scholarly journals Linnaea borealis L. var. borealis—In Vitro Cultures and Phytochemical Screening as a Dual Strategy for Its Ex Situ Conservation and a Source of Bioactive Compounds of the Rare Species

Molecules ◽  
2021 ◽  
Vol 26 (22) ◽  
pp. 6823
Author(s):  
Barbara Thiem ◽  
Dariusz Kruszka ◽  
Natalia Turowska ◽  
Elwira Sliwinska ◽  
Viktor Berge ◽  
...  

Linnaea borealis L. (Twinflower)—a dwarf shrub in the Linnaeeae tribe of Caprifoliaceae family—is distributed across the Northern Hemisphere. By means of this study, a reliable protocol for efficient micropropagation of uniform L. borealis L. var. borealis plantlets has been provided for the first time; callus culture was also established. Different initial explants, types of cultures, media systems, and plant growth regulators in Murashige and Skoog (MS) media were tested. Agitated shoot cultures in the liquid media turned out to be the best system for the production of sustainable plant biomass. After stabilization of the callus lines, the highest growth index (c.a. 526%) was gained for callus maintained on MS enriched with picloram. TLC and UHPLC-HESI-HRMS analysis confirmed the presence of phenolic acids and flavonoids, and for the first time, the presence of iridoids and triterpenoid saponins in this species. Multiplication of L. borealis shoot culture provides renewable raw material, allowing for the assessment of the phytochemical profile, and, in the future, for the quantitative analyses and the studies of the biological activity of extracts, fractions, or isolated compounds. This is the first report on in vitro cultures of traditionally used L. borealis rare taxon and its biosynthetic potential.

REPORTS ◽  
2021 ◽  
Vol 2 (336) ◽  
pp. 18-25
Author(s):  
R. A. Turganova ◽  
E. D. Djangalina ◽  
E. A. Shadenova ◽  
A. I. Kapytina ◽  
G. K. Kamshybayeva

Paulownia sp. are tall and fast-growing perennial plants that grow faster than all woody plants in the world. In many countries, Paulownia sp. are used as a raw material in bioenergy, furniture industry, landscape gardening and technologies for phytoremediation. In this study for the first time in Kazakhstan, conditions of Paulownia tomentosa (P. tomentosa) in vitro cultivation and propagation have been optimized, also the factors influencing the morphogenetic activity of primary explants have been studied. Along with the adaptation potential of Paulownia tomentosa microclones to the ex situ conditions, laboratory standing order for microclonal reproduction have been evaluated. For sterilization of P. tomentosa explants are recommended to use 50% domestos and 0.1% merthiolate. Hormone-free WPM medium was considered as the most optimal for the in vitro propagation. Infrared light is highly recommended for P. tomentosa microclones adaptation, due to its ability to stimulate the formation plants aboveground biomass and root system. For Kazakhstan, the research of this type of tree crops is a relevant, new and promising direction. The development of microclonal propagation of Paulownia tomentosa will accelerate the process of introduction of Paulownia in our Republic.


2017 ◽  
Vol 86 (4) ◽  
Author(s):  
Paweł Kubica ◽  
Agnieszka Szopa ◽  
Halina Ekiert

In vitro cultures of <em>Cistus ×incanus</em> (pink rock-rose) were maintained on two variants of Murashige and Skoog (MS) medium differing in terms of composition of plant growth regulators (PGRs): 6-benzyl­adenine (BA) and 1-naphthaleneacetic acid (NAA) at the following concentrations: 3 mg/L and 0, 3 mg/L and 1 mg/L, respectively, and on a variant without PGRs – a control. Cultures were maintained in a form of agar and agitated shoot cultures. The qualitative and quantitative analyzes of three groups of phenolic compounds (catechins, flavonoids, and free phenolic acids) were performed by using the HPLC-DAD technique in methanolic extracts of in vitro biomasses and of commercial plant raw material. In analyzed extracts from in vitro cultures, the presence of catechin [max. 197.80 mg / 100 g dry weight (DW)], epicatechin gallate (max. 30.74 mg / 100 g DW), gallic acid (max. 83.23 mg / 100 g DW), quercetin (max. 10.15 mg / 100 g DW), and quercitrin (max. 72.89 mg / 100 g DW) was confirmed. The quantities of accumulated compounds varied and depended on the type of in vitro culture and the concentration of PGRs in media. The highest amounts of all estimated compounds were obtained in biomasses from agar cultures cultivated on medium without PGRs in vitro. In extracts obtained from commercial raw material, gallic acid (max. 261.80 mg / 100 g DW) and quercetin (max. 255.96 mg / 100 g DW) were detected as being the dominant compounds.


