Aspects of isolation underpinning mitotic behaviour in lupin protoplasts

2004 ◽  
Vol 52 (5) ◽  
pp. 669 ◽  
Author(s):  
Anupam Sinha ◽  
Peter D. S. Caligari
Keyword(s):  
Protoplast Culture ◽  
Lupinus Albus ◽  
Maturity Level ◽  
Developmental Age ◽  
Early Maturing ◽  
Division Frequency ◽  
Tissue Maceration ◽  
Protoplast Division ◽  
Protoplast Production

This study reports on the influence of critical isolation factors on the subsequent culture of protoplasts of Lupinus albus L. Protoplasts were isolated from in vitro seedling cotyledons of five early maturing accessions in which protoplast yields and division frequencies appeared to be correlated as a high protoplast yield corresponded with a high division frequency. The overall difference among the accessions for mitosis was non-significant, although the highest yield and division frequency were observed in accession LA132, with Alban giving a significantly lower level. Accession Lucrop produced the lowest number of protoplasts, all of which collapsed during culture. Of the enzyme types used for tissue maceration, Pectolyase Y23, was significantly inferior to Macerase in terms of giving way to mitosis. The extent of division in Macerase-isolated protoplast population was 266% higher than that in the Pectolyase Y23-isolated one. The physiological maturity level of the explant, expressed in terms of developmental age, was optimal when 14–18-day-old seedling cotyledons were used for protoplast production and culture, rather than more mature ones, despite higher protoplast yields in the latter. On K8p medium, the protoplast division frequency was 129% greater when 18-day-old seedling cotyledons were used, than that with any other treatment. This work on protoplast culture of the potentially important lupin species, which is a pulse rich in dietary protein, oil and fibre, allows a further understanding of the biology, with an aim to advance lupin biotechnology.

scholarly journals Embedding in Filter-Sterilized Alginate Enhances Brassica Oleracea L. Protoplast Culture

2015 ◽  
Vol 56 (2) ◽  
pp. 20-26
Author(s):  
Agnieszka Kiełkowska ◽  
Adela Adamus
Keyword(s):  
Brassica Oleracea ◽  
Sodium Alginate ◽  
Mitotic Activity ◽  
Protoplast Culture ◽  
Division Frequency ◽  
Hypocotyl Tissue ◽  
Brassica Oleracea L

Abstract The influence of sodium alginate sterilization on the viability and mitotic activity of embedded protoplasts was studied in protoplasts of Brassica oleracea subsp. alba and rubra isolated from hypocotyl tissue and leaves of seedlings or plants grown in vitro. Both leaf and hypocotyl-derived protoplasts were more viable and divided more frequently when embedded in filtrated alginate. Division frequency was highest in cv. Reball F1 and the mitotic activity of its protoplasts was three times higher when embedded in filtrated alginate (36.1 ± 6.8%) than when cultured in autoclaved alginate (10.9 ± 5.0%). Protoplast-derived calli colonies were transferred to solid regeneration media and plants of all tested accessions were obtained.

Effect of biotic factors on the isolation of Lupinus albus protoplasts

2003 ◽  
Vol 51 (1) ◽  
pp. 103 ◽  
Author(s):  
Anupam Sinha ◽  
Andrew C. Wetten ◽  
P. D. S. Caligari
Keyword(s):  
Lupinus Albus ◽  
Protoplast Isolation ◽  
Biotic Factors ◽  
Average Yield ◽  
Fresh Tissue ◽  
Progressive Decline ◽  
Protoplast Yield ◽  
Protoplast Production ◽  
Protoplast Viability

Several tissue types of Lupinus albus L. were investigated as sources for the isolation of protoplasts. Cotyledons from in vitro seedlings were found to yield the highest number of protoplasts compared with leaves, hypocotyls and roots. A combination of the protoplast isolation enzymes, cellulase and Pectolyase Y23, was capable of releasing the highest number of protoplasts compared with a combination of cellulase and Macerase. Protoplast yield increased with increasing cotyledon age but was accompanied by a progressive decline in protoplast viability. The optimal combination of protoplast yield and viability occurred when the protoplasts were isolated from 14- to 18-day-old cotyledons. The ratio between the volume of enzyme solution and the tissue biomass did not affect the protoplast production significantly. This is the first report of the isolation of protoplasts from a lupin cotyledon and, following the procedure described in this paper, an average yield of 1.2 × 106 protoplasts per gram of fresh tissue was obtainable.

