scholarly journals Cytohistological analysis of somatic embryogenesis in cucumber (Cucumis sativus L). I. Comparison of cell suspension containing and lacking natural fluorescence with in vivo developing embryos

2014 ◽  
Vol 63 (3-4) ◽  
pp. 259-263 ◽  
Author(s):  
J. A. Tarkowska ◽  
D. Brzostecka ◽  
W. Burza ◽  
S. Malepszy
Keyword(s):  
Somatic Embryogenesis ◽  
Morphological Difference ◽  
Mature Embryo ◽  
Variable Number ◽  
Histological Structure ◽  
Cucumis Sativus L ◽  
Mature Embryos ◽  
Natural Fluorescence

Under in vivo conditions early-globular embryos occur in cucumber on the 9th day after pollination, heart-shaped ones on the 14th, and morphologically mature embryos appear on the 19th day. Single starch grains already appear in the cells of the globular embryo, and in the heart-shaped one they occur within the forming root cap. In the morphologically mature embryo only the precambium is free from starch. Somatic embryogenesis (SE) in suspension occurs similarly as in vivo, even though the starch localization is somewhat different and torpedo-like embryos occur, which are not observed in vivo. The histological structure of in vitro embryos is similar to in vivo ones, and the greatest morphological difference are the poorly developed cotyledons and their variable number (1 to 3). Aggregates showing fluorescence were found to be composed of cells which differ in morphology from cells not showing fluorescence and appear to be more capable of attaining the mature stages.

scholarly journals Cytohistological analysis of somatic embryogenesis in cucumber (Cucumis sativus L). II. Natural fluorescence and direct somatic embryogenesis from protoplasts

2014 ◽  
Vol 63 (3-4) ◽  
pp. 265-268 ◽  
Author(s):  
W. Burza ◽  
B. Woźniak ◽  
J. A. Tarkowska ◽  
S. Malepszy
Keyword(s):  
Somatic Embryogenesis ◽  
Protoplast Culture ◽  
Direct Somatic Embryogenesis ◽  
Histological Structure ◽  
Cucumis Sativus L ◽  
Natural Fluorescence ◽  
Vacuolated Cells ◽  
Selection Of

The development of protoplast derived from somatic embryos and some of their characteristics were compared with embryos from suspension and in vivo in the same B line. Embryos formed in a protoplast culture differed from others that their younger stages contained vacuolated cells, and older ones had altered morphological and histological structure. Somatic embryogenesis is more regular from suspension then from protoplasts. No distinct differences were observed in the rate of embryo development in vivo and in vitro, and in vitro embryos show a larger variation in size at the same stage. Embryos in vitro with fluorescence are generally larger than zygotic ones at each stage. The use of fluorescence is suggested for the selection of heterokariocytes after protoplast fusion.

scholarly journals REGULATION OF CEREAL EMBRYONIC ORGANOGENESIS IN VITRO CONDITIONS

ÈKOBIOTEH ◽  
2021 ◽  
Vol 4 (1) ◽  
pp. 11-23
Author(s):  
N.N. Kruglova ◽  
Keyword(s):  
Somatic Embryogenesis ◽  
Hormonal Regulation ◽  
Zygotic Embryogenesis ◽  
Review Of The Literature ◽  
Biological Phenomenon ◽  
Organogenesis In Vitro ◽  
Cereal Embryos ◽  
Plant Embryogenesis

The article provides the brief review of the literature and own works devoted to the peculiarities of the cereal embryonic organogenesis at the early stages of ontogenesis in the conditions of in vitro culture (the so-called somatic embryogenesis, or embryoidogenesis in vitro). Particular attention is paid to the issues of hormonal regulation of the development of somatic cereal embryos from initial cells to mature structures in vitro. A comparison of somatic embryogenesis in vitro with similar events in zygotic embryogenesis in vivo confirms the validity of the principle of universality of morphogenesis processes in vivo and in vitro (Batygina, 2014). The prospects of using somatic embryogenesis in vitro as a model for studying the most complex biological phenomenon – zygotic plant embryogenesis in vivo – are discussed.

