AFLP Analysis and Improved Phytoextraction Capacity of Transgenic gshI-Poplar Clones (Populus x canescens L.) for Copper in vitro

2005 ◽  
Vol 60 (3-4) ◽  
pp. 300-306 ◽  
Author(s):  
Gábor Gyulai ◽  
Mervyn Humphreys ◽  
András Bittsánszky ◽  
Kirsten Skøtc ◽  
József Kiss ◽  
...  
Keyword(s):  
Culture Media ◽  
Leaf Disc ◽  
Aflp Analysis ◽  
Polymorphism Analysis ◽  
Tissue Culture Media ◽  
Dna Fragment ◽  
High Concentrations ◽  
Cu Uptake ◽  
Concentration Series

Abstract Clone stability and in vitro phytoextraction capacity of vegetative clones of P. x canescens (2n = 4x = 38) including two transgenic clones (ggs11 and lgl6) were studied as in vitro leaf disc cultures. Presence of the gshI-transgene in the transformed clones was detected in PCR reactions using gshI-specific primers. Clone stability was determined by fAFLP (fluorescent amplified DNA fragment length polymorphism) analysis. In total, 682 AFLP fragments were identified generated by twelve selective primer pairs after EcoRIDMseI digestion. Four fragments generated by EcoAGTDMseCCC were different (99.4% genetic similarity) which proves an unexpectedly low bud mutation frequency in P. \ canescens. For the study of phytoextraction capacity leaf discs (8 mm) were exposed to a concentration series of ZnSO4 (10-1 to 10-5 ᴍ) incubated for 21 days on aseptic tissue culture media WPM containing 1 μᴍ Cu. Zn2+ caused phytotoxicity only at high concentrations (10-1 to 10-2 ᴍ). The transgenic poplar cyt-ECS (ggs11) clone, as stimulated by the presence of Zn, showed elevated heavy metal (Cu) uptake as compared to the non-transformed clone. These results suggest that gshI-transgenic poplars may be suitable for phytoremediation of soils contaminated with zinc and copper.

scholarly journals In vitro tissue culture and genetic analysis of two high-CBD medical cannabis (Cannabis sativa L.) breeding lines

Genetika ◽  
2020 ◽  
Vol 52 (3) ◽  
pp. 925-941 ◽  
Author(s):  
Spela Mestinsek-Mubi ◽  
Sinja Svetik ◽  
Marko Flajsman ◽  
Jana Murovec
Keyword(s):  
Tissue Culture ◽  
Cannabis Sativa ◽  
Therapeutic Potential ◽  
Culture Media ◽  
Shoot Culture ◽  
Medical Cannabis ◽  
Breeding Lines ◽  
In Vitro Tissue Culture ◽  
Tissue Culture Media

The species Cannabis sativa L. has recently witnessed a resurgence of interest all over the world due to its multipurpose applications and the scientific confirmation of its pharmacological properties. Genotypes with high cannabinoid content are appreciated in the pharmaceutical and cosmetic industries due to their therapeutic potential. These genotypes, with predominantly high cannabidiol (CBD) content, are the subject of research and breeding in several programs, but to date, little data is published on the in vitro tissue culture of cannabis. Our study focused on the establishment of an efficient micropropagation method for two high-CBD breeding lines (MX-CBD-11 and MX-CBD-707) as the basis for advanced biotechnological breeding approaches. The results demonstrated that the in vitro culture of medical cannabis can be initiated on different culture media, that cultured plants can be successfully acclimatized, and that nodal position, and especially the genotype, have a significant influence on the success of shoot culture establishment. They showed that the published tissue culture media optimized for one high-THC strain of Mexican cannabis are not as efficient for other genotypes of (medical) cannabis. We complemented this research with a genetic study of 95 plants of the two breeding lines with 16 microsatellite (SSR) markers which clustered the plants based on breeding line. The results demonstrated that only 8 markers are needed for discrimination of all analyzed plants and their usefulness for clonal identification.

scholarly journals PERTUMBUHAN KULTUR TUNAS AKSILAR KENTANG (Solanum tuberosum L.) DENGAN PENAMBAHAN SUPER FOSFAT DAN KNO3 PADA MEDIA AB MIX SECARA IN VITRO

2019 ◽  
Vol 20 (2) ◽  
pp. 71
Author(s):  
Hamami Alfasani Dewanto ◽  
Desi Saraswati ◽  
Oetami Dwi Hadjoeningtijas
Keyword(s):  
Culture Media ◽  
Raw Materials ◽  
Solanum Tuberosum L ◽  
Leaf Color ◽  
Tissue Culture Media ◽  
Randomized Design ◽  
Plantlet Growth ◽  
Number Of Leaves ◽  
Dark Green

