BRIDELIA SCANDENS WILD. A NEW SOURCE FOR PODOPHYLLOTOXIN PRODUCTION IN VITRO BY FEEDING CONIFERYL ALCOHOL

Objective: In the present study, a new method for the production of anticancerous compound podophyllotoxin (PTOX) was developed for Bridelia scandens Wild. by feeding coniferyl alcohol. Methods: The production of anticancerous compound PTOX through leaf explant derived calli of B. scandens. Murashige and Skoog (MS) medium fortified with 0.5 mg/l 6-Benzylaminopurine (BAP) and 0.5 mg/l 2,4-D (2,4-Dichlorophenoxyacetic acid) induced luxuriant mass of callus growth. Suspension culture was initiated by sterile MS media fortified with 0.1–1.0 mg/l BAP and 0.1–1.0 mg/l 2,4-D. and growth product was analyzed by the high-pressure liquid chromatography method. Results: Phytochemical analysis of the B. scandens leaf and leaf calli showed the presence of PTOX at the concentrations of 0.69 and 1.81, respectively. The callus cell suspension was established with the same callogenic media also it is augmented with 10–70 mg/l of coniferyl alcohol to elicit the biosynthesis of PTOX. Successive cultures of the calli suspension yielded stable production of PTOX of 3.91 mg/g dry cell weight at 50 mg/l coniferyl alcohol in the media. The biosynthesis of PTOX was ideal when plant cells were cultivated in the dark with an agitation speed of 100 rpm. Conclusion: The growth and production of PTOX were found to be better with glucose than with sucrose as the medium carbon source. The harvesting of the secondary metabolite from the in vitro grown leaf calli of B. scandens is a better way to stop the exploitation of medicinal plants.

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Induction, biochemical trait and phytochemical screening of calluses of Amburana cearensis (Allemão) A.C. Smith

Acta Scientiarum Biological Sciences ◽

10.4025/actascibiolsci.v42i1.54187 ◽

2020 ◽

Vol 42 ◽

pp. e54187

Author(s):

Jéssica Nascimento Costa Vascocelos ◽

Alone Lima Brito ◽

Andressa Priscila Pianco Santos Lima ◽

Jackson Roberto Guedes da Silva Almeida ◽

Ana Paula de Oliveira ◽

...

Keyword(s):

Growth Curve ◽

High Performance ◽

Reducing Sugars ◽

Dichlorophenoxyacetic Acid ◽

Phytochemical Analysis ◽

Phytochemical Screening ◽

Total Proteins ◽

Biochemical Tests ◽

2,4 Dichlorophenoxyacetic Acid

Amburana cearensis is an arboreal legume of the Fabaceae family, with high phytotherapic and medicinal potential due the presence of secondary metabolites. The objective of this study was to evaluate the effect of 2,4-dichlorophenoxyacetic acid (2,4-D) and 4-amino-2,5,6- trichloro-2-pyridinecarboxylic acid (picloram) on the in vitro induction of callogenesis of A. cearensis and analyze the biochemical and phytochemical potential of these calluses. For callus induction, leaf and cotyledon segments were used as explants, which were inoculated in woody plant medium (WPM) supplemented with different concentrations of 2,4-D (0, 5, 10, 20, 40 μM) or picloram (0, 5, 10, 20, 40, 80 μM). The callus growth curve was estimated based on fresh weight, measured at 7-day intervals until 28 days after inoculation. The calluses were analyzed by biochemical tests to quantify the reducing sugars and total proteins. Phytochemical screening and high-performance liquid chromatography were performed to establish the phytochemical profile of extracts from calluses. The concentrations of 21.94 μM and 26.46 μM of 2,4-D induced the greatest formation of compact and friable calluses from the leaf and cotyledon segments, respectively. The growth curve had two distinct phases (lag and exponential) for both types of calluses evaluated. The maximum levels of reducing sugars and total proteins in the calluses from leaf and cotyledon segments were obtained on the day of inoculation and after 28 days of cultivation, respectively. The results of the phytochemical analysis identified the presence of coumarin in all the extracts evaluated, this secondary metabolite has high pharmacological potential.

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Somatic embryogenesis of Sandalwood (Santalum album L.)

