scholarly journals Conservation and Production of Ipecac (Cephaelis ipecacuanha Rich.) Plants from Long Term Shoot Cultures

1970 ◽  
Vol 18 (2) ◽  
pp. 157-164 ◽  
Author(s):  
Rituparna Kundu Chaudhuri ◽  
Timir Baran Jha
Keyword(s):  
Growth Conditions ◽  
Shoot Cultures ◽  
Shoot Bud ◽  
Regenerated Plants ◽  
One Year ◽  
Bud Initiation ◽  
Cephaelis Ipecacuanha ◽  
Important Medicinal Plant

In vitro germplasms of ipecac (Cephaelis ipecacuanha Rich.), an important medicinal plant, maintained through reduced growth conditions for more than 12-years, were used as source material for micropropagation. MS with different combinations of Kn (2 mg/l), BAP (2 mg/l), 2iP (2-3 mg/l), NAA (0.2 mg/l) and adenine (40-100 mg/l) as additive was used to induce fresh multiplication of shoots from the nodal meristems and direct shoot bud initiation on the internodal segments. Complete plant regeneration has been achieved from such long term cultures. Regenerated plants maintained their phenotypic and chromosomal stability. Eighty per cent hardened plants, survived in the field condition, are growing well and 25% of them produced flowers within one year. Long term preservation through reduced growth conditions and successful regeneration of morphologically stable plants with stable chromosome numbers (2n = 22) from such long term cultures of ipecae plants.  Key words: Long term culture, Ipecac, micropropagation, flowering D.O.I. 10.3329/ptcb.v18i2.3646 Plant Tissue Cult. & Biotech. 18(2): 157-164, 2008 (December)

scholarly journals Clonal fidelity of chrysanthemum regenerated from long term cultures

Genetika ◽  
2006 ◽  
Vol 38 (3) ◽  
pp. 243-249 ◽  
Author(s):  
Sladjana Jevremovic ◽  
Milana Trifunovic ◽  
Marija Nikolic ◽  
Angelina Subotic ◽  
Ljiljana Radojevic
Keyword(s):  
Morphological Characteristics ◽  
Flower Color ◽  
Stem Segment ◽  
Shoot Cultures ◽  
Ms Medium ◽  
Color Changes ◽  
In Vitro Plants ◽  
One Year

Morphological characteristics of flowers of long term regenerated chrysanthemum, cv. "White Spider", after ten years of micropropagation are investigated. Shoot cultures are established and maintained more than ten years by stem segment culture on MS medium supplemented with BAP and NAA (1.0, 0.1 mgL-1, respectively). Rooting of shoots (100 %) has done on MS medium without hormones and it was very successful after ten years, as well as, after two or eight years of micropropagation. Acclimation of rooted chrysanthemum plantlets at greenhouse conditions was excellent and after appropriate photoperiod "in vitro" plants flowered 90.3 % and have the same flower color, shape and size as mother plants. Flower color changes of "in vitro" plants are observed during another flowering cycle one year after acclimatization. Observed variations of chrysanthemum flowers could be attributed to epigenetic factors.

Long-term in vitro Storage Technique of Valuable Birch Genotypes and Plant Production on its Basis

2019 ◽  
pp. 57-67
Author(s):  
T.M. Tabatskaya ◽  
N.I. Vnukova
Keyword(s):  
Plant Production ◽  
High Survival ◽  
Ex Vitro ◽  
Regenerative Capacity ◽  
In Vitro Storage ◽  
Interspecific Differences ◽  
Regenerated Plants ◽  
Growing Conditions

A technique for the long-term (up to 27 years) in vitro storage of valuable birch genotypes under normal (25 °C, 2.0 klx, 16-h day and 8-h night) and low temperature (4 °C, 0.5 klx, 6-h day and 18-h night) growing conditions on hormone-free media has been described. The study explored for the first time the influence of different strategies to store the clones of Betula pubescens and B. pendula var. сarelica (6 genotypes) on the regenerative capacity of collection samples, adaptive potential of regenerated plants and plant production by the in vitro and ex vitro techniques. It was established that both storage strategies provided a persistently high survival rate (82-100%) and regenerative capacity of in vitro shoots (the multiplication coefficient of 4.2-6.3 and rhizogenic activity of 90-100%). The clones retained their characteristics of height growth under the in vitro and ex vitro conditions, and demonstrated intraclonal homogeneity and lack of signs of somaclonal variability. The plants showed substantial interspecific differences at the stage of multiplication and transfer to the greenhouse. The highest percentage of acclimated plants (75-98% depending on the clone genotype) was obtained after planting of micro plants straight in the greenhouse, which simplified the technology and made plant production less costly. long-term in vitro storage, birch, species, genotype, micropropagation, ex vitro adaptation, plant material

scholarly journals Development and test for long-term stability of a synthetic standard for a quantitative Cryptosporidium parvum LightCycler™ PCR assay

