Influence of auxins on somatic embryogenesis and alkaloid accumulation in Leucojum aestivum callus

2013 ◽  
Vol 8 (6) ◽  
pp. 591-599 ◽  
Author(s):  
Agata Ptak ◽  
Anna Tahchy ◽  
Edyta Skrzypek ◽  
Tomasz Wójtowicz ◽  
Dominique Laurain-Mattar
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryo ◽  
Symptomatic Treatment ◽  
Dichlorophenoxyacetic Acid ◽  
Leucojum Aestivum ◽  
Anisic Acid ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Embryogenic Response

AbstractIn vitro cultures of Leucojum aestivum are considered as an alternative for the production of galanthamine, which is used for the symptomatic treatment of Alzheimer’s disease. We studied the effects of auxins 2,4-dichlorophenoxyacetic acid (2,4-D), 4-amino-3,5,6-trichloropicolinic acid (picloram), 3,6-dichloro-o-anisic acid (dicamba) at concentrations of 25 and 50 µM on the induction of embryogenic callus and its capacity to induce somatic embryogenesis and alkaloid accumulation. The embryogenic response of the explants was from 30% for 25 µM of dicamba to 100% for picloram (for both 25 and 50 µM). 2,4-D (50 µM) stimulated greater callus proliferation and somatic embryo induction as compared to the other auxins. Polyethylene glycol (PEG) stimulated somatic embryo maturation. Callus grown on media containing 50 µM of auxins produced fewer phenolic compounds as compared with callus grown on media containing 25 µM of auxins. GC-MS analyses showed seven alkaloids in the in vivo bulbs and two to four in callus culture. Galanthamine was detected in callus cultivated with 2,4-D (25, 50 µM), picloram (25 µM), and dicamba (50 µM). Other alkaloids, trisphaeridine, tazettine, and 11-hydroxyvittatine were accumulated only in callus growing on medium with picloram (50 µM).

scholarly journals Somatic Embryogenesis and Organogenesis in Magnolia dealbata Zucc. (Magnoliaceae), an Endangered, Endemic Mexican Species

HortScience ◽  
2006 ◽  
Vol 41 (5) ◽  
pp. 1325-1329 ◽  
Author(s):  
Martín Mata-Rosas ◽  
Ángel Jiménez-Rodríguez ◽  
Victor M. Chávez-Avila
Keyword(s):  
Somatic Embryogenesis ◽  
Basal Medium ◽  
Direct Somatic Embryogenesis ◽  
Direct Organogenesis ◽  
Limiting Factor ◽  
Zygotic Embryos ◽  
Dichlorophenoxyacetic Acid ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Magnolia Dealbata

Plants of Magnolia dealbata were regenerated from zygotic embryos through somatic embryogenesis and direct organogenesis. Medium and incubation conditions were determinating factors for the development of morphogenetic responses. Photoperiodic exposure was a limiting factor in the general development of the explants, and incubation in darkness allowed their development. The highest formation of shoots per responding explant were obtained on woody plant (WP) medium supplemented with 13.3 μM or 22.2 μM 6-benzylaminopurine (BA) in combination with 2.26 μM or in absence of 2,4-dichlorophenoxyacetic acid (2,4-D) from which 2.5 shoots per explant were induced. Subcultures on WP medium, supplemented with polyvinylpyrrolidone (PUP) 40,000 1 g·L–1) avoided necrosis of explants. Somatic embryos were formed in 85% of explants cultivated on WP medium with 2,4-D (2.3 μM or 4.5 μM); 20% induced indirect embryogenesis and 65% formed direct somatic embryogenesis. The plants were transferred to soil to acclimatize under greenhouse conditions, achieving 90% survival. Somatic embryo conversion to plantlets was obtained with subculture on WP basal medium without growth regulators. In vitro culture can play a key role in the propagation and conservation of this endangered species.