Indonesian Journal of Biotechnology ◽

10.22146/ijbiotech.9311 ◽

2016 ◽

Vol 19 (2) ◽

pp. 168

Author(s):

Toni Herawan ◽

Mohammad Na'iem ◽

Sapto Indrioko ◽

Ari Indrianto

Keyword(s):

Somatic Embryogenesis ◽

Embryo Development ◽

Native Species ◽

Economic Value ◽

Dichlorophenoxyacetic Acid ◽

Santalum Album ◽

The Media ◽

2,4 Dichlorophenoxyacetic Acid ◽

Santalum Album L

Sandalwood (Santalum album L.) is native species of Indonesia, especially in East Nusa Tenggara, is oneof the twenty two species of the genus Santalum in the world. Sandalwood is an important tree because it hashigh economic value can produce sandal oil these can be used for perfumes, cosmetics, pharmaceuticals, andare often used in religious ceremonies. In vitro particularly somatic embryogenesis has been widely appliedin the propagation of sandalwood. The Objective of this research is to obtain regeneration of sandalwoodthrough somatic embryogenesis using leaves explant from various clones. Medium for embryo induction is MS(Murashige and Skoog, 1962) solid medium containing treatment of 2,4-D (2,4-Dichlorophenoxyacetic acid)at various concentrations. To the media 0,15 mg /l kinetin, 40 g/l sucrose, and 2,5 g/l gelrite were added.Culture were incubated in the dark. Medium for Embryo development (maturation) is MS solid mediumcontaining treatment of BAP (Benzyl-amino-purine) at various concentrations. To the media 0,01 mg /l NAA(Napthalene-acetic-acid), 40 g/l sucrose, and 2,5 g/l gelrite were added. Culture were incubated in the light. Tostudy the specifi c structure of sandalwood somatic embryo early detection was conducted using histologicalanalysis. Results of anova showed that the clones, media, and interaction between clones with media did notsignifi cantly affect the development of sandalwood callus percentage. Results of anova showed that the clonesand BAP concentration signifi cantly effect to the embryo development of sandalwood.

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Histology of Microspores of Chile Apple Capsicum pubescens R and P. and in vitro Culture

Biotechnology and Bioprocessing ◽

10.31579/2766-2314/047 ◽

2021 ◽

Vol 2 (7) ◽

pp. 01-06

Author(s):

J. L. Rodríguez-de la O ◽

F. Pérez-Pérez ◽

M. Pérez-Grajales

Keyword(s):

In Vitro Culture ◽

Culture Media ◽

Plant Biotechnology ◽

Pollen Grains ◽

Dichlorophenoxyacetic Acid ◽

The Media ◽

Pure Lines ◽

2,4 Dichlorophenoxyacetic Acid ◽

Short Time

In plant biotechnology, in vitro culture of gametic or sexual cells, microspores or pollen grains, has been described as a successful tool to accelerate genetic improvement, obtaining haploid, homozygotic plants or pure lines in a short time. In chile apple, Capsicum pubescens R and P. Anthers were sown in vitro, and their cytological analysis, locating the meiotic division stage of microspores or pollen grains. Flower buds with diameters from 2.5 to 4.4 mm were pre-incubated at 4°C, in ascorbic and citric acid at 100 and 150 mg-L-1 for 24 h. Five semisolid culture media (A1, A2, A3, A4 and A5) were used, with Murashige and Skoog (1962) salts (MS), modifying iron and vitamin chelates, sucrose, and L-cysteine, 2,4-dichlorophenoxyacetic acid (2,4-D) and Kinetin (Kin). Anthers, in vitro, were plated, in light and dark, for 70 days. Two differentiation media (R1 and R2) were evaluated with 100% MS salts, glycine, kinetin and myo-inositol. The anthers seeded, coincided with the first mitosis of the microspore, the anthers, formed callus in the media (A1) 100 % EDTA-Fe, 0.40 mg-L-1 thiamine, 3 % sucrose) and (A3) 100 % EDTA-Fe, 0.40 mg-L-1 thiamine, 3 % sucrose, 0. 3 mg-L-1 of 2,4-D, and differentiated pro-embryonic structures in (A3) and (A5) 200 % EDTA-Fe, 0.4 mg-L-1 thiamine, 50 mg-L-1 pyridoxine, folic acid, riboflavin and niacin, 0.3 mg-L-1 2,4-D plus 0.3 mg-L-1 Kinetin, as well as roots in (A1). Light influenced the formation of pro-embryos and roots, in the dark callus. The media (R1) and (R2) favored the formation of pro-embryos.

