scholarly journals Carbohydrates stimulated Amaryllidaceae alkaloids biosynthesis in Leucojum aestivum L. plants cultured in RITA® bioreactor

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e8688
Author(s):  
Agata Ptak ◽  
Emilia Morańska ◽  
Edyta Skrzypek ◽  
Marzena Warchoł ◽  
Rosella Spina ◽  
...  
Keyword(s):  
Dry Weight ◽  
Commercial Production ◽  
In Vitro Cultures ◽  
Sod Activity ◽  
Amaryllidaceae Alkaloids ◽  
Leucojum Aestivum ◽  
Interesting Approach ◽  
Important Medicinal Plant

Background Leucojum aestivum L. is an important medicinal plant which produces Amaryllidaceae alkaloids, especially galanthamine and lycorine. Research is currently exploring the possibility of producing these alkaloids using biotechnological methods, including in vitro cultures. The biosynthesis of alkaloids may be affected by the types and concentrations of carbohydrate sources used in the medium. In the present investigation we performed such studies on in vitro cultures of L. aestivum with a view to obtaining plant material of good quality, characterized, in particular, by a high content of valuable Amaryllidaceae alkaloids. Methods We examined the effects of various types of carbohydrate sources—sucrose, glucose, fructose and maltose—at different concentrations (30, 60 and 90 g/L)—on the quality of L. aestivum plants grown in the RITA® bioreactor. The plants’ quality was assessed by their biomass increments, as well by as analysing photosynthetic pigments, endogenous sugar, phenolics and Amaryllidaceae alkaloid content. We also investigated the effect of sugars on the activity of the antioxidant enzymes catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD). Results The highest biomass increments were observed in plants cultivated in the medium containing 90 g/L sucrose. The highest CAT activity was noted in cultures growing in the medium supplemented with 90 g/L maltose, while the highest POD activity was observed in the presence of 90 g/L fructose and 60 g/L maltose. No differences in SOD activity were observed. Moreover, the sugars did not affect the contents of chlorophyll a and carotenoids, whereas the highest amount of chlorophyll b was recorded in plants growing in the medium with 60 g/L maltose. No statistically significant differences were observed in the contents of endogenous sugars and phenolics in any in vitro conditions. However, the addition of sugar had a decisive effect on the biosynthesis of the Amaryllidaceae alkaloids. The highest distribution of alkaloids occurred in plants cultured in the medium containing 60 g/L sucrose. Six Amaryllidaceae alkaloids were detected in the plant tissue. The addition of 30 g/L fructose in the medium resulted in the accumulation of five alkaloids, including ismine, which was not identified in other analysed tissues. The highest concentration of galanthamine was observed in plants cultured in the presence of 30 g/L fructose and 60 g/L sucrose (39.2 and 37.5 µg/g of dry weight (DW), respectively). The plants grown in the medium containing 60 g/L sucrose exhibited the highest lycorine content (1048 µg/g of DW). Conclusions The type and concentration of sugar used in the medium have an essential influence on the biosynthesis of Amaryllidaceae alkaloids in L. aestivum plants cultured in a RITA® bioreactor. The results point to an interesting approach for commercial production of galanthamine and lycorine.

scholarly journals Alkaloid Synthesis and Accumulation in Leucojum Aestivum in Vitro Cultures

2009 ◽  
Vol 4 (3) ◽  
pp. 1934578X0900400 ◽  
Author(s):  
Strahil Berkov ◽  
Atanas Pavlov ◽  
Vasil Georgiev ◽  
Jaume Bastida ◽  
Monique Burrus ◽  
...  
Keyword(s):  
Coefficient Of Variation ◽  
Dry Weight ◽  
High Variability ◽  
In Vitro Cultures ◽  
Intact Plant ◽  
Leucojum Aestivum ◽  
Intact Plants ◽  
Alkaloid Synthesis ◽  
Alkaloid Profiles

