Ultrastructure of conidial germ-tube development in vitro by the insect pathogen Entomophaga aulicae

1989 ◽  
Vol 67 (3) ◽  
pp. 754-762 ◽  
Author(s):  
Faye Murrin ◽  
Richard A. Nolan
Keyword(s):  
Germ Tube ◽  
Culture Medium ◽  
Protoplast Formation ◽  
Insect Pathogen ◽  
Liquid Culture Medium ◽  
Cytoplasmic Membranes ◽  
Development In Vitro ◽  
Germ Tubes ◽  
Infection Hypha

In an effort to understand the factors influencing the formation of infection structures by the insect pathogen Entomophaga aulicae, we examined the ultrastructural development of conidial germ tubes formed in vitro under conditions that resemble those producing appressoria and protoplasts during infection. Conidia germinated on formvar in a nonaqueous environment produced a single viable germ tube, which in turn produced appressorium-like structures or secondary conidia, structures similar to those formed on the host. The formation of appressorium-like structures indicates that stimuli for appressorium formation are relatively nonspecific, whereas development of the infection hypha requires different triggers. Conidia germinated in a liquid culture medium, which supports the growth of the protoplast stage of the fungus, produced a single viable germ tube that continued to grow apically. This apical growth was accompanied by the loss of the outer layer of the germ tube wall and the presence of electron-opaque granules in an extensive system of cytoplasmic membranes. Protoplasts did not form directly from these germ tubes and ultrastructural details of tip growth in this medium did not resemble those previously described in infection hyphae prior to protoplast formation in the host.

Development of embryonic rat eyes in organ culture

Development ◽  
1968 ◽  
Vol 19 (3) ◽  
pp. 407-414
Author(s):  
R. Christy Armstrong ◽  
Joel J. Elias
Keyword(s):  
Organ Culture ◽  
Culture Medium ◽  
Folic Acid Deficiency ◽  
Experimental Conditions ◽  
Acid Deficiency ◽  
Series Of Experiments ◽  
Development In Vitro ◽  
Ocular System

Abnormalities of the ocular system which appear in organ culture in Waymouth's medium with freshly added glutamine (Armstrong & Elias, 1968) resemble those caused by transitory pteryolglutamic acid (PGA or folic acid) deficiency in vivo (Armstrong & Monie, 1966). The configurations of such malformations as lens herniations, retinal diverticula, and rosette-like formations of the retina are remarkably similar in both cases. The experiments reported in this paper were undertaken in an effort to understand the mechanisms involved in the production of similar abnormalities by two very different experimental conditions: the addition of glutamine in vitro and the transitory deficiency of PGA in vivo. One series of experiments involved the effects of manipulation of the PGA and glutamine content of the culture medium on eye development in vitro. Parallel studies on PGA-deficiency in vivo were undertaken in conjunction with organ-culture experiments in order to compare the effects on abnormal eye morphogenesis.

Fate of Safening Agent L-2-Oxothiazolidine-4-Carboxylic Acid in Sorghum (Sorghum bicolor)

Weed Science ◽  
1990 ◽  
Vol 38 (3) ◽  
pp. 201-205 ◽  
Author(s):  
James L. Hilton ◽  
Parthasarathy Pillai ◽  
Helen A. Norman
Keyword(s):  
Carboxylic Acid ◽  
Sorghum Bicolor ◽  
Culture Medium ◽  
In Vitro Assay ◽  
Thiol Compounds ◽  
Vitro Assay ◽  
Herbicide Safener ◽  
Liquid Culture Medium ◽  
Germinating Seeds

The herbicide safener OTC (L-2-oxothiazolidine-4-carboxylic acid) increased the amount of reduced thiol compounds in sorghum [Sorghum bicolor(L.) Moench. ‘DK 427′] seedlings. When seedlings were grown in liquid culture medium containing35S-OTC, the compound was metabolized to radiolabeled cysteine and glutathione. The addition of tridiphane [2-(3,5-dichlorophenyl)-2-(2,2,2-trichloroethyl)oxirane] increased conversion of35S-OTC to cysteine and resulted in the formation of one additional35S-labeled compound. When35S-glutathione was injected into germinating seeds it was converted to35S-cysteine and both thiols were subsequently found in roots and shoots. Seeds injected with35S-OTC both translocated the compound to developing roots and shoots and metabolized35S-OTC to cysteine and glutathione. Excised roots and shoots also metabolized35S-OTC to the thiols. In an in vitro assay the enzyme 5-oxoprolinase converted OTC to cysteine.

scholarly journals 133 INSULIN, TRANSFERRIN AND SELENIUM WITH OR WITHOUT BSA IN A SERUM-FREE CULTURE SYSTEM FOR BOVINE EMBRYO, AND ITS SUITABILITY FOR EMBRYOS CULTURED IN SMALL GROUPS