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Haploid Induction via In Vitro Gynogenesis in Persian Shallot (Allium hirtifolium)

Journal of Horticultural Research ◽

10.2478/johr-2019-0017 ◽

2019 ◽

Vol 27 (2) ◽

pp. 91-98

Author(s):

Jaber Panahandeh ◽

Nasrin Farhadi

Keyword(s):

Somatic Tissue ◽

Dichlorophenoxyacetic Acid ◽

Naphthaleneacetic Acid ◽

Haploid Induction ◽

Breeding Programs ◽

Genetic Studies ◽

The Media ◽

2,4 Dichlorophenoxyacetic Acid ◽

Allium Hirtifolium

AbstractHaploid induction using in vitro cultures of unpollinated flowers has been recognized as an important tool to produce homozygous plants for genetic studies and breeding programs. In this study the potential of gynogenic haploid induction in four ecotypes of Allium hirtifolium under different combinations of benzylaminopurine (BAP) with 2,4-dichlorophenoxyacetic acid (2,4-D), or α-naphthaleneacetic acid (NAA) was investigated. Unpollinated flower buds were excised from an umbel 5 to 3 days before anthesis, and cultured onto B5 medium containing 7.5% sucrose and 2 mg·dm−3 BAP with auxin. The experiments revealed that NAA increased the percentage of gynogenesis induction and number of gynogenic embryos per flower in all ecotypes. Somatic organogenesis from basal callus or other floral parts was most effective on the media containing 2,4-D. Plants obtained by gynogenesis were haploid in 70–77% and plants from somatic tissue were mostly diploid.

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Direct Shoot Regeneration from Vaccinium pahalae (Ohelo) and V. myrtillus (Bilberry) Leaf Explants

HortScience ◽

10.21273/hortsci.31.7.1225 ◽

1996 ◽

Vol 31 (7) ◽

pp. 1225-1228 ◽

Cited By ~ 15

Author(s):

Rida A. Shibli ◽

M.A.L. Smith

Keyword(s):

Callus Formation ◽

Leaf Explants ◽

Shoot Proliferation ◽

Direct Organogenesis ◽

Dichlorophenoxyacetic Acid ◽

Naphthaleneacetic Acid ◽

Direct Shoot Regeneration ◽

The Media ◽

2,4 Dichlorophenoxyacetic Acid

Ohelo (V. pahalae Skottsb.) and bilberry (V. myrtillus L.) shoots were regenerated via direct organogenesis from whole leaves and leaf sections and also from hypocotyl explants of bilberry. Explants preincubated for 1 to 2 weeks in darkness yielded ≈75% regeneration frequencies and the highest number of regenerating shoots/explant on TDZ-supplemented media (0.9 to 2.7 μm). When 2iP or zeatin were substituted as the cytokinin source, frequencies of regeneration and shoot productivity were significantly lower. Explants held under constant illumination (no dark pretreatment) had significantly lower regeneration frequencies in all tested cytokinin-supplemented media. 2,4-D stimulated callus formation, but did not support regeneration from vegetative explants. Cells from callus and suspension cultures did not exhibit regeneration in any of the media that supported organogenesis from leaves. Regenerants were successfully micropropagated, although callus formation caused by zeatin and high 2iP levels interfered with shoot proliferation. Zeatin induced hyperhydricity in shoots from both species, but more severely in ohelo. Ex vitro rooting after treatment with 4.9 μm IBA or 5.4 μm NAA was 95% and 60% successful for bilberry and ohelo, respectively, and plants were readily acclimatized after an interval in a fog chamber. Bilberry microshoots also rooted in vitro in the absence of growth regulator treatment. Chemical names used: 1H-indole-3-butanoic acid (IBA); N-(3-methyl-2-butenyl)-1-H-purine-6-amine (2iP); 6-furfurylaminopurine (kinetin); 1-naphthaleneacetic acid (NAA); thidiazuron=1-phenyl-3-(1,2,3-thiadiazio-5-yl)urea (TDZ); 2,4-dichlorophenoxyacetic acid (2,4-D); 6-(4-hydroxy-3-methylbut-2-enylamino) purine (zeatin).