Isolation, Culture and Plant Regeneration From Protoplasts of Nicotiana debneyi

1980 ◽  
Vol 7 (6) ◽  
pp. 635 ◽  
Author(s):  
WR Scowcroft ◽  
PJ Larkin
Keyword(s):  
Acetic Acid ◽  
Plant Regeneration ◽  
Indole Acetic Acid ◽  
High Efficiency ◽  
Protoplast Culture ◽  
Callus Cultures ◽  
Dna Uptake ◽  
Mesophyll Protoplasts ◽  
Nicotiana Debneyi ◽  
Protoplast Division

Mesophyll protoplasts of two genetically distinct genotypes of N. debneyi were cultured with sustained division following a plating efficiency in excess of 50%. Fully fertile mature plants were regenerated from callus cultures derived from protoplasts. Shoots were induced in medium containing 1 mg/l 6-benzylaminopurine and 0.5 mg/I indole acetic acid. The repeatably high efficiency of protoplast culture was used to evaluate the quantitative effects of two drugs, kanamycin and trimethoprim, which effectively inhibited colony formation at concentrations of 100 and 50 �g/ml, respectively. An enhancer of DNA uptake, poly-L-ornithine, had virtually no effect on sustained protoplast division at a concentration of 7.5 �g/ml or less.

Factors affecting the time of formation of the mouse blastocoele

Development ◽  
1977 ◽  
Vol 41 (1) ◽  
pp. 79-92
Author(s):  
Rosita Smith ◽  
Anne McLaren
Keyword(s):  
Cytochalasin B ◽  
Critical Factor ◽  
Cell Number ◽  
Experimental Manipulation ◽  
Factors Affecting ◽  
Elapsed Time ◽  
Developmental Age ◽  
Culture Formation

In normal mouse embryos developing in vivo, the first appearance of the blastocyst cavity was found to be associated more closely with developmental age, judged by cell number, than with chronological age, i.e. elapsed time since ovulation. When development was slowed by in vitro culture, formation of the blastocoele was delayed. However, cell number itself was not a critical factor, since the number of cells per embryo could be doubled or tripled or halved by experimental manipulation without substantially affecting the timing of blastocoele formation. Experiments in which one cell division was suppressed with cytochalasin-B, leading to tetraploidy, showed that the number of cell divisions since fertilization was also not critical. A possible role is suggested either for nucleocytoplasmic ratio, or for the number of nuclear or chromosomal divisions or DNA replications since fertilization, all of which increase during cleavage.

THE IN VITRO DIGESTIBILITY OF PLANT PARTS OF ORCHARDGRASS CLONES WITH ADVANCING STAGES OF MATURITY

1965 ◽  
Vol 45 (5) ◽  
pp. 503-507 ◽  
Author(s):  
D. N. Mowat ◽  
B. R. Christie ◽  
J. E. Winch
Keyword(s):  
Dry Matter ◽  
In Vitro Digestibility ◽  
Plant Parts ◽  
Dry Matter Digestibility ◽  
Total Leaf ◽  
Early Maturing

The in vitro dry matter digestibility (I.V.D.) of the stem and total leaf fractions of four orchardgrass clones was similar at the preheading stage. Even at 100% head emergence, differences in I.V.D. between stems and living leaves were not great with the early-maturing clones. However, wide differences existed between the values for stems and living leaves in mature plants. Heads were only slightly higher than stems in digestibility.Leaf digestibility was lower for those leaves dead or even partially dead. Nevertheless, in mature plants, dead leaves were higher in I.V.D. than heads or stems. Large differences occurred among clones in stem digestibility. However, the variability among clones in leaf digestibility was small.

Tests by Tissue Culture Methods on the Nature of Immunity Transplanted Skin

1948 ◽  
Vol s3-89 (7) ◽  
pp. 239-252
Author(s):  
P. B. MEDAWAR
Keyword(s):  
Tissue Culture ◽  
Cell Division ◽  
Epidermal Cell ◽  
Cell Suspensions ◽  
Acquired Immunity ◽  
Culture Methods ◽  
Division Frequency

The transplantation of skin from one rabbit to another elicits a reaction that conforms in main outline with that of an actively acquired immunity. The experiments described in this paper were designed to test the hypothesis that the regression of such grafts is secured by the action of antibodies demonstrable in vitro. Skin from adult rabbits has therefore been cultivated in the presence of serum and growing mesenchymal tissues derived solely from rabbits heavily and specifically immunized against it. Immune sera and tissues are without effect on the survival, cell-division frequency and migratory activities of explanted skin, and agglutinins for epidermal cell suspensions are not demonstrable in immune sera. With certain stated qualifications, it has therefore been concluded that the occurrence of free antibodies is not a sufficient explanation of the regression of skin homografts in vivo.