2005 ◽  
Vol 17 (2) ◽  
pp. 217 ◽  
Author(s):  
C. Daniaux ◽  
B. Verhaeghe ◽  
I. Donnay
Keyword(s):  
Small Groups ◽  
Culture Medium ◽  
Culture Media ◽  
Quality Parameters ◽  
Hatching Rate ◽  
Bovine Embryo ◽  
Serum Free ◽  
Abnormal Accumulation ◽  
Development In Vitro

Serum in embryo culture medium may be a potential cause of abnormal accumulation of lipid droplets, which is correlated to a higher sensitivity to cryopreservation. Moreover, serum may introduce pathogens. With the aim of developing a serum-free culture medium, we first (Experiment 1) investigated the effect of adding ITS (5 μg/mL insulin, 5 μg/mL transferrin, 5 ng/mL selenium) as a serum substitute in SOF medium on embryos cultured in large groups (20 embryos per culture drop of 20 μL) and we then (Experiment 2) analyzed the effect of adding BSA. In this second experiment, our serum-free culture media were also tested on embryos cultured in small numbers (5 embryos per drop of 20 μL) in order to mimic ovum pickup (OPU) conditions. Embryos were obtained from slaughterhouse oocytes, matured in vitro for 24 h in a serum-free enriched 199 medium (Donnay et al. 2004 Reprod. Fertil. Dev. 16, 274) containing ITS, and fertilized for 18 h. In experiment 1, embryos were cultured in SOF (Holm et al. 1999 Theriogenology 52, 683–700) supplemented with 0.1 mg/mL polyvinylpyrrolidane (PVP) without (SOF) or with ITS (SOF-ITS), or with 5% FCS (SOF-FCS). Cavitation occurred earlier in presence of serum (Table). Adding ITS to SOF increased blastocyst rates at Day 7 and Day 8 post-insemination (p.i.) and also the hatching rate. In experiment 2, embryos were cultured in SOF-FCS, SOF-ITS, or SOF-ITS supplemented with 4 mg/mL fatty acid free BSA (SOF-ITS-BSA). Within each condition, no differences were observed for blastocyst and hatching rates between embryos cultured in large or in small groups. Adding BSA to SOF-ITS increased blastocyst rate at Day 6 p.i. and also the hatching rate. At Days 7 and 8 p.i., blastocyst rates were higher in SOF-FCS than in SOF-ITS and tended to be higher than in SOF-ITS-BSA, especially for embryos cultured in small groups. Cell numbers of the resulting embryos were unaffected. These results indicate that: (1) ITS as supplement to SOF medium promotes embryo development in vitro. (2) BSA as protein supplement to SOF-ITS medium accelerates blastulation and improves hatching rate. (3) SOF-ITS and SOF-ITS-BSA are two serum-free culture media that can sustain development of embryos, also when cultured in small number, even though SOF-FCS tended to afford better rates of development. Further studies will include evaluation of other quality parameters including resistance to cryopreservation. This work was supported by the Ministery of Agriculture of the Region wallonne de Belgique.

scholarly journals In vitro establishment of Bambusa oldhamii Munro from field-grown matrices and molecular identification of endophytic bactéria

2019 ◽  
Vol 49 ◽  
Author(s):  
Ana Paula de Azevedo Pasqualini ◽  
Gabriela Xavier Schneider ◽  
Hugo Pacheco de Freitas Fraga ◽  
Luiz Antonio Biasi ◽  
Marguerite Quoirin
Keyword(s):  
Molecular Identification ◽  
Culture Medium ◽  
Bacterial Isolate ◽  
Fungal Growth ◽  
Endophytic Bacterium ◽  
Liquid Culture Medium ◽  
Rdna Sequencing ◽  
In Vitro Establishment ◽  
Bambusa Oldhamii

ABSTRACT In plant micropropagation, the establishment stage is difficult, due to the presence of microorganisms in tissues from field-grown matrices, especially for bamboo. This study aimed to establish an efficient asepsis protocol for Bambusa oldhamii explants from field plants, as well as to carry out the molecular identification of a possible endophytic bacterial isolate. The explants were exposed to 70 % alcohol, 1 % sodium hypochlorite (NaOCl), 0.1 % mercuric chloride (HgCl2), thiophanate-methyl (Cercobin®) and chlorhexidine digluconate (2 % Riohex®) in different combinations, and introduced into Murashige and Skoog culture medium (solid or liquid), supplemented or not with 4 mL L-1 of Plant Preservative Mixture (PPMTM), totaling seven treatments. The asepsis and immersion of the explants in the liquid culture medium containing 4 mL L-1 of PPMTM visually inhibited the bacterial and fungal growth, allowed the development of shoots with a mean length of 2.2 cm and posterior subcultures, being the best treatment used for the in vitro establishment of B. oldhamii. The molecular identification of an endophytic bacterium performed by 16S rDNA sequencing allowed to identify the bacterial isolate as Ralstonia sp., with 100 % of similarity, and the phylogenetic analysis grouped it with Ralstonia pickettii. In addition, the bacterial isolate showed to be sensitive to 4 mL L-1 of PPMTM by the minimum inhibitory concentration test.