The alkaloids of intact plants, calli and shoot-clump cultures of L. aestivum were analyzed by GC-MS. Twenty-four alkaloids were detected. Calli appeared to produce sparse alkaloid profiles in stark contrast to shoot-clumps that had similar profiles to those of the intact plant. Seven shoot-clump strains produced galanthamine predominantly whereas another three were dominated by lycorine. Shoot-clump strains cultivated under light accumulated about two-times more galanthamine (an average of 74 μg/g of dry weight) than those cultivated in darkness (an average of 39 μg/g of dry weight). In comparison to intact plants, the shoot-clumps accumulated 5-times less galanthamine. The high variability of both the galanthamine content (67% and 75% of coefficient of variation under light and darkness conditions, respectively) and alkaloid patterns indicates that the shoot-clump cultures initiated from callus could be used as a tool for improvement of the in vitro cultures.

scholarly journals Nickel toxicity induced antioxidant enzyme and phenylalanine ammonia-lyase activities in Jatropha curcas L. cotyledons

2008 ◽  
Vol 54 (No. 7) ◽  
pp. 294-300 ◽  
Author(s):  
R. Yan ◽  
S. Gao ◽  
W. Yang ◽  
M. Cao ◽  
S. Wang ◽  
...  
Keyword(s):  
Jatropha Curcas ◽  
Phenylalanine Ammonia Lyase ◽  
Dry Weight ◽  
Sod Activity ◽  
Nickel Toxicity ◽  
Pal Activity ◽  
High Nickel ◽  
Isoenzyme Patterns ◽  
Negative Link

<I>Jatropha curcas</I> L. embryos were germinated and grown in vitro under nickel concentrations of 100, 200, 400 and 800 &mu;mol to observe the effects of high nickel concentrations on seedling growth. Observed biological makers included biomass, and the activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and phenylalanine ammonia-lyase (PAL) in the cotyledons. The fresh weight of cotyledons in all the tested nickel concentrations was lower than that of the control, but dry weight of cotyledons increased with increasing nickel concentrations up to 200 &mu;mol. SOD activity increased significantly up to 400 &mu;mol and then decreased at 800 &mu;mol nickel. POD activities were induced remarkably at 100 and 200 &mu;mol, but the activity decreased with increasing nickel concentrations. Similarly, a negative link between CAT activity and nickel concentrations was observed in this experiment. PAL activity had a positive correlation to nickel concentrations, and the highest activity was found at 400 &mu;mol nickel. Electrophoresis analysis suggested that a significant correlation between nickel concentrations and isoenzyme patterns of SOD and POD was observed, and these results were consistent with the changes of the activity assayed in solutions.

Rooting of Herbaceous and Semihardwood Cuttings of Guava cv. Século XXI, under Varying Concentrations of Indolebutyric Acid

2019 ◽  
pp. 1-9
Author(s):  
Jussara Cristina Firmino Da Costa ◽  
Rejane Maria Nunes Mendonça ◽  
Gerciane Cabral Da Silva ◽  
Silvanda de Melo Silva ◽  
Walter Esfrain Pereira ◽  
...  
Keyword(s):  
Experimental Design ◽  
Significant Interaction ◽  
Dry Weight ◽  
Psidium Guajava ◽  
Commercial Production ◽  
Control Treatment ◽  
Distilled Water ◽  
Indolebutyric Acid ◽  
Rooting Of Cuttings

In the commercial production of guava seedlings (Psidium guajava L.) the quality of the cuttings, homogeneity, high percentage of rooting are the factors important to be analyzed. Therefore, as the Século XXI cultivar is recent, it is necessary to do more studies regarding the behavior of this guava to the factors that aid in the rhizogenic process. The objective of this work was to verify the concentrations of indolebutyric acid (IBA) that provide the best rooting of cuttings herbaceous and semihardwood, as well as to verify the best kind of cuttings used for vegetative propagation the guava cuttings cultivar ‘Século XXI’. The experimental design was completely randomized in a 5 x 2 + 1 factorial design, beginning with five concentrations of IBA (0, 1000, 1500, 2000 and 2500 mg L-1), two kinds of cuttings (herbaceous and semihardwood) and one control treatment (immersed in distilled water for 12 hours), with four replications and 10 cuttings per plot. In relation of the variables: the percentage of cuttings live without roots, callus, number of roots and dry weight of shoots did not present significant interaction among treatments, not adjusting to any regression model. Therefore, concluding that the herbaceous cutting is the most indicated for the propagation of guava seedlings of ‘Século XXI’; The concentration of 2500 mg L-1 of IBA promotes a higher percentage of rooted semihardwood cuttings without leaf; The hormone solution diluted with alcohol 50% (v/v) resulted in toxicity for herbaceous and semihardwood cuttings cv. Século XXI.

