花生体胚诱导再生体系及基因枪转化条件的初步探讨

Peanut Somatic Embryogenesis and Genetic Transformation via Microprojectile Bombardment

以花生上胚轴为外植体,研究不同Picloram(毒莠定)浓度处理和不同基因型对体细胞胚胎发生及植株再生的影响,并利用基因枪将含GUS基因的pCAMBIA2301质粒载体轰击体胚,对基因枪转化条件进行了初步探索。结果表明:外植体在添加5 mg/L Picloram+1 mg/L Glutamine(谷氨酰胺)的MB培养基上诱导的体胚发生率、产胚数及植株再生率最高。不同基因型以‘中花8号’体胚再生率最高(体胚诱导率为55.36%,植株再生率为56.79%)。在氦气压力为1 100 psi,轰击距离为9 cm时,体细胞胚GUS瞬时表达率可达到22.95%。

Different picloram concentrations and genotypes were used to induce somatic embryo from mature seed epicotyls of peanut.Somatic embryos were transformed with the plant expression vector pCAMBIA2301 containing a β-glucuronidase（GUS） gene via microprojectile bombardment and the transformation conditions were studied.The results demonstrated that the optimal medium was MB medium adding 5 mg/L picloram and 1 mg/L glutamine,in which the ratio of somatic embryogenesis,the number of embryos and the ratio of plant regeneration were the highest;Also,among different genotypes,the best performance was with ＇Zhonghua No.8＇（the ratio of somatic embryogenesis was 55.36%,and the ratio of plant recovery was 56.79%）.Furthermore,the highest GUS transient expression（22.95 %） was obtained when particle bombardment was carried out under the 1 100 psi bombardment pressure and 9 cm distance of bombardment.