Callus, plantlet formation, and polyploidy from cultured anthers of Lotus and Nicotiana

1971 ◽  
Vol 49 (11) ◽  
pp. 2041-2051 ◽  
Author(s):  
Minoru Niizeki ◽  
William F. Grant
Keyword(s):  
Germ Cells ◽  
Developmental Process ◽  
Wild Strain ◽  
Pollen Grains ◽  
Autoradiographic Study ◽  
Plantlet Formation ◽  
Before And After ◽  
Embryoid Formation ◽  
Callus Tissues ◽  
Haploid Plants

Anthers of Lotus corniculatus (a wild strain, cv. Empire and Viking) and L. caucasicus were cultured on agar-solidified media in an attempt to induce haploid plants. Calluses were readily obtained from the anthers, and chromosome number determinations on the callus tissues showed cells of different euploid and aneuploid chromosome numbers but no haploid cells. From these calluses, tetraploid and octoploid plants were regenerated. Except for some pollen grains which hypertrophied, the germ cells produced mature pollen but did not progress beyond this stage to initiate calluses. These results indicate that the calluses were derived from somatic tissues of the anthers rather than from the germ cells. Only a few cell divisions of the pollen grains occurred when they were cultured directly on agar-solidified medium. The induction of haploid plants by means of anther culture was successful for Nicotiana tabacum (cv. Wisconsin 38, Delhi 34, Hicks Broadleaf) but not for N. affinis. The autoradiographic study showed that uninucleate pollen differentiated into embryoids both before and after DNA synthesis for the first mitosis but not in any other developmental stage of the germ cells. The initiation of embryoids was concomitant with an increase in size of the pollen grains and vacuolization. Embryoid formation from pollen grains followed the same developmental process as in normal embryogenesis. Exogenous plant hormones, indole-3-acetic acid and kinetin, were responsible for embryoid formation, although they were not essential for initiation.

Histochemical and autoradiographic study of the cultured rat visceral yolk sac

Development ◽  
1986 ◽  
Vol 97 (1) ◽  
pp. 169-176
Author(s):  
H. Sobis ◽  
J. Goebels ◽  
M. Vandeputte
Keyword(s):  
Organ Culture ◽  
Germ Cells ◽  
Autoradiographic Study ◽  
Yolk Sac ◽  
Differentiated Cells ◽  
Proliferation And Differentiation ◽  
Before And After ◽  
Visceral Yolk Sac ◽  
Mesodermal Origin

The proliferation and differentiation potentiality of the rat visceral yolk sac was investigated both in organ culture and after grafting in vivo. Using alkaline phosphatase as a marker for germ cells, it was shown that these cells are absent in the 12-day-old visceral yolk sac examined before and after organ culture. Therefore, the only cells that proliferate and differentiate must be of endodermal and/or mesodermal origin. By labelling the cells with [3H]thymidine both the endodermal and mesodermal cells were found to proliferate. After 10 days in organ culture or implantation in vivo differentiated tissues (e.g. squamous epidermis, endodermal cysts and giant trophoblast cells) were regularly detected. Several of these differentiated cells contained the radiolabel indicating that they derived from the initial proliferating endodermal and/or mesodermal cells.

Cryopreservation of germ cells from bovine fetal ovaries

1996 ◽  
Vol 8 (2) ◽  
pp. 267 ◽  
Author(s):  
MC Lavoir ◽  
N Rumph ◽  
KJ Betteridge ◽  
SP Leibo
Keyword(s):  
Germ Cells ◽  
Primordial Germ Cells ◽  
Vital Staining ◽  
Cleavage Rate ◽  
Ovarian Cells ◽  
Significant Difference ◽  
Before And After ◽  
Bovine Fetuses ◽  
Future Experimentation ◽  
Logistical Problem

Bovine fetuses at stages required for studies of female germ cells (primordial germ cells and oogonia) become available from the abattoir at unpredictable times. To alleviate this logistical problem, a procedure to cryopreserve these ovarian germ cells has been devised. Fetal ovarian cells were dispersed and suspended in 1.5 M dimethyl sulfoxide (DMSO) prepared in modified TCM 199 medium. The suspensions were aspirated into plastic semen straws, cooled, seeded to induce ice formation at -7 degrees C, and then cooled at 1 degree C min-1 to -70 degrees C before being plunged into liquid nitrogen at -196 degrees C for storage. The straws were thawed at a moderate rate of approximately 250 degrees C min-1, the DMSO was diluted 28-fold with culture medium, and then the cells were cultured for > 2 h before their viability was tested or they were used for nuclear transfer. No statistically significant difference in viability before and after cryopreservation was detected by vital staining with fluorescein diacetate (P > 0.05). When frozen-thawed germ cells were fused to cytoplasts, the cleavage rate of the resultant reconstructed embryos 44 h after fusion was 31%, although none developed into blastocysts. It is concluded that cryopreservation of bovine fetal ovarian germ cells is feasible and can play a major role in facilitating future experimentation.

