Department of Nitrogen Fixation, Institute of Plant Molecular Biology, Czechoslovak Academy of Sciences, Branišovská 31, 37005, České Budějovice, Czechoslovakia.
Plant Cell Rep. 1992 Jul;11(7):363-7. doi: 10.1007/BF00233367.
Kidney vetch seedlings were induced to form hairy roots by inoculating their mesocotyls with the wild-type strain 15834 of Agrobacterium rhizogenes or with the A. tumefaciens strain C58C1 containing a binary vector system (the pRiA4b as a helper and the vector pCB1346 bearing a pTiC58-derived isopentenyl transferase gene (ipt, cytokinin biosynthetic gene) under control of its native regulatory sequences). Transgenic lines of three distinct phenotypes were selected: (i) Typically, the pRi15834-transformed tissues were stabilized in vitro and maintained for long periods as aseptic, fast-growing, hormone-independent, plagiotropic hairy root cultures which never regenerated shoots and lost the ability to synthesize opines. Their genomic DNA contained both the TL- and the TR-DNA. (ii) One of the HR-lines transgenic for the T-DNA of pRi15834 (named 52AV34) started to regenerate spontaneously into teratomous shoots. The shoots were found to produce opines and both the TL and TR parts of T-DNA were found to be partly deleted and/or rearranged. They contained phytohormones in similar levels as those found in seed-born shoots. (iii) A practically identical morphogenic response as in the line 52AV34 was observed in the clone 27AV46. However, its shooty, dark-green, slow-growing teratomas were proven to be kanamycin-resistant, opine-producing, and double-transformed by the pRiA4b sequences and the ipt gene. They over-produced auxins as well as cytokinins (mainly indoleacetylaspartic acid and ribosides of zeatin and isopentenyladenine).
将野油菜黄单胞菌 15834 菌株或含二元载体系统的根癌农杆菌菌株 C58C1（其 pRiA4b 作为辅助因子，载体 pCB1346 携带 TiC58 衍生的异戊烯基转移酶基因（ipt，细胞分裂素生物合成基因），受其天然调控序列控制）接种菜豆中胚轴，诱导菜豆幼苗形成毛状根。选择了三种不同表型的转基因株系：（i）典型情况下，pRi15834 转化组织在体外稳定，并作为无菌、快速生长、激素独立、偏生毛状根培养物长期维持，这些培养物从不再生芽并丧失合成章鱼碱的能力。它们的基因组 DNA 同时含有 TL-DNA 和 TR-DNA。（ii）一株 pRi15834 的 T-DNA 转化的 HR 株系（命名为 52AV34）开始自发地再生为畸胎状芽。发现这些芽产生章鱼碱，并且 T-DNA 的 TL 和 TR 部分被部分缺失和/或重排。它们含有与种子产生的芽中相似水平的植物激素。（iii）在克隆 27AV46 中观察到与 52AV34 几乎相同的形态发生反应。然而，其丛生、深绿色、生长缓慢的畸胎瘤被证明是卡那霉素抗性的、产生章鱼碱的，并被 pRiA4b 序列和 ipt 基因双重转化。它们过度产生生长素和细胞分裂素（主要是吲哚乙酸和玉米素核苷以及异戊烯基腺嘌呤）。
