The in vitro growth and metabolism of Acer saccharum tissue

1971 ◽  
Vol 49 (4) ◽  
pp. 495-500 ◽  
Author(s):  
Martin C. Mathes ◽  
Mariafranca Morselli ◽  
J. W. Marvin
Keyword(s):  
Acer Saccharum ◽  
Sugar Maple ◽  
Culture Medium ◽  
Basal Medium ◽  
Soluble Starch ◽  
Defined Medium ◽  
Tissue Growth ◽  
Starch Digestion ◽  
Callus Tissues

Isolated sugar maple callus was grown on defined medium containing 0.2 kinetin and 2.0 ppm indoleacetic acid. Selected components of the basal medium were varied but did not result in increased tissue growth. The presence of 1, 5, or 25 ppm gibberellic acid in the culture medium did not increase the secretion of starch digesting materials by the tissue. The diameter of the starch digestion zone was not influenced by changing the acidity of the medium in the range of pH which supported tissue growth. The ability of isolated callus tissues from various species to digest extracellular starch was examined. It was found that equal amounts of various tissues do not possess an equal capacity for the in vitro secretion of extracellular materials, most probably enzymes, into the culture medium. It was found that isolated sugar maple tissue metabolized sucrose, galactose, and cellobiose while mannose and rhamnose were not used.Isolated sugar maple tissue which had been subcultured on the same medium for a long period was found to undergo nutritional changes which enabled the tissue to adapt to the medium. The starch-digestion enzyme activity of the tissue was found to increase when sugar maple tissue was placed on soluble starch medium.

scholarly journals Transcriptional Profiling of Porcine Blastocysts Produced In Vitro in a Chemically Defined Culture Medium

Animals ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 1414
Author(s):  
Josep M. Cambra ◽  
Emilio A. Martinez ◽  
Heriberto Rodriguez-Martinez ◽  
Maria A. Gil ◽  
Cristina Cuello
Keyword(s):  
Culture Medium ◽  
Embryo Quality ◽  
Transcriptional Profiling ◽  
Defined Medium ◽  
Platelet Factor ◽  
Defined Media ◽  
Defined Culture ◽  
The Impact

The development of chemically defined media is a growing trend in in vitro embryo production (IVP). Recently, traditional undefined culture medium with bovine serum albumin (BSA) has been successfully replaced by a chemically defined medium using substances with embryotrophic properties such as platelet factor 4 (PF4). Although the use of this medium sustains IVP, the impact of defined media on the embryonic transcriptome has not been fully elucidated. This study analyzed the transcriptome of porcine IVP blastocysts, cultured in defined (PF4 group) and undefined media (BSA group) by microarrays. In vivo-derived blastocysts (IVV group) were used as a standard of maximum embryo quality. The results showed no differentially expressed genes (DEG) between the PF4 and BSA groups. However, a total of 2780 and 2577 DEGs were detected when comparing the PF4 or the BSA group with the IVV group, respectively. Most of these genes were common in both in vitro groups (2132) and present in some enriched pathways, such as cell cycle, lysosome and/or metabolic pathways. These results show that IVP conditions strongly affect embryo transcriptome and that the defined culture medium with PF4 is a guaranteed replacement for traditional culture with BSA.

Continuous in vitro culture of Babesia divergens in a serum-free medium

Parasitology ◽  
1997 ◽  
Vol 115 (1) ◽  
pp. 81-89 ◽  
Author(s):  
N. GRANDE ◽  
E. PRECIGOUT ◽  
M. L. ANCELIN ◽  
K. MOUBRI ◽  
B. CARCY ◽  
...  
Keyword(s):  
Human Serum ◽  
Reduced Glutathione ◽  
Lipid Fraction ◽  
Basal Medium ◽  
Defined Medium ◽  
Specific Lipids ◽  
Serum Free ◽  
Babesia Divergens ◽  
Lipid Fractions

Babesia divergens was cultivated in RPMI 1640 (25 mM HEPES) supplemented with 10% human serum (RPMI-10% HS) with a high percentage of parasitized erythrocytes (PPE) ([ges ]40%). Standardization of in vitro tests, purification of exoantigens, biochemical studies and the safety of the culture handler motivated the development of a serum-free defined medium. Removal of serum greatly reduced the PPE but, after a period of adaptation, the culture was continuous and the parasite was able to develop a 3% routine PPE. Addition of vitamins or reduced glutathione in basal medium (RPMI) did not improve the PPE. The supplementation of basal medium with lipidic carrier (Albumax I or bovine serum albumin–Cohn's fraction V) promoted the growth of B. divergens with high PPE (>30%) close to those obtained in RPMI–10% HS. Neither protein nor lipid fractions alone were able to restore the growth of B. divergens. Nevertheless, the whole lipid fraction from serum or Albumax I added to delipidated albumin partially restored the growth (7% PPE), indicating that the presentation of specific lipids by a carrier is crucial for the parasite. All the data indicate that Albumax I can replace human serum offering the advantages of safety, standardization for chemosensitivity tests, and exoantigen purification.

