drought and ABA and salt stresses.