Murashige&Skoog-based medium Potatoes are one commodity that has the potential to be developed as a resource in the context of food diversification, farmers' income riser, non fossil export commodities and raw materials for processing industry. The objective of this research was to find out the effect of SP-36 fertilizer, KNO3 fertilizer, as well as the interaction between the two fertilizers on the growth of potato nodal culture on AB Mix media in vitro. The results of this study are expected to provide economical potato tissue culture media development. This research used factorial complete randomized design. The treatment were SP-36 concentration: 0 ppm; 50 ppm; 100 ppm; and 200 ppm, in combination with KNO3 concentration: 0 ppm; 100 ppm; 200 ppm; 400 ppm; and 600 ppm, The variables observed included number of leaves, leaf color, length of plantlets, fresh weight of plantlets and percentage of plantlets growth. Based on the results of the analysis of variance (ANOVA) F. Calculate < F. Table with the average success of plantlet growth between 87.5-100%. In addition, there are four types of leaf color produced, namely the color of yellowish green, pale green leaves, green, and dark green. Research showed that the interaction between SP-36 fertilizer and KNO3 fertilizer on AB Mix media had no significant effect on all observed variables.

scholarly journals Metabolomics Guides Rational Development of a Simplified Cell Culture Medium for Drug Screening against Trypanosoma brucei

2013 ◽  
Vol 57 (6) ◽  
pp. 2768-2779 ◽  
Author(s):  
Darren J. Creek ◽  
Brunda Nijagal ◽  
Dong-Hyun Kim ◽  
Federico Rojas ◽  
Keith R. Matthews ◽  
...  
Keyword(s):  
Cell Culture ◽  
Trypanosoma Brucei ◽  
Culture Medium ◽  
Culture Media ◽  
Culture Methods ◽  
Content Type ◽  
Drug Activity ◽  
Rational Development ◽  
High Concentrations

ABSTRACTIn vitroculture methods underpin many experimental approaches to biology and drug discovery. The modification of established cell culture methods to make them more biologically relevant or to optimize growth is traditionally a laborious task. Emerging metabolomic technology enables the rapid evaluation of intra- and extracellular metabolites and can be applied to the rational development of cell culture media. In this study, untargeted semiquantitative and targeted quantitative metabolomic analyses of fresh and spent media revealed the major nutritional requirements for the growth of bloodstream formTrypanosoma brucei. The standard culture medium (HMI11) contained unnecessarily high concentrations of 32 nutrients that were subsequently removed to make the concentrations more closely resemble those normally found in blood. Our new medium, Creek's minimal medium (CMM), supportsin vitrogrowth equivalent to that in HMI11 and causes no significant perturbation of metabolite levels for 94% of the detected metabolome (<3-fold change; α = 0.05). Importantly, improved sensitivity was observed for drug activity studies in whole-cell phenotypic screenings and in the metabolomic mode of action assays. Four-hundred-fold 50% inhibitory concentration decreases were observed for pentamidine and methotrexate, suggesting inhibition of activity by nutrients present in HMI11. CMM is suitable for routine cell culture and offers important advantages for metabolomic studies and drug activity screening.

scholarly journals Cultivation of Clinically Significant Hemoflagellates

2002 ◽  
Vol 15 (3) ◽  
pp. 374-389 ◽  
Author(s):  
Frederick L. Schuster ◽  
James J. Sullivan
Keyword(s):  
Tissue Culture ◽  
Culture Media ◽  
Life Cycles ◽  
Variable Component ◽  
Culture Cells ◽  
Tissue Culture Media ◽  
Oriental Sore ◽  
Clinically Significant ◽  
Cultivation In Vitro

SUMMARY The hemoflagellates, Trypanosoma spp. and Leishmania spp., are causal agents of a number of parasitic diseases having a major impact on humans and domestic animals over vast areas of the globe. Among the diseases are some of the most pernicious and deadly of human afflictions: African sleeping sickness, Chagas' disease, kala-azar, and Oriental sore. The organisms have complex, pleomorphic life cycles typically involving a vertebrate and an invertebrate host, the latter serving as a vector. In the vertebrate host, they are primarily blood and tissue parasites. In their transition from one host to another, the hemoflagellates undergo morphological, physiological, and biochemical changes that facilitate their growth and subsequent transmission. A major goal in the study of the hemoflagellates has been the cultivation in vitro of both vertebrate and invertebrate stages of the organisms. The first types of media used in their cultivation, and still useful for establishment of cultures, were undefined and contained a complex of ingredients. These gave way to semidefined formulations which included tissue culture media as a base and, as a next step, addition of tissue culture cells as a feeder layer to promote parasite growth. More recently developed media are completely defined, having replaced the feeder cells with various supplements. Serum, a sometimes-variable component of the media, can be replaced by various serum substitutes. This review focuses on the hemoflagellates that infect humans, describing stages in the development of media leading to the fully defined formulations that are now available for the cultivation of many of these organisms.