Effects of amino acids on development in vitro of cleavage-stage bovine embryos into blastocysts

1996 ◽  
Vol 8 (5) ◽  
pp. 835 ◽  
Author(s):  
T Pinyopummintr ◽  
BD Bavister
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Embryo Development ◽  
Culture Medium ◽  
Essential Amino Acids ◽  
Bovine Embryo ◽  
Bovine Embryos ◽  
Development In Vitro ◽  
Defined Culture

Effects of amino acids on early bovine embryo development in vitro were examined using a chemically-defined, protein-free culture medium. Bovine embryos produced in vitro were cultured from 18 h to 72 h post insemination in a simple medium containing lactate as the only energy source except for the amino acid treatments. Subsequently, embryos were transferred to TCM-199 supplemented with serum for blastocyst development to substantiate their developmental competence. Treatments were: (1) non-essential amino acids from TCM-199 (NEA); (2) essential amino acids from TCM-199 (EA); (3) NEA+EA; (4) Eagle's minimum essential medium amino acids (MEM AA); (5) 11 amino acids present in HECM-6 (11 AA); and (6) 0.2 mM glutamine (GLN). A higher proportion of embryos (percentage of inseminated ova) cleaved to the > or = 8-cell stage by 72 h post insemination in NEA (56.7%), EA (41.2%), 11 AA (40.3%) and GLN (51.1%) than in either NEA+EA (30.0%) or MEM AA (33.1%). However, after transfer to complex medium, embryos that had developed in EA, as well as those in MEM AA or NEA+EA, produced significantly fewer blastocysts (37.1%, 34.4% and 25.6% respectively) than those in NEA (56.7%), GLN (48.9%) or 11 AA (37.7%). The ability of blastocysts to hatch from their zonae pellucidae was also affected by amino acid treatment during cleavage stages. The present study indicated that the addition of NEA or GLN or 11 AA to a chemically-defined culture medium during the cleavage phase of bovine embryo development increases their subsequent ability to reach the blastocyst stage. These data have implications for understanding the nutritional needs of bovine embryos produced in vitro and for optimizing the composition of culture media to support their development.

Effect of glycine, alanine and calf plasma in serum free culture medium on bovine embryonic development in vitro

1995 ◽  
Vol 43 (1) ◽  
pp. 328 ◽  
Author(s):  
T.K. Suhl ◽  
K.L. White ◽  
T.D. Bunch ◽  
R. Spendlove ◽  
R. Wilkinson
Keyword(s):  
Embryonic Development ◽  
Culture Medium ◽  
Serum Free ◽  
Development In Vitro

THE CYTOLOGY OF INFECTION STRUCTURE DEVELOPMENT IN UREDIOSPORE GERM TUBES OF UROMYCES PHASEOLI VAR. TYPICA (PERS.) WINT.

1967 ◽  
Vol 45 (4) ◽  
pp. 447-450 ◽  
Author(s):  
Ramesh Maheshwari ◽  
A. C. Hildebrandt ◽  
P. J. Allen
Keyword(s):  
Nuclear Division ◽  
In Vitro Differentiation ◽  
Structure Development ◽  
Infection Structures ◽  
Uromyces Phaseoli ◽  
Mother Cells ◽  
Germ Tubes ◽  
Infection Hypha ◽  
Feulgen Technique

Urediospores of Uromyces phaseoli var. typica (Pers.) Wint. race 32 Arth. germinated on mineral oil – nitrocellulose membranes and sequentially developed appressoria, vesicles, and infection hyphae. The nuclear behavior during in vitro differentiation of infection structures was studied by use of the Feulgen technique. The two urediospore nuclei divided in the germ tube before the formation of appressorium. This was followed by a second division of the four daughter nuclei in the appressorium, and occasionally by a third division of the eight nuclei in the vesicle and infection hypha. Haustorial mother cells were formed in infection hyphae in vitro and contained from two to five nuclei. In contrast, nuclear division did not occur in germ tubes where growth continued linearly. Infection structures that developed in vitro resembled those produced during infection of the host by urediospores of other species of rust fungi.

