Iridoid and phenylethanoid glycoside production and phenotypical changes in plants regenerated from hairy roots of Rehmannia glutinosa Libosch.

Abstract

Spontaneous shoot regeneration was obtained from the hairy roots of Rehmannia glutinosa cultured in liquid woody plant medium for 4 weeks in darkness after induction by Agrobacterium rhizogenes strain A4. These shoots were proliferated on solid Murashige and Skoog (MS) medium in 300 mL glass jars and in liquid MS medium in a 5 L nutrient sprinkle bioreactor. These media were supplemented with 0.1 mg L -1 of indole-3-acetic acid (IAA) and 1.0 mg L -1 of 6-benzylaminopurine. All shoots became rooted on MS agar medium containing IAA or indole-3-butyric acid within 4 weeks. The plantlets were then transferred into soil and grew in a greenhouse. The transgenic nature of the plants was confirmed by polymerase chain reaction (PCR) and reverse-transcriptase PCR analyses. The pRi-regenerated transformants demonstrated a few phenotypical alterations. Production of six iridoid (catalpol, aucubin, loganin, catalposide, harpagide, harpagoside) and two phenylethanoid (verbascoside and isoverbascoside) glycosides was determined in shoots cultured in vitro and leaves and roots of 6-and 12-month-old plants by ultra high performance liquid chromatography. The shoot culture was able to synthesize low amounts of catalpol, verbascoside and isoverbascoside. Leaves and roots of pRi-regenerants accumulated similar (harpagoside, isoverbascoside), higher (catalposide, aucubin, harpagide, verbascoside) or lower (catalpol, loganin) levels of the metabolites in comparison with untransformed R. glutinosa plants and their content varied with the age of the plants and the organ analyzed. However, the increase in shoot and root biomass of transformed plants resulted in a significantly greater yield of all analyzed compounds.

Terpenoid and flavonoid spectrum of in vitro cultures of Glycyrrhiza glabra revealed high chemical heterogeneity: platform to understand biosynthesis

“…Many of the researchers are of the view that in vitro cultures of Glycyrrhiza has a better capability of production of flavonoids than the triterpenoids . However, reports about preferential accumulation and age-dependent synthesis of secondary metabolite are available in plants (Piatczak et al 2015).…”

Phenylpropanoid Glycoside and Phenolic Acid Profiles and Biological Activities of Biomass Extracts from Different Types of Verbena officinalis Microshoot Cultures and Soil-Grown Plant

“…Very high concentrations of verbascoside were reported in suspension cultures of Buddleja cordata (11.6 g/100 g DW) [ 47 ], Plantago media (9.16 g/100 g DW) [ 48 ], and cultures of Cistanche salsa (689 mg/L) [ 49 ] and H. procumbens maintained in pulse-aerated column bioreactors (165.42 mg/L/day) [ 50 ]. Depending on the cultivation method, high levels of verbascoside (from 2.2 to 6.0 g/100 g DW) as well as high levels of isoverbascoside (maximum content: from 0.8 to 1.8 g/100 g DW) were obtained in the hairy root cultures of Rehmannia glutinosa [ 51 , 52 , 53 ]. In the microshoot cultures of Scutellaria baicalensis maintained on MS medium, the content of verbascoside reached 830.9 mg/100 g DW [ 54 ], in the microshoot cultures of Scutellaria altissima it was 0.5 g/100 g DW [ 55 ] and in microshoot cultures of Scutellaria lateriflora maximal content on MS medium was 381.73 mg/100 g DW [ 56 ].…”

Production of Phenylpropanoids, Naphthodianthrones and Antioxidant Status of <em>Hypericum perforatum</em> L. Transgenic Shoots

“…The production of phenylpropanoids (total phenolics, flavonoids, flavan-3-ols, proanthocyanidins and phenolic acids) in H. perforatum TS clones was at levels comparable to or even greater than that in control shoots. In this view, various transgenic plant species have shown significantly higher production of secondary metabolites compared to non-transformed shoots [53,55,62] or intact plants [33,54,63]. The importance of phenylpropanoids as defense compounds in H. perforatum shoots has been previously confirmed by using different strategies, such as phytohormone supplementation [64], precursor feeding [65], elicitation [34,35] and genetic transformation [24].…”