scholarly journals Secondary Somatic Embryogenesis in Centaurium erythraea Rafn

Plants ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 199
Author(s):  
Milica D. Bogdanović ◽  
Katarina B. Ćuković ◽  
Angelina R. Subotić ◽  
Milan B. Dragićević ◽  
Ana D. Simonović ◽  
...  
Keyword(s):  
Somatic Embryogenesis ◽  
Histological Analysis ◽  
Developmental Process ◽  
Developmental Pathways ◽  
Dichlorophenoxyacetic Acid ◽  
Embryogenic Cells ◽  
Endangered Medicinal Plant ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Centaurium Erythraea

Somatic embryogenesis (SE) is a developmental process during which plant somatic cells, under suitable conditions, produce embryogenic cells that develop into somatic embryos (se). SE is the most important method for plant propagation in vitro, having both fundamental and applicative significance. SE can be induced from different tissues and organs, but when se are used as explants, the process is recognized as secondary or cyclic SE. We induced secondary SE in Centaurium erythraea by application of 2,4-dichlorophenoxyacetic acid (2,4-D) and N-(2-chloro-4-pyridyl)-N′-phenylurea (CPPU). A medium containing 0.1 mgL−1 2,4-D and 0.25 mgL−1 CPPU was optimal in terms of the number of primary SE explants forming se, the number of well-developed se per explant, and morphological appearance of the obtained se. These concentrations allowed SE to progress through three cycles, whereas at higher concentrations of 0.2 mgL−1 2,4-D and 0.5 mgL−1 CPPU, only two cycles were achieved. Histological analysis revealed that secondary se are formed both directly and indirectly. Secondary SE readily germinated and converted into plantlets. Induction of cyclic SE contributes to the conservation efforts of this endangered medicinal plant and expands the spectrum of in vitro developmental pathways described in centaury—an emerging model in developmental biology.

scholarly journals Somatic embryogenesis of Sandalwood (Santalum album L.)

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.

In vivo and in vitro instability of B chromosomes in palmarosa grass (Cymbopogon martinii var. motia)

Genome ◽  
1988 ◽  
Vol 30 (6) ◽  
pp. 966-973 ◽  
Author(s):  
H. L. Sreenath ◽  
K. S. Jagadishchandra
Keyword(s):  
Germ Line ◽  
Chromosome Instability ◽  
Metaphase Plate ◽  
B Chromosome ◽  
B Chromosomes ◽  
Dichlorophenoxyacetic Acid ◽  
Cymbopogon Martinii ◽  
2,4 Dichlorophenoxyacetic Acid

Meiotic and mitotic instability and elimination of B chromosomes was observed in diploid palmarosa (Cymbopogon martinii var. motia) with 2n = 20 + 1–2 B under in vivo and in vitro conditions. Under in vivo conditions B chromosomes were totally absent from the roots but preferentially transmitted in the germ line tissues. When present as a pair, the B chromosomes formed bivalents showing almost regular orientation, congression, and disjunction. When present singly, the B chromosome formed a univalent and did not pair with any of the A chromosomes and showed nonalignment on the metaphase plate during metaphase I. Immature inflorescences of a race of palmarosa with 2n = 20 + 2 B were cultured on Murashige and Skoog's medium containing 1 mg/L of 2,4-dichlorophenoxyacetic acid to produce embryogenic callus. The cytological analysis of the callus revealed only 20 A chromosomes in nearly all the cells, both the B chromosomes being eliminated. From this callus, plantlets without B chromosomes were regenerated on MS medium without growth regulators and established in the soil. The regenerated plants exhibited 20 A chromosomes with normal meiosis.Key words: Cymbopogon martinii, chromosome instability, palmarosa grass, B chromosomes, tissue culture, plant regeneration.

scholarly journals Study on the presence and influence of phenolic compounds in callogenesis and somatic embryo development of cocoa (Theobroma cacao L.).