scholarly journals Results with the establishment of in vitro culture of Leucojum aestivum

2007 ◽  
Vol 13 (2) ◽  
Author(s):  
E. Kohut ◽  
M. Ördögh ◽  
E. Jámbor-Benczúr ◽  
Á. Máthé
Keyword(s):  
Acetic Acid ◽  
Medicinal Plant ◽  
In Vitro Culture ◽  
In Vitro Cultures ◽  
Naphthalene Acetic Acid ◽  
Benzyl Adenine ◽  
Leucojum Aestivum ◽  
Leaves And Roots ◽  
Bulbous Plant

Leucojum aestivum is a native, protected ornamental and medicinal plant in Hungary and in Ukraine too. The aim of our work was to establish in vitro cultures of this bulbous plant. Prior to surface sterilisation the old leaves and roots were dissected from the bulbs and they were stored in a refrigerator (2-3°C) for different periods (1 week for the first starting experiment and 5 weeks for the second one). After sterilisation, bulbs, bulb scales and leaves of the bulbs were placed on Murashige and Skoog's (1962) medium with 1 mg/1 benzyl-adenine (BA) and 0,1 mg/1 naphthalene acetic acid (NAA). At the first starting experiment 81,3%, and at the second one 92,3% of the explants turned to be sterile. Bulblets and roots were developed on the explants in the case of using bulb plates together with bulb scales and leaves as inoculua. The best result was achieved after 5 weeks chilling and it was possible to gain little bulbs from the bulb leaves too.

scholarly journals In vitro cultures of Salvia officinalis L. as a source of antioxidant compounds

2011 ◽  
Vol 74 (1) ◽  
pp. 17-21 ◽  
Author(s):  
Izabela Grzegorczyk ◽  
Ireneusz Bilichowski ◽  
Elżbieta Mikiciuk-Olasik ◽  
Halina Wysokińska
Keyword(s):  
Cell Suspension ◽  
Rosmarinic Acid ◽  
Dry Weight ◽  
Suspension Cultures ◽  
In Vitro Cultures ◽  
Salvia Officinalis ◽  
Carnosic Acid ◽  
Shoot Cultures ◽  
Undifferentiated Callus

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.

LC-MS analysis of alkaloids in bulbs and in vitro cultures of Leucojum aestivum (Amaryllidaceae)

Planta Medica ◽  
2008 ◽  
Vol 74 (09) ◽  
Author(s):  
A Ptak ◽  
AE Tahchy ◽  
F Dupire ◽  
M Boisbrun ◽  
M Henry ◽  
...  
Keyword(s):  
In Vitro Cultures ◽  
Leucojum Aestivum ◽  
Ms Analysis

scholarly journals The effect of cytokinins on shoot proliferation, biochemical changes and genetic stability of Rhododendron ‘Kazimierz Odnowiciel’ in the in vitro cultures

2021 ◽  
Author(s):  
Karolina Nowakowska ◽  
Anna Pińkowska ◽  
Ewa Siedlecka ◽  
Andrzej Pacholczak
Keyword(s):  
Efficient Method ◽  
Plant Material ◽  
Soluble Sugars ◽  
Shoot Proliferation ◽  
Issr Markers ◽  
In Vitro Cultures ◽  
Biochemical Changes ◽  
Stock Plant

