生物技术通报2024，Vol.40Issue(7)：207-215,9.DOI:10.13560/j.cnki.biotech.bull.1985.2024-0152
新麦草异戊烯基转移酶PjIPT基因的功能验证
Functional Verification of Isopentenyl Transferases PjIPT Gene in Psathyrostachys juncea
摘要
Abstract
[Objective]The isopentenyl transferase(IPT)enzyme plays a crucial role in the biosynthesis of trans-zeatin(t Z),a cytokinin that regulates plant growth,development,and resistance to stress.It significantly contributes to bud growth promotion and regulation of lateral branch development in plants.Exploring the function of IPT provides a theoretical basis for the subsequent improvement of new wheat grass yield traits.[Method]Through phylogeny and multiple sequence alignment,the features and homology of the coding amino acid sequence were explored.PjIPT-overexpressed Arabidopsis thaliana plants were obtained by pollen tube passage,and the transgenic plants were verified by reverse PCR(RT-PCR)and protein blot(Western blot,WB),their tissue expression patterns were analyzed by real-time quantitative reverse PCR(RT-qPCR),and trans-zeatin was determined by liquid mass combination system.[Result]The IPT proteins in Psathyrostachys juncea with Triticum dicoccoides and other wheat crops had high homology,and AtIPT9 is a homologous protein of PjIPT.PjIPT-overexpressed in A.thaliana plants was achieved through the pollen tube pathway method,demonstrating an upregulation in the number of branches.RT-PCR and Western blot analysis confirmed the normal expressions of PjIPT at both transcriptional and protein levels.Spatial-specific expression analysis using RT-qPCR indicated that PjIPT demonstrated specific expression patterns in the tillering nodes and leaves of Arabidopsis.Furthermore,quantitative analysis of tZ revealed that PjIPT regulated the number of branches in Arabidopsis by participating in the biosynthesis of tZ.[Conclusion]PjIPT is a key gene involved in upregulating plant branching.关键词
新麦草/异戊烯基转移酶/反式玉米素/分蘖/功能验证Key words
Psathyrostachys juncea/isopentenyl transferase/trans-zeatin/tillering/functional verification引用本文复制引用
任晓敏,云岚,艾芊,赵乔..新麦草异戊烯基转移酶PjIPT基因的功能验证[J].生物技术通报,2024,40(7):207-215,9.基金项目
国家自然科学基金项目(32371762),内蒙古自治区自然科学基金重点项目(2023ZD07),内蒙古自治区揭榜挂帅项目(2022JBGS00400303) （32371762）
