海马齿再生体系的优化及GUS基因的转化  

Optimization of Plant Regeneration System and GUS Transient Transformation of Sesuvium portulacastrum L.

作　　者：申龙斌[1,2] 段瑞军[1] 郭建春[1] 符少萍[1] 

机构地区：[1]中国热带农业科学院热带生物技术研究所,海口571101 [2]海南大学农学院,海南儋州571737

出　　处：《热带作物学报》2009年第6期822-826,共5页Chinese Journal of Tropical Crops

基　　金：国家重点基础研究发展计划(973计划)项目(No.92007CB108903);中国热带农业科学院院基金项目(No.RKy0725);中央级公益性科研院所基本科研项目(No.ITBBYB072)资助

摘　　要：以海马齿无菌实生小苗的叶片为外植体,对不同激素浓度和组合以及不同pH值等培养条件进行实验比较。结果表明:适于愈伤组织诱导的培养基为MS+2,4-D2.0mg/L+6-BA0.5mg/L+蔗糖3%,诱导率达100%;适于芽分化的培养基为MS+NAA3.0mg/L+6-BA0.5mg/L,分化率达到77.4%;诱导愈伤组织形成培养基的最适pH值为4.5,而诱导愈伤组织分化培养基的最适pH值为5.0。以小苗叶片为外植体材料和农杆菌介导的方法进行GUS基因的转化,组织化学染色检测发现共培养2d后的外植体中有蓝色斑点产生,在转化2个月后的外植体上形成的愈伤组织也存在点状的蓝色斑点,说明GUS基因已转入海马齿外植体中。Sesuvium portulacastrum shows strong resistance to salt, drought and heavy metals. Rapid propagation and salt-resistance gene transformation are of significance. Leaves of S. portulacastrum were used as explants and cultured on MS media with different hormone combinations at various concentrations and pH value to induce calli and shoots. The results showed that the optimal media for callus and shoot induction were MS＋2, 4-D 2.0 mg/L＋6-BA 0.5 mg/L＋3% sucrose and MS＋NAA 3.0 mg/L＋6-BA 0.5 mg/L, respectively, on which all the explants were induced to form calli and 77.4% of calli differentiated into shoots. The optimal pH values for the callus and shoot culture were 4.5 and 5.0 respectively. GUS transient transformation of the calli with the leaves as the explants was conducted via agrobacterium mediation, and clear blue dots were identified in the transformed calli after 2 d of culture and 60 d of subculture through GUS histochemical staining. This indicates that GUS gene has been transferred into the calli of S. portulacastrum.

关 键 词：海马齿 离体再生 瞬时表达 GUS基因 

分 类 号：Q78[生物学—分子生物学] Q949.745.5