1山西农业大学农学院,太谷 030800;2中国农业科学院作物科学研究所,北京 100081;3安徽荃银高科种业股份有限公司,合肥 230000
研究方向为食用豆类遗传育种,E-mail: 1587032644@qq.com;
国家食用豆产业技术体系(CARS-08);中国农业科学院科技创新工程
1College of Agriculture, Shanxi Agricultural University, Taigu 030800;2Institute of Crop Sciences, Chinese Academy of Agricultural Sciences, Beijing 100081;3Anhui Win-All Hi-Tech Seed Co., Ltd., Hefei 230000
China Agriculture Research System of MOF and MARA-Food Legumes (CARS-08);Science and Technology Innovation Project of Chinese Academy of Agricultural Sciences
建立并完善普通菜豆遗传转化体系,对促进普通菜豆基因资源的发掘与利用、分子育种发展等都具有重要意义。以高再生率的普通菜豆种质资源小白豆为材料,通过单一变量试验研究,明确了侵染、分化再生、抗性筛选3个关键阶段的最适遗传转化条件,为建立普通菜豆高效遗传转化体系提供参考。研究结果证实在制备外植体时切除子叶、顶芽和胚根,保留胚轴与子叶连接部位的方法效果较好,同时对外植体进行30 s超声处理能显著提升农杆菌的侵染与转化效率;在培养基中添加天冬酰胺与谷氨酰胺两种氨基酸,可提高遗传转化的成功率。同时,为进一步优化外植体再生状态,确定了再生培养基中添加激素或调节剂的最佳浓度,分别为6-苄氨基嘌呤(6-BA)8 mg/L、腺嘌呤60 mg/L、肌醇200 mg/L、尿囊素20 mg/L、蔗糖90 g/L,在最优浓度组合下,普通菜豆外植体的再生率、再生芽长度及生长状态均达到最佳。最后,明确了草铵膦的最佳浓度为5.0 mg/L,可区分阳性与阴性转化苗。研究结果为建立普通菜豆高效、稳定的遗传转化体系提供了技术指导和实践依据,也为其他食用豆类遗传转化体系的构建提供了参考。
The establishment and improvement of an efficient genetic transformation system for common bean is crucial for the exploration and utilization of its genetic resources, as well as for advancing the progress of molecular breeding. In the present study, the high-regeneration common bean germplasm small white common bean was selected as the experimental material. By conducting single-factor experiments, the optimal transformation conditions for three critical procedures, namely Agrobacterium infection, differentiation and regeneration, and resistance screening, were identified, which lays a foundation for developing a high-efficiency genetic transformation system for common bean.The results demonstrated that during explant preparation, removal of cotyledons, apical buds and radicles while preserving the connecting region between hypocotyls and cotyledons yielded the best effect. In addition, 30-second ultrasonic treatment of explants markedly enhanced the infection capacity and transformation efficiency of Agrobacterium tumefaciens. Supplementing the medium with asparagine and glutamine also greatly improved the genetic transformation success rate. To further improve the regeneration performance of explants, the optimal concentrations of plant growth regulators and additives in the regeneration medium were determined: 8 mg/L 6-benzylaminopurine (6-BA), 60 mg/L adenine, 200 mg/L inositol, 20 mg/L allantoin, and 90 g/L sucrose. Under this optimal combination, the explants exhibited the highest regeneration rate, the longest regenerated shoot length and the best overall growth status. Moreover, 5.0 mg/L glufosinate was confirmed as the optimal screening concentration to effectively separate positive transgenic seedlings from negative ones.This study provides reliable technical guidance and a practical theoretical basis for constructing an efficient and stable genetic transformation system for common bean, and also offers an important reference for the establishment of transformation systems in other edible legume species.
姚佳莹,程媛,常玉洁,等.普通菜豆遗传转化体系的优化[J].植物遗传资源学报,2026,27(8):1673-1683.
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