Enhancement by Osmotic Treatment of Hairy Root Transformation of Alfalfa Suspension Cultures, and Chromosomal Variation in the Transformed Tissues

1997 ◽  
Vol 24 (3) ◽  
pp. 345 ◽  
Author(s):  
Zi-Qin Xu ◽  
Jing-Fen Jia ◽  
Zhi-De Hu
Keyword(s):  
Chromosomal Abnormalities ◽  
Paper Electrophoresis ◽  
Suspension Cultures ◽  
Transformation Frequency ◽  
Dichlorophenoxyacetic Acid ◽  
Structural Alterations ◽  
Root Transformation ◽  
Selection Medium ◽  
Osmotic Treatment ◽  
2,4 Dichlorophenoxyacetic Acid

Seeds of alfalfa were germinated on Murashige and Skoog medium without phytohormones. The hypocotyls ofseedlings were excised and cultured on the same medium with 2,4- dichlorophenoxyacetic acid (9.05 mM) to induce callus. Granular calluses were suspended and cultured in Schenk and Hildebrandt medium supplemented with 2.26 mM 2,4-dichlorophenoxyacetic acid on a shaker at 80 rpm. Alfalfa suspension cultures were treated with 0.45 M mannitol for 1 h, and washed twice with 0.16 M CaCl2 .2H2 O. After pretreatment, the pellets were resuspended in Schenk and Hildebrandt medium without phytohormone (10 mL g-1 suspension cultures), and 0.2 mL of Agrobacterium rhizogenes suspension was added. A mixture of alfalfa cells and Agrobacterium was co-cultured at 25 ± 2˚C for 2 days. Transformants (transformed calluses and hairy roots) were obtained by hormone-free selection. Several factors, such as culturing stage of suspension cultures, phytohormone constitution of suspension medium and basal selection medium of transformants, affected the transformation frequency remarkably. Paper electrophoresis revealed that over 70% of transformants could synthesise agropine and mannopine. A comparative cytological analysis revealed the number and structural alterations of chromosomes in the resulting materials. The transformed tissue was not able to be regenerated, possibly due to chromosomal abnormalities.

scholarly journals Effects of Plant Growth Regulators and Sugars on Ehretia asperula Zoll. et Mor. Cell Cultures

2021 ◽  
Author(s):  
P.T.M. Tram ◽  
N.K. Suong ◽  
L.T.T. Tien
Keyword(s):  
Cell Suspension ◽  
Callus Induction ◽  
Malignant Tumors ◽  
Single Cells ◽  
Cell Suspension Cultures ◽  
Suspension Cultures ◽  
Dichlorophenoxyacetic Acid ◽  
Human Immune System ◽  
Friable Callus ◽  
2,4 Dichlorophenoxyacetic Acid

Background: Belonging to the Boraginacae family, Ehretia asperula Zoll. et Mor., called “Xa den”, is a precious medicinal plant also known as the “cancer tree” by the Muong ethnic group in Hoa Binh Province, Vietnam. Xa den has been demonstrated to inhibit the development of malignant tumors, reduce oxidation and enhance the human immune system. This research focused on examining friable callus induction from young stems of Ehretia asperula Zoll. et Mor. Methods: Samples of Xa den were less than two years old. Young stems with 2 to 6 leaves served as explants for callus induction. Explants placed on autoclaved B5 nutrients incubated at 25oC, in the dark. The testing factors were concentrations of 2,4-Dichlorophenoxyacetic acid (2,4-D) and Benzyl adenine (BA), types and concentrations of sugars.Result: Friable callus was induced on B5 medium with 0.4 mg/L of 2,4-D, 0.1 mg/L of BA and 30 g/L of glucose at the highest rate (100%). Additionally, callus grew best after 5 weeks of culture weighing 0.194 g. Friable callus was used as material for cell suspension cultures. After two weeks, the Xa den cell suspension cultures contained single cells and small cell clumps. The liquid medium had changed from dark yellow to light brown.

Culture methods and detection of glucanases in suspension cultures of wheat and barley

1968 ◽  
Vol 46 (5) ◽  
pp. 417-421 ◽  
Author(s):  
Oluf L. Gamborg ◽  
D. E. Eveleigh
Keyword(s):  
Hordeum Vulgare ◽  
Culture Media ◽  
Early Period ◽  
Suspension Cultures ◽  
Dichlorophenoxyacetic Acid ◽  
Hordeum Vulgare L ◽  
Culture Methods ◽  
Mineral Salts ◽  
Triticum Vulgare ◽  
2,4 Dichlorophenoxyacetic Acid

Suspension cultures of Triticum monococcum L., Triticum vulgare Vill. var. Thatcher, Hordeum vulgare L. var. Gateway, and Hordeum vulgare L. var. Gateway mutant yv2 have been established. The cultures were derived from root sections of seedlings and cultured in a denned medium consisting of mineral salts, sucrose, B vitamins, and 2,4-dichlorophenoxyacetic acid, with nitrate and ammonia as the sources of nitrogen. In the early period of the cultures the cell aggregates readily, differentiated to form roots, but this characteristic diminished after several generations of subculture. The cells and medium contained a number of glucanases. The presence of a laminaranase (endo-β-(1 → 3)-D-glucan glucanohydrolase (EC 3.2.1.99)) that did not attack lichenan was established. The culture media of the wheat contained an oligosaccharide which on acid hydrolysis yielded galactose, arabinose, and xylose. Hydrolysis of a cell-wall fraction yielded the same sugars in addition to glucose and mannose.

