Cui M, Takayanagi K, Kamada H, Nishimura S, Handa T
Institute of Agriculture and Forestry, University of Tsukuba, Tsukuba, Ibaraki 305-8572, Japan e-mail:
Institute of Biological Sciences, University of Tsukuba, Tsukuba, Ibaraki 305-8572, Japan, , , , , , JP.
Plant Cell Rep. 2001 Jan;20(1):55-59. doi: 10.1007/s002990000277.
Eleven independent GUS-positive hairy roots were induced by co-cultivation of leaf explants of Antirrhinum majus L. with Agrobacterium tumefaciens strain GV2260 containing the rol type MAT vector pNPI702. The MAT vector pNPI702 possesses a GUS gene under the 35 S promoter and a removal element in which the 7.6-kb DNA fragments containing the rolA, B, C and D genes and recombinase gene with a 35 S promoter are located between two directly oriented recombination site sequences. A total of 326 adventitious shoots regenerated from 11 independent hairy root lines cultured on 1/2MS medium without plant growth regulators at 25 °C under a 16/8 h (day/night) photoperiod after 8 weeks of stock-culture of hairy roots and 4 weeks of culture of the green segments of hairy roots. Regenerated plants showed either a normal or dwarf morphology. GUS activity was observed in the hairy roots and regenerated shoots. The presence of the GUS gene in the regenerated, morphologically normal plants was confirmed by PCR analysis.
通过将金鱼草（Antirrhinum majus L.）的叶片外植体与携带rol型MAT载体pNPI702的根癌农杆菌（Agrobacterium tumefaciens）菌株GV2260共培养，诱导出了11条独立的GUS阳性毛状根。MAT载体pNPI702在35S启动子下拥有一个GUS基因和一个去除元件，其中含有rolA、B、C和D基因以及带有35S启动子的重组酶基因的7.6 kb DNA片段位于两个同向的重组位点序列之间。在毛状根继代培养8周和毛状根绿色切段培养4周后，将11条独立的毛状根系在不含植物生长调节剂的1/2MS培养基上于25°C、16/8小时（白天/黑夜）光周期下培养，共再生出326个不定芽。再生植株表现出正常或矮小的形态。在毛状根和再生芽中观察到了GUS活性。通过PCR分析证实了形态正常的再生植株中存在GUS基因。
