The Relative Amounts and Identification of Some 2,4-Dichlorophenoxyacetic Acid Metabolites Isolated from Soybean Cotyledon Callus Cultures

Sections of callus of Ranunculus sceleratus reveal its organization into cellular aggregates, the superficial cells of which are highly cytoplasmic and the inner cells of which are larger, have less-dense cytoplasm and are more highly vacuolated. Expansion and ultimate death of some of the internal cells leads to cell separation and break up of the aggregates. Many of the superficial cells are involved in the initiation of embryoids in the callus and they correspond in structure with the embryogenic cells of the seedling stem epidermis. The embryoids are retarded in their development in presence of 2,4-dichlorophenoxyacetic acid (2,4-D) and embryoid development becomes more rapid and more prolific when cultures are transferred to a medium without 2,4-D. The similarity between the embryogenic cells of the callus cultures and those of the seedling stem epidermis extends to cell size, nuclear size, degree of vacuolation, abundance of ribosomes and mitochondria, presence of amyloplasts and prominence of spherosomes. The various forms of spherosome are described and their possible function discussed. Amyloplasts differentiating into chloroplasts are observed in the more advanced embryoids. There is evidence that embryoids can arise from single cells but it is uncertain whether all are of single-cell origin. The embryogenic cells are in protoplasmic continuity with surrounding cells when they embark upon embryogenesis. Some of the superficial cells, also clearly undergoing active division, are of rather different structure; characteristically their nuclei show a high degree of chromatin condensation and their cytoplasm contains bundles of fibrous material. It is suggested that these cells do not function directly in embryogenesis. The internal cells of the aggregates have a low density of ribosomes and very few ER profiles or normal mitochondria. Extremely elongated mitochondrial structures following the outline of the nucleus are observed in these cells. An unidentified structure is frequently observed in cells in which cytoplasmic disorganization appears to be occurring.

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EFFICIENT CALLUS INDUCTION AND PLANT REGENERATION VIA SOMATIC EMBRYOGENESIS FROM IMMATURE LEAF-DERIVED PROTOPLASTS OF GROUNDNUT (ARACHIS HYPOGAEA L.)

Israel Journal of Plant Sciences ◽

10.1080/07929978.1996.10676660 ◽

1996 ◽

Vol 44 (4) ◽

pp. 387-396 ◽

Cited By ~ 3

Author(s):

Perumal Venkatachalam ◽

Narayanasamypillai Jayabalan

Keyword(s):

Somatic Embryogenesis ◽

Plant Regeneration ◽

Arachis Hypogaea ◽

Callus Cultures ◽

Alternative Methods ◽

Naphthalene Acetic Acid ◽

Dichlorophenoxyacetic Acid ◽

Ms Medium ◽

High Yields ◽

2,4 Dichlorophenoxyacetic Acid

High yields of protoplasts were obtained from immature leaves of aseptically grown plants of Arachis hypogaea using an enzyme solution containing cellulase 2.0% (w/v) and Macerozyme 1.0% (w/v) in 0.6 M mannitol. Isolated protoplasts were cultured in Kao's medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzylaminopurine (BAP). The protoplasts started to divide after 3–5 days of culture. Sustained divisions resulted in mass production of cell colonies and mini calli in 4 weeks. After 4 weeks, protoplast colonies were transferred to the Murashige and Skoog (MS) medium supplemented with a-naphthalene acetic acid (NAA) and BAP. Colonies proliferated into actively growing calli. Further attempts to regenerate plants from such calli were not successful. However, protoclones differentiated roots on the same medium. Alternative methods for plant regeneration from protoplast derived callus cultures were tried through somatic embryogenesis. Protoplast-derived calli treated with 2,4-D and BAP formed somatic embryos. Somatic embryogenesis began in the proembryo stage and proceeded from globular to dicotyledonary stage. Embryos were then transferred onto hormone-free MS medium for germination. Five to ten percent of these embryoids germinated and grew to plantlets. Regenerated plants were transferred to plastic cups and grown to maturity.