Atrial natriuretic peptide regulates release of Na+-K+-ATPase inhibitor from rat brain

1988 ◽  
Vol 254 (6) ◽  
pp. F912-F917 ◽  
Author(s):  
M. Crabos ◽  
D. A. Ausiello ◽  
G. T. Haupert ◽  
H. F. Cantiello
Keyword(s):  
Atrial Natriuretic Peptide ◽  
Rat Brain ◽  
Natriuretic Peptide ◽  
Culture Media ◽  
Electrolyte Balance ◽  
Atpase Inhibitor ◽  
Tissue Culture Media ◽  
The One ◽  
Atrial Natriuretic

Tissue culture media from incubations of fragments of rat brain were collected and partially purified. These supernatants were effective in inhibiting the Na+-K+ pump as indicated by a 77% reduction of ouabain-sensitive 86Rb+ uptake into human erythrocytes. Release of the Na+-K+-ATPase inhibitor depended on the amount of tissue, the temperature, and the length of incubation. Atrial natriuretic peptide (ANP) injected intravenously, or included (10(-8) M) in the in vitro incubation of brain tissue, decreased the release of the Na+-K+-ATPase inhibitor by 74 and 42%, respectively. Control experiments using the neuropeptide arginine vasopressin showed no effect on release of the inhibitor. These studies indicate that ANP is capable of regulating the release from brain of a Na+-K+-ATPase inhibitor with similar chromatographic characteristics to the one previously obtained from extraction of bovine hypothalamus and raise the possibility that the two factors are interrelated in the regulation of fluid and electrolyte balance.

Tetracycline-Based MMP Inhibitors Can Prevent Fibroblast-Mediated Collagen Gel Contraction In Vitro

1998 ◽  
Vol 12 (1) ◽  
pp. 86-93 ◽  
Author(s):  
S.A. Myers ◽  
R.G. Wolowacz
Keyword(s):  
Culture Media ◽  
Collagen Gel ◽  
Collagen Gels ◽  
Cytotoxic Effects ◽  
Chemically Modified ◽  
Tissue Culture Media ◽  
Collagen Gel Contraction ◽  
Chemically Modified Tetracyclines

Collagen gels in vitro can be contracted by fibroblasts. The role of matrix metalloproteinases (MMPs) in the contraction of collagen lattices by human neonatal foreskin fibroblasts (HuFFs) was investigated in tissue culture media supplemented by various doses of known gelatinase inhibitors. Fluorescent assays with model gelatinase substrates and media conditioned by fibroblasts apparently confirmed the ability of chemically modified tetracyclines (CMTs) to act as inhibitors of MMP2, and zymography demonstrated that this was the major cell-derived MMP activity. There were no observable effects on the rate of contraction of attached FPCLs containing 6 x 104 HuFFs (passages 18-25) with either CMT-5 or CMT-2 at all concentrations tested (0-100 μg/mL). However, at greater than 20 μg/mL doxycycline and greater than 5 μg/mL CMT-3, FPCL contraction was completely abolished. Quantitative assessment of cell viability by means of the MTT assay in monolayer and qualitatively within the FPCLs with CalceinAM suggested that differences were not due to cytotoxic effects. Seeding FPCLs with lower-passage fibroblasts produced identical trends. These results may implicate the involvement of MMPs in the process of gel contraction, although tetracyclines have effects additional to their ability to inhibit MMPs directly.

scholarly journals Urease Produced by Coccidioides posadasii Contributes to the Virulence of This Respiratory Pathogen

2006 ◽  
Vol 74 (1) ◽  
pp. 504-515 ◽  
Author(s):  
Fariba Mirbod-Donovan ◽  
Ruth Schaller ◽  
Chiung-Yu Hung ◽  
Jianmin Xue ◽  
Utz Reichard ◽  
...  
Keyword(s):  
Culture Media ◽  
Host Tissue ◽  
Respiratory Pathogen ◽  
Coccidioides Posadasii ◽  
Additional Material ◽  
Urease Gene ◽  
Predicted Signal Peptide ◽  
High Concentrations ◽  
Host Inflammatory Response