Establishment of Fine Suspension Cultures of Triticum tauschii ([Coss.] Schmal.) which remain Embryogenic for Several Years

1999 ◽  
Vol 47 (4) ◽  
pp. 611
Author(s):  
Shoukat Afshar-Sterle ◽  
James F. Kollmorgen ◽  
Geoffrey B. Fincher
Keyword(s):  
Suspension Cultures ◽  
Dichlorophenoxyacetic Acid ◽  
D Genome ◽  
Triticum Tauschii ◽  
Initiation Phase ◽  
Genome Donor ◽  
Embryogenic Suspension ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Term Maintenance

Immature embryos of 10 accessions of Triticum tauschii, the D genome donor of Triticum aestivum, were used to produce embryogenic callus for the initiation of suspension cultures. For the long-term maintenance of embryogenicity of these suspensions, it was necessary to use different media for the initiation, establishment and maintenance phases. The initiation phase required media supplemented with L-proline, L-asparagine and L-glutamine, together with Dicamba at 12 mg L −1 and maltose. In the establishment phase, it was essential to reduce the concentration of Dicamba to 6 mg L −1 for the rapid production of fine suspension cultures. Finally, the long-term maintenance of a capacity for regeneration depended on the inclusion of 1.1 mg L −1 2,4-dichlorophenoxyacetic acid and 30 g L −1 sucrose in the medium. By the use of these procedures, long-term embryogenic fine suspension cultures were established from two accessions, while non-embryogenic fine suspension cultures were produced from five accessions. Over 90% of plants regenerated from fine suspensions of 1-year-old embryogenic cultures were fertile, and embryogenic suspension cultures retained their regeneration capacity for more than 3 years.

scholarly journals Influence of Growth Regulators on Callogenesis and Somatic Embryo Development in Date Palm (Phoenix dactyliferaL.) Sahelian Cultivars

2012 ◽  
Vol 2012 ◽  
pp. 1-8 ◽  
Author(s):  
Djibril Sané ◽  
Frédérique Aberlenc-Bertossi ◽  
Léopold Ibrahima Djitiningo Diatta ◽  
Badara Guèye ◽  
Abdourahman Daher ◽  
...  
Keyword(s):  
Nuclear Dna ◽  
Leaf Explants ◽  
Suspension Cultures ◽  
Naphthalene Acetic Acid ◽  
Dichlorophenoxyacetic Acid ◽  
Benzyl Adenine ◽  
Embryogenic Cell ◽  
Significant Difference ◽  
2,4 Dichlorophenoxyacetic Acid

This study provides a physiological analysis of somatic embryogenesis in four elite cultivars of date palms: Ahmar, Amsekhsi, Tijib, and Amaside, from the initial callogenesis to establishment and proliferation of embryogenic suspension cultures. Somatic embryos development and in vitro plants rooting were also studied. For each step, auxins and cytokinins concentrations were optimised. The primary callogenesis from leaf explants of seedlings appeared highly dependent on genotype. Ahmar (80%) and Amsekhsi (76%) appeared highly callogenic, whereas Tijib (10%) and Amaside (2%) produced low amounts of calluses. 2,4-Dichlorophenoxyacetic acid appeared favorable to the induction of primary callogenesis and its effect was enhanced by the addition of benzyl adenine or adenine sulfate. Secondary friable calli obtained from chopped granular calli were used to initiate embryogenic cell suspensions in media supplied with 2,4-dichlorophenoxyacetic acid. Suspension cultures showed a growth rate of fourfold after four subcultures in presence of 2,4-dichlorophenoxyacetic acid 2 mg/L. Our results showed that a seven-day transitory treatment with benzyl adenine 0,5 mg/L was necessary to optimize embryos development. Naphthalene acetic acid induced the development of primary orthogravitropic roots during embryos germination. The comparison with cytofluorometry of nuclear DNA amounts showed no significant difference in ploidy level between regenerated plants and seedlings.

Effect of either 2,4-dichlorophenoxyacetic acid or kinetin on production and composition of various fractions from extracellular polysaccharides produced by cotyledon cell suspension cultures of bush bean (Phaseolus vulgaris cv. Contender)

1978 ◽  
Vol 56 (15) ◽  
pp. 1816-1822 ◽  
Author(s):  
S. Mante ◽  
W. G. Boll
Keyword(s):  
Cell Suspension ◽  
Cell Suspension Cultures ◽  
Extracellular Polysaccharide ◽  
Suspension Cultures ◽  
Extracellular Polysaccharides ◽  
Dichlorophenoxyacetic Acid ◽  
Neutral Sugar ◽  
Bush Bean ◽  
Cotyledon Cell ◽  
2,4 Dichlorophenoxyacetic Acid

Cotyledon cell suspension cultures of bush bean required 2, 4-dichlorophenoxyacetic acid (2,4-D) for growth. Kinetin was not essential but was required for optimum growth. Both of the regulators were required for optimum production of extracellular polysaccharide (EP).The two regulators had different effects upon the production of three polysaccharide fractions (two pectins and a neutral polysaccharide) isolated from the EP at various stages of the culture cycle.The neutral sugar composition of the pectin fractions from all treatments, including regulator treatments, showed considerable fluctuation during the culture cycle and could be changed markedly by regulator treatments. Changes in composition of the neutral polysaccharide were slight. As a consequence of these results it is now possible to obtain cells differing in the nature of the systems synthesizing, or controlling the synthesis of, the polysaccharides.

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