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Production of Anthocyanins in Callus Cultures of Angelica archangelica

Natural Product Communications ◽

10.1177/1934578x1801301219 ◽

2018 ◽

Vol 13 (12) ◽

pp. 1934578X1801301

Author(s):

Tomáš Siatka

Keyword(s):

Woody Plant ◽

Callus Cultures ◽

Attractive Alternative ◽

Dichlorophenoxyacetic Acid ◽

Angelica Archangelica ◽

Callus Proliferation ◽

Order Of Magnitude ◽

Culture Growth ◽

2,4 Dichlorophenoxyacetic Acid ◽

Production Characteristics

Anthocyanins have been used as food color additives, but they also possess many properties beneficial to health. Plant tissue culture technology is an attractive alternative for obtaining these valuable natural pigments. In this work, dark-grown anthocyanin producing callus cultures of Angelica archangelica were established. They were cultured on a Murashige and Skoog medium supplemented with 2 mg/L 2,4-dichlorophenoxyacetic acid and 0.4 mg/L benzylaminopurine. Anthocyanin contents in cultures were around 2%, i.e. one order of magnitude higher than in the intact plant that contains up to 0.17% anthocyanins. Growth and production characteristics of the culture were determined – fresh and dry biomass as well as anthocyanin levels reached a maximum on day 30. Effects of basal nutrient media on callus proliferation and anthocyanin accumulation were tested. Culture growth (fresh weight) achieved 105%, 102%, 141%, 129%, 54%, and 26%, and anthocyanin contents attained 114%, 41%, 33%, 31%, 25%, and 15% on Linsmaier and Skoog, Gamborg B5, Schenk and Hildebrandt, Woody plant, Nitsch and Nitsch, and Heller medium, respectively, in comparison with that of Murashige and Skoog.

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Isolation and cultivation of protoplasts from morphogenetic callus cultures of Lilium

Canadian Journal of Botany ◽

10.1139/b79-066 ◽

1979 ◽

Vol 57 (5) ◽

pp. 512-516 ◽

Cited By ~ 8

Author(s):

John A. Simmonds ◽

Daina H. Simmonds ◽

Bruce G. Cumming

Keyword(s):

Cell Wall ◽

Culture Media ◽

Continuous Light ◽

Callus Cultures ◽

Sodium Fluorescein ◽

Dichlorophenoxyacetic Acid ◽

Naphthaleneacetic Acid ◽

Cell Wall Formation ◽

Wall Formation ◽

2,4 Dichlorophenoxyacetic Acid

Protoplasts isolated from Lilium callus which was maintained on media containing 2% sucrose contained large deposits of starch granules and lysed during isolation and washing procedures. Stable protoplast preparations could be obtained from callus which had been subcultured on sucrose-free medium for 3 weeks. Maximum protoplast yield (1.5 × 106 per gram fresh weight) was obtained when KCl (0.3 M) was the osmotic stabilizer. Inclusion of CaCl2 (25 mM) and MgSO4 (25 mM) in the isolation and wash media decreased protoplast lysis. Viability of protoplasts isolated in the high salts medium was determined by their ability to accumulate sodium fluorescein in the cytoplasm. No cell-wall formation occurred when salts were used as the osmoticum in various culture media. Continuous light (5000 lx) was inhibitory to protoplast survival. When protoplasts were transferred, via a series of wash solutions, to culture media using sugars as the osmoticum and cultured in darkness, cell-wall formation was detected after 3 days and cell divisions after 21 days. Zeatin (10−6 M), was needed for cell-wall formation. Cell division was stimulated by a combination of zeatin (10−6 M), naphthaleneacetic acid (10−5 M), and 2,4-dichlorophenoxyacetic acid (10−7 M) in the basic nutrient medium.

