Multiple shoot formation and phenotypic changes of R<sub>0</sub> regenerants in <i>Hypericum perforatum</i> L.

Čellárová, Eva; Kimáková, Katarína; Brutovská, R.

doi:10.1002/abio.370120602

Cited by 52 publications

(10 citation statements)

References 9 publications

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“…The formation of both shoots and roots on the same medium eliminates one step of medium transfer in the propagation of this species, thus reducing the total time and cost required for plant regeneration. Similar results were reported for H. perforatum (Cellá rová et al 1992;McCoy and Camper 2002) and H. brasiliense (Cardoso and Oliveira 1996).…”

Section: Resultssupporting

confidence: 93%

“…Explants cultured on control medium without growth regulators did not respond satisfactorily. Similar effect of BAP on shoot development in H. perforatum and H. foliosum has been reported previously (Cellá rová et al 1992;Moura 1998). The absence of rooting in the presence of NAA, as mentioned by the same studies, was also confirmed for H. polyanthemum.…”

Section: Resultssupporting

confidence: 84%

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Benzopyrans in Hypericum polyanthemum Klotzsch ex Reichardt cultured in vitro

et al. 2007

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Hypericum polyanthemum Klotzsch ex Reichardt, an endemic species of Southern Brazil, was micropropagated on MS medium supplemented with 1.78 lM BAP. Shoot proliferation and rooting was achieved on hormone-free medium and plantlets survived acclimatization. The bioactive compounds: 6-isobutyryl-5,7-dimethoxy-2,2-dimethylbenzopyran (HP1), 7-hydroxy-6-isobutyryl-5-methoxy-2,2-dimethyl-benzopyran (HP2) and 5-hydroxy-6-isobutyryl-7-methoxy-2,2-dimethyl-benzopyran (HP3) were quantified in the leaves, stems and roots of propagated and acclimatized plantlets and compared with the field-grown plants. The HPLC analysis revealed that the three benzopyrans are accumulated in the aerial parts and the concentration varied with the age of the plant whereas the roots were capable of accumulating only HP3. Greatest yield of HP1 (7.12 mg/g DW) was quantified in the leaves of the acclimatized plantlets, whereas the flowers of the plants from natural habitat displayed higher amounts of HP2 (11.04 mg/g DW) and HP3 (13.99 mg/g DW).

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Section: Resultssupporting

confidence: 93%

Section: Resultssupporting

confidence: 84%

Benzopyrans in Hypericum polyanthemum Klotzsch ex Reichardt cultured in vitro

et al. 2007

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“…As a result, many investigators have discussed how one might obtain H. perforatum shootlet meristem cultures in vitro from which hypericins and hyperforin can be detected [10,47,66,68,86,87]. Callus and cell‐suspension cultures of H. perforatum are usually established from the shoots of surface‐sterilized seeds that germinate in culture.…”

Section: Challenges and Strategies To Enhance Production Of Hypericinmentioning

confidence: 99%

“…Having more leaf surface in comparison with stem tissue in the sample would shift the proportion of hypericin recovered, since the glands containing hypericin are generally located along the leaf margins. Cellárová et al [66] demonstrated that several morphological characteristics of H. perforatum cv. 'Topas' regenerants varied considerably without a clear trend when grown on media with a range of kinetin concentrations.…”

Section: Genetic and Environmental Effects On Secondary Metabolismmentioning

confidence: 99%

The production of hypericins and hyperforin by in vitro cultures of St. John's wort (Hypericum perforatum)

Kirakosyan

Sirvent

Gibson

et al. 2004

Biotech and App Biochem

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St. John's wort ( Hypericum perforatum L.) is a herbaceous perennial distributed throughout the World that has been widely used in traditional medicine. H. perforatum produces several types of biologically active compound, including the hypericins--a family of light-activated anthraquinones, localized within specialized glands found predominantly on flowers and leaves--and the hyperforins--a family of prenylated acylphloroglucinols localized in the reproductive structures of the plant. Hypericins are known to be toxic to mammals and display antiviral and anticancer activity, but the role of these compounds within the plant is unknown. Hyperforins display potent antimicrobial activity and are thought to be the primary bioactive ingredient for anti-depressive effects of the herb. The introduction of H. perforatum from Europe into the U.S.A. occurred in the 17th Century. Since the plant is considered a noxious weed, few efforts have been carried out to analyse populations in the context of secondary-metabolite concentrations. But in terms of secondary-metabolite studies, H. perforatum is an ideal model system to study the biosyntheses of aromatic polyketides and regulation of those pathways by environmental and genetic influences. This is due, in part, to the ease of conducting these studies in plant tissue culture. This review describes the progress of secondary-metabolite studies currently underway using H. perforatum. Specifically, this Review focuses on the production and regulation of the hypericins and the hyperforin in wild populations, field cultivation, greenhouse studies and plant tissue culture. Additionally, factors optimizing compound production--particularly in in vitro cultures--are presented and reviewed.

