Hairy root cultures of Rehmannia glutinosa and production of iridoid and phenylethanoid glycosides

Piątczak, Ewelina; Królicka, Aleksandra; Wielanek, Marzena; Wysokińska, Halina

doi:10.1007/s11738-012-1022-y

Cited by 23 publications

(18 citation statements)

References 37 publications

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“…Our earlier study has shown that catalposide, a 6-O-ester of catalpol, is the main iridoid in hairy roots of R. glutinosa (Piątczak et al 2012). In contrast, the transformed roots did not produce catalpol.…”

Section: Production Of Iridoid Glycosidesmentioning

confidence: 90%

“…UHPLC analyses of the methanolic extracts revealed the presence of catalpol, aucubin, loganin, catalposide, harpagide and harpagoside as well as verbascoside and isoverbascoside in tested samples. The compounds were identified by comparison of their retention times, UV spectra and ion mass spectra with authentic standards, as described in our previous work (Piątczak et al 2012). Additionally, in the present study, LC-ESI-MS was performed to confirm the identity of the peaks of harpagide and harpagoside.…”

Section: Secondary Metabolite Productionmentioning

confidence: 95%

“…The materials were lyophilized and powdered. The procedure of methanolic extract preparation was described in our earlier study (Piątczak et al 2012). UHPLC analysis was used to determine iridoid (catalpol, aucubin, loganin, harpagide, harpagoside, catalposide) and phenylethanoid glycoside (verbascoside, isoverbascoside) contents.…”

Section: Uhplc Methodsmentioning

confidence: 99%

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Shoot organogenesis, molecular analysis and secondary metabolite production of micropropagated Rehmannia glutinosa Libosch.

Piątczak

Kuźma

Sitarek

et al. 2014

Plant Cell Tiss Organ Cult

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This study is the first description of a procedure for shoot organogenesis from calli derived from various seedling explants. The frequency of shoot induction and number of shoots per explant were measured on Murashige and Skoog (MS) medium with 0.2-3.0 mg L -1 6-benzylaminopurine (BAP) alone or combined with auxin [0.1 mg L -1 indole-3-acetic acid (IAA) or 1-naphthalenacetic acid]. The highest organogenic response where 90 % of explants formed a regenerative callus with a mean number of nine shoots was achieved after 6 weeks of culture when hypocotyl explants and 1.0 mg L -1 BAP was used together with 0.1 mg L -1 IAA. The shoots regenerated from hypocotyl-derived calli were rooted for 6 weeks on MS medium lacking growth regulators or containing auxin (IAA or indole-3-butyric acid). The plantlets were successfully acclimatized in a greenhouse. Micropropagated plants subjected to random amplified polymorphic DNA and inter simple sequence repeat (ISSR) marker based profiling revealed a uniform banding pattern identical with that of donor plants. The iridoid and phenylethanoid glycoside content of the organogenic callus culture and in vitro-raised plants was determined using UHPLC. In the tested samples catalpol, aucubin, harpagid, harpagoside, verbascoside, isoverbascoside and traces of loganin and catalposide were identified. The leaves produced the highest levels of catalpol (43-45 mg g -1 DW). Organogenic callus and 6-week-old in vitro grown plantlets were characterized by the highest accumulation of aucubin, verbascoside and isoverbascoside. The work also demonstrated the regenerative ability of hypocotyl-derived calli and production of bioactive metabolites to be stable for 4 years of culture.

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Section: Production Of Iridoid Glycosidesmentioning

confidence: 90%

Section: Secondary Metabolite Productionmentioning

confidence: 95%

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Shoot organogenesis, molecular analysis and secondary metabolite production of micropropagated Rehmannia glutinosa Libosch.

Piątczak

Kuźma

Sitarek

et al. 2014

Plant Cell Tiss Organ Cult

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“…The extraction procedure was performed according to Sesterhenn et al (2007) with slight modifications described earlier (Piątczak et al 2012). Extraction yields (% w/w) were calculated by the following formula: weight of the dried extract (g)/weight of the original sample (g) 9 100 %.…”

Section: Preparation Of Extractsmentioning

confidence: 99%

Micropropagation of Rehmannia glutinosa Libosch.: production of phenolics and flavonoids and evaluation of antioxidant activity

2014

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Rehmannia glutinosa Libosch., a valuable medicinal plant, was successfully propagated in vitro using shoot tip explants. Shoot multiplication was performed in glass tubes and in a nutrient sprinkle bioreactor. A mixture of 0.1 mg L -1 indole-3-acetic acid (IAA) and 1.0 mg L -1 of 6-benzylaminopurine in Murashige and Skoog (MS) agar-solidified medium proved the best combination for multiple shoot induction, yielding 8.2 shoots per explant after 4 weeks of culture in glass tubes. The number of shoots increased to 21 per explant when the same combination of growth regulators was used in a nutrient sprinkle bioreactor. The shoots rooted with a frequency of 93 % after 6 weeks of culture on MS agar medium supplemented with IAA (0.1 mg L -1 ) before being acclimatized in the greenhouse. The antioxidant activities of methanolic extracts from the leaves and roots of the in vitro-regenerated plants of R. glutinosa cultivated in the greenhouse were evaluated using four in vitro assays: scavenging of free radicals (DPPH and ABTS), transition metal reduction and total antioxidant activity phosphomolybdenum test. In all cases, the methanolic extract from leaves demonstrated better antioxidant activity than those taken from roots. A strong correlation was found between total phenolic and flavonoid content, and the antioxidant capacity of the studied extracts.

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“…The pharmacological properties of the roots have been attributed mainly to the presence of iridoid (catalpol, aucubin) and phenylethanoid (verbascoside, isoverbascoside, echinacoside) glycosides. Our previous paper has described the establishment of hairy root cultures of R. glutinosa and production of iridoid (catalposide, loganin, catalpol and aucubin) and phenylethanoid (verbascoside and isoverbascoside) glycosides in ten hairy root lines transformed with Agrobacterium rhizogenes strain A4 (Piątczak et al 2012). These hairy root lines exhibited differences in secondary metabolite contents.…”

mentioning

confidence: 99%

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

Piątczak

Kuźma

Skała

et al. 2015

Plant Cell Tiss Organ Cult

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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.

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Hairy root cultures of Rehmannia glutinosa and production of iridoid and phenylethanoid glycosides

Cited by 23 publications

References 37 publications

Shoot organogenesis, molecular analysis and secondary metabolite production of micropropagated Rehmannia glutinosa Libosch.

Shoot organogenesis, molecular analysis and secondary metabolite production of micropropagated Rehmannia glutinosa Libosch.

Micropropagation of Rehmannia glutinosa Libosch.: production of phenolics and flavonoids and evaluation of antioxidant activity

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

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