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The occurrence of polar structures in callus cultures from mature lodgepole pine (Pinus contorta var. latifolia)

Canadian Journal of Botany ◽

10.1139/b88-351 ◽

1988 ◽

Vol 66 (12) ◽

pp. 2595-2596

Author(s):

Susan C. MacDougall ◽

Shona M. Ellis ◽

Iain E. P. Taylor

Keyword(s):

Pinus Contorta ◽

Zygotic Embryo ◽

Lodgepole Pine ◽

Leaf Explants ◽

Callus Cultures ◽

Small Cells ◽

Dichlorophenoxyacetic Acid ◽

Pine Trees ◽

Embryo Cells ◽

2,4 Dichlorophenoxyacetic Acid

A somatic polar structure was observed in white callus cultured, in the presence of 2,4-dichlorophenoxyacetic acid (10−6 M) and benzylaminopurine (4 × 10−6 M), from leaf explants taken from mature lodgepole pine trees. The structure contained elongate, vacuolate cells and small cells arranged with some resemblance to the first zygotic embryo cells. We were not able to induce further development.

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Prenylated Flavanone Production in Callus Cultures of Sophoraßavescens var. angustifolia

Zeitschrift für Naturforschung C ◽

10.1515/znc-1991-3-402 ◽

1991 ◽

Vol 46 (3-4) ◽

pp. 172-176 ◽

Cited By ~ 15

Author(s):

Hirobumi Yamamoto ◽

Sumiko Kawai ◽

Junko Mayumi ◽

Toshiyuki Tanaka ◽

Munekazu Iinuma ◽

...

Keyword(s):

Cell Growth ◽

Inverse Relationship ◽

Time Course ◽

Callus Cultures ◽

The Other ◽

Dichlorophenoxyacetic Acid ◽

Skoog Medium ◽

Other Hand ◽

Murashige Skoog ◽

2,4 Dichlorophenoxyacetic Acid

Callus cultures of Sophora favescens var. angustifolia established on Murashige-Skoog medium containing 1 μм 2,4-dichlorophenoxyacetic acid and 1 μм kinetin produced the prenylated flavanones (2S)-5,7,2′,4′-tetrahydroxy-8-lavandulylflavanone (sophoraflavanone G) and (25)-7,2′,4′-trihydroxy-8-lavandulylflavanone (lehmannin). In addition, maackiain and its 3-O-β-glucoside (trifolirhizin) were also produced in the callus. Up on transfer to White’s medium or M9 medium, the content of prenylated flavanones, in particular lehmannin, was increased, whereas that of pterocarpans was decreased. Time-course experiments indicated that the production of pterocarpans was closely related with cell growth. On the other hand, an inverse relationship existed between cell growth and the production of prenylated flavanones.

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Somatic embryogenesis and plant regeneration from callus cultures of Cleome rosea Vahl

Brazilian Archives of Biology and Technology ◽

10.1590/s1516-89132010000300024 ◽

2010 ◽

Vol 53 (3) ◽

pp. 679-686 ◽

Cited By ~ 7

Author(s):

Claudia Simões ◽

Norma Albarello ◽

Cátia Henriques Callado ◽

Tatiana Carvalho de Castro ◽

Elisabeth Mansur

Keyword(s):

