Curtis I S, Nam H G
Department of Life Science, Pohang University of Science and Technology, Kyungbuk, Republic of Korea.
Transgenic Res. 2001 Aug;10(4):363-71. doi: 10.1023/a:1016600517293.
Transgenic radish (Raphanus sativus L. longipinnatus Bailey) plants were produced from the progeny of plants which were dipped into a suspension of Agrobacterium carrying both the beta-glucuronidase (gusA) gene and a gene for resistance to the herbicide Basta (bar) between T-DNA border sequences. The importance of development of the floral-dipped plant and presence of surfactant in the inoculation medium were evaluated in terms of transgenic plant production. Plants dipped at the primary bolt stage of growth, into a suspension of Agrobacterium containing 0.05% (v/v) Silwet L-77 resulted in optimum transformation efficiency, with 1.4% from 1110 seeds. The presence of Pluronic F-68 or Tween 20 in the inoculation medium was beneficial towards transgenic plant output compared to treatments without surfactant. Putative transformed T1 plants were efficiently selected by spraying with 0.03% (v/v) Basta and all herbicide-resistant plants tested positive for GUS activity when analysed both histochemically and fluorometrically. Southern analysis revealed that both the gusA and bar genes integrated into the genome of transformed plants and segregated as dominant Mendelian traits. These results demonstrate that radish can be genetically modified for the improvement of this important vegetable crop.
转基因萝卜(Raphanus sativus L. longipinnatus Bailey)植株是由浸蘸于携带β-葡萄糖醛酸酶(gusA)基因和抗除草剂Basta(bar)基因且位于T-DNA边界序列之间的农杆菌悬浮液中的植株后代培育而成。就转基因植株的培育而言,评估了花期浸蘸植株的发育情况以及接种培养基中表面活性剂的存在情况的重要性。在生长的初花阶段,将植株浸蘸于含有0.05%(v/v)Silwet L-77的农杆菌悬浮液中,转化效率最佳,1110粒种子中有1.4%获得转化。与不含表面活性剂的处理相比,接种培养基中存在普流罗尼克F-68或吐温20有利于转基因植株的产出。通过用0.03%(v/v)Basta喷雾有效地筛选出了假定转化的T1植株,并且当对所有抗除草剂植株进行组织化学和荧光测定分析时,它们的GUS活性均呈阳性。Southern分析表明,gusA和bar基因均整合到了转化植株的基因组中,并作为显性孟德尔性状分离。这些结果表明,萝卜可通过基因改造来改良这种重要的蔬菜作物。