Media Of Known Composition Research Articles

MICROPROPAGATION OF MATURE TERMINALIA CATAPPA VARIETIES

Micropropogation is used to multiply plants such as those that been genetically conventional plant breeding with the multiplying progeny plants. Plant tissue culture is a collection of techniques used to maintain or grow plant cells, tissues or organs under sterile conditions on a nutrient culture medium of known composition. Plant tissue culture is widely used to produce clones of a plant in a method known as micropropagation. The success of Plant tissue culture in recent years approaches have been used as an efcient tool for micropropagation of trees in short time. [1][2]. Propagation of woody trees through tissue culture has many advantages over conventional propagation with different methods like fast growing, season independent of production of the plant, germ plasm conservation and facilitating their growth. This review provides an overview of the success achieved on in vivo work done for a number of important forest trees.Rapid culturing of producing clonal plants this review states that invitro work has been done for no of important forest trees [3]. Both the varieties were subjected, nodal explants were subjected with MS Media and Growth hormones with the dilution of kinetin with 1-1.5 mg/ml and with the mixture of both Kinetin, BAP with the dilution of 1-3 mg/ml with an increase of time for 3-5 weeks the growth was 3-6 cm length. The study of micropropagation in both the varieties, all the values of the work are Mean of at least 3 independent experiment. The MEAN±SEM and the extract number of experiments are given in legends. The Signicance of difference between control and each trial treatment was analysed using ANOVA showed statistical results as and ANOVA is p≤0.05*is signicant

ISOLATION AND CHARACTERIZATION OF NATURAL HERBICIDAL COMPOUND FROM Drechslera rostrata

ABSTRACT In agriculture, weeds have inevitable importance because of reducing the crop growth and also affecting food quality. Although, synthetic herbicides are available to combat these weeds but during recent years there is a trend of using ecofriendly herbicides extracted from natural resources. Present investigation is a continuity of the research, which reported a natural herbicidal compound named as Ophiobolin A from a fungus, Drechslera rostrata. The fungus was incubated in a growth medium of known composition (minimal medium) up to 28 days and its metabolites were extracted with organic solvents. The compound showing bioactivity was purified with the help of Reversed Phase High Performance Liquid Chromatography (RPHPLC) and identified with the help of Spectroscopic techniques viz. Mass Spectroscopy (MS) (LRESIMS and HRESIMS) and Nuclear Magnetic Resonance Spectroscopy (NMR). The purified compound caused death of superficial leaf cells of Chenopodium album, a noxious weed of wheat. Present investigation concludes that the identified compound could be used as structural analogue alternative to synthetic herbicides to synthesize natural herbicides.

Bioactive secondary metabolites produced in the author's laboratory by tissue culture techniques

SUMMARY Plant tissue culture techniques used to maintain or grow plant cells, tissues or organs under sterile conditions on a nutrient culture medium of known composition. Plant tissue culture relies on the fact that many plant cells have the ability to regenerate a whole plant. Single cells, plant cells without cell walls (protoplasts), pieces of leaves, stems or roots can often be used to generate a new plant on culture media given the required nutrients and plant hormones. Cultured plant cells can be offered alternative sources over whole plant cultivation for the production of secondary metabolites. The explants, such as plant leaves, stems, roots, meristems etc. were used for cultivation and establishment of plant cell and tissue cultures. Unfortunately, most often plant cell cultures technologies not lead to an enhancement in secondary metabolites production. Therefore, Strain improvement, methods for the selection of high-producing cell lines, and medium optimizations must be considered to improve the production of secondary metabolites. Several types of cell culture methods are being used to produce bioactive compounds. Cell suspension cultures are preferred for production large amounts of cells for quantitative or qualitative analytical studies. This review refers to certain bioactive secondary metabolites produced in the author’s laboratory by different tissue cultures techniques.

A defined glycosaminoglycan-binding substratum for human pluripotent stem cells

To exploit the full potential of human pluripotent stem cells for regenerative medicine, developmental biology, and drug discovery, defined culture conditions are needed. Media of known composition that maintain human embryonic stem (hES) cells have been developed, but finding chemically-defined, robust substrata has proved difficult. We employed an array of self-assembled monolayers to identify peptide surfaces that sustain pluripotent stem cell self-renewal. The effective substrates display heparin-binding peptides, which can interact with cell surface glycosaminoglycans, and can be used with a defined medium to culture hES cells for more than 3 months. The resulting cells maintain a normal karyotype and display high levels of pluripotency markers. The peptides are able to support growth of multiple (eight) pluripotent cell lines on a variety of scaffolds. Our results indicate that synthetic substrates that recognize cell surface glycans can facilitate the long-term culture of pluripotent stem cells.

