Wounding by bombardment yields highly efficient Agrobacterium-mediated transformation of carnation (Dianthus caryophyllus L.)
Wounding by bombardment yields highly efficient Agrobacterium-mediated transformation of carnation (Dianthus caryophyllus L.)
复制标题
轰击损伤产生高效的农杆菌介导的康乃馨转化（Dianthus caryophyllus L.）
作者：
A. Zuker;A. Ahroni;T. Tzfira;H. Ben;A. Vainstein
农杆菌介导法高效转化香石竹是通过微弹轰击首先伤害茎外植体获得的。然后在黑暗中与解除武装的农杆菌共培养，转化频率-基于瞬时GUS表达-增加到用其他方法伤害的外植体和在恒光下共培养的10倍以上。为了获得稳定转化的康乃馨植株和消除嵌合体，在卡那霉素上进行了两轮再生/选择：首先，从接种的茎段外植体再生植株，然后用这些植株的叶片在第二轮不定芽再生的选择周期中产生转基因。农杆菌Aglo菌株携带含有uidA和nptII基因的双元载体pCGN7001进行稳定转化实验。在所分析的3个香石竹品种中，轰击伤伤、黑暗共培养和卡那霉素筛选两轮组合的总转化效率为每10个茎段有1-2个转基因。对T0代和T1代标记基因的组织化学和分子分析证实了所选植株的转基因性质。
Highly efficient Agrobacterium-mediated transformation of carnation (Dianthus caryophyllus L.) was obtained by first wounding stem explants via microprojectile bombardment. When this was followed by cocultivation with disarmed Agrobacterium in the dark, the transformation frequency-based on transient GUS expression-increased to over 10-fold that of explants wounded by other means and cocultivated under constant light. Two cycles of regeneration/selection on kanamycin were employed to generate stably transformed carnation plants and eliminate chimeras: first, plantlets were regenerated from inoculated stem explants and then leaves from these plantlets were used to generate transgenes in a second selection cycle of adventitious shoot regeneration. Agrobacterium strain AGLO, carrying the binary vector pCGN7001 containing uidA and nptII genes, was used in the stable transformation experiments. The combination of wounding via bombardment, cocultivation in the dark and two cycles of kanamycin selection yielded an overall transformation efficiency of 1–2 transgenes per 10 stem explants for the three carnation varieties analyzed. Histochemical and molecular analyses of marker genes in T0 and T1 generations confirmed the transgenic nature of the selected plants.
“一种将 DNA 限制性内切酶片段放射性标记至高比活性的技术”。
影响因子：
2.9
作者：
Feinberg,AP;Vogelstein,B
通讯作者：
Vogelstein,B