STUDIES ON THE FAILURE OF HYBRID GERM CELLS TO FUNCTION IN WHEAT SPECIES CROSSES

1932 ◽  
Vol 6 (4) ◽  
pp. 362-373 ◽  
Author(s):  
W. P. Thompson ◽  
J. M. Armstrong
Keyword(s):  
Germ Cells ◽  
Chromosome Numbers ◽  
Pollen Grains ◽  
Wheat Species ◽  
Experimental Conditions ◽  
Degree Of Reduction ◽  
Random Segregation ◽  
Nuclear Development

Chromosome numbers were determined in numerous male gametophytes of F1 between 21- and 14-chromosome species of wheat. The results show that pollen grains with various chromosome numbers from 14 to 21 are actually formed and in approximately the theoretically expected proportions. The lack of plants in later generations which should result from the functioning of pollen grains with intermediate numbers is therefore not due to the failure of such grains to be formed because of a lack of random segregation at the second reduction division.Grains with intermediate numbers are retarded in their nuclear development, so that counts made on stamens in which division is most active give a smaller proportion of grains with intermediate numbers and a higher proportion with parental numbers than is expected theoretically. Retardation in nuclear development is correlated with a deficiency in cytoplasmic contents, 10 to 15% of the grains showing little or no cytoplasm, and another 15 or 20% showing some degree of reduction in cytoplasm. AH grains with reduced cytoplasm and some of those with normal contents are so retarded in nuclear development (having only one or two nuclei or no organized male cells) that they could not function when the normal ones are mature and the stamen dehisces. Unfavorable chromosome conditions in grains with intermediate numbers cause a complete abortion of some grains and retardation of nuclear development in others.Under the best available experimental conditions only 11 or 12% of F1 pollen grains germinate, in contrast to 70 or 80% for parental pollen. No grains with reduced cytoplasm germinate, and at least 50% of those with apparently normal cytoplasm fail to germinate.

scholarly journals Optimization of In Vitro Culture Conditions of Sturgeon Germ Cells for Purpose of Surrogate Production

Animals ◽  
2019 ◽  
Vol 9 (3) ◽  
pp. 106 ◽  
Author(s):  
Xuan Xie ◽  
Ping Li ◽  
Martin Pšenička ◽  
Huan Ye ◽  
Christoph Steinbach ◽  
...  
Keyword(s):  
Germ Cell ◽  
Germ Cells ◽  
Somatic Cells ◽  
Culture Conditions ◽  
Specific Gene ◽  
Acipenser Gueldenstaedtii ◽  
Acipenser Ruthenus ◽  
Before And After ◽  
Optimal Culture Condition

To expand germ cell populations and provide a consistent supply for transplantation, we established basal culture conditions for sturgeon germ cells and subsequently increased their mitotic activity by eliminating gonad somatic cells, supplementing with growth factor, and replacing fetal bovine serum (FBS). The initial basal culture conditions were Leibovitz’s L-15 medium (pH 8.0) supplemented with 5% FBS (p < 0.001) at 21 °C. Proliferation of germ cells was significantly enhanced and maintained for longer periods by elimination of gonad somatic cells and culture under feeder-cell free conditions, with addition of leukemia inhibitory factor and glial-cell-derived neurotrophic factor (p < 0.001). A serum-free culture medium improved germ cell proliferation compared to the L-15 with FBS (p < 0.05). Morphology remained similar to that of fresh germ cells for at least 40 d culture. Germline-specific gene expression analysis revealed no significant changes to germ cells before and after culture. Sterlet Acipenser ruthenus germ cells cultured more than 40 days showed development after transplant into Russian sturgeon Acipenser gueldenstaedtii. Polymerase chain reaction showed 33.3% of recipient gonads to contain sterlet cells after four months. This study developed optimal culture condition for sturgeon germ cells. Germ cells after 40 d culture developed in recipient gonads. This study provided useful information for culture of sturgeon germ cells.