2018 ◽  
Vol 30 (2) ◽  
pp. 259-267 ◽  
Author(s):  
Agnieszka Wojtania ◽  
Bożena Matysiak

Abstract The aim of the study was to develop an efficient micropropagation system for Rosa ‘Konstancin’, an interspecific hybrid between R. rugosa and R. beggeriana, whose fruits have high pro-health value. Shoot cultures were initiated from shoot buds collected in May and August from 15-year-old field-grown Rosa ‘Konstancin’ shrubs. The effect and interaction of different concentrations of phytohormones, sucrose and iron sources on in vitro initiation, multiplication and rooting of shoots were studied. The time of collecting explants from donor plants significantly affected the initiation of shoot culture of Rosa ‘Konstancin’. Considerably higher frequency of bud break (100%) was obtained in explants isolated in August as compared to those collected at the end of May (30%). All buds developed into single shoots after 2-4 weeks of growing on the basal Murashige and Skoog medium containing 2.2 µM BAP, 0.3 µM GA3 and 88 mM of sucrose. The highest multiplication rate (4.8 shoots/explant) in a 5-week period was obtained on MS medium containing 50% of nitrogen salts, 3.1 µM BAP, 0.9 µM GA3 and 58 mM sucrose. High rooting frequency (100%) and quality of rooted plantlets was obtained on a medium containing 0.5 µM IBA, 138 µM Fe-EDDHA and 88 mM sucrose. Fe-EDDHA had a beneficial effect on the growth and photosynthetic activity of Rosa ‘Konstancin’ plantlets, which were successfully acclimatized ex vitro, with a more than 90% survival rate.


Weed Science ◽  
1998 ◽  
Vol 46 (1) ◽  
pp. 24-29 ◽  
Author(s):  
Terry R. Wright ◽  
Donald Penner

Acetolactate synthase (ALS)-inhibiting herbicide carryover in soil can severely affect sugarbeets grown in the year(s) following application. Two newly developed imidazolinone-resistant (IMI-R) sugarbeet somatic cell selections (Sir-13 and 93R30B) were examined for magnitude of resistance and extent of cross-resistance to other classes of ALS inhibitors and compared to a previously developed sulfonylurea-resistant (SU-R) selection, Sur. In vitro shoot culture tests indicated Sir-13 resistance was specific to imidazolinone (IMI) herbicides at approximately a 100-fold resistance compared to the sensitive control sugarbeet. Sur was 10,000-fold resistant to the sulfonylurea (SU) herbicide, chlorsulfuron, and 40-fold resistant to the triazolopyrimidine sulfonanilide (TP) herbicide, flumetsulam, but not cross-resistant to the IMI herbicides. 93R30B was selected for IMI-R from a plant homozygous for the SU-R allele,Sur, and displayed similar in vitro SU-R and TP-R as Sur, but also displayed a very high resistance to various IMI herbicides (400- to 3,600-fold). Compared to the sensitive control, Sir-13 was 300- and > 250-fold more resistant to imazethapyr and imazamox residues in soil, respectively. Response by whole plants to postemergence herbicide applications was similar to that observed in shoot cultures. Sir-13 exhibited > 100-fold resistance to imazethapyr as well as imazamox, and 93R30B showed > 250-fold resistance to both herbicides. 93R30B showed great enough resistance to imazamox to merit consideration of imazamox for use as a herbicide in these sugarbeets. Sir-13 showed a two- to threefold higher level of resistance in the homozygous vs. heterozygous state, indicating that like most ALS-inhibitor resistance traits, it was semidominantly inherited.


2014 ◽  
Vol 9 (1) ◽  
pp. 1934578X1400900 ◽  
Author(s):  
Sibylle Kümmritz ◽  
Christiane Haas ◽  
Atanas I. Pavlov ◽  
Doris Geib ◽  
Roland Ulber ◽  
...  

