碳离子辐照对菘蓝药性品质和分子水平的诱变效应

以野生菘蓝种子为材料,以不同剂量的碳离子(12 C6+)进行辐照处理(辐照剂量分别为30Gy、60Gy、90Gy、120Gy),分析12 C6+辐照对菘蓝种子萌发、幼苗生长、主要药用成分含量及其基因和蛋白质多态性变化的影响,为菘蓝品质育种、分子生物学研究和重离子辐照诱变的应用提供依据。结果显示:(1)12 C6+辐照处理后菘蓝的成苗率和根鲜重均随辐照剂量增加而逐步显著降低,其中30Gy处理对菘蓝生长抑制程度最小,但处理后菘蓝根中的主要药效成分4(3H)喹唑酮和靛玉红的含量增加幅度最大且最高,分别为野生型的2.2倍和2.3倍。(2)SRAP分子标记分析表明,菘蓝基因组的变异度随着辐照强度的增强而增大,其中30Gy处理的突变体与野生型相比有33.59%的多态性变异。(3)SDS-PAGE考马斯亮蓝染色和磷酸化染色分析表明,菘蓝的总蛋白和磷酸化蛋白表达水平均随辐照剂量变化出现了不同程度的改变,但并不与辐照强度呈正相关,说明植物在防御重离子辐照伤害时存在补偿机制。研究发现,30Gy的12 C6+辐照是菘蓝诱变的最佳剂量,能够显著提高其有效成分的含量,为优质菘蓝诱变育种奠定了基础。

Impact of Carbon Ion Irradiation on Potency and Molecular Level Variation of Isatis indigotica

In this study,the wide type seeds of Isatis indigotica were used as research materials,and carbon ions (irradiation doses:30Gy,60Gy,90Gy and 120Gy) were used to irradiate the seeds.The impact of different irradiation dosages on seed germination,seeding growth,main effective ingredients,genome and total protein polymorphism of I.indigotica were also comprehensively evaluated,which can provide a reference for I.indigotica breeding,molecular biology researching and the application of heavy ion irradiation mutagenesis.The results showed that:(1)the seedling surviving rate and root fresh weight of I.indigotica gradually decreased with enhanced irradiation intensity.Compared to other dosages,30Gy exhibited the minimum inhibition of growth and maximum increase of main effective ingredients 4(3H) quinoxalinol and indirubin in roots,by 2.2 and 2.3 fold compared to wide type,respectively.(2)SRAP analysis results indicated that the degree of genome variation was increased with enhanced irradiation intensity.Compared to wide type,mutants after using 30Gy had recovered 33.59% polymorphic variations for DNA.(3)SDS-PAGE gels stained by CBB and Pro-Q showed that total protein and phosphoprotein had been changed,but they were not positively correlated with the irradiation intensity,which indicates that plants may have a compensatory mechanism for the damage caused by heavy ion irradiation.After all,for I.indigotica mutagenesis,30Gy is the best carbon ion irradiation dosage which can induce a high content of active ingredients,and these results also laid a basis for mutation breeding of high quality I.indigotica.