scholarly journals 3H-thymidyne incorporation into the microspores and pollen grains nuclei in excised Tradescantia stamens

2015 ◽  
Vol 47 (1–2) ◽  
pp. 163-172 ◽  
Author(s):  
Maria Charzyńska ◽  
Joanna Maleszka
Keyword(s):  
Dna Synthesis ◽  
Generative Cell ◽  
Pollen Grains ◽  
Vegetative Nucleus ◽  
Generative Nucleus ◽  
Tetrad Stage ◽  
Pollen Grain Wall ◽  
Medium Culture

The development of microspores and pollen grains lasts in <i>Tradescantia bracteata in vivo</i> from the tetrad stage to pollen shedding about 14 days. This including 7 days of the microspore life cycle. In stamens excised and placed on a medium the microspores and pollen grains develop normally for at least 3 days. <sup>3</sup>H-thymidine is added into medium culture. DNA synthesis m the microspore nucleus is demonstrated 6 days after tetrad formation so at the end of microspore interphase. During synthesis the nucleus lies at one end of the long axis of the vacuolated microspore. Synthesis ends before migration of the nucleus to the proximal pole of the microspore where mitosis begins. Incorporation of <sup>3</sup>H-thymidine into the generative nucleus is noted in two-celled pollen grains as early as about 24h after the end of microspore division. During DNA synthesis the generative cell is rounded and is still adjacent to the pollen grain wall. DNA synthesis ends before separation of the generative cell from the sporoderm, before the generative nucleus starts to elongate. <sup>3</sup>H-thymidine is not incorporated into the vegetative nucleus in stamens developing <i>in vitro</i>.

scholarly journals Tradescantia bracteata pollen in vitro: pollen tubes development and mitosis

2015 ◽  
Vol 46 (4) ◽  
pp. 587-598 ◽  
Author(s):  
E. Lewandowska ◽  
M. Charzyńska
Keyword(s):  
Pollen Tube ◽  
Generative Cell ◽  
Metaphase Plate ◽  
Pollen Grains ◽  
Neutral Red ◽  
Generative Nucleus ◽  
Mitotic Spindles ◽  
Germinating Pollen ◽  
Vacuolar System

About 90 per cent of <i>Tradescantia bracteata</i> pollen germinates <i>in vitro</i> after 15 min. Mitosis starts in the pollen tube after about 3 h. The mitotic trans-formations of chromosomes within the generative nucleus are not synchronized. They involve succesively the linearly arranged chromosomes in the elongated generative nucleus. In metaphase the chromosomes are arranged tandem-like linearly along the pollen tube. The chromatides translocate in anaphase from various distances to the poles in a plane parallel to the metaphase plate. This suggests that chromosomes have individual mitotic spindles and that coordination of the chromosome transformations in the generative cell is much less strict than in a typical somatic mitosis. Starch is the storage material of pollen grains. In the vegetative cytoplasm of mature pollen grains minute reddish-orange vesicular structures are visible after staining with neutral red. They do not fuse with the vacuoles proper arising in germinating pollen grains to form the vacuolar system of the pollen tube.

Pollen germination of Larix sibirica (Siberian larch) in vitro

1970 ◽  
Vol 48 (2) ◽  
pp. 213-215 ◽  
Author(s):  
Ronghui Ho ◽  
Glenn E. Rouse
Keyword(s):  
Pollen Tube ◽  
Pollen Germination ◽  
Generative Cell ◽  
Siberian Larch ◽  
Pollen Grains ◽  
The Body ◽  
Stalk Cell ◽  
Body Cell

Mature pollen grains of Siberian larch were successfully germinated in vitro for the first time. Mature grains, containing two prothallial cells, a generative cell, and a tube cell, were incubated in stock solution B for periods up to 5 days. At the 2-day stage, the generative cell had divided into a stalk cell and body cell. In 5 days, the body nucleus had divided into two male nuclei contained within the body cell. Pollen germination was limited to elongation of the intine towards the distal pole, accompanied by a splitting of the exine, but without any suggestion of the formation of a pollen tube. This supports the suggestion of previous studies in vivo on other species of Larix pollen that a pollen tube does not form before the pollen grain reaches the nucellus of the ovule.

Pollen germination of Tsuga heterophylla in vitro

1971 ◽  
Vol 49 (1) ◽  
pp. 117-119 ◽  
Author(s):  
RongHui Ho ◽  
Oscar Sziklai
Keyword(s):  
Male Gametophyte ◽  
Generative Cell ◽  
Calcium Nitrate ◽  
Pollen Grains ◽  
Potassium Nitrate ◽  
Tsuga Heterophylla ◽  
The Body ◽  
Stalk Cell

The development of the male gametophyte from western hemlock (Tsuga heterophylla (Raf.) Sarg.) pollen was complete after 5 days of incubation. This normally takes at least 6 weeks in vivo. The pollen was cultured in a solution containing boron, calcium nitrate, magnesium sulfate, and potassium nitrate. In 2 days the generative cell divided into the body cell and the stalk cell and after a further 3 days the body nucleus divided into two sperm nuclei. Morphological descriptions and measurements of the germinating pollen grains were made.

