Department of Experimental Biology, Masaryk University, Brno, Czech Republic.
Department of Plant Sciences, Oxford University, Oxford, UK.
BMC Plant Biol. 2021 Oct 9;21(1):461. doi: 10.1186/s12870-021-03241-w.
Inducible systems for transgene expression activated by a chemical inducer or an inducer of non-plant origin are desirable tools for both basic plant research and biotechnology. Although, the technology has been widely exploited in dicotyledonous model plants such as Arabidopsis, it has not been optimised for use with the monocotyledonous model species, namely rice. We have adapted the dexamethasone-inducible pOp6/LhGR system for rice and the results indicated that it is fast, sensitive and tightly regulated, with high levels of induction that remain stable over several generations. Most importantly, we have shown that the system does not cause negative growth defects in vitro or in soil grown plants. Interestingly in the process of testing, we found that another steroid, triamcinolone acetonide, is a more potent inducer in rice than dexamethasone. We present serious considerations for the construct design to avoid undesirable effects caused by the system in plants, leakiness and possible silencing, as well as simple steps to maximize translation efficiency of a gene of interest. Finally, we compare the performance of the pOp6/LhGR system with other chemically inducible systems tested in rice in terms of the properties of an ideal inducible system.
诱导型转基因表达系统可被化学诱导物或非植物来源的诱导物激活,这是基础植物研究和生物技术的理想工具。尽管该技术已在拟南芥等双子叶模式植物中得到广泛应用,但尚未针对单子叶模式物种(如水稻)进行优化。我们已将地塞米松诱导型 pOp6/LhGR 系统适配于水稻,结果表明,该系统快速、敏感且调控严格,诱导水平高,且在数代中保持稳定。最重要的是,我们表明该系统在体外或在土壤中生长的植物中不会引起负生长缺陷。有趣的是,在测试过程中,我们发现另一种甾体化合物,曲安奈德,在水稻中比地塞米松更具诱导能力。我们提出了对构建设计的认真考虑,以避免系统在植物中引起不良影响、漏泄和可能的沉默,以及最大化感兴趣基因翻译效率的简单步骤。最后,我们根据理想诱导系统的特性,将 pOp6/LhGR 系统与在水稻中测试的其他化学诱导系统进行了性能比较。