Establishment ofanAgrobacterium mediated transformation protocol forthedetection ofcytokinin intheheterophyllous plant Hygrophila difformis (Acanthaceae)
Establishment ofanAgrobacterium mediated transformation protocol forthedetection ofcytokinin intheheterophyllous plant Hygrophila difformis (Acanthaceae)
复制标题
农杆菌介导的转化方法用于检测异叶植物异形水螅（棘科）细胞分裂素的建立
DOI：
10.1007/s00299-020-02527-x
复制
发表时间：
2020
影响因子：
6.2
通讯作者：
Hongwei Hou
中科院分区：
文献类型：
--
作者：
Gaojie Li;Shiqi Hu;Jingjing Yang;Xuyao Zhao;Elizabeth Schultz;Seisuke Kimura;Hongwei Hou
植物在响应周围环境的变化时，在叶形上表现出惊人的形态差异，这是一种被称为异叶性的现象。以往的研究表明，水生植物异形水葫芦（Hydrophila difformis）是研究异叶性的理想模式。然而，遗传转化效率低、重复性差，限制了H。作为模式植物。本研究以根癌农杆菌LBA 4404为介导剂，成功地诱导了愈伤组织和芽，并建立了一套高效、稳定的遗传转化体系。结果表明：1 mg/L的1-萘乙酸（NAA）和2 mg/L的6-苄基腺嘌呤（6-BA）对愈伤组织的诱导效果最好; 0.1 mg/L的NAA和0.1 mg/L的6-BA对芽的诱导效果最好;鉴于植物激素在植物异株生长调节中的重要作用，以及目前对细胞分裂素（CK）在这一过程中的作用认识不足，本研究通过外源CK的施用和内源CK的检测，分析了CK在植物异株生长调节中的作用。利用我们新开发的转化系统检测CK信号，分析了H. difformis的研究结果表明，CK在环境介导的异叶性中起着重要作用。
Plants show amazing morphological differences in leaf form in response to changes in the surrounding environment, which is a phenomenon called heterophylly. Previous studies have shown that the aquatic plant Hygrophila difformis (Acanthaceae) is an ideal model for heterophylly study. However, low efficiency and poor reproducibility of genetic transformation restricted H. difformis as a model plant. In this study, we reported successful induction of callus, shoots and the establishment of an efficient stable transformation protocol as mediated by Agrobacterium tumefaciens LBA4404. We found that the highest callus induction efficiency was achieved with 1mg/L 1-Naphthaleneacetic acid (NAA) and 2mg/L 6-benzyladenine (6-BA), that efficient shoot induction required 0.1mg/L NAA and 0.1mg/L 6-BA and that high transformation efficiency required 100µM acetosyringone. Due to the importance of phytohormones in the regulation of heterophylly and the inadequate knowledge about the function of cytokinin (CK) in this process, we analyzed the function of CK in the regulation of heterophylly by exogenous CK applicationand endogenous CK detection. By using our newly developed transformation system to detect CK signals, contents and distribution in H. difformis, we revealed an important role of CK in environmental mediated heterophylly.