The purpose of the present study was to establish an efficient and stable callus genetic transformation system for Pyrus armeniacaefolia. Fruits 15 days after flowering were used for callus inducing. Major factors that influence growth state of callus (concentrations of plant growth regulators, subculture cycle) and transformation (Agrobacterium tumefaciens concentration, infection time, co-culture time, AS and kanamycin concentrations) were examined. The results showed that the optimum mediums for callus induction was MS supplemented with 3.0 mgL⁻¹ 2,4-Dichlorophenoxyacetic acid (2, 4-D) and 0.5 mgL⁻¹ 6-Benzylaminopurine (6-BA). The appropriate proliferation medium was MS containing 2.5 mgL⁻¹ 2,4-D and 0.5 mgL⁻¹ 6-BA with 21 d subculture cycle. Transformation was successfully achieved using the following protocol: callus were soaked in A. tumefaciens strain EHA105 (OD600 = 0.8) harboring pBI121 plasmid for 15 min. After 3 d co-culture on proliferation medium supplemented with 20 mgL⁻¹ AS, the callus were then screened on the selection medium (proliferation medium containing 25 mgL⁻¹ kanamycin and 200 mgL⁻¹ cefotaxime) to select the resistant callus. The GUS-positive rate was 96%. And the GUS-positive callus were further identification at DNA and transcription levels. We believe that the present study provides technical support for functional researches on fruit quality related genes.

本研究的目的是建立一种高效稳定的山梨愈伤组织遗传转化体系。开花后15天的果实用于诱导愈伤组织。检查了影响愈伤组织生长状态（植物生长调节剂浓度、继代培养周期）和转化（根癌农杆菌浓度、感染时间、共培养时间、AS 和卡那霉素浓度）的主要因素。结果表明，诱导愈伤组织的最佳培养基是添加有 3.0 mgL ^-1 2,4-二氯苯氧乙酸 (2, 4-D) 和 0.5 mgL ^-1 6-苄氨基嘌呤 (6-BA) 的MS 。合适的增殖培养基是含有 2.5 mgL ^-12,4-D 和 0.5 mgL ^-1 6-BA 具有 21 天的继代培养周期。使用以下方案成功实现了转化：将愈伤组织浸泡在含有 pBI121 质粒的根癌土壤杆菌菌株 EHA105 (OD600 = 0.8) 中 15 分钟。在补充有20 mgL ^-1 AS的增殖培养基上共培养3 d后，然后在选择培养基（含有25 mgL ^-1卡那霉素和200 mgL ^-1头孢噻肟的增殖培养基）上筛选愈伤组织以选择抗性愈伤组织。GUS 阳性率为 96%。GUS阳性愈伤组织在DNA和转录水平得到进一步鉴定。相信本研究为果实品质相关基因的功能研究提供了技术支持。