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In Vitro Shoot Regeneration from Callus Derived from Marubakaido Apple Rootstock under Different Aluminium Concentrations

HortScience ◽

10.21273/hortsci.33.3.460e ◽

1998 ◽

Vol 33 (3) ◽

pp. 460e-460 ◽

Cited By ~ 1

Author(s):

Marisa F. de Oliveira ◽

Gerson R. de L. Fortes ◽

João B. da Silva

Keyword(s):

Shoot Regeneration ◽

Dichlorophenoxyacetic Acid ◽

Apple Rootstock ◽

Under Five ◽

Significant Difference ◽

Previously Treated ◽

2,4 Dichlorophenoxyacetic Acid ◽

In Vitro Shoot Regeneration

The aim of this work was to evaluate the organogenesis of Marubakaido apple rootstock under different aluminium concentratons. The explants were calli derived from apple internodes treated with either 2,4-dichlorophenoxyacetic acid or pichloram at 0.5 and 1.0 μM and under five different aluminium concentrations (0, 5, 10, 15, 20 mg/L). These calli were then treated with aluminium at 0, 5, 10, 15, and 20 mg/L. It was observed shoot regeneration only for those calli previously treated with pichloram. There were no significant difference among the aluminium concentrations.

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Characterization of the Reproductive Mode in Guayule In Vitro

HortScience ◽

10.21273/hortsci.33.3.483a ◽

1998 ◽

Vol 33 (3) ◽

pp. 483a-483

Author(s):

Roy N. Keys ◽

Dennis T. Ray ◽

David A. Dierig

Keyword(s):

Field Experiments ◽

Reproductive Mode ◽

Dichlorophenoxyacetic Acid ◽

Initial Field ◽

Breeding Lines ◽

Reproductive Modes ◽

Desert Southwest ◽

2,4 Dichlorophenoxyacetic Acid

Guayule (Parthenium argentatum Gray, Asteraceae) is a latex-producing perennial desert shrub that is potentially of economic importance as an industrial crop for the desert Southwest. It is known to possess complex reproductive modes. Diploids are predominantly sexual and self-incompatible, while polyploids show a range of apomictic potential and self-compatibility. This paper describes the development of a relatively rapid and simple technique for characterizing reproductive modes of breeding lines of P. argentatum. Initial field experiments were based on an auxin test used successfully to characterize reproductive mode in the Poaceae. The application of 2,4-dichlorophenoxyacetic acid inhibited embryo formation in P. argentatum, but this was not the case with other auxins tested. Results of field experiments were ambiguous because: 1) the floral structure of P. argentatum is such that auxins might not have penetrated to the ovules, and 2) there was potential self-fertilization by pollen released within isolation bags. Therefore, in vitro culture of flower heads was tested because it provided much better control of environmental conditions, growth regulator application, and pollen release. Auxin alone, or in combination with gibberellic acid or kinetin, inhibited parthenogenesis in vitro. Embryo production did not vary using two substantially different nutrient media. In vitro flower head culture using a (Nitsch and Nitsch) liquid nutrient medium without growth regulators, enabled characterization of the reproductive mode of seven breeding lines, ranging from predominantly sexual to predominantly apomictic. The results of this technique were substantiated using RAPD analyzes of progeny arrays from controlled crosses.

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Initiation and morphological development of somatic embryoids from barley cell cultures

Canadian Journal of Botany ◽

10.1139/b84-167 ◽

1984 ◽

Vol 62 (6) ◽

pp. 1245-1249 ◽

Cited By ~ 41

Author(s):

L. S. Kott ◽

K. J. Kasha

Keyword(s):

Somatic Embryogenesis ◽

Abscisic Acid ◽

Gibberellic Acid ◽

Cell Cultures ◽

Dichlorophenoxyacetic Acid ◽

Morphological Development ◽

Low Concentrations ◽

The Media ◽

2,4 Dichlorophenoxyacetic Acid ◽

Barley Cell

Somatic embryogenesis was induced in callus previously initiated from immature embryos of barley. These cultures ranged in age from 6 weeks to 30 months. Embryoids were readily initiated from homogenized suspension-grown aggregates when plated on modified B5 media with 2,4-dichlorophenoxyacetic acid. Low concentrations (0.1 and 0.05 mg∙L−1) of abscisic acid promoted further maturation of embryoids, while gibberellic acid (1 mg∙L−1) and kinetin (0.1 mg∙L−1) were used in the media to encourage embryoid germination. The development of somatic embryoids from initiation through maturation and germination is described.

