以野蔷薇(Rosa multiflora)当年生带芽茎段为试材, 建立了野蔷薇组培快繁体系。结果表明, 最佳外植体是带腋芽的茎段, 外植体最佳消毒方法是以75%的乙醇浸泡30秒, 再以10%的次氯酸钠溶液浸泡20分钟, 成活率可达96%左右; 带芽茎段萌芽最佳诱导培养基为MS+1.0 mg∙L–1 6-BA+0.01 mg∙L–1 NAA+0.1 mg∙L–1 GA3, 培养30天, 萌芽率可达98%左右; 无菌苗增殖最佳基础培养基为WPM, 增殖系数为2.87; 无菌苗生根最佳培养基为1/2MS+1.0 mg∙L–1 6-BA+0.1 mg∙L–1 NAA, 生根率可达93%左右; 无菌苗移栽成活率达到98%。在此基础上, 以野蔷薇无菌苗为受体, 建立了野蔷薇瞬时表达体系。结果表明, 瞬时表达最佳转化条件是菌液OD600为0.8, 负压为–0.10 MPa, 真空抽吸2次, 每次15分钟, 瞬时表达效率可达96%左右。研究结果为建立野蔷薇再生及遗传转化体系奠定了基础, 也为鉴定蔷薇属植物的基因功能提供了技术支持。
A tissue culture rapid propagation system of Rosa multiflora was established using the stem segments with buds of the current-year as the experimental material. The results showed that the best explants were stem segments with axillary buds. The best disinfection method was to soak the explants in 75% ethanol for 30 seconds, and then soak them in 10% sodium hypochlorite solution for 20 minutes. The survival rate can reach around 96%. The optimal germination induction medium was MS+1.0 mg∙L–1 6-BA+0.01 mg∙L–1 NAA+0.1 mg∙L–1 GA3. The germination rate can reach around 98% after 30 days of cultivation. WPM was the best basal medium for the proliferation of sterile seedlings, and the proliferation coefficient was 2.87. The best medium for rooting was 1/2MS+1.0 mg∙L–1 6-BA+0.1 mg∙L–1 NAA, and the rooting rate can reach around 93%. The transplanting survival rate of sterile seedlings was 98%. On this basis, the transient expression system of Rosa Multiflora was established. The results showed that the optimal transformation conditions for transient expression were OD600 of 0.8, vacuum negative pressure of –0.10 MPa and vacuum suction twice for 15min each time. The transient expression efficiency can reach around 96%. The results of this study laid a foundation for the establishment of regeneration and genetic transformation system of Rosa multiflora, and also provided technical support for identifying the gene function of Rosa plants.
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