2005 ◽  
Vol 3 (1) ◽  
pp. 15-25 ◽  
Author(s):  
Renata Filkorn-Kaiser ◽  
Konrad Botzenhart ◽  
Albrecht Wiedenmann
Keyword(s):  
Cryptosporidium Parvum ◽  
Surrogate Marker ◽  
Target Sequence ◽  
Positive Trend ◽  
Long Term Stability ◽  
Synthetic Standard ◽  
One Year

A recently described quantitative rapid cycle real time PCR (LightCycler™) assay detects Cryptosporidium parvum after in vitro excystation, which is a surrogate marker for the viability of the organisms. In the original assay the quantification standard is a dilution series of C. parvum oocysts with a microscopically determined excystation rate. The need to keep suspensions of viable oocysts in stock and to continuously monitor their excystation rate, however, renders the assay impracticable for routine application. A synthetic standard was developed to replace the in vivo standard and was calibrated using oocysts with known excystation rates. The standard consists of a 486 bp DNA segment ranging from 229 bp upstream to 79 bp downstream of the actual PCR target site. Aliquots of the standard were frozen and stored at −20 °C and at −70 °C or lyophilised and stored at room temperature in the dark. For a period of one year samples preserved with each of the three methods were restored every four or five weeks. They were amplified in the LightCycler™ and the crossing points (CP) were monitored. No significant trend in the raw CP values could be observed for any of the three storage methods. However, when the methods were compared to each other by calculating the CP ratios (−20 °C/−70 °C; −20 °C/lyophilised; −70 °C/lyophilised) at the 10 monitoring dates, the CP ratios −20 °C/−70 °C and −20 °C/lyophilised showed a highly significant positive trend (p<0.0001) while the CP ratio −70 °C/lyophilised did not differ from the null hypothesis (p=0.53). It can be concluded that the latter two preservation methods are both appropriate, while storage at −20 °C is less advisable. Calculations based on the molecular weight of the standard and on the assumption of an average yield of three sporozoites per oocyst led to the conclusion that the target sequence is probably located on a double copy gene

Stroma-free long-term growth of primary acute myeloid leukemia (AML) cells.

2017 ◽  
Vol 35 (15_suppl) ◽  
pp. e18535-e18535
Author(s):  
Jing Chen ◽  
Glorymar Ibanez Sanchez ◽  
Paul Calder ◽  
Mark G. Frattini
Keyword(s):  
Growth Factors ◽  
Conditioned Medium ◽  
Stromal Cells ◽  
Drug Sensitivity ◽  
Growth Conditions ◽  
Leukemic Cells ◽  
Sensitivity Testing ◽  
Drug Sensitivity Testing

e18535 Background: To individualize therapy for relapsed/refractory AML patients, optimal in-vitro culture conditions to support primary leukemic cells are essential for drug sensitivity testing. Our lab has validated a high throughput chemosensitivity assay with primary AML cells maintained by growth factors (cytokines); however, growth factors have not been shown to support long-term assays of primary AML cells. Stromal cells of the tumor microenvironment are crucial to maintain normal hematopoiesis and leukemic cells and have been shown to support long term in-vitro expansion of primary AML cells. However, there is little information characterizing these growth conditions. The aim of this study was to compare long-term proliferation and phenotypes of primary AML cells with growth factors or stromal support to best determine their utility as a platform for drug sensitivity testing in functional assays. Methods: Patient-derived AML cells were cultured in 96-well plates in: 1) cell culture medium only 2) Human HS5 or HS27 stromal cells 3) HS5 or HS27 stroma-conditioned media or 4) cytokine cocktail. Viability readout by Guava ViaCount and leukemic cell surface phenotypes by fluorescently-conjugated antibodies were performed weekly over 3 weeks. Results: Primary AML cells cultured with only cytokines maintained proliferation at 3 weeks. In comparison, AML cells cultured in HS5 stroma-conditioned medium also maintained proliferation at a similar rate at 3 weeks, while co-culture with HS5 stromal cells demonstrated significantly higher proliferation. Leukemic immunophenotypes were maintained for all growth conditions over 3 weeks. Conclusions: Contrary to known data, primary AML cells with cytokines continued to expand at 3 weeks, at a similar rate to HS5 stroma-conditioned medium, a finding that has not been reported. Consistent with previous studies, we confirmed that stromal cells such as HS5 can provide long-term in-vitro expansion of primary AML cells, which cannot be substituted by stroma-conditioned medium. The ability to maintain long-term expansion of primary AML cells by both cytokines and stromal cells sets up a platform for a direct comparison of high throughput drug sensitivity testing under these growth conditions.

