Agrobacterium rhizogenes-transformed roots of Astragalus membranaceus (Fisch. Ex Link) Bunge as a source of valuable secondary metabolites

Tatyana Novikova,Elena Ambros,Tatyana Zheleznichenko,Yungeree Oyunbileg,Yulianna Zaytseva,E.V Banaev,M.A Tomoshevich,Y.G Zaytseva

doi:10.1051/bioconf/20202400064

Tatyana Novikova, Elena Ambros + Show 6 more

Open Access

https://doi.org/10.1051/bioconf/20202400064

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Abstract

In this study the results of agrobacterial transformation of Astragalus membranaceus (Fisch. Ex Link) Bunge, a medicinal plant from Mongolia and China with a wide spectrum of pharmacological action were presented. As explants for transformation, in vitro derived microshoots as well as hypocotyls, cotyledons and primary shoots isolated from seedlings were used. The most effective from two tested Agrobacterium rhizogenes strains (A4-RT, 15834 SWISS) was 15834 SWISS, causing the transformation of various types of explants, including primary shoots as a more responsive type. The optimal co-cultivation time of explants with bacterial suspension (48 hours) and the highest transformation rate of primary shoots (36.4%) were established. PCR analysis confirmed the transgenic nature of the roots transformed with strain 15834 SWISS and the absence of bacterial contamination.

Highlights

In the past decades the growing demand for natural, renewable sources of medicinal substances has significantly increased interest in tissue culture techniques as an attractive alternative that allows enhancing the yield of target metabolites or inducing biosynthesis of new compounds without damaging the natural plant populations
The proven therapeutic activities of the secondary metabolites of A. membranaceus stimulate the creation of biotechnological platforms for their production in tissue culture systems
Results on the frequency of transformation of explants isolated from A. membranaceus seedlings and in vitro derived microshoots showed a variation in this parameter depending on the type of explant, A. rhizogenes strain used and the duration of co-cultivation (Fig. 1)

Summary

Introduction

In the past decades the growing demand for natural, renewable sources of medicinal substances has significantly increased interest in tissue culture techniques as an attractive alternative that allows enhancing the yield of target metabolites or inducing biosynthesis of new compounds without damaging the natural plant populations. The advantages of the biotechnological approach are obvious: high productivity of important plant pharmaceuticals, ease of extraction of target metabolites, guaranteed yield of target products regardless of seasonal and weather conditions, the absence of toxic substances (pesticides, herbicides and other pollutants) in crops, the ability to scale the process by the use of bioreactors [1, 2] Despite these advantages nowadays the commercial list of producer cultures is limited to dozens [3]. Research efforts are focused on obtaining drugs with antitumor and antiviral activities that increase the duration and quality of life (adaptogens, biostimulants, including immunostimulants) as well as cardiological and antimicrobial medicines [5, 7] According to these demands, we chose Astragalus membranaceus Despite the progress made, the developed protocols for the cultivation of A. membranaceous HR have not reached the high level of the target metabolites production (astralogosides, isoflavonoids) sufficient for their commercial application [12]

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