Missing pieces in protein deposition and mobilization inside legume seed storage vacuoles: calcium and magnesium ions

2012 ◽  
Vol 22 (4) ◽  
pp. 249-258 ◽  
Author(s):  
Cláudia N. Santos ◽  
Marta M. Alves ◽  
Isabel T. Bento ◽  
Ricardo B. Ferreira
Keyword(s):  
Alkaline Earth ◽  
Storage Proteins ◽  
Lupinus Albus ◽  
Physiological Role ◽  
Experimental Conditions ◽  
Protein Storage Vacuoles ◽  
Aggregation And Disaggregation ◽  
Alkaline Earth Cations

AbstractDuring the maturation of dicotyledonous seeds, organic carbon, nitrogen and sulphur are stored in protein storage vacuoles (PSVs) as storage globulins. Several studies point to the coexistence of storage proteins with proteases responsible for their degradation inside PSVs. Different mechanisms have been proposed to explain why there is no proteolysis during this period. Protein aggregation to form large supramolecular structures resistant to proteolytic attack could be the reason. However, during germination, and particularly following its completion, the globulin aggregates must undergo disintegration to allow protease attack for protein reserve mobilization. Based on the well-described concentration-dependent ability of Ca2+ and Mg2+ to promote in vitro aggregation and disaggregation of globulins, we explored a possible role for these alkaline earth cations in globulin packaging and mobilization. Ca2+ and Mg2+ measurements in purified PSVs [6.37 μmol and 43.9 μmol g− 1 dry weight (DW) of cotyledons, respectively] showed the presence of these two alkaline earth cations within this compartment. To our knowledge, this is the first time that Ca2+ and Mg2+ have been quantified in purified PSVs from Lupinus albus seeds. Considering the importance of these two alkaline earth cations inside PSVs, which represent 14.6% and 60.7% of the total seed Mg2+and Ca2+, respectively, globulin aggregation and disaggregation profiles were assayed using experimental conditions closer to those that are physiologically present (proportion of Ca2+ and Mg2+, and acidic pH). Based on: (1) the high in vivo abundance of Ca2+ and Mg2+ inside PSVs; and (2) globulin aggregation and disaggregation profiles, together with structural and physiological evidence already reported in the literature, an important physiological role for Ca2+ and Mg2+ in globulin packaging and mobilization inside PSVs is suggested.

scholarly journals Germinated and Ungerminated Seeds Extract from TwoLupinusSpecies: Biological Compounds Characterization and In Vitro and In Vivo Evaluations

2016 ◽  
Vol 2016 ◽  
pp. 1-8 ◽  
Author(s):  
Bogdan Andor ◽  
Corina Danciu ◽  
Ersilia Alexa ◽  
Istvan Zupko ◽  
Elena Hogea ◽  
...  
Keyword(s):  
Cinnamic Acid ◽  
Therapeutic Potential ◽  
Lupinus Albus ◽  
Biological Evaluation ◽  
Lupinus Angustifolius ◽  
Cinnamic Acid Derivatives ◽  
Natural Agents ◽  
Germinated Seeds

In recent years, nutraceuticals attracted a great amount of attention in the biomedical research due to their significant contribution as natural agents for prevention of various health issues. Ethanolic extracts from the ungerminated and germinated seeds ofLupinus albusL. andLupinus angustifoliusL. were analyzed for the content in isoflavones (genistein) and cinnamic acid derivatives. Additionally, the extracts were evaluated for antimicrobial, antiproliferative, and anti-inflammatory properties, using in vitro and in vivo tests. Germination proved to be a method of choice in increasing the amount of genistein and cinnamic acid derivatives in bothLupinus albusL. andLupinus angustifolius L.seeds. Biological evaluation of all vegetal extracts revealed a weak therapeutic potential for both ungerminated and germinated seeds.

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