Abstract
Salvia bulleyana is a rare Chinese medicinal plant that due to the presence of polyphenols lowers the risk of some chronic diseases especially those related to the cardiovascular system. The present study examines the organogenic competence of various combinations and concentrations of plant growth regulators to develop an efficient protocol for in vitro regeneration of S. bulleyana via leaf explants, maintaining the high production of active constituents. The purpose of the study was also to assess the possibilities of using a cytokinin-based regeneration to effectively produce therapeutic compounds. The adventitious shoot formation was observed through direct organogenesis on media with purine derivatives (meta-topolin, mT and benzylaminopurine, BAP), and through indirect organogenesis on media with urea derivatives (tidiazuron, TDZ and forchlorfenuron, CPPU). The highest regeneration frequency (95%) with 5.2 shoots per explant was obtained on leaves cultured on Murashige and Skoog (MS) medium containing 0.1 mg/L naphthalene-1-acetic acid (NAA) and 2 mg/L BAP. Following inter simple sequence repeat (ISSR) marker-based profiling, the obtained organogenic shoot lines revealed a similar banding pattern to the mother line, with total variability of 4.2–13.7%, indicating high level of genetic stability. The similar genetic profile of the studied lines translated into similar growth parameters. Moreover, HPLC analysis revealed no qualitative differences in the profile of bioactive metabolites; also, the total polyphenol content was similar for different lines, with the exception of the shoots obtained in the presence of CPPU that produced higher level of bioactive compounds. This is the first report of an effective and rapid in vitro organogenesis protocol for S. bulleyana, which can be efficiently employed for obtaining stable cultures rich in bioactive metabolites.
Highlights
IntroductionPlant tissue culture (PTC) is unquestionably a powerful tool that can be used to continuously reproduce commercially-important crops [1]
Previous reports on the in vitro propagation of Salvia species have usually focused on regeneration from apical buds of nodal explants
The present study aims to find an effective method of regeneration of S. bulleyana shoots using the organogenic potential of the plant
Summary
Plant tissue culture (PTC) is unquestionably a powerful tool that can be used to continuously reproduce commercially-important crops [1]. In vitro culture allows fast and efficient multiplication of disease-free, sometimes endangered, plant species. With the increasing reliance on plant-derived compounds and materials, the value of in vitro culture has grown exponentially. Crops are sources of a range of non-food goods, such as nutrients, fiber, dyes, pharmaceuticals, insecticides or cosmetics; as such, there is a need for more efficient strategies of boosting their production while conserving their natural pool [1]. Even when metabolic engineering is valuable and gives
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