大豆GmGSTa基因启动子克隆及表达调控功能分析OA

GmGSTa is a protein that interacts with both the seed high-vigor annexin GmANN and the calcium-dependent protein kinase GmCDPK SK5.To investigate the stress response mechanism and regulatory function of the soybean GmGSTa promoter,DNA was extracted from soybean leaves using the CTAB method,and the GmGSTa promoter sequence was cloned via PCR.The obtained promoter sequence was analyzed for cis-acting elements,identifying core elements as well as stress-and hormone-responsive elements.A fusion vector,GmGSTaPRO::GUS,was constructed and introduced into tobacco leaves and Arabidopsis via Agrobacterium-mediated transformation.Subsequently,abiotic stress and ABA induction experiments were conducted to observe the tissue-specific and developmental stage-dependent expression patterns of this promoter in Arabidopsis.Gene expression driven by the promoter was analyzed qualitatively and quantitatively through GUS histochemical staining and GUS enzyme activity assays.The results showed that a 2 316 bp GmGSTa promoter sequence was isolated.Analysis revealed that it contains core elements such as TATA box and CAAT box,as well as various stress-and hormone-responsive elements including DRE1,MYB,and ABRE.The GmGSTa promoter was significantly induced in tobacco by 150 mmol·L-1 NaCl,250 mmol·L-1 mannitol,and high temperature and humidity(40℃,95%relative humidity),with GUS staining intensity increasing with the severity of the stress.In Arabidopsis,its activity exhibited tissue specificity and developmental stage dependence,with significant expression in roots,leaves,stamens,and mature seeds.Treatment with 50-150 mmol·L-1 NaCl,150-250 mmol·L-1 mannitol,and 8-10 μmol·L-1 ABA significantly enhanced promoter activity,with the highest increase in GUS enzyme activity observed under 250 mmol·L-1 mannitol induction.The findings indicate that the GmGSTa promoter is a functional regulatory element capable of responding to various abiotic stresses and ABA signaling,with its activity showing concentration dependence and tissue-and developmental-stage specificity.This promoter can serve as a potential target for inducible and efficient promoters,offering new regulatory element options for crop stress resistance genetic engineering and breeding.