Minlong C, Takayanagi K, Kamada H, Nishimura S, Handa T
Institute of Agriculture and Forestry, University of Tsukuba, Tsukuba, 305-8572, Ibaraki, Japan
Plant Sci. 2000 Nov 6;159(2):273-280. doi: 10.1016/s0168-9452(00)00351-4.
A total of 11 independent beta-glucuronidase (GUS) positive hairy roots were induced following co-cultivation of leaf explants of Antirrhinum majus L. with Agrobacterium tumefaciens strain GV2260 containing rol-type multi-auto-transformation (MAT) vector pNPI702. A total of 326 adventitious shoots were regenerated from the hairy root lines on 1/2 MS medium without plant growth regulators at 25 degrees C under a 16 h/day photoperiod condition 4 months after infection of the A. tumefaciens GV2260. The absence of the rol genes in five plants was verified by polymerase chain reaction (PCR) and Southern blot analysis. Acclimatized transformants exhibited normal phenotypes in height and in the morphology of leaves and flowers. Furthermore, the GUS gene was strongly expressed in the leaves, inflorescence of the transformed plant, and the progeny. This result demonstrates that the rol-type MAT vector can be used to study gene functions controlling the morphogenesis of Antirrhinum majus plants.
将金鱼草（Antirrhinum majus L.）的叶片外植体与携带rol型多自转化（MAT）载体pNPI702的根癌农杆菌（Agrobacterium tumefaciens）菌株GV2260共培养后，共诱导出11个独立的β-葡萄糖醛酸酶（GUS）阳性毛状根。在感染根癌农杆菌GV2260 4个月后，在25℃、16小时/天光照周期条件下，于不含植物生长调节剂的1/2 MS培养基上，从毛状根系再生出总共326个不定芽。通过聚合酶链反应（PCR）和Southern印迹分析验证了5株植物中rol基因的缺失。驯化后的转化体在株高以及叶片和花朵形态方面表现出正常的表型。此外，GUS基因在转化植株的叶片、花序及其后代中强烈表达。这一结果表明，rol型MAT载体可用于研究控制金鱼草植株形态发生的基因功能。