AbstractShoot proliferation is a very important micropropagation phase, decisive for economic efficiency of this method for a given taxon. To obtain a high multiplication ratio and a good quality of microshoots a detailed propagation protocol must be developed for particular species or even cultivars. Rhododendron ‘Kazimierz Odnowiciel’ is a relatively new cultivar distinguished by large, beautiful flowers and high frost resistance so there is a need to develop an efficient method of its propagation to satisfy a growing demand for this plant. The aim of the experiment was to evaluate effects of cytokinins: meta-Topolin (mT), zeatin (ZEA), 6-benzyladenine (BA), thidiazuron (TDZ), 2-isopentenyladenine (2iP), or the combination of 2iP+ZEA on proliferation of shoots in R. ‘Kazimierz Odnowiciel’ cultured on Anderson’s medium (AN). Biochemical changes in plant material affected by cytokinins during this phase of micropropagation were determined and occurrence of genetical changes was followed using ISSR markers. TDZ, ZEA or the combination of ZEA+2iP resulted in 100% explant regeneration. On the medium with TDZ or ZEA over two new shoots per explant were produced but the highest proliferation was attained on the medium containing ZEA+2iP – over three shoots per explant. Microshoots developed in this treatment had also the highest contents of chlorophyll, carotenoids and soluble sugars as well as the highest catalase activity. Microshoots formed on the medium with zeatin showed the lowest polymorphism (below 4%) relative to a stock plant.

Exogenous melatonin stimulated Amaryllidaceae alkaloid biosynthesis in in vitro cultures of Leucojum aestivum L

2019 ◽  
Vol 138 ◽  
pp. 111458 ◽  
Author(s):  
Agata Ptak ◽  
Magdalena Simlat ◽  
Emilia Morańska ◽  
Edyta Skrzypek ◽  
Marzena Warchoł ◽  
...  
Keyword(s):  
In Vitro Cultures ◽  
Alkaloid Biosynthesis ◽  
Exogenous Melatonin ◽  
Leucojum Aestivum

scholarly journals In vitro shoot cultures of pink rock-rose (Cistus ×incanus L.) as a potential source of phenolic compounds

2017 ◽  
Vol 86 (4) ◽  
Author(s):  
Paweł Kubica ◽  
Agnieszka Szopa ◽  
Halina Ekiert
Keyword(s):  
Phenolic Compounds ◽  
Gallic Acid ◽  
Dry Weight ◽  
In Vitro Cultures ◽  
Raw Material ◽  
Naphthaleneacetic Acid ◽  
Shoot Cultures ◽  
Epicatechin Gallate ◽  
Cistus Incanus

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.

scholarly journals Virus-Induced Flowering by Apple Latent Spherical Virus Vector: Effective Use to Accelerate Breeding of Grapevine

Viruses ◽  
2020 ◽  
Vol 12 (1) ◽  
pp. 70 ◽  
Author(s):  
Kiyoaki Maeda ◽  
Teppei Kikuchi ◽  
Ichiro Kasajima ◽  
Chungjiang Li ◽  
Noriko Yamagishi ◽  
...  
Keyword(s):  
In Vitro Cultures ◽  
F1 Hybrids ◽  
Experimental Conditions ◽  
Apple Latent Spherical Virus ◽  
Virus Rna ◽  
Biolistic Inoculation ◽  
Viable Seeds ◽  
Effective Use

Apple latent spherical virus (ALSV) was successfully used in promoting flowering (virus-induced flowering, VIF) in apple and pear seedlings. In this paper, we report the use of ALSV vectors for VIF in seedlings and in vitro cultures of grapevine. After adjusting experimental conditions for biolistic inoculation of virus RNA, ALSV efficiently infected not only progeny seedlings of Vitis spp. ‘Koshu,’ but also in vitro cultures of V. vinifera ‘Neo Muscat’ without inducing viral symptoms. The grapevine seedlings and in vitro cultures inoculated with an ALSV vector expressing the ‘florigen’ gene (Arabidopsis Flowering locus T, AtFT) started to set floral buds 20–30 days after inoculation. This VIF technology was successfully used to promote flowering and produce grapes with viable seeds in in vitro cultures of F1 hybrids from crosses between V. ficifolia and V. vinifera and made it possible to analyze the quality of fruits within a year after germination. High-temperature (37 °C) treatment of ALSV-infected grapevine disabled virus movement to newly growing tissue to obtain ALSV-free shoots. Thus, the VIF using ALSV vectors can be used to shorten the generation time of grapevine seedlings and accelerate breeding of grapevines with desired traits.

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