Liu Lingyun, Sun Mengying, Qi Xiaokun, Zhang Jun, Jiang Min, Cai Hongyu, Yang Minsheng, Wang Jinmao
Institute of Forest Biotechnology, Forestry College, Agricultural University of Hebei, Baoding, 071000, People's Republic of China.
Hebei Key Laboratory for Tree Genetic Resources and Forest Protection, Baoding, 071000, People's Republic of China.
BMC Plant Biol. 2025 Aug 19;25(1):1100. doi: 10.1186/s12870-025-07091-8.
has significant ecological and economic value and is widely used. This study developed an efficient regeneration system using sterile leaves as explants to directly induce adventitious buds. The effects of exogenous hormone concentration (6-Benzylaminopurine: 0, 1.0, and 2.0 mg/L), type, Thidiazuron pre-treatment duration, culture medium type, and leaf position (upper, middle, or lower) on leaf regeneration were investigated. Endogenous hormone content, physiological indices, microscopic observations, and transcriptome sequencing analyses were conducted.Results showed that the highest adventitious bud induction was achieved in Murashige and Skoog medium with 1.0 mg/L 6-BA, 0.05 mg/L indole-3-acetic acid, and 0.01 mg/L TDZ, reaching an 85% adventitious bud adventitious bud induction rate. Upper leaves pre-treated with TDZ for 6 days were most effective. Endogenous hormone content and microscopic properties under different treatments matched the regeneration system. Transcriptomic analyses indicated that differentially expressed genes were enriched in pathways related to phenylpropanoid biosynthesis, starch and sucrose metabolism, and plant hormone signal transduction. Core genes identified were , , and for 6-BA, TDZ, and leaf position treatments, respectively. Cellular status during pre-culture showed the highest Synthesis phase cell proportion on day 3, with Gap 2/Mitosis phase cells appearing from day 4. Key cell cycle S-phase gene expression peaked on day 3. β-Glucuronidase staining was highest on days 0, 2, and 3. Day 3 of pre-culture was identified as the optimal time for genetic transformation, with a transient transformation efficiency of up to 68%. This study provides a theoretical and technical basis for poplar regeneration and genetic transformation, offering insights for future forestry biotechnology applications.
The online version contains supplementary material available at 10.1186/s12870-025-07091-8.
具有重要的生态和经济价值且被广泛应用。本研究以无菌叶片为外植体建立了一种高效再生系统，直接诱导不定芽。研究了外源激素浓度（6-苄氨基嘌呤：0、1.0和2.0毫克/升）、类型、噻苯隆预处理时间、培养基类型以及叶片位置（上部、中部或下部）对叶片再生的影响。进行了内源激素含量、生理指标、微观观察和转录组测序分析。结果表明，在含有1.0毫克/升6-苄氨基嘌呤、0.05毫克/升吲哚-3-乙酸和0.01毫克/升噻苯隆的Murashige和Skoog培养基中不定芽诱导率最高，达到85%。用噻苯隆预处理6天的上部叶片效果最佳。不同处理下的内源激素含量和微观特性与再生系统相匹配。转录组分析表明，差异表达基因富集在与苯丙烷生物合成、淀粉和蔗糖代谢以及植物激素信号转导相关的途径中。分别鉴定出与6-苄氨基嘌呤、噻苯隆和叶片位置处理相关的核心基因、和。预培养期间的细胞状态显示，第3天合成期细胞比例最高，第4天出现G2/有丝分裂期细胞。关键细胞周期S期基因表达在第3天达到峰值。β-葡萄糖醛酸酶染色在第0、2和3天最高。预培养第3天被确定为遗传转化的最佳时间，瞬时转化效率高达68%。本研究为杨树再生和遗传转化提供了理论和技术基础，为未来林业生物技术应用提供了见解。
在线版本包含可在10.1186/s12870-025-07091-8获取的补充材料。
