TRANSFORMATION OF Lotus corniculatus L.MEDIATED BY Agrobacterium tumefaciens

A simple and effective system for the transformation and regeneration of a leguminous plant has been developed. The cotyledon explants of Lotus corniculatus (var. Leo) were infected by Agrobacterium tumefaciens that contained a non-oncogenic Ti plasmid vector. The vector carried a chimaeric npt-Ⅱ gene and a nopaline synthase gene (nos). On the selective medium that contained kanamycin, 40% of the explants formed buds within 3 weeks. Developed shoots were cut off and transferred to rooting medium. Normal-looking plants were recovered and grew well after being transplanted into soil, bloomed and set seeds. Foreign genes were integrated onto the L. corniculatus genome, expressed, and inherited through sexual reproduction, which was proven by nopaline detection, NPT-Ⅱ enzyme activity detection and DNA hybridization test of the transformed plants and the nopaline detection of the progeny.     

根癌农杆菌(Agrobacterium tumefaciens)介导的百脉根(Lotus corniculatus L.)的转化

本文报道一个简单有效的豆科植物转化再生实验系统。百脉根(品种:里澳)子叶外植体,被含非致瘤性Ti质粒载体的根癌农杆菌感染。该载体带有一个嵌合的npf-Ⅱ基因和一个胭脂碱合成酶基因。在合卡那霉素的选择培养基上,有40％的外植体在3周内出芽。长大的芽可在生根培养基上生根并移栽成活,开花结实。从一个切块得到了16个抗性再生植株。对再生植株的胭脂碱检测、NPT-Ⅱ活性检测及DNA分子杂交实验及种子后代的胭脂碱检测结果表明,外源基因整合到百脉根基因组上,获得表达并能通过有性生殖过程传递。