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“…Such universal type medium can be a good choice for large-scale continuous biomass production systems. Shoot cultures of H. perforatum require exogenous cytokinins alone or in combination with auxins for good growth and multiplication (Čellárová et al 1992, Zdunek and Alfermann 1992, Murch et al 2000. Proper cytokinin/auxin balances is also required for the establishment and shoot regeneration of H. perforatum callus tissues (Čellárová et al 1995, Preto andSantarém 2000).…”

mentioning

confidence: 98%

Shoot and root culture of Hypericum perforatum L. transformed with Agrobacterium rhizogenes A4M70GUS

Vinterhalter¹,

Ninković²,

Cingel³

et al. 2006

Biologia plant.

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Hairy root cultures of Hypericum perforatum were obtained following inoculation of aseptically germinated seedlings with A. rhizogenes strain A4M70GUS. Effect of sucrose on the growth and biomass production of hairy root cultures was investigated. Hairy root cultures spontaneously regenerated shoots buds from which a number of shoot culture clones was established. Transformed shoot cultures exhibited good shoot multiplication, elongation and rooting on a hormone-free woody plant medium. Plants regenerated from hairy roots were similar in appearance to the normal, nontransformed plants.Additional key words: GUS activity, hairy roots, shoot regeneration, St. John's wort, sucrose. ⎯⎯⎯⎯ Hypericum perforatum L. (St. John's wort) is an important medicinal plant rich in secondary metabolites considered as an interesting drug source for the pharmaceutical industry. Most of the in vitro culture studies on this species are usually dedicated to cell and tissue cultures and their ability to produce various secondary metabolites. The aim of our investigation was to obtain and study hairy root cultures of H. perforatum. For this purpose we selected A. rhizogenes strain A4M70GUS, previously used in our laboratory to obtain hairy root cultures in several species of Gentiana (Momčilović et al. 1997, Vinterhalter et al. 1999), Lotus corniculatus (Nikolić et al. 2003/04), Centaurium erythrea (Subotić et al. 2003/04), Aesculus hippocastanum (Zdravković-Korać et al. 2004) and Blackstonia perfoliata (Bjelović et al. 2004). So far there are no papers published on Agrobacterium mediated transformation of H. perforatum.Hypericum perforatum L. seeds collected on locations around Belgrade were washed with 70 % ethanol for 1 min and than surface sterilized in 15 % commercial bleach (4 -5 % NaOCl) for 20 min. Seeds were rinsed 3 × 5 min in autoclaved water and aseptically germinated. Skoog (1962; MS) microsalts, iron and vitamins. Media were supplemented with 0.62 % agar and 2 % sucrose. Media pH was adjusted to 5.8 prior to autoclaving performed 20 min at 114 °C. Conditions in the growth room were: temperature of 25 ± 2 °C, 16-h photoperiod and irradiance 46.5 μmol m -2 s -1 . Basal media contained woody plant medium (Lloyd and McCown 1981; WPM) macrosalts and Murashige andEpicotyls were excised from seedling and cultured on basal media supplemented with 0.2 -0.5 mg dm -3 kinetin. Shoot cultures derived from epicotyls were subculture in 5 -6 weeks intervals.A. rhizogenes strain A4M70GUS contains a GUS construct integrated into the TL region of the cointegrative plasmid pRiA4 (Tepfer and Casse-Delbart 1987). GUS construct contains uidA sequence under the 70S promoter (enhancer-doubled 35S Ca MV promoter), followed by NOS polyadenylation sequence. Bacterial strains were maintained according to Van Larebake et al. (1977).Shoots comprising 3 -4 internodes were inoculated by wounding with a needle dipped in bacterial suspension. Wounding was done at the first node above ⎯⎯⎯⎯

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Multiple shoot formation and phenotypic changes of R₀ regenerants in Hypericum perforatum L.

Cited by 52 publications

References 9 publications

Benzopyrans in Hypericum polyanthemum Klotzsch ex Reichardt cultured in vitro

Benzopyrans in Hypericum polyanthemum Klotzsch ex Reichardt cultured in vitro

The production of hypericins and hyperforin by in vitro cultures of St. John's wort (Hypericum perforatum)

Shoot and root culture of Hypericum perforatum L. transformed with Agrobacterium rhizogenes A4M70GUS

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Multiple shoot formation and phenotypic changes of R0 regenerants in Hypericum perforatum L.

Cited by 52 publications

References 9 publications

Benzopyrans in Hypericum polyanthemum Klotzsch ex Reichardt cultured in vitro

Benzopyrans in Hypericum polyanthemum Klotzsch ex Reichardt cultured in vitro

The production of hypericins and hyperforin by in vitro cultures of St. John's wort (Hypericum perforatum)

Shoot and root culture of Hypericum perforatum L. transformed with Agrobacterium rhizogenes A4M70GUS

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Multiple shoot formation and phenotypic changes of R₀ regenerants in Hypericum perforatum L.