Acta Agrestia Sinica

Acta Agrestia Sinica ›› 2026, Vol. 34 ›› Issue (10): 3875-3881.DOI: 10.11733/j.issn.1007-0435.2026.10.032

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Establishment of A Seed Induction Regeneration System for Agropyron mongolicum Keng and Study of Agrobacterium-mediated Transformation

FAN Bo-bo1, LIU Jie1, WU Jing2, YAN Xiu-xiu3, MA Yan-hong4   

  1. 1. College of Ecology and Environment of Baotou Teacher's College, Baotou, Inner Mongolia 014030, China;
    2. College of Life Sciences of Inner Mongolia Agricultural University, Hohhot, Inner Mongolia 010029, China;
    3. Urban Forestry and Grassland Bureau of Wulanchabu City, Ulanqab, Inner Mongolia 012000, China;
    4. Agricultural University college of Agriculture, Hohhot, Inner Mongolia 010019, China
  • Received:2026-01-13 Revised:2026-03-26 Published:2026-10-10

蒙古冰草种子诱导再生体系的建立及农杆菌介导遗传转化研究

范菠菠1, 刘杰1, 吴婧2, 闫秀秀3, 马艳红4   

  1. 1. 包头师范学院生态环境学院, 内蒙古 包头 014030;
    2. 内蒙古农业大学生命科学学院, 内蒙古 呼和浩特 010029;
    3. 乌兰察布市林业和草原局, 内蒙古 乌兰察布 012000;
    4. 内蒙古农业大学农学院, 内蒙古 呼和浩特 010019
  • 通讯作者: 马艳红, E-mail: mayanhong80@126.com
  • 作者简介:范菠菠（1995-），女，汉族，内蒙古呼和浩特人，博士，讲师，主要从事牧草遗传育种及耐逆性研究，E-mail：fanbobo19@126.com
  • 基金资助:
    包头师范学院高层次人才引进科研启动基金项目（BTTCRCQD2025-207）；内蒙古自治区自然科学基金项目（2025QN08116）资助

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Abstract: Agropyron mongolicum Keng is a perennial diploid forage within the Poaceae. Due to the lack of an efficient and stable regeneration system as well as a transformation system， studies on its gene functions have largely relied on model plants such as Arabidopsis， tobacco， or rice. Hence， establishing highly efficient and stable regeneration and genetic transformation systems for A. mongolicum is of significant importance. the induction of embryogenic callus represents a critical step in the development of a reliable regeneration system. The key to establishing a stable genetic transformation system lies in determining the optimal concentration of the infection solution and the appropriate infection duration. The optimum 2，4-D concentration and frequency of embryonic callus formation were studied using A.mongolicum seeds as induction materials. Embryogenic calluses were infected via Agrobacterium tumefaciens-mediated transformation， and the optimal transformation conditions were investigated using GUS staining.The results showed that when callus was induced from seeds of A.mongolicum， the callus induction rates were 38.33% and 58.33% at 2，4-D concentrations of 1.0 mg·L-1 and 3.0 mg·L-1， respectively， while at a 2，4-D concentration of 2.0 mg·L-1， the induction rate reached 81.66%， indicating that the optimal 2，4-D concentration for callus induction was 2.0 mg·L-1. Through two batches of callus induction from A. mongolicum seeds， the embryogenic callus rates were 52% and 50.5%， respectively. Agrobacterium infection of embryonic callus and GUS staining showed that the optimum bacterium concentration was OD600 =0.5， and the optimum infestation time was 10 min. the positive rate of GUS staining was 87.27%. This laid the foundation for the rapid， stable propagation and gene function studies of A. mongolicum.

Key words: Agropyron mongolicum Keng, Regeneration system, Genetic transformation, GUS staining

摘要： 蒙古冰草（Agropyron mongolicum Keng）是一种多年生二倍体禾本科（Poaceae）牧草。因缺乏高效、稳定的再生体系和遗传转化体系，研究其基因功能大多借助拟南芥、烟草或水稻完成，所以建立高效、稳定的蒙古冰草再生体系和遗传转化体系尤为重要。而胚性愈伤的诱导是再生体系建立的关键，稳定遗传转化体系建立的关键是对侵染液最佳浓度和侵染时间的确定。本研究以蒙古冰草种子作为诱导材料，研究了胚性愈伤诱导最适2，4-D浓度及胚性愈伤率；以农杆菌介导法侵染胚性愈伤组织，通过GUS染色研究蒙古冰草的遗传转化侵染条件，发现蒙古冰草种子诱导愈伤，2，4-D浓度为1.0 mg·L-1和3.0 mg·L-1时，出愈率分别为38.33%和58.33%，而2，4-D浓度为2.0 mg·L-1时，出愈率为81.66%，表明诱导愈伤最适2，4-D浓度为2.0 mg·L-1；通过2批次种子诱导愈伤的胚性愈伤率分别为52%和50.5%。农杆菌侵染胚性愈伤及GUS染色结果表明最适侵染液OD600值为0.5，最适侵染时间为5 min，GUS染色阳性率为87.27%。这为蒙古冰草快速、稳定繁殖及基因功能验证奠定重要基础。

关键词: 蒙古冰草, 再生体系, 遗传转化, GUS染色

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