Plant in vitro cultures are a prospective alternative for biochemicals production, for example the triterpenes oleanolic and ursolic acid present in plants and cell cultures of Salvia sp. Our objective was to develop a suitable analysis protocol for evaluation of triterpenic acid yield in plant raw material and in vitro cultures supporting selection processes. Moreover, valuable bioactive compounds had to be revealed. Thus, different strategies enhancing the separation for a sensitive and effective HPLC-UV method were investigated and the developed method was validated for linearity, precision, accuracy, limits of detection and quantification. A baseline separation of these isomers enabled detection limits of below 0.4 μg/mL and quantification limits of about 1.2 μg/mL. Over the tested concentration range a good linearity was observed (R2 > 0.9999). The variations in the method were below 6% for intra- and inter-day assays of concentration. Recoveries were between 85–98% for both compounds using ethanol as extraction solvent. Additionally, metabolite profiling of cell suspension culture extracts by GC-MS has shown the production variability of different plant metabolites and especially the presence of plant phenols and sterols. These studies provide a method suitable for screening plant and cell culture productivity of triterpenic acids and highlighted interesting co-products of plant cell cultures.


1970 ◽  
Vol 16 (1) ◽  
pp. 5-9
Author(s):  
MH Kabir ◽  
PK Roy ◽  
Golam Ahmed

In vitro plant regeneration of Thuja occidentalis was obtained in apical shoot cultures from field grown plants. Hormone free MS medium 100% explants produced shoots. The average number of shoots per explant was 6.57 ± 0.45 and the average shoot length of 4.5 ± 0.27 cm were recorded in this medium. Shoots rooted well when they were transferred into half strength MS with 1.0 mg/l IBA. The average number of root per shoot was 3.92 ± 0.28 and the average root length of 3.64 ± 0.38 cm were observed in this medium. No morphological variants were observed during the passage of in vitro culture.Key words: In vitro, Propagation, Thuja occidentalis, Apical shootDOI = 10.3329/ptcb.v16i1.1099Plant Tissue Cult. & Biotech. 16(1): 5-9, 2006 (June)


2011 ◽  
Vol 74 (1) ◽  
pp. 17-21 ◽  
Author(s):  
Izabela Grzegorczyk ◽  
Ireneusz Bilichowski ◽  
Elżbieta Mikiciuk-Olasik ◽  
Halina Wysokińska

The concentrations of carnosic acid, carnosol and rosmarinic acid in different materials from differentiated (multiple shoot cultures and regenerated plants) and undifferentiated (callus and cell suspension) in vitro cultures of <em>Salvia officinalis</em> were determined by HPLC. The results suggested that diterpenoid (carnosic acid and carnosol) production is closely related to shoot differentiation. The highest diterpenoid yield (11.4 mg g<sup>-1</sup> for carnosic acid and 1.1 mg g<sup>-1</sup> for carnosol) was achieved in shoots of 10-week-old micropropagated plants. The levels were comparable to those found in shoots of naturally growing plants. Undifferentiated callus and cell suspension cultures produced only very low amounts of carnosol (ca. 0.05 mg g<sup>-1</sup> of dry weight). In contrast, content of rosmarinic acid in callus and suspension cultures as well as shoots growing in vitro and in vivo was similar and ranged between 11.2 and 18.6 mg g<sup>-1</sup> of dry weight.


2016 ◽  
Vol 58 (1) ◽  
pp. 43-56 ◽  
Author(s):  
Małgorzata Kikowska ◽  
Barbara Thiem ◽  
Elwira Sliwinska ◽  
Monika Rewers ◽  
Mariusz Kowalczyk ◽  
...  