2015 ◽  
Vol 31 (1) ◽  
pp. 14-20
Author(s):  
Sulistyani Pancaningtyas
Keyword(s):  
Tissue Culture ◽  
Somatic Embryogenesis ◽  
Phenolic Compounds ◽  
Somatic Embryo ◽  
Phenolic Compound ◽  
Embryogenic Callus ◽  
Theobroma Cacao ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Theobroma Cacao L

Cocoa (Theobroma cacao L.) like most tropical trees is recalcitrant in tissue culture. Somatic embryogenesis is generally efficient micropropagation technique to multiply elite material. However, Somatic embryogenesis in cocoa is difficult and this species is considered as recalcitrant. One of the factors often considered as a component of in vitro recalsitrance is a high phenolic content and oxidation of these compounds. In cocoa tissue culture accumulate large amounts of poliphenolics compounds which probably impair further development. This study was conducted to investigate the composition of phenolic compounds in cocoa flower and leaves, and their changes troughout the somatic embryogenesis process. Calli were induced in cacao floral and leaves explants on a half-strenght Murashige and Skoog medium containing 30 g/L Glucose and combination of 2,4 dichlorophenoxyacetic acid (2,4 D) with kinetin (kin). Total polyphenol content was observed on Sulawesi 1 cocoa clone. Embryogenic and non-embryogenic callus were also compared. The percentage of callus production from flower tissue is 85%, percentage of embryogenic callus 40 %, although  the percentage of somatic embryo production from embryogenic callus callus is 70%. The conservation of callus into somatic embryos followed by decline in phenol content and an increase in peroxidase. The synthesis kinetics for these compounds in calli, under different somatic embryogenesis conditions, revealed a higher concentration under non-embryogenic conditions. So that, phenolic compound can influence the production of calli and an absence the phenolic compound can enhance production of somatic embryo.Kata kunci: Theobroma cacao L., polifenol, embrio somatik, kalus, flavonoid, katekin, in vitro recalcitance

scholarly journals Somatic Embryogenesis and Plant Regeneration in Viola canescens Wall. Ex. Roxb.: An Endangered Himalayan Herb

Plants ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 761
Author(s):  
Arun Kumar Khajuria ◽  
Christophe Hano ◽  
Narendra Singh Bisht
Keyword(s):  
Somatic Embryogenesis ◽  
Culture Medium ◽  
Casein Hydrolysate ◽  
Dichlorophenoxyacetic Acid ◽  
Ms Medium ◽  
Mature Embryos ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Root Axis ◽  
Somatic Embryo Induction

Viola canescens Wall. ex. Roxb. is an important but threatened medicinal herb found at 1500–2400 m above mean sea level in the Himalayas. Overexploitation and habitat preference have put the plant under serious threat. Thus, the present study was undertaken to develop an efficient protocol for in vitro propagation via somatic embryogenesis. The results revealed that plant can be regenerated successfully through somatic embryogenesis using leaf derived calli. Regular subculturing of calli on Murashige and Skoog (MS) medium with 2,4-dichlorophenoxyacetic acid (2,4-D)/indole-3-butyric acid (IBA)/kinetin (Kn) and varying combinations of 2,4-D+Kn induced somatic embryogenesis. The maximum average number of somatic embryos (SE) (19.15 ± 2.66) was induced on the medium with 0.15 + 0.05 mg L−1 of 2,4-D and Kn, respectively, and this medium was used as a control. To enhance somatic embryo induction, the control MS medium was supplemented with l-glutamine (200–400 mg L−1) and casein hydrolysate (1–4%). The maximum average number of SE (27.66 ± 2.67) and average mature SE (13.16 ± 3.48) were recorded on the medium having 2 % l-glutamine and 50 mg L−1 casein hydrolysate. The induced SE were asynchronous, so, to foster their maturation, the culture medium (free from growth regulators) was supplemented with abscisic acid (ABA) and silver nitrate (AgNO3). The maximum average number (35.96 ± 3.68) of mature SE was noticed on MS medium supplemented with 1.5 mg L−1 ABA. Mature embryos had two well-developed cotyledons and an elongated hypocotyl root axis. The development of SE into plantlets was significant for embryos matured on the medium with AgNO3 and ABA, with 86.67% and 83.33% conversion on the medium with 0.20 mg L−1 6-benzylaminopurine (BAP). The plantlets thus produced acclimatized in a growth chamber before being transferred to the field, which showed 89.89% survival. The plants were morphologically similar to the mother plant with successful flowering.

scholarly journals The miR396–GRF Regulatory Module Controls the Embryogenic Response in Arabidopsis via an Auxin-Related Pathway