scholarly journals In Vitro Regeneration of Phyllanthus amarus Schum. and Thonn.: An Important Medicinal Plant

Our Nature ◽  
1970 ◽  
Vol 7 (1) ◽  
pp. 110-115 ◽  
Author(s):  
A. Sen ◽  
M.M. Sharma ◽  
D. Grover ◽  
A. Batra
Keyword(s):  
In Vitro Regeneration ◽  
Multiple Shoots ◽  
Shoot Elongation ◽  
Nodal Segments ◽  
Nodal Segment ◽  
Phyllanthus Amarus ◽  
Regenerated Plants ◽  
Half Strength ◽  
Important Medicinal Plant

An efficient in vitro plant regeneration protocol was developed for the medicinally potent plant species Phyllanthus amarus Schum. and Thonn. (Euphorbiaceae) using nodal segment as explant. Maximum multiplication of shoots (15.275±0.96) was achieved on Murashige and Skoog’s medium supplemented with BAP (0.5 mg/l) after 3-4 weeks of inoculation. The shoots were separated from cluster and subcultured for their elongation on the same medium. In vitro flowering was also observed on the elongated shoots after 3–4 weeks of sub culturing on the shoot elongation medium. In vitro rooting was obtained on half strength MS medium supplemented with IBA (0.5 mg/l).  Regenerated plants were successfully hardened and acclimatized, 80 % of plantlets survived well under natural conditions after transplantation.Key words: In vitro regeneration, multiple shoots, nodal segments, Phyllanthus amarusDOI: 10.3126/on.v7i1.2557Our Nature (2009) 7:110-115

Changing chemosusceptibility in the second-stage larvae of Toxocara canis by long-term incubation

1993 ◽  
Vol 67 (2) ◽  
pp. 145-150 ◽  
Author(s):  
Nobuaki Akao ◽  
Yoshihisa Goto ◽  
Kaoru Kondo ◽  
Yoshisuke Tsuda
Keyword(s):  
Decanoic Acid ◽  
Toxocara Canis ◽  
Second Stage ◽  
One Year

AbstractSecond-stage larvae of Toxocara canis were maintained in vitro for one year. Susceptibility of the larvae to drugs was evaluated by means of minimal larvicidal concentration (MLC) and larval bursting percentage. MLCs of citral and decanoic acid were almost constant throughout all stages of incubation. However, bursting percentage markedly varied within the first 20 weeks of incubation. Therefore, while larvae are available for use in the MLC assay at any stage of incubation, those beyond the first 20 weeks after incubation should be used for the bursting assay to obtain reproducible results.

scholarly journals Long-term in Vitro Regeneration of Hamelia patens

HortScience ◽  
1995 ◽  
Vol 30 (4) ◽  
pp. 873G-874
Author(s):  
D. Sankhla ◽  
T.D. Davis ◽  
N. Sankhla ◽  
A. Upadhyaya
Keyword(s):  
Culture Medium ◽  
In Vitro Regeneration ◽  
Shoot Formation ◽  
Shoot Tips ◽  
Shoot Cultures ◽  
Ms Medium ◽  
Prolific Shoot ◽  
Ornamental Shrub

This report describes an efficient in vitro regeneration protocol for H. patens (firebush), a heat-tolerant ornamental shrub native to tropical and subtropical America. Shoot cultures were initially established using shoot tips placed on MS-revised medium containing 2.3 μM 2,4-D, 2.3 μM kinetin, and 0.25% polyvinylpyrrolidone. Other types of explants (nodal and internodal segments, leaf pieces, floral buds) did not regenerate shoots when placed on this medium. Two-month-old plantlets derived from the shoot tips were subcultured on MS medium supplemented with 0.5 μM thidiazuron (TDZ), and within 3 to 4 weeks, some callus was produced at the root–shoot junction. When this callus, with a small portion of the root and shoots, was placed on MS medium with 0.05 μM TDZ and 0.01 μM ABA, prolific shoot formation occurred within 3 to 4 weeks followed by root formation. By regular subculturing every 5 to 6 weeks, hundreds of plantlets have been obtained over the past 3 years with no apparent decline in regeneration potential. Addition of activated charcoal (0.5%) to the culture medium has greatly improved growth of the plantlets.

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