scholarly journals Three sympatrically occurring species of Metarhizium show plant rhizosphere specificity

Microbiology ◽  
2011 ◽  
Vol 157 (10) ◽  
pp. 2904-2911 ◽  
Author(s):  
Michael Wyrebek ◽  
Cristina Huber ◽  
Ramanpreet Kaur Sasan ◽  
Michael J. Bidochka
Keyword(s):  
Tree Species ◽  
Acer Saccharum ◽  
Panicum Virgatum ◽  
Sugar Maple ◽  
Root Exudate ◽  
Pathogenic Fungus ◽  
Plant Roots ◽  
Metarhizium Robertsii ◽  
Grass Roots

Here we tested the hypothesis that species of the soil-inhabiting insect-pathogenic fungus Metarhizium are not randomly distributed in soils but show plant-rhizosphere-specific associations. We isolated Metarhizium from plant roots at two sites in Ontario, Canada, sequenced the 5′ EF-1α gene to discern Metarhizium species, and developed an RFLP test for rapid species identification. Results indicated a non-random association of three Metarhizium species (Metarhizium robertsii, Metarhizium brunneum and Metarhizium guizhouense) with the rhizosphere of certain types of plant species (identified to species and categorized as grasses, wildflowers, shrubs and trees). M. robertsii was the only species that was found associated with grass roots, suggesting a possible exclusion of M. brunneum and M. guizhouense. Supporting this, in vitro experiments showed that M. robertsii conidia germinated significantly better in Panicum virgatum (switchgrass) root exudate than did M. brunneum or M. guizhouense. M. guizhouense and M. brunneum only associated with wildflower rhizosphere when co-occurring with M. robertsii. With the exception of these co-occurrences, M. guizhouense was found to associate exclusively with the rhizosphere of tree species, predominantly Acer saccharum (sugar maple), while M. brunneum was found to associate exclusively with the rhizosphere of shrubs and trees. These associations demonstrate that different species of Metarhizium associate with specific plant types.

scholarly journals In vitro answer of Bulgarian pepper (Capsicum annuum L.)

Genetika ◽  
2006 ◽  
Vol 38 (2) ◽  
pp. 129-136
Author(s):  
Velichka Rodeva ◽  
Stanislava Grozeva ◽  
Velichka Todorova
Keyword(s):  
Culture Medium ◽  
Culture Media ◽  
Casein Hydrolysate ◽  
Basal Medium ◽  
Regeneration Ability ◽  
Stem Explants ◽  
Media Composition ◽  
Breeding Lines ◽  
High Level

Callusogenesis and regeneration ability of cotyledon and hypocotyl explants from three Bulgarian pepper varieties in MS basal medium supplemented with l-3mg/l BAP. l.0mg/1 IAA and 0.5mg/l GA3 was studied. In the different variants of culture medium was registered high level of callusogenesis and organogenesis in both type of explants from the all varieties. The highest percentage of plant-regenerants is established in cotyledon explants (from 3.3 to 18.3) in variant 3 of the culture medium containing 3mg/l BA. In the process of micropropagation by stem explants of the same studied pepper varieties the addition of the vitamins C. B12. Casein hydrolysate and Ferulic acid had a stimulation effect on the plant growth in height and rooting. In result of anther cultivation from three pepper varieties and four breeding lines the highest percentage of embryo structure formation was registered in varieties Albena and Strjama (12.0 and 13.8 respectively). The Bulgarian peppers are recalcitrant and their in vitro answer is different depending from the explants type, genotype and the culture media composition.

scholarly journals 319IN VITRO MATURATION OF EQUINE OOCYTES IN A COMPLETELY DEFINED MEDIUM SUPPLEMENTED WITH PROGESTERONE

2004 ◽  
Vol 16 (2) ◽  
pp. 279
Author(s):  
B. Merlo ◽  
E. Iacono ◽  
F. Prati ◽  
G. Mari
Keyword(s):  
Polar Body ◽  
Basal Medium ◽  
Defined Medium ◽  
Chi Square ◽  
Nuclear Maturation ◽  
Maturation Rate ◽  
Chi Square Test ◽  
The Media ◽  
Cytoplasmic Maturation

