Heenatigala P P M, Yang Jingjing, Bishopp Anthony, Sun Zuoliang, Li Gaojie, Kumar Sunjeet, Hu Shiqi, Wu Zhigang, Lin Wei, Yao Lunguang, Duan Pengfei, Hou Hongwei
The State Key Laboratory of Freshwater Ecology and Biotechnology, The Key Laboratory of Aquatic Biodiversity and Conservation of Chinese Academy of Sciences, Institute of Hydrobiology, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Wuhan, China.
Inland Aquatic Resources and Aquaculture Division, National Aquatic Resources Research and Development Agency, Colombo, Sri Lanka.
Front Chem. 2018 Jun 21;6:227. doi: 10.3389/fchem.2018.00227. eCollection 2018.
Members of the genus are fascinating plants for many biologists as they are the smallest flowering plants on Earth and exhibit a reduced body plan that is of great interest to developmental biologists. There has also been recent interest in the use of these species for bioenergy or biorefining. Molecular and developmental studies have been limited in species due to the high genome complexity and uncertainties regarding the stable genetic transformation. In this manuscript we present new protocols for both stable and transient genetic transformation for using . For the transient transformation, we used fronds whereas we used clusters for the stable transformation. As proof of concept we transformed two synthetic promoter constructs driving expression of the GUS marker gene, that have previously been used to monitor auxin and cytokinin output in a variety of species. Using these approaches we obtained a Transformation Efficiency (TE) of 0.14% for the stable transformation and 21.8% for the transient transformation. The efficiency of these two methods of transformation are sufficient to allow future studies to investigate gene function. This is the first report for successful stable transformation of .
该属的成员对许多生物学家来说是迷人的植物，因为它们是地球上最小的开花植物，展现出一种简化的身体结构，这对发育生物学家来说极具吸引力。最近人们也对将这些物种用于生物能源或生物精炼产生了兴趣。由于基因组高度复杂以及稳定遗传转化存在不确定性，该属物种的分子和发育研究受到限制。在本手稿中，我们展示了针对[该属物种名称未给出]的稳定和瞬时遗传转化的新方案。对于瞬时转化，我们使用[具体植物部分未给出]叶状体，而对于稳定转化我们使用丛生体。作为概念验证，我们转化了两个驱动GUS标记基因表达的合成启动子构建体，这两个构建体先前已用于监测多种物种中的生长素和细胞分裂素输出。使用这些方法，我们获得了稳定转化的转化效率（TE）为0.14%，瞬时转化的转化效率为21.8%。这两种转化方法的效率足以让未来的研究去探究基因功能。这是关于[该属物种名称未给出]成功稳定转化的首次报告。
