福建农业学报2025，Vol.40Issue(11)：1110-1116,7.DOI:10.19303/j.issn.1008-0384.2025.11.002
木薯MeABF1基因的克隆及功能
Cloning and Function of MeABF1 in Cassava
摘要
Abstract
[Objective]The gene of transcription factor ABF1 from Manihot esculenta Crantz(MeABF1)was cloned,and its prokaryotic expression and disease resistance-related function analyzed.[Method]Coding sequence(CDS)of MeABF1 was amplified from the M.esculenta cultivar,'South China 124'('SC124').MeABF1-pET32a fusion expression vector was constructed and transformed into DE3 competent cells for protein induction.Expression of the gene was determined by SDS-PAGE and western blotting.Cassava leaves were sprayed with a solution of purified MeABF1 protein and tested for burst of reactive oxygen species(ROS),expression of pathogenesis-related gene(PR),and disease resistance to Xanthomonas axonopodis pv.manihotis(Xam).[Result]The CDS sequence of MeABF1 was 1 491 bp long.The MeABF1-Myc fusion protein with increasing band intensity over time was detected after 1 mmol·L-1 isopropyl β-D-thiogalactoside(IPTG)induction at 37℃for 1 h.The peak ROS burst in the treated leaves was 3-fold of control,and the MePR1 and MePR2 expressions upregulated 3.5-fold and 1.75-fold,respectively.A 2 d treatment significantly enhanced the disease resistance to Xam of the cassava leaves.The bacterial counts and lesion area on the leaves 6 d post inoculation(dpi)were significantly reduced with improved phenotypic outcomes over control.[Conclusion]MeABF1-pET32a was successfully expressed in cassava leaves with the induction by 1 mmol·L-1 IPTG at 37℃.The foliar MeABF1 application significantly enhanced the resistance of the leaves to Xam infection with burst of ROS and heightened PR expression.关键词
木薯/ABF1基因/蛋白/免疫反应Key words
Cassava/ABF1 gene/protein/immunity reaction分类
农业科技引用本文复制引用
孙志元,赵惠萍,韦运谢..木薯MeABF1基因的克隆及功能[J].福建农业学报,2025,40(11):1110-1116,7.基金项目
国家自然科学基金项目(32372563) （32372563）
海南省教育厅项目(Hnky2024-10) （Hnky2024-10）
海南大学协同创新中心科研项目(XTCX2022NYC13) （XTCX2022NYC13）