Improved Microassays Used to Test Natural Product-Based and Conventional Fungicides on Plant Pathogenic Fungi.

Seven important plant pathogenic fungi (Botrytis cinerea, Colletotrichum acutatum, C. fragariae, C. gloeosporioides, Fusarium oxysporum, Phomopsis obscurans, and P. viticola) valuable in screening fungicides were tested. Our procedure included washing conidia to reduce germination times, incorporating Roswell Park Memorial Institute 1640 as a medium of known composition, and using coverslips in the 24-well cell culture clusters to document the effect of fungicides on fungal morphology. The natural product-based fungicide, sampangine, a sampangine analog, 4-bromosampangine, plus seven conventional fungicides (benomyl, captan, cyprodinil, fenbuconazole, fenhexamid, iprodione, and kresoxim-methyl) were tested in vitro for their ability to inhibit germination and growth of the seven fungal species. Sampangine inhibited germination in all fungi except C. acutatum. Comparison of results of germination and morphology microbioassays with results of microtiter assays suggests that some fungicides stop fungal germination, whereas others only slow down fungal growth. We hypothesize that sampangine, except against C. acutatum, has the same physical mode of action, germination inhibition, as the conventional fungicides captan, iprodione, and kresoxim-methyl. 4-Bromosampangine caused morphological anomalies including excessive branching of germ tubes of C. fragariae and splaying and branching of germ tubes of B. cinerea.

Detection of NO-synthase activity of lactobacilli by fluorescent staining

-argin-ine, and functions as one of the universal regulators ofcell and tissue metabolism. It also functions as the maincytotoxic/cytostatic effector of the cellular immunitysystem. In mammalian cells, NO production is cata-lyzed by the family of NO synthase (NOS) isoenzymes[1]. The process of bacterial NO synthesis, in contrastto the eukaryotic one, is still far from being fully under-stood. An NO synthase mechanism of nitric-oxide pro-duction alternative to denitrification was initiallyrevealed in the microorganisms of the genus Nocardia[2]. Later, genes that encode proteins homologous tothe mammalian NOS oxygenase domain were identi-fied in the genomes of a number of gram-positive bac-teria [3]. As yet, there are few works on the isolation,purification, and study of the properties of bacterial NOsynthases [4–6].NO synthase is supposed to be present in Lactoba-cillus plantarum [7]. Previously, we have applied EPR(electron paramagnetic resonance) to establish thecapability of L. plantarum 8P-A3 to synthesize nitricoxide. This strain was shown not to produce NO viadenitrification but to have a NO-synthase activity, likemammalian cells [8].This work presents the first evidence of NO-syn-thase nitric-oxide production in L. plantarum obtainedby the method of fluorescent staining.The culture of L. plantarum 8P-A3 [8] isolated fromthe Lactobacterin Dried preparation (Biomed, Russia)was a test subject. MRS medium of known composition[8] was used for bacterial growth. One ml of 18-h cul-ture was added into 50-ml test tubes containing 30 mlof the medium with or without L-arginine (100 µM)and incubated without mixing at 37°e. The growthkinetics was determined at 590 nm on a Lambda 35double-beam spectrophotometer, Perkin Elmer Instru-ments (USA). For fluorescent detection of nitric oxide,30 ml of the culture from the stationary growth phase(48 h) was precipitated by centrifugation at 4000 g for20 min in an Avanti

Medium requirements for neuritic outgrowth from goldfish retinal explants and the trophic effect of taurine

The use of culture media of known composition are necessary for studying the role of trophic molecules. Since most of the in vitro research on regeneration of the optic nerve has been performed in the presence of fetal calf serum, the aim of this study was to obtain a medium in which the neuritic outgrowth from post-crush goldfish retinal explants could take place without adding fetal calf serum. After the lesion of the optic nerve (10 days), the retina of goldfish was dissected and explants were cultured for 5 and 10 days in the absence or in the presence of fetal calf serum, at which time the neuritic outgrowth was determined. Various concentrations and combinations of glucose, albumin, calcium, HEPES and taurine were used. The highest neuritic outgrowth was observed in the presence of fetal calf serum, in which condition the amino acid taurine increased length and density of neurites. Media supplemented with albumin, calcium or HEPES did not modify the outgrowth of neurites from the explants. However, glucose favored the neuritic outgrowth in a bell-shaped manner, although fibers were thinner than those observed in the presence of fetal calf serum. Taurine did not stimulate outgrowth of neurites from explants growing in a medium with optimal concentrations of glucose, indicating that elements of the fetal calf serum are determinant for the trophic effect of taurine. The present results contribute to further studies, such as those related to the effect of taurine and of trophic factors derived from the optic tectum, which would be performed in the presence of a medium free of fetal calf serum.

