Accumulation of Saponarin by in Vitro Cultures of Gypsophila trichotoma in Respect of Mineral Additives

Background: For a long time, the researchers have been looking for new efficient methods to enhance production and obtain valuable plant secondary metabolites, which would contribute to the protection of the natural environment through the preservation of various plant species, often rare and endangered. These possibilities offer plant in vitro cultures which can be performed under strictly-controlled conditions, regardless of the season or climate and environmental factors. Biotechnological methods are promising strategies for obtaining the valuable plant secondary metabolites with various classes of chemical compounds including caffeoylquinic acids (CQAs) and their derivatives. CQAs have been found in many plant species which are components in the daily diet and exhibit a wide spectrum of biological activities, including antioxidant, immunomodulatory, antihypertensive, analgesic, anti-inflammatory, hepato- and neuroprotective, anti-hyperglycemic, anticancer, antiviral and antimicrobial activities. They have also been found to offer protection against Alzheimer’s disease, and play a role in weight reduction and lipid metabolism control, as well as modulating the activity of glucose-6-phosphatase involved in glucose metabolism. Methods: This work presents the review of the recent advances in use in vitro cultures of various plant species for the alternative system to the production of CQAs and their derivatives. Production of the secondary metabolites in in vitro culture is usually performed with cell suspension or organ cultures, such as shoots and adventitious or transformed roots. To achieve high production of valuable secondary metabolites in in vitro cultures, the optimization of the culture condition is necessary with respect to both biomass accumulation and metabolite content. The optimization of the culture conditions can be achieved by choosing the type of medium, growth regulators or growth conditions, selection of high-productivity lines or culture period, supplementation of the culture medium with precursors or elicitor treatments. Cultivation for large-scale in bioreactors and genetic engineering: Agrobacterium rhizogenes transformation and expression improvement of transcriptional factor or genes involved in the secondary metabolite production pathway are also efficient strategies for enhancement of the valuable secondary metabolites. Results: Many studies have been reported to obtain highly productive plant in vitro cultures with respect to CQAs. Among these valuable secondary metabolites, the most abundant compound accumulated in in vitro cultures was 5-CQA (chlorogenic acid). Highly productive cultures with respect to this phenolic acid were Leonurus sibiricus AtPAP1 transgenic roots, Lonicera macranthoides and Eucomia ulmoides cell suspension cultures which accumulated above 20 mg g-1 DW 5-CQA. It is known that di- and triCQAs are less common in plants than monoCQAs, but it was also possible to obtain them by biotechnological methods. Conclusion: The results indicate that the various in vitro cultures of different plant species can be a profitable approach for the production of CQAs. In particular, an efficient production of these valuable compounds is possible by Lonicera macranthoides and Eucomia ulmoides cell suspension cultures, Leonurus sibiricus transformed roots and AtPAP1 transgenic roots, Echinacea angustifolia adventitious shoots, Rhaponticum carthamoides transformed plants, Lavandula viridis shoots, Sausera involucrata cell suspension and Cichorium intybus transformed roots.

Abstract Unorganized callus and leaf/root-differentiating callus cultures of Pimpinella major have been established in liquid nutrient medium. Their capacity to accumulate rare phenylpropanoids such as epoxy-pseudoisoeugenol tiglate, epoxy-anol tiglate and anol tiglate was compared with that of seedlings and whole plants. The unorganized callus cultures were not able to accumulate any phenylpropanoids. In comparison, the leaf/root-differentiating callus culture promoted the accumulation of epoxy-pseudoisoeugenol tiglate (up to 90 mg/100 g fr.wt.) but not that of anol-derivatives. The accumulated amount of EPT in PMD-SH was comparable with that in plant seedlings.

Flavonoids represent a popular class of industrially important bioactive compounds. They possess valuable health-benefiting and disease preventing properties, and therefore they are an important component of the pharmaceutical, nutraceutical, cosmetical and medicinal industries. Moreover, flavonoids possess significant antiallergic, antihepatotoxic, anti-inflammatory, antioxidant, antitumor, antiviral, and antibacterial as well as cardio-protective activities. Due to these properties, there is a rise in global demand for flavonoids, forming a significant part of the world market. However, obtaining flavonoids directly from plants has some limitations, such as low quantity, poor extraction, over-exploitation, time consuming process and loss of flora. Henceforth, there is a shift towards the in vitro production of flavonoids using the plant tissue culture technique to achieve better yields in less time. In order to achieve the productivity of flavonoids at an industrially competitive level, elicitation is a useful tool. The elicitation of in vitro cultures induces stressful conditions to plants, activates the plant defense system and enhances the accumulation of secondary metabolites in higher quantities. In this regard, nanoparticles (NPs) have emerged as novel and effective elicitors for enhancing the in vitro production of industrially important flavonoids. Different classes of NPs, including metallic NPs (silver and copper), metallic oxide NPs (copper oxide, iron oxide, zinc oxide, silicon dioxide) and carbon nanotubes, are widely reported as nano-elicitors of flavonoids discussed herein. Lastly, the mechanisms of NPs as well as knowledge gaps in the area of the nano-elicitation of flavonoids have been highlighted in this review.