摘要
为获得香石竹最佳转化体系及稳定表达的转基因植株,以香石竹组培苗叶片为材料,研究农杆菌菌种、香石竹基因型、预培养天数、侵染方式、共培养方式和时间等多个因素对香石竹遗传转化的影响。结果表明:香石竹转化的最佳方式是预培养2d,采用LBA4404农杆菌缓冲液摇床摇12min(200r/min),静置3min的侵染方式。侵染后叶片外植体接种于50μmol/L乙酰丁香酮(AS)溶液浸湿的无菌滤纸(3层),进行黑暗共培养5d。不同基因型对比结果表明:‘小桃红'的抗性芽分化率优于‘马斯特'和‘锦葵钛合金'。60mg/L卡那霉素(Kan)浓度对3个基因型的香石竹品种均有较好的抗性芽筛选效果,0.5mg/L吲哚丁酸(IBA)+0.22mg/L噻苯隆(TDZ)培养基是分化效率高、丛芽多、生长状态好的直接出芽培养基。本研究建立的的香石竹遗传转化体系,重复性好、转化效率高、基因型依赖较小,该体系能显著提高DcaPIP1;1基因的转化效率。
In order to optimize transformation system and obtain stable transgenic plants,the effects of Agrobacterium tumefaciens strains,carnation genotype, the number of pre-culture days,the way of infection, the mode and time of co- culture on the leaf genetic transformation of carnation were studied. The optimized carnation transformation system was as follows：Shaking with Agrobacterium tumefaciens buffer for 12 rain （200 r/min） and standing for 3 min after 2 d pre- culture;Then co-culture for 5 d on three-layer sterile filter paper soaked with 50 mol/L AS solution, The results of different genotype comparion showed that the differentiation rate of resistant buds of ＇Mallow Titanium Alloy＇ was higher than ＇ MasteP and ＇ Pink Shirt＇ . Treatment of 60 mg/L Kan had good select effect for all carnation genotypes. Medium of 0.5 mg/L IBA ＋ 0.22 mg/L TDZ was direct germination medium with high differentiation efficiency, high budding rate and good growth status. Optimum genetic transformation system of carnation was established with high reproducibility, high transformation efficiency and Low genotype dependence in this study. DcaPIPT; 7 gene transformation efficiency was improved by using this system.
出处
《中国农业大学学报》
CAS
CSCD
北大核心
2018年第4期34-44,共11页
Journal of China Agricultural University
基金
国家自然科学基金项目(31000918)
中央高校基本科研业务费专项(2662015PY052
2662016PY041)
关键词
香石竹
农杆菌介导
遗传转化体系
高效
carnation
agrobacterium-mediated
genetic transformation system
high efficiency
