The first report on the induction of hairy roots in Trapa natans , a unique aquatic plant with photosynthesizing roots
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RESEARCH NOTE
The first report on the induction of hairy roots in Trapa natans, a unique aquatic plant with photosynthesizing roots Elena V. Mikhaylova1 · Alexander Artyukhin1 · Khalit Musin1 · Maria Panfilova1 · Gulnar Gumerova1 · Bulat Kuluev1 Received: 31 July 2020 / Revised: 26 October 2020 / Accepted: 27 October 2020 © Springer Nature B.V. 2020
Abstract Hairy root cultures are often used to produce valuable metabolites. They are grown on sucrose-rich medium, which is highly susceptible to contamination. Trapa natans is a unique plant with photosynthesizing roots. It is a promising object to obtain photoautotrophic hairy root culture. Protocols for transformation of this species are yet unknown. We report that hairy roots can be induced in aquarium and in vitro cultures of T. natans by agrobacterium-mediated and biolistic transformation. 64 roots were induced by Agrobacterium rhizogenes strain 15834, two roots were obtained using strain K599. Strain A4 was not effective. Biolistics with either amplicons of rol genes and 1301 pCAMBIA plasmid carrying rol genes resulted in the formation of six roots. All these roots contained chloroplasts. This achievement opens a prospect for genetic transformation of T. natans and use of its green photosynthesizing hairy root cultures in production of bioactive substances and in phytoremediation. Key message Protocols for the Agrobacterium-mediated and biolistic transformation of Trapa natans to induce hairy roots have been developed for the first time. All induced roots contained chloroplasts. Keywords Water caltrop · Agrobacterium rhizogenes · Biolistics · In vitro · Browning · Chloroplasts Agrobacterium rhizogenes (Rhizobium rhizogenes) are natural genetic engineers. They induce the formation of hairy roots, which are characterized by indefinite growth on hormone-free medium. Cultures of such tissues are often used to produce valuable metabolites, both natural (Danphitsanuparn et al. 2012; Kochan et al. 2019) and exogenous (Ekouna et al. 2017). Hairy roots are usually cultured as heterotrophic systems on sucrose-rich medium, which is highly susceptible to contamination. Mixotrophic or photoautotrophic tissues and reduction of sugar concentration could be a key to solve this problem (Kodym and Leeb 2019). The water caltrop Tapa natans L. (Lythraceae) is a unique aquatic plant which has photosynthesizing roots. T. natans has two types of roots: brown roots with typical structure Communicated by Amita Bhattacharya. * Elena V. Mikhaylova [email protected] 1
Institute of Biochemistry and Genetics UFRC RAS, Prospect Oktyabrya, 71, 450054 Ufa, Russia
which absorb nutrients, and green photosynthetic pinnately branched roots. Therefore, T. natans is a promising object for generation of photoautotrophic hairy roots. In vitro culture of the water caltrop is not easy. Only two research groups have accomplished it. Embryo and anther explants from Japanese (Hoque et al. 2009) and Indian (Agrawal and Ram 1995) populations of T. natans have been recovered in spite of phenolic accu
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