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Selectivity of auxin for induction and growth of callus from excised embryo of spring and winter wheat

Canadian Journal of Botany ◽

10.1139/b84-189 ◽

1984 ◽

Vol 62 (7) ◽

pp. 1393-1397 ◽

Cited By ~ 6

Author(s):

M. D. Zhou ◽

T. T. Lee

Keyword(s):

Winter Wheat ◽

Chinese Spring ◽

Shoot Growth ◽

Callus Formation ◽

Triticum Aestivum L ◽

Dichlorophenoxyacetic Acid ◽

Naphthaleneacetic Acid ◽

2,4 Dichlorophenoxyacetic Acid ◽

Methoxybenzoic Acid

The callus-promoting activity of most commonly known as well as some rarely tested auxins was compared with that of 2,4-dichlorophenoxyacetic acid (2,4-D) for in vitro culture of the excised embryo of spring and winter wheat (Triticum aestivum L.), cv. Chinese Spring and cv. Fredrick. Different auxins in a concentration range from 1 to 50 μM showed widely different activities. Also the two wheat cultivars responded differently to the auxins. When rapid callus formation with limited root growth was used as the basis for comparison, 2-(2-methyl-4-chlorophenoxy)propionic acid (2-MCPP), α-naphthaleneacetic acid, 3,6-dichloro-2-methoxybenzoic acid (dicamba), 4-amino-3,5,6,trichloropicolinic acid (picloram), γ-(2,4-dichlorophenoxy)butyric acid, 2,4,5-trichlorophenoxyacetic acid, and 2,4,5-trichlorophenoxypropionic acid, in the order of effectiveness, were superior to 2,4,-D for callus induction from the embryo of 'Chinese Spring,' although the concentration required was higher than that of 2,4-D. For the winter wheat 'Fredrick,' however, only picloram, dicamba, and 2-MCPP performed as well as 2,4-D. All auxins tested promoted shoot growth; 2-methyl-4-chlorophenoxypropionic acid was most effective for 'Chinese Spring,' whereas picloram was most effective for 'Fredrick.'

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Impact of Ionic Liquids on Induction of Wheat Microspore Embryogenesis and Plant Regeneration

Agronomy ◽

10.3390/agronomy10060839 ◽

2020 ◽

Vol 10 (6) ◽

pp. 839

Author(s):

Dorota Weigt ◽

Idzi Siatkowski ◽

Magdalena Magaj ◽

Agnieszka Tomkowiak ◽

Jerzy Nawracała

Keyword(s):

Ionic Liquids ◽

Plant Regeneration ◽

Positive Impact ◽

Microspore Embryogenesis ◽

Green Plant ◽

Induction Medium ◽

Dichlorophenoxyacetic Acid ◽

2,4 Dichlorophenoxyacetic Acid ◽

The Impact

Ionic liquids are novel compounds with unique chemical and physical properties. They can be received based on synthetic auxins like 2,4-dichlorophenoxyacetic acid or dicamba, which are commonly used hormones in microspore embryogenesis. Nevertheless, ionic liquids have not been adapted in plant in vitro culture thus far. Therefore, we studied the impact of ionic liquids on the ability to undergo microspore embryogenesis in anther cultures of wheat. Two embryogenic and two recalcitrant genotypes were used for this study. Ten combinations of ionic liquids and 2,4-dichlorophenoxyacetic acid were added to the induction medium. In most cases, they stimulated induction of microspore embryogenesis and green plant regeneration more than a control medium supplemented with only 2,4-dichlorophenoxyacetic acid. Two treatments were the most favorable, resulting in over two times greater efficiency of microspore embryogenesis induction in comparison to the control. The effect of breaking down the genotype recalcitrance (manifested by green plant formation) was observed under the influence of 5 ionic liquids treatments. Summing up, ionic liquids had a positive impact on microspore embryogenesis induction and green plant regeneration, increasing the efficiency of these phenomena in both embryogenic and recalcitrant genotypes. Herbicidal ionic liquids can be successfully used in in vitro cultures.

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