Alleles of the bovine DGAT1 variable number of tandem repeat associated with a milk fat QTL at chromosome 14 can stimulate gene expression

2006 ◽  
Vol 25 (1) ◽  
pp. 116-120 ◽  
Author(s):  
Rainer Fürbass ◽  
Andreas Winter ◽  
Ruedi Fries ◽  
Christa Kühn
Keyword(s):  
Gene Expression ◽  
Tandem Repeat ◽  
Milk Fat ◽  
Variable Number ◽  
Chromosome 14 ◽  
Fat Percentage ◽  
Vntr Polymorphism ◽  
Causal Mutation

A quantitative trait locus (QTL) affecting milk fat percentage has been mapped to the centromeric end of the bovine chromosome 14 (BTA14). This genomic area includes the DGAT1 gene, which encodes acyl-CoA:diacylglycerol acyltransferase 1, the key enzyme of triglyceride biosynthesis. Genetic and biochemical studies led to the identification of the nonconservative DGAT1-K232A polymorphism as a causal mutation for the QTL. In addition to this, another polymorphism in the 5′-regulatory region of this gene, the DGAT1 variable number of tandem repeat (VNTR), also showed a strong association with milk fat percentage. This promoter VNTR polymorphism affects the number of potential Sp1 binding sites and therefore might have an impact on DGAT1 expression and also milk fat content. Hence, the DGAT1 VNTR polymorphism might be another causal mutation for the BTA14 QTL. However, evidence for Sp1 binding to this polymorphic site and for the capability of DGAT1 VNTR alleles to stimulate gene expression was lacking. In the current work Sp1-VNTR interactions were analyzed by EMSA. In addition, effects of DGAT1 VNTR alleles on gene expression were measured with reporter gene analyses. Conclusions from the results are that 1) the DGAT1 VNTR sequence is indeed a target for Sp1 binding; 2) DGAT1 VNTR alleles can stimulate gene expression in vitro and probably in vivo as well; and 3) although the stimulating effects of the different DGAT1 VNTR alleles did not show significant differences in vitro, their effects on transcription might be different in the chromatin context existing in vivo.

In Vitro Shoot Regeneration of NAA-Pulse Treated Plumular Leaf Explants of Cowpea

2010 ◽  
Vol 2 (2) ◽  
pp. 60-63 ◽  
Author(s):  
Muhammad AASIM
Keyword(s):  
Somatic Embryogenesis ◽  
Shoot Regeneration ◽  
Leaf Explants ◽  
Leaf Explant ◽  
Grain Legume ◽  
Ms Medium ◽  
Pulse Treatment ◽  
Mature Embryos ◽  
Legume Crop

Cowpea (Vigna unguiculata L.) is an economically important grain legume crop and is an important source of dietary protein in many of the developing countries. The present study reports the effect of pulse treatment duration, concentration of NAA and presence of NAA in the culture medium on shoot regeneration from plumular leaf explant of Turkish cowpea cv. ‘Akkiz’ and ‘Karagoz’. Pulse treatment of mature embryos with 20 mg l-1 NAA for 1 and 3 weeks followed by culturing of plumular leaf explant on MS medium containing 0.25, 0.50 and 1.0 BAP with 1.0, 2.0 and 4.0 mg l-1 NAA promoted somatic embryogenesis in both cultivars. Longer duration of pulse treatment was deleterious resulting in browning and consequently death of the embryos on explants. Pulse treatment with 20 mg l-1 NAA for one week was less deleterious and developed two plantlets after the explants were transferred to MS0 medium after 6 weeks through somatic embryogenesis in cv. ‘Akkiz’. Pulse treatment with 10 mg l-1 NAA for 1 week showed 33.33-50.00% and 25.00-50.00% shoot regeneration frequency in cv. ‘Akkiz’ and ‘Karagoz’ respectively on MS medium containing 0.25-1.00 mg l-1 BAP. Maximum number of 2.50 shoots each per explant were recorded in cv. ‘Akkiz’ and ‘Karagoz’ on MS medium containing 1.00 and 0.50 mg l-1 BAP respectively. Contrarily, maximum shoot length of 8.98 cm of cv. ‘Akkiz’ and 9.42 cm of cv. ‘Karagoz’ was recorded on MS medium containing 0.50 mg l-1 BAP and 1.00 mg l-1 BAP respectively. Regenerated shoots were rooted on MS medium containing 0.5 mg l-1 IBA and and acclimatized in growth room at room temperature where they produced viable seeds.

Evidence for Nucleosomal Phasing and a Novel Protein Specifically Binding to Cucumber Satellite DNA

1994 ◽  
Vol 49 (1-2) ◽  
pp. 79-86 ◽  
Author(s):  
Thilo C. Fischer ◽  
Sabine Groner ◽  
Ulrike Zentgraf ◽  
Vera Hemleben
Keyword(s):  
Satellite Dna ◽  
Specific Binding ◽  
Apparent Molecular Weight ◽  
Micrococcal Nuclease ◽  
Type I ◽  
Dna Repeats ◽  
Cucumis Sativus L ◽  
Nuclease Digestion

The nucleosomal organization and the protein-binding capability of highly repeated and methylated satellite DNA of cucumber (Cucumis sativus L.), comprising approx. 30% of the genome, were analyzed. Nucleosomal core DNA from satellite type I was prepared after micrococcal nuclease digestion of chromatin and sequenced. Most of the core sequences obtained could be grouped in two main (A and B) and two minor groups (C and D) indicating a specific and complex phasing of nucleosomes on this satellite DNA. In vitro, gel retardation assays with cloned satellite DNA repeats (types I-IV) demonstrated a specific binding of nuclear proteins. These specific binding effects are also obtained with genomic, in vivo methylated and sequence heterogeneous (1 to 10% diversity) satellite type I DNA. For the first time in plants, a satellite DNA-binding protein with an apparent molecular weight of 14 kDa (SAT 14) was identified.

