串叶松香草高效再生体系的建立与兔出血症病毒YL株外壳蛋白基因VP60对其遗传转化

通过对串叶松香草(Silphium perfoliatumL.)不同激素浓度配比的诱导分化实验,建立了串叶松香草离体培养高效再生体系,结果表明MS 6-BA(2.0mg/L) NAA0.1(mg/L)培养基可高效诱导愈伤组织和芽的分化,1/2MS IBA(0.1mg/L)培养基可快速诱导根的生成,形成再生植株。构建了植物表达载体pBI121-VP60,利用根癌农杆菌(Agrobacterium tumefaciens)介导叶盘法转化串叶松香草以研究高效的串叶松香草转化体系,结果显示以农杆菌LBA4404为介导菌株、以叶片为转化外植体、3d预培养时间和3～4d共培养时间、400mg/L羧苄青霉素和40mg/L卡那霉素筛选浓度转化效果较好,并已筛选到两株拟转基因植株,为利用串叶松香草生产兔出血症病毒动物可食用疫苗建立了初步的技术基础。

Tissue Culture and High-efficient Plant Regeneration of Perfoliate Rosinweed and its Transformation with VP60 gene

The research had established high-efficient regeneration system of Perfoliate Rosinweed by different phytohormone combination experiments. The result showed that the medium of MS 6-BA(2.0mg/L) NAA(0.1mg/L)could efficiently induce the differentiation of callus and buds and the medium of 1/2MS IBA(0.1mg/L)could induce the regeneration of roots quickly. The vector pBI121-VP60 was constructed and then transformed to Perfoliate Rosinweed by agrobacterium-mediated method. The regenerated resistant plants could be obtained by pre-culturing for 3d,co-culturing with Agrobacterium tumefaciens LBA4404 for 3~4d,and then transfered to the callus inducement media containing 400mg/L Carb and 40mg/L Kan for 28d. Under this condition,the rate of plant transformation was relatively higher and we had gotten two putative transgenic plants. This system was an elementary technology for production of oral plant vaccine for rabbit hemorrhagic disease with Perfoliate Rosinweed.