Nutrition research with lactic acid bacteria: a retrospective view.

From the above discussion, it is apparent that one or another of the lactic acid bacteria requires each of the B vitamins required by animals and that assay methods developed during study of nutrition of bacteria and yeasts have played a large role in the initial or independent discovery, isolation, and characterization of vitamins, vitamin derivatives, and functionally similar substances. Clear examples include biotin, biocytin, lipoic acid, nicotinic acid, pantothenic acid, pantetheine, folic acid and tetrahydrofolic acid (and their derivatives), pyridoxal, pyridoxamine, and pyridoxamine phosphate. Improved assay methods that use these organisms also have provided much of the currently available information concerning distribution and stability of the vitamins in natural products, while quantitative inconsistencies between assays, when traced to their origin, have frequently revealed previously unknown metabolic precursors, products, or functions of the vitamins and have provided explanations of the mechanisms by which certain peptide growth factors act. Extension of such studies to organisms that cannot yet be grown in media of known composition should provide additional insights into currently obscure areas of nutrition.

Ultrastructural Characteristics of Platelets Separated from Plasma by ADP-Induced Aggregation

The utilization of blood platelets in experimentation frequently requires their separation from blood and subsequent resuspension in media of known composition. Several methods are available for preparation of isolated platelets (1-3) by differential centrifugation or gel filtration, but most methods are tedious and time consuming. Often platelets obtained by use of such methods are in a state different functionally and ultrastructurally from that of platelets in plasma (4).Recently Mohammad, Reddick, and Mason (5) reported a method in which platelets were separated from plasma by ADP-induced aggregation, washed several times, and then incubated in a carefully selected medium that resulted in deaggregation of platelets.

Cell electrophoretic mobilities in Friend virus disease.

CHANGES in cell surface charge density have been reported several times in various types of malignancy1–4. One of the problems of applying the cell electrophoretic method to such problems is the need to obtain a quantity of single cells in suspension in a medium of known composition. The spleen, however, can be dispersed mechanically with ease, and we have chosen to investigate such spleen cell suspensions electrophoretically during the course of Friend virus disease, where gross pathological changes occur in the spleen.

THE ROLE OF CYTOKININS IN THE “FASCIATION” DISEASE CAUSED BY CORYNEBACTERIUM FASCIANS

The disease caused by Corynebacterium fascians can be imitated by treatment with kinetin. Cultures of the bacteria were grown on a purine‐free medium of known composition. They require thiamine. Two assays for cytokinin were developed; one depends on the release of lateral pea buds from apical dominance and is highly specific but of only moderate sensitivity. The other depends on the retention of chlorophyll in senescing oat leaves and is very sensitive (detecting 2 × 10–4 μg of kinetin equivalents), though it is somewhat less specific and, therefore, requires care in usage. Both give results in 3–4 days and are thus far more rapid than tissue culture methods. With these assays C. fascians has been shown to produce a chloroform‐soluble cytokinin active in both tests. The substance is stable to heat in acid or basic media, is soluble in non‐polar solvents, and behaves as a base. It is precipitated from water solution by Ag ions and may, therefore, be a purine derivative. Pea tissue infected with C. fascians, but not uninfected tissue, yields a compound of similar solubilities and biological activity. Reasons are given for believing that the synthesis of cytokinins may be important, not only for C. fascians, but also for many other plant parasites.

The biosynthesis of protein amino acids in plant tissue culture I. Isotope competition experiments using glucose-U-C14 and the protein amino acids.

