Genetic transformation of Ruta graveolens L. by Agrobacterium rhizogenes: hairy root cultures a promising approach for production of coumarins and furanocoumarins

Sidwa-Gorycka, Matylda; Królicka, Aleksandra; Orlita, Aleksandra; Maliński, Edmund; Gołębiowski, Marek; Kumirska, Jolanta; Chromik, Agnieszka; Biskup, Edyta; Stepnowski, Piotr; Łojkowska, Ewa

doi:10.1007/s11240-009-9498-x

Cited by 38 publications

(14 citation statements)

References 29 publications

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“…Light is known to regulate not only plant growth and development but also the biosynthesis of both primary and secondary metabolites (Vázquez-Flota and De Luca 1998). Our findings are in line with the results of Sander (2009) and Sidwa-Gorycka et al (2009) who stated that light adaptation decreases some metabolite levels by increasing the flux out of the metabolite pools in hairy roots of C. roseus and Ruta graveolens, respectively.…”

Section: Plant Cell Repsupporting

confidence: 93%

Establishment of transgenic Rhazya stricta hairy roots to modulate terpenoid indole alkaloid production

et al. 2015

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Transgenic hairy roots of R. stricta were developed for investigation of alkaloid accumulations. The contents of five identified alkaloids, including serpentine as a new compound, increased compared to non-transformed roots. Rhazya stricta Decne. is a rich source of pharmacologically active terpenoid indole alkaloids (TIAs). In order to study TIA production and enable metabolic engineering, we established hairy root cultures of R. stricta by co-cultivating cotyledon, hypocotyl, leaf, and shoot explants with wild-type Agrobacterium rhizogenes strain LBA 9402 and A. rhizogenes carrying the pK2WG7-gusA binary vector. Hairy roots initiated from the leaf explants 2 to 8 weeks. Transformation was confirmed by polymerase chain reaction and in case of GUS clones with GUS staining assay. Transformation efficiency was 74 and 83% for wild-type and GUS hairy root clones, respectively. Alkaloid accumulation was monitored by HPLC, and identification was achieved by UPLC-MS analysis. The influence of light (16 h photoperiod versus total darkness) and media composition (modified Gamborg B5 medium versus Woody Plant Medium) on the production of TIAs were investigated. Compared to non-transformed roots, wild-type hairy roots accumulated significantly higher amounts of five alkaloids. GUS hairy roots contained higher amounts two of alkaloids compared to non-transformed roots. Light conditions had a marked effect on the accumulation of five alkaloids whereas the composition of media only affected the accumulation of two alkaloids. By successfully establishing R. stricta hairy root clones, the potential of transgenic hairy root systems in modulating TIA production was confirmed.

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Section: Plant Cell Repsupporting

confidence: 93%

Establishment of transgenic Rhazya stricta hairy roots to modulate terpenoid indole alkaloid production

et al. 2015

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“…In order to confirm that the examined hairy roots were truly transformed, PCR was performed using virG genes (gene present in Ri plasmid but beyond the transferred T-DNA) to detect the presence of any contaminating Agrobacterium in the hairy roots. Oligonucleotide primers and procedure was previously described by Sidwa-Gorycka et al (2009).…”

Section: Pcr Analysismentioning

confidence: 99%

Genetic transformation of Harpagophytum procumbens by Agrobacterium rhizogenes: iridoid and phenylethanoid glycoside accumulation in hairy root cultures

et al. 2010

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A genetic transformation method using Agrobacterium rhizogenes was developed for Harpagophytum procumbens. The influence of three factors on hairy root formation was tested: bacterial strains (A4 and ATCC 15834), various types of explants and acetosyringone (AS) (200 lM). The highest frequency of transformation (over 50% of explants forming roots at the infected sites after 6 weeks of culture on Lloyd and McCown (WP) medium) was achieved using a combination of nodal stem explants and A. rhizogenes strain A4. The addition of 200 lM AS to root induction medium was found to enhance hairy root induction but its effect varied depending on bacterial strain and explant type. Three of the most vigorously growing hairy root clones of H. procumbens were chosen and analyzed for accumulation of iridoid and phenylethanoid glycosides. The transgenic nature of these root clones was confirmed by PCR amplification; they were positive for rolB and rolC genes. Harpagoside, verbascoside and isoverbascoside were identified by HPLC and LC-ESI-MS as the major compounds from all analyzed hairy root clones. The Hp-3 root clone showed the higher harpagoside content (0.32 mg g -1 dry wt.) compared with other analyzed transformed and non-tuberized untransformed roots of H. procumbens. However, the level of the compound in the hairy root clone was lower than that detected in a sample of commercially available root tubers of H. procumbens. The Hp-3 root clone also produced high amounts of verbascoside and isoverbascoside (8.12 mg g -1 dry wt. and 9.97 mg g -1 dry wt., respectively) comparable to those found in root tubers.

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“…Plant cell cultures produce higher quantities of secondary metabolites, often with different profiles compared to their parent plants. Several strategies have been developed to increase the productivity of these compounds: media optimization (Hanchinal et al 2008), selection of a suitable plant tissue culture (Sidwa-Gorycka et al 2009;Krolicka et al 2010), elicitation (Pawar et al 2011;Coste et al 2011;Rhee et al 2010) and addition of precursors (Cheng et al 2008). The use of biotic or abiotic elicitors to stimulate product formation has become an important progress strategy and has been very useful in reducing the process time required to attain high product concentrations and increased volumetric productivity.…”

Section: Introductionmentioning

confidence: 99%

Polysaccharide elicitors enhance anthocyanin and phenolic acid accumulation in cell suspension cultures of Vitis vinifera

Cai¹,

Kastell²,

Mewis

et al. 2011

Plant Cell Tiss Organ Cult

107

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The effects of yeast extract and selected polysaccharide elicitors on secondary metabolite production, particularly of anthocyanin and phenolic acid, in cell suspension cultures of Vitis vinifera were investigated. All elicitors either maintained or promoted cell growth in culture. Overall, secondary metabolite production in V. vinifera cell suspension cultures responded differently to different elicitors. Chitosan, pectin, and alginate enhanced production of anthocyanin within 13 days of culture with levels of 2.5-, 2.5-, and 2.6-fold increase, respectively, over that of control. Chitosan, alginate, and gum arabic significantly promoted accumulation of phenolic acids, particularly 3-O-glucosyl-resveratrol, in V. vinifera cultures, as well as in the culture medium. Intracellular phenolic acid production was significantly enhanced by alginate and chitosan, with 1.7-and 1.5-fold levels, respectively, of that of control. Extracellular phenolic acid production was also significantly increased in the presence of chitosan and gum arabic, with levels of 3.3-and 1.7-fold higher, respectively, than those of control. In addition, DPPH (1,1-diphenyl-2-picrylhydrazyl) radical scavenging activity was enhanced in the presence of elicitors, and this was positively correlated with increased accumulation of anthocyanin in V. vinifera cell suspension cultures.

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Genetic transformation of Ruta graveolens L. by Agrobacterium rhizogenes: hairy root cultures a promising approach for production of coumarins and furanocoumarins

Cited by 38 publications

References 29 publications

Establishment of transgenic Rhazya stricta hairy roots to modulate terpenoid indole alkaloid production

Establishment of transgenic Rhazya stricta hairy roots to modulate terpenoid indole alkaloid production

Genetic transformation of Harpagophytum procumbens by Agrobacterium rhizogenes: iridoid and phenylethanoid glycoside accumulation in hairy root cultures

Polysaccharide elicitors enhance anthocyanin and phenolic acid accumulation in cell suspension cultures of Vitis vinifera

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