scholarly journals Effects of a commercial biocide, kasugamycin and consistency of the culture medium on the in vitro establishment of bamboo1

2019 ◽  
Vol 49 ◽  
Author(s):  
Daniela Werner Ribeiro dos Santos ◽  
Théo Piucco Rocker ◽  
Thiago Sanches Ornellas ◽  
Miguel Pedro Guerra
Keyword(s):  
Liquid Culture ◽  
Culture Medium ◽  
Liquid Culture Medium ◽  
Bambusa Vulgaris ◽  
Phyllostachys Bambusoides ◽  
In Vitro Establishment ◽  
Dendrocalamus Asper

ABSTRACT The contamination by microorganisms and oxidation of explants in the in vitro establishment of bamboo are recurrent problems for its micropropagation. In the present study, effects of the biocide Plant Preservative Mixture (PPM™), the antibiotic kasugamycin and the consistency of the culture medium were evaluated in the in vitro establishment of Bambusa vulgaris,Phyllostachys bambusoides and Dendrocalamus asper. The presence of PPM™ in the culture medium had a significant effect using 2 mL L-1 or 4 mL L-1 concentrations, as well as in the liquid culture medium, increasing the plants established in the autumn. Kasugamycin promoted variable responses for all the three species, depending on the season. There was interaction among the factors, demonstrating that higher rates of viable plants can be obtained by combining different strategies to reduce the oxidation and contamination. For the in vitro establishment of B. vulgaris,P. bambusoides and D. asper, it is recommended to add 2 mL L-1 or 4 mL L-1 of PPM™ to the liquid culture medium.

scholarly journals Influence of genotype on protein and oil concentration of soybean seeds

2010 ◽  
Vol 53 (4) ◽  
pp. 793-799 ◽  
Author(s):  
Esmael Lopes dos Santos ◽  
Antonio Eduardo Pípolo ◽  
Ricardo Tadeu de Faria ◽  
Cássio Egidio Cavenaghi Prete
Keyword(s):  
Protein Concentration ◽  
Culture Medium ◽  
Seed Oil ◽  
Soybean Seeds ◽  
Glutamine Concentration ◽  
Liquid Culture Medium ◽  
Immature Seeds ◽  
Oil Concentration

Aiming at evaluating genotype influence on the concentration of protein and oil, immature seeds of cultivars CD 202 and CD 206 were removed from the mother-plant, in the stage R5, and were grown in vitro, in a liquid culture medium which contained 20, 40 and 60 mM of glutamine, during eight days. Afterwards, the concentrations of oil and protein were compared to the contents of the seeds cultivated in vivo. With a higher availability of glutamine for the seed, there was an increase of protein content. The genotypes were statistically different as far as the protein concentration was concerned,which confirmed that the genotype had influence on the concentration of protein in the seed. Oil and protein concentrations were inversely related when a variation of glutamine concentration occurred.

scholarly journals Early studies on the effect of peptide growth factor phytosulfokine-α on Brassica oleracea var. capitata L. protoplasts

2017 ◽  
Vol 86 (3) ◽  
Author(s):  
Agnieszka Kiełkowska ◽  
Adela Adamus
Keyword(s):  
Brassica Oleracea ◽  
Shoot Regeneration ◽  
Liquid Culture ◽  
Culture Medium ◽  
Breeding Lines ◽  
Liquid Culture Medium ◽  
Peptide Growth Factor ◽  
The Media ◽  
Cells Cultured

<span>Phytosulfokines (PSK) are peptidyl growth factors with the potential of inducing cell proliferation. We examined the effect of supplementation of liquid culture medium with 0.1 µM phytosulfokine-α (PSK-α) on protoplast viability and division frequencies in seven accessions of <em>Brassica oleracea</em> var. <em>capitata</em> L., including cultivars and breeding lines. Protoplasts were isolated from leaves and hypocotyls of in vitro grown plants and immobilized in calcium-alginate layers. Cabbage protoplast-derived cells cultured in medium supplemented with 0.1 µM of PSK-α had higher viability and division frequencies compared to cells cultured in PSK-α-free control medium. The effect of PSK-α was more pronounced in low-responding accessions (‘Sława z Gołębiewa’, ‘Ramkila F1’, LM, and LM98); however, in two cultivars with very low response (‘Badger Shipper’ and ‘Oregon 123’), although the division frequencies in the media supplemented with PSK-α were increased over the control, the differences were not significant. Obtained callus colonies were subjected to regeneration. PSK-α supplemented into the liquid culture medium had an indirect effect on shoot regeneration by inducing sustained cell divisions leading to an increase in shoot regeneration in Sława z Gołębiewa and both breeding lines.</span>

Sign in / Sign up

Export Citation Format

Share Document