农业生物技术学报2026，Vol.34Issue(2)：254-270,17.DOI:10.3969/j.issn.1674-7968.2026.02.003
青枯菌胁迫下番茄叶片cDNA文库的构建及SlMYB86-like互作蛋白筛选鉴定
Construction of Tomato(Solanum lycopersicum)Leaf cDNA Library Under Ralstonia solanacearum Stress and Screening and Identification of SlMYB86-like Interacting Proteins
摘要
Abstract
The MYB transcription factor is one of the largest transcription factor families in plants and plays a significant role in the response of plants to both biotic and abiotic stresses.Previous research has indicated that the tomato(Solanum lycopersicum)SlMYB86-like(Solyc06g071690)transcription factor can be induced by the Ralstonia solanacearum and participates in the resistance response process of bacterial wilt.Nonetheless,the regulatory mechanism remains to be elucidaded.To screen the protein that interacted with the MYB transcription factor SlMYB86-like in tomato under the infection with R.solanacearum,the tomato inbred line BY 1-2(susceptible to bacterial wilt)was used as the material.Total RNA was extracted from tomato leaves at 0,3,6,9 and 24 h post-inoculation with R.solanacearum and subsequently mixed equally to create a tomato cDNA yeast two-hybrid library.The library capacity and recombination efficiency were calculated.Meanwhile,the constructed library was utilized to explore the interaction proteins during the infection process of R.solanacearum by using SlMYB86-like as the bait protein,and the potential proteins that might be involved in the defense response to pathogen were verified for interaction in yeast.The yeast library was constructed with a capacity of 1.9×107 CFU and showed complete recombination,corresponding to a 100%recombination rate.Furthermore,the average insert size was confirmed to be over 1 000 bp.Screening of the yeast two-hybrid system identified a total of 44 potential proteins that interacted with SlMYB86-like.Further verification through yeast rotation showed that SlMYB86-like interacted with the transcription factors vascular plant one-zinc finger 1(VOZ1),teosinte branched 1,cycloidea,and proliferating cell factors 15(TCP15),E3 ubiquitin protein ligase ring finger protein 14(RNF14),serine/threonine protein kinase D6 protein kinase(D6PK),ethylene-responsive transcription factor 1(ERF1),and calcium-dependent protein kinases(CDPK).In conclusion,these results offer significant experimental evidence for further investigation into the molecular mechanisms underlying SlMYB86-like's resistance to bacterial wilt in tomato.关键词
青枯菌/酵母双杂交/cDNA文库/SlMYB86-like/互作蛋白Key words
Ralstonia solanacearum/Yeast two-hybrid/cDNA library/SlMYB86-like/Interacting protein分类
农业科技引用本文复制引用
陈娜,温逸俊,邵勤..青枯菌胁迫下番茄叶片cDNA文库的构建及SlMYB86-like互作蛋白筛选鉴定[J].农业生物技术学报,2026,34(2):254-270,17.基金项目
国家自然科学基金(32260776) （32260776）
江西省自然科学基金(20232BAB215041) （20232BAB215041）
