浙江农林大学学报2026，Vol.43Issue(3)：496-503,8.DOI:10.11833/j.issn.2095-0756.20250347
预培养对尾巨桉DH32-29遗传转化效率的影响
Effect of preculture on genetic transformation efficiency of Eucalyptus urophylla×E.grandis DH32-29
摘要
Abstract
[Objective]To obtain a stable and efficient genetic transformation rate,generate a large number of transgenic lines,and create improved varieties,the effects of different preculture treatment durations on genetic transformation of Eucalyptus urophylla×E.grandis DH32-29 were studied.[Method]This study aimed to fine-tune different preculture durations to determine the optimal timing for stable genetic transformation of E.urophylla×E.grandis DH32-29,5-ethynyl-2'-deoxyuridine(EdU)staining and flow cytometry were used to quantify the proportion of cells in the DNA synthesis(S)phase of the cell cycle,and RT-qPCR was used to m easure the expression levels of S phase-related genes EgrCDKD3,EgrCYCD3;3,EgrCYCD1;1,and EgrCYCH1;1.[Result]The duration of preculture of leaf explants significantly affected both the transient and stable transformation efficiency of E.urophylla×E.grandis DH32-29.After 3 days of preculture,the proportion of explant cells in the S phase was the highest,and the expression levels of key S-phase genes EgrCDKD3,EgrCYCD3;3,EgrCYCD1;1,and EgrCYCH1;1 peaked,indicating that explants precultured for 3 days are optimal for Agrobacterium tumefaciens infection.Further stable transformation assays demonstrated that a 3-day preculture substantially improved the efficiency of Agrobacterium-mediated genetic transformation in eucalyptus,increasing the transformation efficiency from 0.77%to 4.67%.[Conclusion]Preculture is crucial for the efficiency of Agrobacterium-mediated genetic transformation in eucalyptus.A 3-day preculture period can effectively enhance the genetic transformation efficiency of eucalyptus.关键词
尾巨桉/转化效率/流式细胞术/EdU染色/细胞周期/预培养Key words
Eucalyptus urophylla×E.grandis/transformation efficiency/flow cytometry/EdU staining/cell cycle/preculture分类
农业科技引用本文复制引用
王学君,张双,于茹恩,杨珺,曾炳山,范春节..预培养对尾巨桉DH32-29遗传转化效率的影响[J].浙江农林大学学报,2026,43(3):496-503,8.基金项目
国家重点研发计划项目(2021YFD2200102-05) （2021YFD2200102-05）
中国林业科学研究院基本科研业务费专项资金项目(CAFYBB2020ZB004) （CAFYBB2020ZB004）