Most investigators have used excised tissue, cellfree preparations, or whole plants for studies of metabolism in angiosperms. Each of these systems represents a different state of the cells or tissues. Another state of plant cells, that of exponential growth, has been obtained by tissue culture techniques (2, 5). Studies of the metabolism of cells growing exponentially would provide information which is complementary to that already available for plants. Such studies would allow direct comparison of metabolism of higher plant cells with that of microorganisms also growing exponentially. When specific information is not available, it is generally assumed that the metabolism in plant cells is the same as that in micro-organisms. This assumption may be tested using exponentially growing plant cells. For use in studies of metabolism a culture system in which plant cells are growing exponentially should have several features. The media used should have a known composition. The tissue should be grown in suspension as very small clumps or single cells. The use of media of known composition makes it possible to manipulate the nutrients at will and to avoid the presence of unrecognized compounds which occur in media supplemented with natural extracts. The growth of tissues as suspensions of single cells or small clumps of cells prevents the formation of the diffusion gradients which occur in larger pieces of tissue. Rapid agitation of the suspension keeps the cells in equilibrium with the medium and the medium in equilibrium with its gaseous atmosphere. Such desirable features can now be found in at least 2 plant tissue cultures. These are cultures established from Paul's Scarlet Rose by Dr. W. Tulecke (Tulecke, private communication) and from Nicotiana tabaccumn var. Xanthi by Dr. P. Filner (22). Methods established for growth of microbial cultures have been successfully adapted to the culture of plant cells. For example, plant tissues have been maintained on solid media as callus. Single plant cells have been plated in agar to yield visible colonies (8). Also, plant tissues have been maintained as suspensions of cells and clumps in liquid media of known or unknown composition (2) in shake flasks or large carboys (29). The close parallels in methods of growth suggest that methods developed to study the nietabolism of microbial cells in culture might also be readily adapted to the study of plant cell metabolism. Several studies (10, 18, 34) of plant metabolism have employed the isotope competition method (25). This paper is to report the application of the isotope competition method to the study of amino acid biosynthesis in suspensions of plant cells growing exponentially in media of known composition.

In vitro Culture of the Mosquito Phase of Avian Malaria

In vitro Culture of the Mosquito Phase of Avian Malaria

STUDIES ON OESTROGENS IN CATTLE

ABSTRACT A review is given of previous in vitro investigations on oestrogen metabolism. In the present investigation use has been made of the tissue culture technique, whereby possible blood or serum effects on oestrogen transformations could be excluded. The conversion products were characterized by chemical methods. In the presence of bovine kidney cells grown on a medium of known composition, the following conversions were recorded: Oestrone to oestradiol-17β, oestradiol-17β to oestrone, oestradiol-17α to oestrone. Control incubations of the hormones with medium only showed that the transformations must be due to the presence of the living cells. The rate of conversion to oestrone was markedly higher for oestradiol-17β than for oestradiol-17α. As previous in vivo experiments have shown oestradiol-17α to be an important end product in the bovine, following injections of both oestrone and oestradiol-17β, the free interconvertibility of oestrone and oestradiol-17β demonstrated in the present investigation becomes significant. The findings are discussed in relation to recent observations on hormoneenzyme interrelations.

In vitro cultivation of the histotrophic stages of Haemonchus contortus and Ostertagia spp.

LAPAGE1 working with Haemonchus contortus reported in 1933 that: “Attempts have been made to cultivate the parasitic larval stages, a method having been found by which the free-living infective third stages may be exsheathed and sterilized, so that the first parasitic stages may be obtained and studied further in sterile media of known composition. Although these first parasitic stages have been studied in hanging drops in more than 200 media designed to imitate environmental factors to which parasites are presumably exposed in the body of the host no success has been obtained”.

Production of unesterified fatty acids from isolated rat adipose tissue incubated in vitro.

SummaryAdipose tissue from rats, incubated in vitro in a medium of known composition, was found to release unesterified fatty acids (UFA). The rate of evolution of UFA was increased by the addition of epinephrine to the medium or by fasting the donor animal. It was decreased by the addition of glucose and insulin to the medium, or by feeding the donor animal with carbohydrate.

An Aerobic Medium for Fungus Growth Studies

THE aerobic nature of mushroom composts plays an essential part in the production of good mushroom crops, so that it is desirable, in laboratory growth-experiments on mushroom mycelium, to use an aerobic medium of known composition in which growth measurements can be made. The only fully aerobic medium of this nature in general use, so far as I am aware, is milled filter paper1,2,3. Unfortunately, this material loses its aerobio qualities when only a small amount of nutrient solution has been absorbed on it. A medium based on milled transparent cellulose shavings, dyed black to enable mycelial growth to be observed, has now been found to be more satisfactory, and is prepared as follows.