2019 ◽  
Vol 20 (20) ◽  
pp. 5221 ◽  
Author(s):  
Szczygieł-Sommer ◽  
Gaj
Keyword(s):  
Positive Control ◽  
Dichlorophenoxyacetic Acid ◽  
Regulatory Relationship ◽  
Gus Reporter ◽  
Regulatory Module ◽  
Genes Encoding ◽  
Cell Transcriptome ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Embryogenic Response

In plants, microRNAs have been indicated to control various developmental processes, including somatic embryogenesis (SE), which is triggered in the in vitro cultured somatic cells of plants. Although a transcriptomic analysis has indicated that numerous MIRNAs are differentially expressed in the SE of different plants, the role of specific miRNAs in the embryogenic reprogramming of the somatic cell transcriptome is still poorly understood. In this study, we focused on performing a functional analysis of miR396 in SE given that the transcripts of MIR396 genes and the mature molecules of miR396 were found to be increased during an SE culture of Arabidopsis [1]. In terms of miR396 in embryogenic induction, we observed the SE-associated expression pattern of MIR396b in explants of the β-glucuronidase (GUS) reporter line. In order to gain insight into the miR396-controlled mechanism that is involved in SE induction, the embryogenic response of mir396 mutants and the 35S:MIR396b overexpressor line to media with different 2,4-Dichlorophenoxyacetic acid (2,4-D) concentrations was evaluated. The results suggested that miR396 might contribute to SE induction by controlling the sensitivity of tissues to auxin treatment. Within the targets of miR396 that are associated with SE induction, we identified genes encoding the GROWTH-REGULATING FACTOR (GRF) transcription factors, including GRF1, GRF4, GRF7, GRF8, and GRF9. Moreover, the study suggested a regulatory relationship between miR396, GRF, and the PLETHORA (PLT1 and PLT2) genes during SE induction. A complex regulatory relationship within the miR396–GRF1/4/8/9–PLT1/2 module that involves the negative and positive control of GRFs and PLT (respectively) by miR396 might be assumed.

Physiologically Based Pharmacokinetic Modeling of Salivary Concentrations for Noninvasive Biomonitoring of 2,4-Dichlorophenoxyacetic Acid (2,4-D)

2019 ◽  
Vol 172 (2) ◽  
pp. 330-343
Author(s):  
Alice A Han ◽  
Charles Timchalk ◽  
Zana A Carver ◽  
Thomas J Weber ◽  
Kimberly J Tyrrell ◽  
...  
Keyword(s):  
Pbpk Model ◽  
Pbpk Modeling ◽  
Dichlorophenoxyacetic Acid ◽  
Diffusion Limited ◽  
Physiologically Based Pharmacokinetic ◽  
Physiologically Based ◽  
Dynamic Time ◽  
2,4 Dichlorophenoxyacetic Acid

Abstract Saliva has become a favorable sample matrix for biomonitoring due to its noninvasive attributes and overall flexibility in collection. To ensure measured salivary concentrations reflect the exposure, a solid understanding of the salivary transport mechanism and relationships between salivary concentrations and other monitored matrices (ie, blood, urine) is needed. Salivary transport of a commonly applied herbicide, 2,4-dichlorophenoxyacetic acid (2,4-D), was observed in vitro and in vivo and a physiologically based pharmacokinetic (PBPK) model was developed to translate observations from the cell culture model to those in animal models and further evaluate 2,4-D kinetics in humans. Although apparent differences in experimental in vitro and in vivo saliva:plasma ratios (0.034 and 0.0079) were observed, simulations with the PBPK model demonstrated dynamic time and dose-dependent saliva:plasma ratios, elucidating key mechanisms affecting salivary transport. The model suggested that 2,4-D exhibited diffusion-limited transport to saliva and was additionally impacted by protein binding saturation and permeability across the salivary gland. Consideration of sampling times post-exposure and potential saturation of transport mechanisms are then critical aspects for interpreting salivary 2,4-D biomonitoring observations. This work utilized PBPK modeling in in vitro to in vivo translation to explore benefits and limitations of salivary analysis for occupational biomonitoring.

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