A completely defined medium for in vitro maturation (IVM) of equine oocytes has not yet been developed, since most of the media used for IVM are supplemented with serum or BSA. Furthermore, in this species there is no report about the influence of progesterone on maturation, although it has already been used as supplement (500ngmL−1) in EMMI (Maclellan LJ et al., 2001, Theriogenolgy 55, 310 abst). The aims of this study were to develop a completely defined medium for equine oocyte maturation and to investigate the effect of progesterone on nuclear maturation. Equine oocytes were collected by follicular scraping of abattoir-derived ovaries between April and June. The basal medium for maturation was SOFaa supplemented with pFSH-LH 0.1IUmL−1 (Pluset, Laboratorios Calier, Barcelona, Spain), EGF* 50ngmL−1, ITS (Insulin, Transferrin, Sodium selenite), L-cysteine 1.2mM, Maturation SOF (MSOF). Compact cumulus-oocyte complexes were selected, washed three times in H-SOF and matured in one of the following media (15–20 oocytesmL−1): (1) MSOF+FCS 10% (MSOF-FCS), (2) MSOF+progesterone 100ngmL−1 (MSOF-P4), (3) MSOF. After 24h of culture in 5% CO2 in air at 38.5°C, the oocytes were denuded by gently pipetting in a 0.25% trypsin solution, washed and stained with Hoechst 33258 (10μgmL−1 in PBS) for 30min at room temperature. Oocytes were examined under a fluorescent microscope to assess nuclear maturation. Only oocytes with an evident polar body and metaphase II plate (MII) were considered mature. The experiment was done in 6 replicates. Chi Square test was used for statistical analysis (Statistica for Windows – Stat Soft Inc., Tusla, OK, USA). Significance was assessed for P<0.05. The results of this study show that MSOF can be considered a suitable completely defined medium for IVM of equine oocytes. Adding progesterone significantly (P<0.05) increases the nuclear maturation rate at 24h of culture. It can be speculated that although cumuls cells produce this hormone, supplementation is useful to reach progesterone concentrations similar to those present in follicular fluid (early dominant 63.4±19.3ngmL−1, healthy preovulatory follicle 1094.3±170.9ngmL−1; Gerard N et al., 2002, Reproduction 124, 241–248). Further studies are needed to investigate the influence of progesterone on cytoplasmic maturation and to test the effect of different progesterone concentrations and time of maturation in a completely defined system.*All chemicals were purchased from Sigma, St. Louis, MO, USA, unless otherwise stated. Table 1 Maturation of equine oocytes in different media

Effect of Inorganic Nutrients on Production of Steroid Glycoalkaloids by Phytophthora infestans Race 1.2.4 In Vitro

1979 ◽  
Vol 42 (1) ◽  
pp. 27-30 ◽  
Author(s):  
MELANIE R. MAAS ◽  
FREDERICK J. POST ◽  
D. K. SALUNKHE
Keyword(s):  
Phytophthora Infestans ◽  
Culture Medium ◽  
Basal Medium ◽  
Maximum Growth ◽  
Phosphate Concentration ◽  
Sodium Salts ◽  
Cobalt Nickel ◽  
Magnesium Salts ◽  
The Individual

The influence of sodium salts of the macronutrients nitrate, phosphate, sulfate, and chloride; and the micronutrients iron, calcium, copper, cobalt, nickel, and manganese on growth of Phytophthora infestans and synthesis of glycoalkaloids by the fungus was investigated. Maximum growth levels were demonstrated when 0.04% phosphate, 0.04% chloride, or 2–5 mg of iron/liter were employed in the culture medium. Results indicate that upon substitution of the individual sodium salts of the macronutrients for the potassium and magnesium salts or addition of sodium chloride to the basal medium, the concentration of glycoalkaloids synthesized by the fungus decreased significantly. Regression analysis showed that the concentration of phosphate in the medium had the most influence on the amount of glycoalkaloids produced by P. infestans. Increasing the phosphate concentration in the medium resulted in increasing amounts of glycoalkaloids being produced by the fungus. As the mass of mycelium increased in media containing different amounts of phosphate, the quantity of glycoalkaloids synthesized by the fungus decreased.

Brassinolide on In Vitro Growth of Apical Meristem of Bananas

2014 ◽  
Vol 8 (1) ◽  
Author(s):  
FLORENDA C. BALLESTEROS-TEMANEL
Keyword(s):  
Plant Growth ◽  
In Vitro Propagation ◽  
Apical Meristem ◽  
Basal Medium ◽  
Tissue Growth ◽  
Shoot Length ◽  
New Class ◽  
Randomized Design ◽  
Plant Growth Substances

