商陆愈伤组织的筛选及其对TMV抗性的研究

Selection the Calli Lines of High PAP Production from Phylacca america and Research in the Resistence of PAP to TMV

商陆抗病毒蛋白(PAP)是一种广谱的植物抗病毒蛋白.本试验以商陆叶片为外植体,在附加0.5 mg/L 2,4-D(2,4-二氯苯氧乙酸)和0.5 mg/L 6-BA(6-苄氨基嘌呤)的MS培养基上诱导愈伤组织,通过继代筛选培养获得3个稳定的愈伤组织株系:黄色系(Y-型)、粉色系(P-型)、红色系(R-型),不同株系具有不同的生长特性.Northern杂交结果证明不同株系中PAP基因转录水平有明显差异.离体烟草叶片摩擦接种实验表明,不同株系的蛋白提取液对TMV均具有显著抗性,其中黄色株系具有较明显的优势.商陆离体愈伤组织的研究为生产安全、高效的植物源抗病毒制剂提供了一定基础.

PAP is a kind of broadspectrum antivirus protein. A callus induction system was developed for Phytolacca americana on MS medium supplement with 0.5 mg/L 2, 4-D and 0.5 mg/L 6-BA. Tirade stable types specialized in morphology, color, and growth characteristics were obtained after 2-years subculture and selection： type Y was yellow-green and compact, and grew relatively slow. Type P was white pink and watery. Type R was dark red and crisp, and grew fastest. The results Northern blot also revealed the different transcription level of PAP gene in different lines. Disease challenge test of detached tobacco leaves with TMV showed that the resistance was dramatically enhanced when TMV treated by protein of different calli lines, especially type Y. The result implicated that callus culture of Phylacca america in vitro had a potential application in the production of the safe, efficiency plant viral inhibitor.