Oropeza-Aburto A, Cervantes-Pérez S A, Albert V A, Herrera-Estrella L
1Laboratorio Nacional de Genómica para la Biodiversidad, Centro de Investigación y de Estudios Avanzados del Instituto Politécnico Nacional, 36824 Irapuato, Guanajuato Mexico.
2Department of Biological Sciences, University at Buffalo, Buffalo, NY 14260 USA.
Plant Methods. 2020 Apr 10;16:50. doi: 10.1186/s13007-020-00592-7. eCollection 2020.
The genus belongs to Lentibulariaceae, the largest family of carnivorous plants, which includes terrestrial, epiphytic and aquatic species. The development of specialized structures that evolved for carnivory is a feature of this genus that has been of great interest to biologists since Darwin's early studies. is itself an aquatic plant with sophisticated bladder traps having one of the most complex suction mechanisms for trapping prey. However, the molecular characterization of the mechanisms that regulate trap development and the biophysical processes involved in prey trapping are still largely unknown due to the lack of a simple and reproducible gene transfer system.
Here, we report the establishment of a simple, fast and reproducible protocol for genetic transformation of based on the T-DNA of . An in vitro selection system using Phosphinotricin as a selective agent was established for . Plant transformation was confirmed by histochemical GUS assays and PCR and qRT-PCR analyses. We report on the expression pattern of the 35S promoter and of the promoter of a trap-specific ribonuclease gene in transgenic plants.
The genetic transformation protocol reported here is an effective method for studying developmental biology and functional genomics of this genus of carnivorous plants and advances the utility of as a model system to study developmental processes involved in trap formation.
该属植物属于狸藻科，狸藻科是食虫植物中最大的科，包含陆生、附生和水生种类。自达尔文早期研究以来，为食肉习性而进化出的特化结构的发育一直是该属植物的一个特征，备受生物学家关注。该植物本身是一种水生植物，具有复杂的囊状捕虫器，拥有捕获猎物的最复杂的抽吸机制之一。然而，由于缺乏简单且可重复的基因转移系统，调节捕虫器发育的机制以及猎物捕获所涉及的生物物理过程的分子特征仍基本未知。
在此，我们报告了基于发根农杆菌的T-DNA建立一种简单、快速且可重复的该植物遗传转化方案。建立了一种以草铵膦作为选择剂的体外选择系统用于该植物。通过组织化学GUS分析、PCR和qRT-PCR分析证实了植物转化。我们报告了转基因该植物中35S启动子和捕虫器特异性核糖核酸酶基因启动子的表达模式。
本文报道的遗传转化方案是研究这种食虫植物属发育生物学和功能基因组学的有效方法，并提升了该植物作为研究捕虫器形成所涉及发育过程的模型系统的效用。