Brassinosteroids (BRs) are new class of hormones noted to perform multiplephysiological functions in plant growth and development and have the potentialof influencing cell and tissue growth in vitro. Many naturally occurring BRs,including brassinolide, have been discovered, their mode of action and their growthpromoting activities on plants. The use of brassinolide in in vitro propagation isnew. The Murashige and Skoog (1962) medium was used as basal medium. Plantgrowth regulators - IAA, BA and BR - were added to the medium. The study usedthe Completely Randomized Design (CRD) in factorial with three replications.The cultivar of banana and plant growth substances affected the number of budsproduced, shoot length, root length, and stem girth. The interaction of thesetwo factors (cultivar x PGR) influenced the number of buds produced in vitroand the shoot length of the meriplants. The study shows that brassinolide has aninfluence on shoot induction, proliferation, and elongation of bananas in in vitro propagation.Keywords: Agriculture, in vitro propagation, induction, proliferation, elongation, apical meristem, plant growth regulators, cultivars, Isabela, Philippines

Growth substance requirements and major lipid constituents of tissue cultures of Euphorbia esula and E. cyparissias

1972 ◽  
Vol 50 (4) ◽  
pp. 723-726 ◽  
Author(s):  
T. T. Lee ◽  
A. N. Starratt
Keyword(s):  
Callus Tissue ◽  
Defined Medium ◽  
Growth Substance ◽  
Tissue Growth ◽  
Euphorbia Esula ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Callus Tissues ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Root Tissues

The root tissues of Euphorbia esula and E. cyparissias form callus on chemically defined medium. Both species require an exogenous supply of auxin for growth, but the appearance and color of the tissue and their responses to kinetin, 2,4-dichlorophenoxyacetic acid (2,4-D) and indoleacetic acid (IAA) are different. The tissue growth is more satisfactory with α-naphthaleneacetic acid (NAA) than with 2,4-D, IAA, or 4-amino-3,5,6-trichloropicolinic acid (picloram). Gibberellic acid has no effect. The callus tissues of E. esula become intensely green under light but are not autotrophic.Triglycerides, palmitic acid, and β-sitosterol are the major lipid constituents of the callus tissue of E. esula. Chromatographic analysis reveals no significant differences in the composition of extracts from the non-green and green tissues. Long-chain aldehydes, alcohols, and triterpenes found in the plant are not detected in the cultures.

scholarly journals In vitro production of cattle blastocysts in chemically defined medium with or without insulin supplementation

1996 ◽  
Vol 5 (5) ◽  
pp. 509-514 ◽  
Author(s):  
Kristiina Bredbacka ◽  
Peter Bredbacka
Keyword(s):  
Bovine Serum Albumin ◽  
Culture Medium ◽  
Defined Medium ◽  
Blastocyst Stage ◽  
Chemically Defined Medium ◽  
In Vitro Production ◽  
Cell Numbers ◽  
Vitro Production ◽  
Bovine Oocytes

In this study we evaluated the use of a chemically defined medium in the production of blastocysts from bovine oocytes fertilized in vitro. As culture medium we used CRI-PVP, a modification of CRlaa medium with bovine serum albumin replaced by polyvinylpyrrolidone. After 168 h of culture (192 h after insemination) 8.7%, 10.5 and 12.8% of the cleaved embryos developed to the blastocyst stage in the presence of 0, 2 or 200 nM insulin, respectively. The supplementation of 200 nM insulin tended to increase cell numbers in morulae and blastocysts (P=0.10). It is concluded that CRI-PVP can be used as a chemically defined medium in the production of blastocysts from bovine 1-cell embryos. However, further modifications are needed, and the insulin concentrations used may be below the optimum for blastocyst production.

scholarly journals In vitro organogenesis in Albizia guachapele, Cedrella odorata and Swietenia macrophylla (Fabaceae, Meliaceae)

2015 ◽  
pp. 225-228
Author(s):  
Lisette Valverde Cerdas ◽  
Magali Dufour ◽  
Víctor Villalobos
Keyword(s):  
Latin American ◽  
Culture Medium ◽  
Basal Medium ◽  
Petri Dish ◽  
Adventitious Buds ◽  
Swietenia Macrophylla ◽  
Half Strength ◽  
Aseptic Conditions ◽  
Spanish Cedar

Regeneration of adventitious buds was achieved from hypocotyl explanls of Albizia guachapele (Guayaquil) and Cedrella odorata (Spanish cedar), and from epicotyl explants from Swietenia macrophylla (Honduran Mahogany). Seeds were obtained from CATIE's Latin American Fores! Seed Bank and genninated under aseptic conditions .. Four explants were cultured in each Petri dish on half strength modified Murashige and Skoog basal medium, and five concentrations of BA (benzyladenine) were studied; A. guachapele and S. macrophylla responded positively lo the presence of BA in the culture medium. Otherwise, Cedrella odorata requíred media supplemented with citokinin and auxin combinations lo induce adventitious buds.

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