Somatic Embryogenesis ◽

Callus Formation ◽

Callus Cultures ◽

Dichlorophenoxyacetic Acid ◽

Naphthaleneacetic Acid ◽

Stem Explants ◽

Ms Medium ◽

Fold Reduction ◽

2,4 Dichlorophenoxyacetic Acid ◽

Embryogenic Callus Formation

This paper describes a protocol for the efficient vegetative propagation of Cleome rosea by somatic embryogenesis. Leaf and stem explants from nursery-grown seedlings of C. rosea were cultivated on Murashige and Skoog (MS) medium supplemented with indole-3-acetic acid (IAA), a -naphthaleneacetic acid (NAA), 4-amino-3,5,6-trichloropicolinic acid (picloram) or 2,4-dichlorophenoxyacetic acid (2,4-D). Nodular calli were produced from both explant types in the presence of 4.5 and 9.0 µM 2,4-D. Embryo development and maturation were achieved when calli from stem explants were transferred to media containing a ten-fold reduction of 2,4-D concentration initially used (0.45 and 0.90 µM). Leaf-derived calli did not form embryos with the same treatments. The highest frequency of embryogenic callus formation (85%) and number of embryo per callus (13.45 ± 2.8) were achieved during the first subculture on medium supplemented with 0.90 µM 2,4-D. Embryo conversion into plantlets was achieved following transfer to growth regulator-free MS medium solidified with 2 g.L-1 phytagel. An acclimatization rate of 53% was found three months after transfer to ex vitro conditions and the recovered plants presented a normal phenotypic aspect.

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Growth, ferulic acid synthesis, and histochemistry of calli of Pouteria caimito (Ruiz & Pav.) Radlk under different light qualities

Research Society and Development ◽

10.33448/rsd-v9i8.5822 ◽

2020 ◽

Vol 9 (8) ◽

Author(s):

Anielly Monteiro Melo ◽

Márcio Rosa ◽

Luciana Arantes Dantas ◽

Paulo Sérgio Pereira ◽

Sebastião Carvalho Vasconcelos Filho ◽

...

Keyword(s):

Ferulic Acid ◽

Light Quality ◽

Acid Synthesis ◽

Callus Cultures ◽

Dichlorophenoxyacetic Acid ◽

Liquid Chromatograph ◽

Histochemical Tests ◽

Ferulic Acid Production ◽

2,4 Dichlorophenoxyacetic Acid

Interest in harnessing biological processes for the production of bioactive compounds from natural sources has increased considerably. The manipulation of light quality in callus culture is considered a promising strategy for in vitro metabolite production. The objective of this study was to investigate the influence of light quality on the growth, histochemistry, and ferulic acid production of callus cultures of P. caimito. For in vitro callus induction, 1-cm2 leaf fragments were cultured in 50% MS medium supplemented with 2,4-dichlorophenoxyacetic acid and benzylaminopurine in the absence or presence of light (white, blue, green, yellow, or red). Methanol extraction was performed with partitioning of the extract and subsequent quantification of ferulic acid using a liquid chromatograph coupled to a mass spectrometer. The presence of light promoted greater growth than the absence of light. In the interaction between light quality and culture time, linear biomass growth until 28 days was observed under yellow, red, and blue lights and in the dark. The highest callus biomass values were observed under yellow and red lights. The histochemical tests showed the presence of phenolic compounds, alkaloids, flavonoids, and terpenes. The exposure of calli cultured under white light to different light qualities and culture times did not result in significant differences in the concentration or yield of ferulic acid.

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Promotion by tryptophan of growth and root formation in lowbush blueberry pericarp callus cultures

Canadian Journal of Botany ◽

10.1139/b80-112 ◽

1980 ◽

Vol 58 (8) ◽

pp. 881-885 ◽

Cited By ~ 3

Author(s):

Nancy L. Nickerson

Keyword(s):

Growth Rates ◽

Root Formation ◽

Indoleacetic Acid ◽

Callus Cultures ◽

Callus Growth ◽

Dichlorophenoxyacetic Acid ◽

Naphthaleneacetic Acid ◽

Indolebutyric Acid ◽

Lowbush Blueberry ◽

2,4 Dichlorophenoxyacetic Acid

Lowbush blueberry (Vaccinium angustifolium Ait.) pericarp callus grew slowly and formed normal tetraploid roots on Nitsch's medium containing L-tryptophan and kinetin. Both growth and rooting depended on the levels of these two substances in the medium. Rooting declined but callus growth rates changed little over successive subcultures. When tryptophan was replaced by indoleacetic acid, indolebutyric acid, 2,4-dichlorophenoxyacetic acid, or naphthaleneacetic acid, callus growth rates increased but no roots formed. Tryptophan medium did not support callus growth or induce rooting unless the tryptophan was autoclaved with the rest of the medium; thus suggesting that an active substance is produced by reaction of the tryptophan with some other constitutent(s) of the medium during heating.

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