Schroeder H. E., Schotz A. H., Wardley-Richardson T., Spencer D., Higgins TJV.
Division of Plant Industry, Commonwealth Scientific and Industrial Research Organization, GPO Box 1600, Canberra, ACT 2601, Australia.
Plant Physiol. 1993 Mar;101(3):751-757. doi: 10.1104/pp.101.3.751.
A reproducible transformation system was developed for pea (Pisum sativum L.) using as explants sections from the embryonic axis of immature seeds. A construct containing two chimeric genes, nopaline synthase-phosphinothricin acetyl transferase (bar) and cauliflower mosaic virus 35S-neomycin phosphotransferase (nptII), was introduced into two pea cultivars using Agrobacterium tumefaciens-mediated transformation procedures. Regeneration was via organogenesis, and transformed plants were selected on medium containing 15 mg/L of phosphinothricin. Transgenic peas were raised in the glasshouse to produce flowers and viable seeds. The bar and nptII genes were expressed in both the primary transgenic pea plants and in the next generation progeny, in which they showed a typical 3:1 Mendelian inheritance pattern. Transformation of regenerated plants was confirmed by assays for neomycin phosphotransferase and phosphinothricin acetyl transferase activity and by northern blot analyses. Transformed plants were resistant to the herbicide Basta when sprayed at rates used in field practice.
利用未成熟种子胚轴的切段作为外植体,为豌豆(Pisum sativum L.)开发了一种可重复的转化系统。使用根癌农杆菌介导的转化程序,将含有两个嵌合基因——胭脂碱合酶-膦丝菌素乙酰转移酶(bar)和花椰菜花叶病毒35S-新霉素磷酸转移酶(nptII)的构建体导入两个豌豆品种。通过器官发生进行再生,并在含有15 mg/L膦丝菌素的培养基上筛选转化植株。转基因豌豆在温室中培育以产生花朵和有活力的种子。bar和nptII基因在第一代转基因豌豆植株及其下一代后代中均有表达,在后代中呈现典型的3:1孟德尔遗传模式。通过新霉素磷酸转移酶和膦丝菌素乙酰转移酶活性测定以及Northern印迹分析,证实了再生植株的转化。当按照田间实际使用的剂量喷洒时,转化植株对除草剂Basta具有抗性。