Uptake and biochemical analysis of 2,4-D in cultured zygotic embryos of zea mays L.

Bronsema, F.B.F.; Redig, Pascale; Oostveen, W.J.F. van; Onckelen, Henri A. Van; Lammeren, A.A.M. van

doi:10.1016/s0176-1617(96)80135-0

Cited by 11 publications

(11 citation statements)

References 14 publications

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“…Although the quantification of uptake per mg of dry weight should be more precise, uptake per mg of fresh weight seems to show more exact physiological status. Both methods are generally used in similar uptake experiments (Bronsema et al, 1996;Ceccarelli et al, 2000). In our experiments the level of 14 C-labelled compounds per embryo seemed to bee usable compromise.…”

Section: Resultsmentioning

confidence: 78%

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Uptake and metabolism of 2,4-dichlorophenoxyacetic acid during induction of Norway spruce (Picea abies (L.) Karst.) somatic embryogenesis

Vlašínová¹,

Havel²,

Klemš³

et al. 2014

Acta Univ. Agric. Silvic. Mendelianae Brun.

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Uptake and metabolism of 2,4-dichlorophenoxyacetic acid (2,4-D) into zygotic embryos of Norway spruce (Picea abies (L.) Karst.) were studied on medium used for induction of somatic embryogenesis. The main task of this work was to study effect of longitudinal bisection of embryos, which was found as increasing the subsequent induction of embryogenic cultures. The maximal uptake of 2,4-D per one embryo was detected after 16 hours on medium in bisected embryos. The bisection increased 2,4-D uptake per embryo in first 16 th hours, but then increased its release back to the medium. The metabolism of 2,4-D in bisected embryos was lower than in intact ones during first two days of culture.

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Section: Resultsmentioning

confidence: 78%

“…3A,B). On the contrary Bronsema et al (1996) detected in immature maize ZEs the first metabolised 2,4-D from 2 hours onwards. Formation of polar metabolites (conjugates with amino acid and glucose) started in maize after 16 hours of culture as well as in intact ZEs of Norway spruce in our experiments (Fig.…”

Section: Resultsmentioning

confidence: 82%

Uptake and metabolism of 2,4-dichlorophenoxyacetic acid during induction of Norway spruce (Picea abies (L.) Karst.) somatic embryogenesis

Vlašínová¹,

Havel²,

Klemš³

et al. 2014

Acta Univ. Agric. Silvic. Mendelianae Brun.

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“…Cobs were hand-pollinated and immature embryos were excised at 11-12 days after pollination. They were either directly processed for immunocytochemical labelling, or first cultured in vitro on modified N6 medium containing 2 mg of 2,4-D per liter (for details, see Bronsema et al 1996) for 5-7 days and then sampled. Mature caryopses were imbibed in tap water for 6 h, and germinated at 25 ~ on moist filter paper in the dark.…”

Section: Plant Materialsmentioning

confidence: 99%

“…Now we ask whether there is a relation between the localisa-tion of 2,4-D and the localisation of ABP-1 in the cultured embryo. During culture, 2,4-D was taken up by the embryo (Bronsema et al 1996) and its distribution was studied by autoradiography on median sections after 16 h of pulse and 72 h of chase on 14C-2,4-D-containing medium (Bronsema et al 1998). 2,4-D was observed in the basal part of the scutellum, in the area around the coleorhiza, and in the scutellum opposite the shoot meristem but not with preference in the epidermis.…”

Section: Localisation Of Auxin-binding Proteins In Cultured Embryos Omentioning

confidence: 99%

Immunocytochemical localisation of auxin-binding proteins in coleoptiles and embryos ofZea mays L.