ABSTRACT Urease activity during in vitro growth in the saprobic and parasitic phases of Coccidioides spp. is partly responsible for production of intracellular ammonia released into the culture media and contributes to alkalinity of the external microenvironment. Although the amino acid sequence of the urease of Coccidioides posadasii lacks a predicted signal peptide, the protein is transported from the cytosol into vesicles and the central vacuole of parasitic cells (spherules). Enzymatically active urease is released from the contents of mature spherules during the parasitic cycle endosporulation stage. The endospores, together with the urease and additional material which escape from the ruptured parasitic cells, elicit an intense host inflammatory response. Ammonia production by the spherules of C. posadasii is markedly increased by the availability of exogenous urea found in relatively high concentrations at sites of coccidioidal infection in the lungs of mice. Direct measurement of the pH at these infection sites revealed an alkaline microenvironment. Disruption of the urease gene of C. posadasii resulted in a marked reduction in the amount of ammonia secreted in vitro by the fungal cells. BALB/c mice challenged intranasally with the mutant strain showed increased survival, a well-organized granulomatous response to infection, and better clearance of the pathogen than animals challenged with either the parental or the reconstituted (revertant) strain. We conclude that ammonia and enzymatically active urease released from spherules during the parasitic cycle of C. posadasii contribute to host tissue damage, which exacerbates the severity of coccidioidal infection and enhances the virulence of this human respiratory pathogen.

scholarly journals PENGARUH EKSTRAK Sargassum cristaefolium PADA MULTIPLIKASI Dendrobium antennatum Rchb.f SECARA IN VITRO

2020 ◽  
Vol 8 (1) ◽  
pp. 18
Author(s):  
Faisal Ansyarif ◽  
Mursal Ghazali ◽  
Aida Muspiah ◽  
Rina Kurnianingsih
Keyword(s):  
Culture Media ◽  
Positive Control ◽  
Negative Control ◽  
Growth Media ◽  
Extract Concentration ◽  
Tissue Culture Media ◽  
Randomized Design ◽  
Completely Randomized Design ◽  
Natural Cytokinin

The purpose of this study was to determine the potential and concentration of Sargassum cristaefolium extract as a natural cytokinin in tissue culture media of Dendrobium antennatum Rchb.f. This study is experimental with a completely randomized design, using several extract concentrations compared with the positive control (BAP 1.5 ppm) and negative control (MS0 media). Extract concentrations used 5 ppm, 10 ppm, 15 ppm, 20 ppm, and 25 ppm. Based on analysis of variance (ANOVA), the effect of Sargassum cristaefolium extract on the growth media significant on all parameters. Sargassum cristaefolium extracts caused different responses at certain levels of concentration. Extract concentration of 10 ppm was able to initiate the highest number of shoots and leaves compared to other extract concentrations, where as the concentration 20 ppm was able to accelerate and increase root growth.

scholarly journals A Comparison between Synthetic and Organic Iron Chelators on in Vitro Growth of Nicotiana tabacum Callus Cultures

HortScience ◽  
1996 ◽  
Vol 31 (4) ◽  
pp. 630g-631
Author(s):  
Lisa C. Berg ◽  
Henry R. Owen
Keyword(s):  
Nicotiana Tabacum ◽  
Culture Media ◽  
Basal Medium ◽  
Callus Cultures ◽  
Callus Growth ◽  
Fresh Weight ◽  
Iron Chelate ◽  
Tissue Culture Media ◽  
The Media

Nicotiana tabacum callus growth (fresh weight) was measured after culture in the light (16-hour photoperiod) or in darkness for four different culture media, differing in iron chelate type or concentration. All media contained MS basal medium supplemented with 30 g·L–1 sucrose, 2 mg·L–1 IAA, 0.2 mg·L–1 KIN, and 7 g·L–1 agar, pH 5.8. Three of the media contained iron-metalosate (Albion Laboratories), an organic iron chelate, at 100, 200, and 400 micromolar concentrations, and the fourth medium contained 100 μm Fe-EDTA. Twenty-five culture tubes were prepared for each of the 4 different media concentrations and 2 light treatments (8 treatments total). A 1-cm3 callus explant was used for each treatment and cultured for 56 days at 20°C. About 20-fold increases in callus fresh weight were observed for cultures incubated in light or in darkness. In addition, callus growth was not significantly affected by iron chelate type, suggesting the potential utility of this organic chelator in tissue culture media to alleviate potential problems of light-induced EDTA instability and subsequent IAA inactivation. These cultures are being maintained to examine the influence of iron chelate type on organogenesis.

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