核农学报2026，Vol.40Issue(5)：884-896,13.DOI:10.11869/j.issn.1000-8551.2026.05.0884
小黑麦转录因子TwNAC01互作蛋白筛选及其抗旱性验证
Screening of Interacting Proteins of Transcription Factor TwNAC01 in Triticale and Its Function under Drought Stress
摘要
Abstract
The NAC transcription factor plays a crucial regulatory role in plant drought stress responses.Our research group previously cloned the full-length cDNA of the triticale transcription factor TwNAC01.This study is aimed to identify interacting proteins with TwNAC01 and investigate its molecular mechanisms under drought stress.At cotyledon stage,triticale seedlings were subjected to various treatments including high temperature stress(40℃),low temperature stress(4℃),Osmotic stress(20%PEG6000),Salinity stress(200 mmol·L-1 NaCl),and hormonal foliar spray treatment[100 μmmol·L-1 methyl jasmonate(MeJA)and 100 μmol·L-1 abscisic acid(ABA)].Total RNA of these seedlings was extracted,reverse-transcribed into double-stranded cDNA,and a normalized cDNA library was constructed.Yeast two-hybrid(Y2H)screening was performed to identify interacting proteins with TwNAC01 as bait.The drought resistance conferred by selected stress-related proteins was substantiated through bimolecular fluorescence complementation(BiFC)and yeast stress phenotype assays.A normalized triticale cDNA library was constructed,yielding a titer of 1.436×108 CFU·mL-1 and 2.872×108 total recombinant clones.A total of 33 potential TwNAC01-interacting proteins were identified,which are involved in various biological processes including photosynthesis,redox reactions,defense responses,and ubiquitination modification.BiFC analysis corroborated the interaction between TwNAC01 and two key proteins:ubiquitin-conjugating enzyme E2(UbcE2)and serine carboxypeptidase 1-like protein(SCPL1).Yeast strains expressing TwNAC01,UbcE2,or SCPL1 exhibit enhanced tolerance to high osmotic stress compared to controls.Additionally,TwNAC01,UbcE2,and SCPL1 were predominantly expressed in the roots,with high expression levels under drought stress.Both TwNAC01 and its interacting partners,SCPL1 and UbcE2,contribute to enhanced plant drought resistance,thereby providing a theoretical foundation for elucidating the molecular mechanisms of drought resistance in triticale.关键词
小黑麦/干旱胁迫/转录因子/酵母双杂/TwNAC01Key words
triticale/drought stress/transcription factor/yeast two-hybrid/TwNAC01引用本文复制引用
甘长波,卫小勇,宗金和,孔广超..小黑麦转录因子TwNAC01互作蛋白筛选及其抗旱性验证[J].核农学报,2026,40(5):884-896,13.基金项目
国家自然科学基金地区科学基金项目(32160478),兵团重点领域科技攻关项目(2024AB006),石河子大学自然科学研究与技术创新项目(校20200209) （32160478）