1998

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Summary. The auxin-binding protein ABP-1 was localisedimmunocytochemically in coleoptiles and immature embryos of Zea mays. Two primary polyclonal antibodies raised against ABP-1 and secondary antibodies were either labelled with FITC or 10 nm gold particles for light microscopy, and with 10 nm gold particles for transmission electron microscopy. Light microscopy revealed that ABP-1 was localised in the epidermal cells of etiolated maize coleoptiles, in subepidermal parenchymatic mesophyll cells of the coleoptile and in the companion cells of the vascular bundles. Most labelling was found in the cytoplasm, less in nuclei and vacuoles and cell walls appeared negative. The region of the plasma membrane exhibited prominent labelling. Embryos showed low labelling throughout their tissues just after excision, but after culture for 7 days intensive labelling was found in the epidermis of the scutellum. Quantitative electron microscopy confirmed that ABP-1 was present in the cytoplasm of epidermal, mesophyll, and companion cells of coleoptiles. Gold particles were neither found in cell walls nor in the cuticle. Areas with ER and dictyosomes within epidermal and mesophyll ceils of coleoptiles had a denser labelling with gold particles than elsewhere. Labelling at the plasma membrane, being the site where the auxin binds to the ABP, was observed at low levels in all cells examined, which is due to the method applied. Epidermal cells of embryos cultured for 5 days exhibited high levels of gold particles in ER and nuclei, and lower levels in the cytoplasm. The distribution is only partly in accordance with the model in which ABP is thought to cycle through the plant cell from the ER via the Golgi system towards the plasma membrane.

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“…in the inbred line A188. In contrast, the inbred line A632 is unable to regenerate somatic embryos (Bronsema et al 1996, 1997, Van Lammeren 1988. In both inbreds, the 1st day of culture is characterized by cytologic changes, such as an increase in the number of organelles, and changes in vacuolation and nuclear morphology.…”

mentioning

confidence: 99%

Distribution of [14C] Dichlorophenoxyacetic Acid in Cultured Zygotic Embryos of Zea mays L.

Bronsema

Oostveen

Prinsen

et al. 1998

J Plant Growth Regul

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The uptake of 2,4-dichlorophenoxyacetic acid (2,4-D), necessary for the in vitro induction of callus formation and somatic embryogenesis in cultured immature maize embryos, was quantified after culture on nutrient medium with [ 14 C]2,4-D. The identity of the 14 C label in the embryos was determined by high performance liquid chromatography (HPLC), and its distribution within embryos was visualized on sections of plastic embedded material. Quantification of the 14 C label after a pulse label of 16 h showed a hundredfold accumulation of 2,4-D in the embryos with respect to the initial medium concentration. During tissue processing for in situ detection of 14 C, however, up to 70% of the label disappeared because of the embedding process. The best structural preservation was obtained after ethanol-mediated infiltration of Technovit 7100. Water-mediated infiltration of Technovit 7100 gave the highest retention of 14 C. HPLC analysis showed that more than 95% of the residual 14 C label found in embryos was still 2,4-D. Autoradiography showed that the embryogenic inbred line A188 contained 14 C label in distinct regions of the scutellum, coleoptile, and suspensor. The nonembryogenic inbred line A632 contained more label after 16 h of culture in a different distribution compared with A188. Subculture of the embryos for 24 and 72 h and histologic analysis showed that cell proliferation and callus formation were restricted to specific regions of the embryo in both inbred lines. The pattern of 2,4-D distribution did not codistribute with regions of proliferation, indicating that 2,4-D is not the only trigger for proliferation.

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Uptake and biochemical analysis of 2,4-D in cultured zygotic embryos of zea mays L.

Cited by 11 publications

References 14 publications

Uptake and metabolism of 2,4-dichlorophenoxyacetic acid during induction of Norway spruce (Picea abies (L.) Karst.) somatic embryogenesis

Uptake and metabolism of 2,4-dichlorophenoxyacetic acid during induction of Norway spruce (Picea abies (L.) Karst.) somatic embryogenesis

Immunocytochemical localisation of auxin-binding proteins in coleoptiles and embryos ofZea mays L.

Distribution of [14C] Dichlorophenoxyacetic Acid in Cultured Zygotic Embryos of Zea mays L.

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