Cytology of exceptional development of the male gametophyte in Ophiorrhiza mungos

1975 ◽  
Vol 53 (18) ◽  
pp. 2032-2037 ◽  
Author(s):  
Omana Philip ◽  
P. M. Mathew
Keyword(s):  
Male Gametophyte ◽  
Pollen Grains ◽  
Nuclear Bodies ◽  
Vegetative Nucleus ◽  
Generative Nucleus ◽  
Sperm Nuclei ◽  
Normal Division

In Ophiorrhiza mungos L. the first division of the microspore nucleus gives rise to a large vegetative nucleus and a smaller generative nucleus. The vegetative nucleus subsequently fragments into a number of irregularly sized particles. These vegetative nuclear bodies move to and flow out of the three germ pores to form three spherical buds on the developing pollen grains. The 'pollen buds' are shed off from the pollen proper before dehiscence. The uninucleate grains readily germinate in vitro, and the single generative nucleus undergoes normal division to give rise to two sperm nuclei.

Thrombopoietin-Induced Stimulation of Megakaryocyte- Enriched Bone Marrow Cultures

1979 ◽  
Author(s):  
K. L. Kellar ◽  
B. L. Evatt ◽  
C. R. McGrath ◽  
R. B. Ramsey
Keyword(s):  
Bone Marrow ◽  
Dna Synthesis ◽  
Bone Marrow Cells ◽  
Rabbit Plasma ◽  
Bone Marrow Cultures ◽  
Plasma Fractions ◽  
Marrow Cultures ◽  
Stimulation Of

Liquid cultures of bone marrow cells enriched for megakaryocytes were assayed for incorporation of 3H-thymidine (3H-TdR) into acid-precipitable cell digests to determine the effect of thrombopoietin on DNA synthesis. As previously described, thrombopoietin was prepared by ammonium sulfate fractionation of pooled plasma obtained from thrombocytopenic rabbits. A control fraction was prepared from normal rabbit plasma. The thrombopoietic activity of these fractions was determined in vivo with normal rabbits as assay animals and the rate of incorporation of 75Se-selenomethionine into newly formed platelets as an index of thrombopoietic activity of the infused material. Guinea pig megakaryocytes were purified using bovine serum albumin gradients. Bone marrow cultures containing 1.5-3.0x104 cells and 31%-71% megakaryocytes were incubated 18 h in modified Dulbecco’s MEM containing 10% of the concentrated plasma fractions from either thrombocytopenic or normal rabbits. In other control cultures, 0.9% NaCl was substituted for the plasma fractions. 3H-TdR incorporation was measured after cells were incubated for 3 h with 1 μCi/ml. The protein fraction containing thrombopoietin-stimulating activity caused a 25%-31% increase in 3H-TdR incorporation over that in cultures which were incubated with the similar fraction from normal plasma and a 29% increase over the activity in control cultures to which 0.9% NaCl had been added. These data suggest that thrombopoietin stimulates DNA synthesis in megakaryocytes and that this tecnique may be useful in assaying thrombopoietin in vitro.

scholarly journals RuvAB and RecG Are Not Essential for the Recovery of DNA Synthesis Following UV-Induced DNA Damage in Escherichia coli

Genetics ◽  
2004 ◽  
Vol 166 (4) ◽  
pp. 1631-1640 ◽  
Author(s):  
Janet R Donaldson ◽  
Charmain T Courcelle ◽  
Justin Courcelle
Keyword(s):  
Dna Damage ◽  
Dna Synthesis ◽  
Dna Lesions ◽  
Replication Forks ◽  
Wild Type ◽  
Replication Machinery ◽  
Dna Junctions ◽  
Fork Regression

Abstract Ultraviolet light induces DNA lesions that block the progression of the replication machinery. Several models speculate that the resumption of replication following disruption by UV-induced DNA damage requires regression of the nascent DNA or migration of the replication machinery away from the blocking lesion to allow repair or bypass of the lesion to occur. Both RuvAB and RecG catalyze branch migration of three- and four-stranded DNA junctions in vitro and are proposed to catalyze fork regression in vivo. To examine this possibility, we characterized the recovery of DNA synthesis in ruvAB and recG mutants. We found that in the absence of either RecG or RuvAB, arrested replication forks are maintained and DNA synthesis is resumed with kinetics that are similar to those in wild-type cells. The data presented here indicate that RecG- or RuvAB-catalyzed fork regression is not essential for DNA synthesis to resume following arrest by UV-induced DNA damage in vivo.

scholarly journals Prunus necrotic ringspot virus Early Invasion and Its Effects on Apricot Pollen Grain Performance