Plantlet formation in callus tissues of Anthurium andraeanum Lind.

1974 ◽  
Vol 2 (2) ◽  
pp. 193-198 ◽  
Author(s):  
R.L.M. Pierik ◽  
H.H.M. Steegmans ◽  
J.A.J. Van Der Meys
Keyword(s):  
Plantlet Formation ◽  
Anthurium Andraeanum ◽  
Callus Tissues

Influence of Fe concentration in the medium on multicellular pollen grains and haploid plants induced by mannitol pretreatment in barley (Hordeum vulgare L.)

PROTOPLASMA ◽  
2006 ◽  
Vol 228 (1-3) ◽  
pp. 101-106 ◽  
Author(s):  
A. Pulido ◽  
F. Bakos ◽  
A. Castillo ◽  
M. P. Vallés ◽  
B. Barnabás ◽  
...  
Keyword(s):  
Hordeum Vulgare ◽  
Pollen Grains ◽  
Hordeum Vulgare L ◽  
Haploid Plants

Development of wheat (Triticum aestivum) pollen wall before and after effect of a gametocide

1990 ◽  
Vol 68 (11) ◽  
pp. 2509-2516 ◽  
Author(s):  
Gamal A. El-Ghazaly ◽  
William A. Jensen
Keyword(s):  
Electron Microscopy ◽  
Triticum Aestivum ◽  
Key Words ◽  
Light And Electron Microscopy ◽  
Pollen Grains ◽  
Pollen Wall ◽  
Seed Plants ◽  
Before And After ◽  
After Effect

Light and electron microscopy studies show that pollen wall development in plants treated with the gametocide RH0007 and untreated plants was similar until the stage at which sporopollenin is normally deposited on the wall. At this stage, the pollen wall of treated plants is 80% thinner than that of the control. Shortly after this stage, the pollen grains in the treated plants collapse and abort. We conclude that the gametocide clearly acts through the inhibition of sporopollenin formation, which results in pollen death. As sporopollenin is found only in the pollen wall of seed plants and the spores of nonseed plants, harm to other parts of the plant is not expected to occur. Key words: pollen wall development, Triticum aestivum, gametocide.

scholarly journals XY oocytes of sex-reversed females with a Sry mutation deviate from the normal developmental process beyond the mitotic stage†

2018 ◽  
Vol 100 (3) ◽  
pp. 697-710 ◽  
Author(s):  
Akihiko Sakashita ◽  
Takuya Wakai ◽  
Yukiko Kawabata ◽  
Chiaki Nishimura ◽  
Yusuke Sotomaru ◽  
...  
Keyword(s):  
Germ Cells ◽  
Developmental Process ◽  
Sex Differentiation ◽  
Primordial Germ Cells ◽  
Primordial Follicle ◽  
Genetic Ablation ◽  
Female Mice ◽  
Meiotic Progression ◽  
Molecular Etiology ◽  
Mitotic Proliferation

Abstract The fertility of sex-reversed XY female mice is severely impaired by a massive loss of oocytes and failure of meiotic progression. This phenomenon remains an outstanding mystery. We sought to determine the molecular etiology of XY oocyte dysfunction by generating sex-reversed females that bear genetic ablation of Sry, a vital sex determination gene, on an inbred C57BL/6 background. These mutant mice, termed XYsry− mutants, showed severe attrition of germ cells during fetal development, resulting in the depletion of ovarian germ cells prior to sexual maturation. Comprehensive transcriptome analyses of primordial germ cells (PGCs) and postnatal oocytes demonstrated that XYsry− females had deviated significantly from normal developmental processes during the stages of mitotic proliferation. The impaired proliferation of XYsry− PGCs was associated with aberrant β-catenin signaling and the excessive expression of transposable elements. Upon entry to the meiotic stage, XYsry− oocytes demonstrated extensive defects, including the impairment of crossover formation, the failure of primordial follicle maintenance, and no capacity for embryo development. Together, these results suggest potential molecular causes for germ cell disruption in sex-reversed female mice, thereby providing insights into disorders of sex differentiation in humans, such as “Swyer syndrome,” in which patients with an XY karyotype present as typical females and are infertile.

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