Preliminary material

Stern 'chips' from large-diameter stern regions of Larix laricina were produced free of contaminating organisms. These chips, consisting of dormant vascular cambium sandwiched intact between mature xylem and phloem, were grown as in vitro cultures on the surface of an agar medium that supports apparently normal cambial cell division and xylogenesis. It was determined that auxin (l-naphthaIene acetic acid) together with nutrients at low concentrations in the medium was essential for cambial growth, whether chips were grown in continuous light or darkness. Fusiform cambial cells underwent successive periclinal divisions to produce radial files of enlarged, bordered-pitted, secondary-walled, lignified and autolysed earlywood tracheids. As many as 20 new tracheids per radial file were produced, and > 95% of these were autolysed after 35 days of culture.

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Comparative Analysis of Therapeutically Important Indole Compounds in in vitro Cultures of Hypericum perforatum Cultivars by HPLC and TLC Analysis Coupled with Densitometric Detection

Natural Product Communications ◽

10.1177/1934578x1400901009 ◽

2014 ◽

Vol 9 (10) ◽

pp. 1934578X1400901 ◽

Cited By ~ 2

Author(s):

Bożena Muszyńska ◽

Halina Ekiert ◽

Inga Kwiecień ◽

Anna Maślanka ◽

Rawad Zodi ◽

...

Keyword(s):

Acetic Acid ◽

Growth Regulators ◽

Hypericum Perforatum ◽

Dry Mass ◽

In Vitro Cultures ◽

Indole Compounds ◽

Densitometric Detection ◽

Tlc Analysis

Five indole compounds (5-hydroxy-L-tryptophan, L-tryptophan, indole-3-acetic acid, melatonin, serotonin) and hypericin were identified and quantified in methanolic extracts of shoot cultures of three Hypericum perforatum cultivars (Helos, Elixir, Topas) growing on two variants of Murashige -Skoog medium differing in concentrations of growth regulators (naphthalene-1-acetic acid and 6-benzylaminopurine). Extracts of the aboveground parts of field-grown plants ( Hyperici herba) were also analyzed by HPLC and TLC analysis coupled with densitometric detection. Determination of four compounds was based on our assay described earlier. The methods of determination of 5-hydroxy-L-tryptophan and hypericin were developed and validated in this study. The composition and contents of the metabolites under study differed between the cultivars cultured in vitro and between medium variants containing diverse contents of growth regulators. The contents of individual indole compounds in the biomass from in vitro cultures ranged from 39.6 to 343.2 mg/100 g dry mass. 5-Hydroxy-L-tryptophan was the dominating metabolite (from 78.2 to 343.2 mg/100 g dry mass). Extracts from shoots of the cultivar Helos also contained high contents of serotonin (319.9 and 197.4 mg/100 g dry mass). The contents of indole compounds in Hyperici herba were also diverse (from 7.1 to 55.3 mg/100 g dry mass). 5-Hydroxy-L-tryptophan was the dominating metabolite as well. Hypericin content of Hyperici herba, equaling 12.2 mg/100 g dry mass was from 3.3 to 10 times higher than in extracts from shoots cultured in vitro. The present report is the first analysis of endogenous accumulation of indole compounds in Hyperici herba which involves, apart from melatonin, four other compounds.

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Results with the establishment of in vitro culture of Leucojum aestivum

International Journal of Horticultural Science ◽

10.31421/ijhs/13/2/723 ◽

2007 ◽

Vol 13 (2) ◽

Cited By ~ 1

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.

