Department of Plant Production and Genetics, Faculty of Agriculture, University of Kurdistan, Sanandaj, Iran.
Research Center for Medicinal Plant Breeding and Development, University of Kurdistan, Sanandaj, Iran.
Sci Rep. 2022 Feb 2;12(1):1784. doi: 10.1038/s41598-022-05776-w.
Dragon's head plant (Lallemantia iberica), is a flowering species belongs to the mint family (Lamiaceae). The species contains valuable essential oils, mucilage and oil which are used in pharmaceutical and food industries. Tissue culture is a feasible strategy to attain large-scale production of plantlets with a huge potential to produce plants with superior quality. The objective of this study was to develop a simple and efficient method for regeneration and transformation of L. iberica. To reach this goal, the regeneration ability of various explants including leaf, cotyledonary node, hypocotyl and cotyledon segments was investigated in MS medium supplemented with diverse concentrations of NAA (Naphthalene acetic acid) and BAP (6-Benzyl Amino Purine). According to the results, cotyledonary nodes showed the best regeneration response. The maximum rate of regeneration (and number of induced shoots was achieved in 1 mg l BAP in combination with 0.05 mg l NAA from the cotyledonary nodes. Additionally, through the optimized regeneration technique Agrobacterium-mediated transformation of L. iberica was successfully accomplished. Gene transfer was assessed on leaf samples from regenerated plantlets under a fluorescent microscope to detect the GFP signals. Moreover, transgene integration and its expression were confirmed by PCR and RT-PCR analysis, respectively. The establishment of these efficient regeneration and genetic transformation methods paved the way for further application such as plant improvement, functional analysis and gene editing.
龙舌兰（Lallemantia iberica），是一种开花物种，属于薄荷科（Lamiaceae）。该物种含有有价值的精油、粘液和油，用于制药和食品工业。组织培养是一种可行的策略，可以实现大量植物的大规模生产，具有生产优质植物的巨大潜力。本研究的目的是开发一种简单有效的方法，用于再生和转化 L. iberica。为了实现这一目标，研究了叶片、子叶节、下胚轴和子叶切段等各种外植体在添加不同浓度 NAA（萘乙酸）和 BAP（6-苄基氨基嘌呤）的 MS 培养基中的再生能力。结果表明，子叶节具有最佳的再生响应。在 1mg·l BAP 与 0.05mg·l NAA 的组合下，子叶节的再生率（和诱导芽数）最高。此外，通过优化的再生技术，成功地实现了 L. iberica 的农杆菌介导转化。通过荧光显微镜检测再生植株叶片样本中的 GFP 信号，评估基因转移。此外，通过 PCR 和 RT-PCR 分析分别确认了转基因的整合和表达。这些高效的再生和遗传转化方法的建立为进一步的应用铺平了道路，如植物改良、功能分析和基因编辑。
