Agrobacterium tumefaciens mediated transformation of the aquatic carnivorous plant Utricularia gibba

Abstract

Background: The genus Utricularia 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. Utricularia gibba 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. Results:Here, we report the establishment of a simple, fast and reproducible protocol for genetic transformation of U. gibba based on the T-DNA of Agrobacterium tumefaciens. An in vitro selection system using Phosphinotricin as a selective agent was established for U. gibba. 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 trapspecific ribonuclease gene in transgenic U. gibba plants. Conclusions: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 U. gibba as a model system to study developmental processes involved in trap formation.

Transformed tissue of Dionaea muscipula J. Ellis as a source of biologically active phenolic compounds with bactericidal properties

“…To the best of our knowledge, five articles about genetic transformation of carnivorous plants are available (Hirsikorpi et al 2002 ; Królicka et al 2010 ; Blehova et al 2015 ; Miguel et al 2019 ; Oropeza-Aburto et al 2020 ), while only one is about transformation with wild strains of R. rhizogenes bacteria (Królicka et al 2010 ). Hirsikorpi et al ( 2002 ) and Oropeza-Aburto et al ( 2020 ) used Rhizobium tumefaciens (former: Agrobacterium tumefaciens ) as a vector organism, while in the work of Blehova et al ( 2015 ), R. rhizogenes with green fluorescent protein gene served for Drosera rotundifolia L. transformation. Furthermore, Miguel et al ( 2019 ) explored virus-based plant transformation to create transgenic sundew and pitcher plants, for the overproduction of recombinant proteins.…”

“…To the best of our knowledge, five articles about genetic transformation of carnivorous plants are available (Hirsikorpi et al 2002;Królicka et al 2010;Blehova et al 2015;Miguel et al 2019;Oropeza-Aburto et al 2020), while only one is about transformation with wild strains of R. rhizogenes bacteria (Królicka et al 2010). Hirsikorpi et al (2002) and Oropeza-Aburto et al 2020 It was shown by Franklin et al (2008) on the Hypericum perforatum L. model that the most limiting factor in successful genetic transformation of plants using Rhizobium bacteria is plant's recalcitrance.…”

Genetic Basis of Carnivorous Leaf Development

“…The tremendous progress seen in Arabidopsis is, in part, attributed to the ease at which this plant can be genetically transformed, allowing various genes for leaf development to be functionally validated. Recent progress in this direction, such as the establishment of Agrobacterium -mediated transformation protocols in N. mirabilis ( Miguel et al, 2020 ) and U. gibba ( Oropeza-Aburto et al, 2020 ), offers much-needed hope for a complete genetic dissection of these complex leaf shapes.…”

Rapid movements in plants

“…Biological research in this field has long been hampered by the lack of powerful, efficient tools to analyze gene function in vivo. Recent advances in transgenic techniques (Lee et al 2019;Mano et al 2014;Oropeza-Aburto et al 2020;Suda et al 2020)…”