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Growth and Secondary Product Formation of in vitro Cultures from the Liverwort Reboulia hemisphaerica

Zeitschrift für Naturforschung C ◽

10.1515/znc-1991-1-205 ◽

1991 ◽

Vol 46 (1-2) ◽

pp. 28-32 ◽

Cited By ~ 7

Author(s):

Rui M. S. C. Morais ◽

Hans Becker

Keyword(s):

Agar Medium ◽

Product Formation ◽

Suspension Cultures ◽

Light Regime ◽

In Vitro Cultures ◽

Secondary Product ◽

Naphthylacetic Acid

Abstract Reboulia hemisphaerica has been cultivated on Gamborg B5 medium with various sugars. Differentiated cultures grew best on agar medium with the addition of 0.5% sucrose and a light regime of 18 h light/6h dark . Callus was induced either by a combination of phytohormones (2 mg/l α-naphthylacetic acid + 1 mg/l kinetin) or by 4% glucose. The differentiated cultures on agar produced sesquiterpenes in a tenfold increased amount compared to undiffer entiated suspension cultures. Cultures kept in da rk produced no sesquiterpenes, but 5-hydroxy-7,8,4′-trimethoxyflavone was present whereas apigenin-7,4′-dimethylether was the only flavonoid in cultures cultivated in the light.

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A bioassay for systemic fungicides using axenic cultures of the carnation rust fungus, Uromyces dianthi

Canadian Journal of Botany ◽

10.1139/b73-260 ◽

1973 ◽

Vol 51 (10) ◽

pp. 2004-2006 ◽

Cited By ~ 1

Author(s):

D. R. Jones

Keyword(s):

Inhibitory Action ◽

Agar Medium ◽

Rust Fungus ◽

Screening Programs ◽

Systemic Fungicides ◽

Low Concentrations ◽

Wide Range ◽

Axenic Cultures ◽

High Concentrations

The inhibitory action of three systemic fungicides (Benlate, Hoe 6053, and Milstem) on the growth of axenic cultures of the carnation rust fungus, Uromyces dianthi, was assessed. The rust- and smut-specific fungicide (Hoe 6053) was found to inhibit growth when present in the agar medium in very low concentrations, while moderate concentrations of the wide range fungicide (Benlate), and high concentrations of the powdery mildew-specific fungicide (Milstem) were needed for inhibition. The results indicated the possible use of U. dianthi in future in vitro fungicide screening programs and mode-of-action studies.

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REGENERASIRUMPUT LAUT Kappaphycus alvarezii (Doty) MELALUI INDUKSI KALUS DAN EMBRIO DENGAN PENAMBAHAN HORMON PERANGSANG TUMBUH SECARA IN VITRO

Jurnal Riset Akuakultur ◽

10.15578/jra.4.1.2009.39-45 ◽

2009 ◽

Vol 4 (1) ◽

pp. 39

Author(s):

Emma Suryati ◽

Sri Rejeki Hesti Mulyaningrum

Keyword(s):

Acetic Acid ◽

Growth Regulator ◽

Culture Medium ◽

Solid Medium ◽

Agar Medium ◽

Kappaphycus Alvarezii ◽

Indol Acetic Acid ◽

High Quality Seed ◽

Medium Culture

Regenerasi rumput laut Kappaphycus alvarezii dilakukan dalam rangka penyediaan benih yang bermutu dan mempunyai keunggulan melalui induksi kalus dan embrio dengan penambahan hormon pertumbuhan yang diintroduksi ke dalam media kultur yang dapat memacu induksi kalus dan penebalan pigmen rumput laut. Media kultur yang digunakan adalah media Conwy padat dengan penambahan agar 0,8%-1,6%. Hormon perangsang tumbuh yang digunakan untuk memacu pertumbuhan kalus dan filamen embrio yaitu IAA (Indol acetic acid), kinetin, dan auxilin dengan konsentrasi berkisar 0,4-1 mg/L. Embrio yang dihasilkan merupakan anakan yang mempunyai sifat yang sama dengan induknya. Sintasan dan perkembangan embrio yang paling baik yaitu dengan penambahan IAA dengan konsentrasi 0,4 mg/L pada media padat. Pembentukan anakan dilakukan dengan mengiris embrio dan menumbuhkan pada media cair yang diperkaya dengan hormon yang sama. Pemeliharaan anakan pada media kultur dilakukan hingga mencapai ukuran 2-3 cm.Regeneration of seaweed Kappaphycus alvarezii was done to provide high quality seed through callus and embryo induction using plant growth regulator which was introducted to the culture medium. This growth regulator can stimulate the callus induction procces and thickening the seaweed pigment. Applied medium culture was agar medium with 0.8%-1.6% concentration enriched with Conwy and the applied growth regulators were IAA (Indol acetic acid), kinetin dan auxilin with 0.4-1 mg/L concentration range. Resulted embryo has the same characteristics with the stock. The best survival rate and embryo growth was IAA treatment with 0.4 mg/L concentration. Formation of embryo was done by transferring them from solid medium to the liquid one with the same growth regulator treatment. The nursery of the seed in culture medium was carried out until it has reached 2-3 cm in size.