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 Evaluation of the artherogenic potential of homocysteine and its cytotoxic effect on HUVECs

2017 ◽  
Vol 1 (2) ◽  
pp. 33-34
Author(s):  
Chérifa AGGOUN ◽  
Ibtissem ELOUAR ◽  
Souad LAMDA ◽  
Dalila NAIMI
Keyword(s):  
Endothelial Cells ◽  
Biochemical Study ◽  
Histological Study ◽  
Liver Steatosis ◽  
In Vitro Study ◽  
Vitro Study ◽  
Histological Structure ◽  
Senescent Phenotype

Background: In recent decades, several studies have shown that hyperhomocysteinemia is an independent risk factor for atherosclerosis. The objective of this work is, first, to assess the atherogenic potential of homocysteine in rats and and appraise its effect on endothelial cells: the main components of the blood vessel. Methods: In the in vivo study, hyperhomocysteinemia is induced in rats by a diet supplemented with L-methionine (1 g/kg/day). After 21 days of treatment, a blood sample is performed for biochemical study (total cholesterol, triglycerides, HDL, LDL cholesterol, ALT, AST, CRP, and total protein) and a removal of organs (aorta, liver, and heart) for the assay MDA and histological study. For the in vitro study, HUVECs were cultured for 24 h then exposed to various concentrations of homocysteine (2 mM, 4 mM, 8 mM). After 24 hours, we confirm the cellular phenotype and its number. Results: The angiotoxic dose of methionine induces a significant increase for all biochemical parameters (except HDL-C, which is diminished). In addition, an alteration of the histological structure of the aorta (appearance of foam cells) of the liver (steatosis) and of the heart. The in vitro study revealed a senescent phenotype of HUVECs and an inhibition of cell proliferation. Conclusion: Our results suggest that homocysteine is involved in the genesis of atherosclerosis by affecting directly the endothelial cells.

scholarly journals (79) Direct Bulblet Regeneration from Mature Embryo: A Rapid, Efficient and Genotype-independent In Vitro Morphogenesis Pathway for Preservation of Endangered Wild Populations of Fritillaria imperialis and Fritillaria persica

HortScience ◽  
2006 ◽  
Vol 41 (4) ◽  
pp. 1066C-1066
Author(s):  
Manijeh Mohammadi-Dehcheshmeh ◽  
Ahmad Khalighi ◽  
Esmaeil Ebrahimie ◽  
Manoochehr Sardari ◽  
Rohangiz Naderi
Keyword(s):  
Tissue Culture ◽  
Mature Embryo ◽  
Direct Organogenesis ◽  
Wild Populations ◽  
In Vitro Morphogenesis ◽  
Culture Techniques ◽  
Mature Embryos ◽  
Best Response ◽  
Heterozygous Plant

Wild populations of Fritillaria sp. have dramatically decreased in Iran because of pest overflow and continual grazing. Previous studies have shown that Fritillaria cannot rapidly and efficiently propagate by traditional methods. In vitro tissue culture techniques have shown high potential for micropropagation of endangered plants. The use of bulb-scale pieces for tissue culture can result in the destruction of the endangered parent plant. Fritillaria is a heterozygous plant in which the genetic content of each embryo is different from others, even on the same plant. In this study, mature embryos of F. imperialis and F. persica were used as explant for the first time. Embryos were extracted from seeds and cultured on B5 medium supplemented with various combinations of BAP (0, 0.1, 1 mg/L), NAA (0, 0.4, 4 mg/L), and IAA (0, 0.4, 4 mg/L). Embryo explant showed low genotype dependency between different heterogenous and heterozygote populations of both F. imperialis and F. persica. The best response of bulblet regeneration in both F. imperialis and F. persica was obtained from 1 mg/L BAP + 0.4 mg/L NAA+4 mg/L IAA and direct organogenesis pathway, with 15 bulblets per explant for F. imperialia and 20 for F. persica. Because of the large number of embryos in a plant and their different genetic contents, established in vitro propagation by using embryo explant in this study can provide broad genetic resources and variations. As explained above, in vitro protocols can play a major role in rescuing F. imperialis and F. persica from extinction.

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