GROWTH IN VITRO OF EXCISED TOBACCO AND SUNFLOWER TISSUE WITH DIFFERENT TEMPERATURES, HYDROGEN‐ION CONCENTRATIONS AND AMOUNTS OF SUGAR

FACTORS RESPONSIBLE for the initiation and continuation of pathological growth have been studied with plant materials because they offer certain opportunities for research (Riker, 1939) not provided by animal materials. Some advantages of plant materials include (a) large numbers, (b) low cost, (c) ease of experimental use, (d) physiological variability, (e) genetic purity, and (f) the possibilitv of tissue culture on a medium of known composition (Riker, 1942). Among the plant materials the crown gall disease caused by Phytomonas tumefaciens (Smith and Town.) Bergey et al. has received special attention because of certain similarities between this pathological growth in plants and cancer of higher animals (e.g., Smith, 1922; Levine, 1936; White and Braun, 1912). The early literature on crown gall was reviewed by Riker and Berge (1935). The anatomy of the host tissue and physiology of the bacteria have been extensively studied by various investigators. The physiology of the host tissue, however, has remained relatively obscure because of certain technical difficulties, some of which mav be overcome by cultivation of tissue on a synthetic medium in vitro. Plant tissue culture has been accomplislhed only recently. The idea was probably first suggested by Haberlandt (1902). Limited growth of excised root tips was reported by Kotte (1922) and by Robbins (1922), while unlimited growth was reported by White (1934) with tomato roots. Since root tips contain several kinds of tissue, their cultivation may be considered organ culture. True tissue cultures capable of unlimited growth were first reported by White (1939) with tobacco callus and by Gautheret (1939) and Nobecourt (1939) with carrot callus. Recently, White and Braun (1941) have cultivated tissue from secondary crown galls on sunflower. The extensive history and applications of plant tissue culture have been reviewed by White (1943). In tissue culture simple, somatic cells may be isolated from the influence of neighboring cells, tissues, and organs and may be studied under controlled conditions on a synthetic medium containing only ingredients whose chemical formulae can be written. Such a technique offers a tool for the study of critical vital processes of tissue metabolisn. While many basic requirements of excised root tips in vitro have been determined by various investi-

The In Vitro Metabolism of Plasmodium Gallinaceum

Within the past few years the metabolism of malarial parasites has been studied in several laboratories. Christophers and Fulton' measured the respiration of Plasmodium knowlesi and Maier and Coggeshall2 have demonstrated the utilization of glucose and other substrates during respiration of the same organism. The latter workers also found that the rate of respiration was a function of the size of the parasite, i.e., it increased with the development of the parasite from merozoite to segmenter. Wendel,3 also working with P. knowlesi, has demonstrated that both the respiration and glycolysis of infected red blood cells are greater than those of unparasitized erythrocytes.4 Velick5 has reported that the respiratory quotient of P. cathemerium increases when nuclear division of the parasite occurs. In all these experiments the intact red cells containing the parasite were studied. However, Christophers and Fulton' have obtained P. knowlesi free from red cells by saponin cytolysis and report that such organisms also respire in the presence of glucose. The present report deals with the glucose metabolism of P. gallinaceum. This organism exhibits little synchronism, but has particular advantages in that its natural host, the chicken, is readily available and infections can be obtained which yield large amounts of parasitic material. The study of this organism is made difficult by its presence within the nucleated red blood cell of the host. It would be desirable to lyse the cell and then to study the parasite suspended in a medium of known composition. In view of our lack of any convenient way of doing this, we have been forced to study the organism together with the red cell, hoping by the use of proper controls to arrive at a picture of the activities of the organism itself. A large portion of our experiments, therefore, are concerned with parasitized red blood cells suspended in an inorganic medium containing glucose.

Nutrition of Tetrahymena geleii (Protozoa, Ciliata)

All attempts to grow Tetrahymena geleii in a medium of known composition have, so far, failed. Failure to obtain growth in anything but peptones led Lwoff1 to formulate the hypothesis that the ciliate requires polypeptides and cannot utilize free amino acids. Earlier2 he had hinted that the failure of growth in a mixture of silk peptone, fibrin peptone and gelatin, which contains all the known amino acids, is the result of the lack of some supplementary substance. Gelatin3 will support growth if supplemented with a small amount of yeast extract; indeed, it is stated4 that transplantable growth is obtained in gelatin plus thiamin and riboflavin. Silk peptone gives slight, but transplantable, growth if thiamin is added.5It is clear, then, that the question of the amino acid requirement of T. geleii cannot be answered until more is known of the supplementary needs.In order to discover the vitamin (or supplement) requirements, a basal medium is necessary. Since it had been found that the ciliate will grow wel...