Biotechnology Bulletin ›› 2024, Vol. 40 ›› Issue (7): 207-215.doi: 10.13560/j.cnki.biotech.bull.1985.2024-0152

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Functional Verification of Isopentenyl Transferases PjIPT Gene in Psathyrostachys juncea

REN Xiao-min1(), YUN Lan1,2(), AI Qian1, ZHAO Qiao3()   

  1. 1. College of Grassland, Resources and Environment, Inner Mongolia Agricultural University, Hohhot 010018
    2. Key Laboratory of Grassland Resources of Education Ministry, Hohhot 010011
    3. Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055
  • Received:2024-02-11 Online:2024-07-26 Published:2024-05-24
  • Contact: YUN Lan, ZHAO Qiao E-mail:2937923090@qq.com;yunlan@imau.edu.cn;qiao.zhao@siat.ac.cn

Abstract:

【Objective】 The isopentenyl transferase(IPT)enzyme plays a crucial role in the biosynthesis of trans-zeatin(tZ), 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.

Key words: Psathyrostachys juncea, isopentenyl transferase, trans-zeatin, tillering, functional verification