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Coniferin, a biochemical indicator of commitment to tracheid differentiation in conifers

Canadian Journal of Botany ◽

10.1139/b89-343 ◽

1989 ◽

Vol 67 (9) ◽

pp. 2663-2668 ◽

Cited By ~ 39

Author(s):

Rodney Arthur Savidge

Keyword(s):

Acetic Acid ◽

Picea Glauca ◽

Pinus Banksiana ◽

High Performance ◽

Wood Formation ◽

Defined Medium ◽

Naphthalene Acetic Acid ◽

Larix Laricina ◽

Tracheid Differentiation

Endogenous E-coniferin was characterized chemically and found to accumulate in cambia of Picea glauca, Pinus banksiana, Pinus strobus, and Larix laricina at the time of resumption of springtime cell-division activity and well before the initiation of lignification in differentiating tracheids. Coniferin content, quantified by high performance liquid chromatography, was highest on the centripetal side of the cambium, in developing xylem, at time of radial expansion of cambial derivatives. Stem ringing resulted in coniferin accumulation above the ring and disappearance below the ring. Coniferin was absent from leaves and dormant cambia. Exogenous coniferin was metabolized rapidly by in vitro cultures of P. strobus cambium grown on a defined medium containing the auxins 1-naphthalene acetic acid and indol-3-yl acetic acid and the cytokinin benzylaminopurine. The evidence suggests coniferin content of the cambium will be of practical value for determining commitment to and competence for wood formation in conifers.

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Polyurethane Foam Rafts Supported In Vitro Cultures of Rindera graeca Roots for Enhanced Production of Rinderol, Potent Proapoptotic Naphthoquinone Compound

International Journal of Molecular Sciences ◽

10.3390/ijms23010056 ◽

2021 ◽

Vol 23 (1) ◽

pp. 56

Author(s):

Mateusz Kawka ◽

Irena Bubko ◽

Mirosława Koronkiewicz ◽

Beata Gruber-Bzura ◽

Konstantia Graikou ◽

...

Keyword(s):

In Vitro Cultures ◽

Efficient Production ◽

Anticancer Properties ◽

Enhanced Production ◽

Product Removal ◽

Low Concentrations ◽

Cervical Cancer Cells ◽

Fast Acting

Unique phytochemical profile of plants belonging to Boraginaceae family provides a prolific resource of lipophilic pigments from the group of naphthoquinone derivatives. To overcome low compound content, the major obstacle of plant-based production, immobilization of Rindera graeca roots in in vitro cultures was implemented for efficient production of rinderol, novel furanonaphthoquinone derivative with anticancer properties. Chromatographic procedures revealed rinderol presence in extracts of all investigated root lines, derived both from root biomass and post-culture medium. Unexpectedly, in the second stage of the experiment, rinderol production was ceased in control, unmodified culture systems. On the contrary, roots immobilized on PUF rafts uniformly and stably produced rinderol, and its highest amount was noted for transformed root lines after 42 days of cultivation (222.98 ± 10.47 µg/flask). PUF occurred to be the main place of compound accumulation. Moreover, investigation of rinderol biological activity revealed its fast-acting cell death induction in HeLa cervical cancer cells at relatively low concentrations. Presented results revealed successful application of R. graeca roots immobilization on PUF rafts for production and in situ product removal of rinderol, novel lipophilic furanonaphthoquinone with suggested proapoptotic activity.

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