AbstractAn efficient micropropagation protocol for production of genetically uniform clones ofEryngium campestreL. was developed. To determine the effect of nutritional and hormonal factors on shoot and root development and bioactive compounds production, three variants of media differing in the content of macro- and micronutrients, as well as plant growth regulators of various types and concentrations were tested. The highest regeneration (100%), with over 13 shoots per explant, was induced on Murashige and Skoog (MS) medium with 1.0 mg l−1benzyladenine (BA) and 0.1 mg l−1indole-3-acetic acid (IAA). The in vitro derived shoots multiplied through axillary bud formation were rooted and transferred to an experimental plot with 78% frequency of survival. Flow cytometry showed no variation in nuclear DNA between the seedlings and micropropagated plants. Preliminary thin layer chromatography (TLC) analysis indicated that phenolic acids, saponins, flavonoids and acetylenes were present in plant biomass. Ultra high performance liquid chromatography (UHPLC) analysis revealed that shoots and roots from in vitro derived plants and root cultures maintained the ability to produce rosmarinic acid (RA), rosmarinic acid hexoside (RA-HEX) and chlorogenic acid (CGA). The highest phenolic acid content was detected in roots of in vitro regenerated plants. The extract from those roots expressed the highest inhibitory effect against bacteriaStaphylococcus aureus, as well as dermatophytesTrichophyton mentagrophytesandT. rubrum.


2013 ◽  
Vol 82 (4) ◽  
pp. 313-320 ◽  
Author(s):  
Wojciech J. Szypuła ◽  
Paulina Mistrzak ◽  
Olga Olszowska

This study presents a protocol for a fast and effective in vitro axenic culture of <em>Huperzia selago</em> (Huperziaceae Rothm.) sporophytes, a club moss which is a source of huperzine A, an alkaloid of a considerable therapeutic potential extensively investigated for its uses as treatment for some neurodegenerative diseases. The proposed procedure allowed approximately tenfold shortening of the species developmental stages with the omission of the gametophyte stage while the sporophyte mass could be increased tenfold within a 6-month period. The cultures were established using vegetative propagules (bulbils) procured from sporophytes growing in the wild without degrading the habitats of this endangered plant species. Explants underwent surface and internal disinfection to eliminate the epiphytic and endophytic bacteria and fungi. In in vitro cultures, the optimum results were achieved using Moore (Mr) medium without growth regulators or supplemented with 0.015 mg/l IBA and 0.3 mg/l kinetin. These media ensured both viability of the propagules and their further development. The biomass growth index for <em>H. selago</em> sporophytes grown from propagules, determined at 3 months of culture (1 passage) on Mr medium with IBA and kinetin was 650%. At 6 months, the biomass growth index increased to 1114%. Vigorous growth of adventitious roots, especially on Mr medium with the addition of 0.25 mg/l NAA, and callus formation on shoot apices were observed. At 6 months of culture, some sporophytes obtained from the bulbils were used as the initiating material for shoot subcultures, which developed best on Mr medium with IBA and kinetin.


Plants ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 206 ◽  
Author(s):  
Michał P. Maliński ◽  
Małgorzata Anna Kikowska ◽  
Agata Soluch ◽  
Mariusz Kowalczyk ◽  
Anna Stochmal ◽  
...  

Lychnis flos-cuculi L., a species with potential medicinal value, contains flavonoids, phenolic acids, triterpenoid saponins and ecdysteroids. In this study, the antioxidant activity of plant material of L. flos-cuculi obtained from in vitro cultures compared to that of intact plants from the natural site has been evaluated for the first time. Phytochemical screening of the in-vitro-derived material by ultra-high-performance liquid chromatography mass spectrometry (UHPLC-MS) confirmed the presence of the aforementioned metabolite classes. The aqueous methanolic extracts from in-vitro-derived plant material and the organs of intact plants were analyzed using spectrophotometric methods to quantify total phenolics, phenolic acids and flavonoids, and determine the preliminary antioxidant activity by ferric reducing antioxidant potential (FRAP) and DPPH radical scavenging activity assays. The results showed that the inflorescence (Ns-F), and flowering herb of both plants gathered from natural habitat (Ns-H) and in-vitro-derived plants from the experimental plot (ExV-H) are the materials richest in polyphenols (195.4, 113.47, 112.1 mg GAE g−1 d.w., respectively), and demonstrate the highest antioxidant activity (20.14, 11.24, and 11.46 mg AAE g−1 d.w.). The extract from callus exhibited the lowest polyphenol content and antioxidant potential. The contents of total phenolics, flavonoids and phenolic acids correlate with the results of the antioxidant capacity of L. flos-cuculi extracts.


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