草地学报 ›› 2026, Vol. 34 ›› Issue (10): 3853-3861.DOI: 10.11733/j.issn.1007-0435.2026.10.030

• 技术研发 • 上一篇    下一篇

杂交狼尾草‘邦得1号’遗传转化体系的建立及应用

李甜1, 王婷婷2, 李薇1, 管聪2, 王慧1   

  1. 1. 中国农业大学/草业科学与技术学院, 北京 100193;
    2. 山东省农业科学院/休闲农业研究所, 山东 济南 250100
  • 收稿日期:2026-06-04 修回日期:2026-07-09 出版日期:2026-10-15 发布日期:2026-10-10
  • 通讯作者: 王慧, E-mail: huiwang211@cau.edu.cn
  • 作者简介:李甜(1998-),女,汉族,陕西宝鸡人,硕士研究生,主要从事草类植物分子育种研究,E-mail:ltbwcx9@163.com

Establishment and Application of the Genetic Transformation System for Hybrid Pennisetum‘Bangde No.1’

LI Tian1, WANG Ting-ting2, LI Wei1, GUAN Cong2, WANG Hui1   

  1. 1. College of Grassland Science and Technology, China Agricultural University, Beijing 100193, China;
    2. Institute of Leisure Agriculture, Shandong Academy of Agricultural Sciences, Jinan, Shandong Province 250100, China
  • Received:2026-06-04 Revised:2026-07-09 Online:2026-10-15 Published:2026-10-10

摘要: 杂交狼尾草(Pennisetum glaucum×Pennisetum purpureum)具有生物量大、生长快、适应性强等优点,是我国畜牧生产中重要的多年生牧草。然而,其稳定高效的遗传转化体系尚未建立,严重制约了其抗逆基因功能解析与定向分子育种进程。为建立稳定高效的杂交狼尾草遗传转化体系,本研究以 ‘邦得1号’为材料,系统筛选外植体及其消毒方式、愈伤组织诱导条件、农杆菌菌株、菌液浓度、乙酰丁香酮(Acetosyringone, AS)浓度及侵染方式。结果表明,种子经75%乙醇和7.5%次氯酸钠处理后可获得无菌外植体并诱导胚性愈伤组织;采用EHA105、菌液OD600为0.8、AS浓度150 μmol·L-1,并结合谷氨酰胺(Glutamine, Gln)预处理和负压侵染,转化效率达33%。利用该体系成功获得导入ZmRR1、PpICE1-1等耐寒基因的阳性转化株。综上,优化侵染条件、提高胚性愈伤组织质量和抗性愈伤分化率,是提升转化效率的关键。该体系为杂交狼尾草抗逆基因功能研究和抗寒新品种选育提供技术支撑。

关键词: 杂交狼尾草, 胚性愈伤组织, 遗传转化, 根癌农杆菌

Abstract: Hybrid Pennisetum (Pennisetum glaucum×Pennisetum purpureum) is an important perennial forage crop in China’s livestock production with advantages such as high biomass yield, rapid growth, and strong adaptability. However, the absence of a stable and efficient genetic transformation system severely restricts the functional verification of stress-resistant genes and molecular breeding. In this study, hybrid Pennisetum ‘Bangde No. 1’ was used as material. Through the systematic screening of explants and surface disinfection methods, callus induction, and the optimization of key genetic transformation parameters including Agrobacterium strain, bacterial concentration, acetosyringone (AS) dosage, and infection methods, an Agrobacterium-mediated genetic transformation system was established. The results showed that surface sterilization with 75% ethanol combined with 7.5% sodium hypochlorite was optimal for seed disinfection, The optimal transformation conditions consisted of Agrobacterium tumefaciens strain EHA105, a bacterial suspension at OD600=0.8, 150 μmol·L-1 AS, glutamine pretreatment, and vacuum-assisted infection, resulting in a transformation efficiency of 33%. Using this system, positive transformants carrying the cold-tolerance genes ZmRR1 and PpICE1-1 were successfully obtained. These results indicate that optimizing infection conditions and improving embryogenic callus quality and resistant callus differentiation are critical for enhancing transformation efficiency. The established system provides technical support for functional studies of stress-resistance genes and the breeding of cold-tolerant hybrid Pennisetum cultivars.

Key words: Hybrid Pennisetum, Embryogenic callus tissue, Genetic transformation, Agrobacterium tumefaciens

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