2007 ◽  
Vol 97 (8) ◽  
pp. 892-899 ◽  
Author(s):  
Khalid Amari ◽  
Lorenzo Burgos ◽  
Vicente Pallas ◽  
María Amelia Sanchez-Pina
Keyword(s):  
Developmental Stages ◽  
Generative Cell ◽  
Pollen Grains ◽  
Growth Zone ◽  
Pollen Tubes ◽  
Climatic Conditions ◽  
Pollen Grain ◽  
Ringspot Virus ◽  
Prunus Necrotic Ringspot Virus

The route of infection and the pattern of distribution of Prunus necrotic ringspot virus (PNRSV) in apricot pollen were studied. PNRSV was detected both within and on the surface of infected pollen grains. The virus invaded pollen during its early developmental stages, being detected in pollen mother cells. It was distributed uniformly within the cytoplasm of uni- and bicellular pollen grains and infected the generative cell. In mature pollen grains, characterized by their triangular shape, the virus was located mainly at the apertures, suggesting that PNRSV distribution follows the same pattern as the cellular components required for pollen tube germination and cell wall tube synthesis. PNRSV also was localized inside pollen tubes, especially in the growth zone. In vitro experiments demonstrated that infection with PNRSV decreases the germination percentage of pollen grains by more than half and delays the growth of pollen tubes by ≈24 h. However, although PNRSV infection affected apricot pollen grain performance during germination, the presence of the virus did not completely prevent fertilization, because the infected apricot pollen tubes, once germinated, were able to reach the apricot embryo sacs, which, in the climatic conditions of southeastern Spain, mature later than in other climates. Thus, infected pollen still could play an important role in the vertical transmission of PNRSV in apricot.

scholarly journals Effect of incorporation of cidofovir into DNA by human cytomegalovirus DNA polymerase on DNA elongation.

1997 ◽  
Vol 41 (3) ◽  
pp. 594-599 ◽  
Author(s):  
X Xiong ◽  
J L Smith ◽  
M S Chen
Keyword(s):  
Dna Synthesis ◽  
Dna Polymerase ◽  
Human Cytomegalovirus ◽  
Chain Elongation ◽  
Exonuclease Activity ◽  
Dna Chain ◽  
Alternate Substrate

Cidofovir (CDV) (HPMPC) has potent in vitro and in vivo activity against human cytomegalovirus (HCMV), CDV diphosphate (CDVpp), the putative antiviral metabolite of CDV, is an inhibitor and an alternate substrate of HCMV DNA polymerase. CDV is incorporated with the correct complementation to dGMP in the template, and the incorporated CDV at the primer end is not excised by the 3'-to-5' exonuclease activity of HCMV DNA polymerase. The incorporation of a CDV molecule causes a decrease in the rate of DNA elongation for the addition of the second natural nucleotide from the singly incorporated CDV molecule. The reduction in the rate of DNA (36-mer) synthesis from an 18-mer by one incorporated CDV is 31% that of the control. However, the fidelity of HCMV DNA polymerase is maintained for the addition of the nucleotides following a single incorporated CDV molecule. The rate of DNA synthesis by HCMV DNA polymerase is drastically decreased after the incorporation of two consecutive CDV molecules; the incorporation of a third consecutive CDV molecule is not detectable. Incorporation of two CDV molecules separated by either one or two deoxynucleoside monophosphates (dAMP, dGMP, or dTMP) also drastically decreases the rate of DNA chain elongation by HCMV DNA polymerase. The rate of DNA synthesis decreases by 90% when a template which contains one internally incorporated CDV molecule is used. The inhibition by CDVpp of DNA synthesis by HCMV DNA polymerase and the inability of HCMV DNA polymerase to excise incorporated CDV from DNA may account for the potent and long-lasting anti-CMV activity of CDV.

scholarly journals Interaction between the DrosophilaCAF-1 and ASF1 Chromatin Assembly Factors

2001 ◽  
Vol 21 (19) ◽  
pp. 6574-6584 ◽  
Author(s):  
Jessica K. Tyler ◽  
Kimberly A. Collins ◽  
Jayashree Prasad-Sinha ◽  
Elizabeth Amiott ◽  
Michael Bulger ◽  
...  
Keyword(s):  
Dna Synthesis ◽  
Polytene Chromosomes ◽  
Normal Growth ◽  
Chromatin Assembly ◽  
Truncated Form ◽  
Assembly Factor ◽  
Development And Differentiation ◽  
Growth Development

ABSTRACT The assembly of newly synthesized DNA into chromatin is essential for normal growth, development, and differentiation. To gain a better understanding of the assembly of chromatin during DNA synthesis, we identified, cloned, and characterized the 180- and 105-kDa polypeptides of Drosophila chromatin assembly factor 1 (dCAF-1). The purified recombinant p180+p105+p55 dCAF-1 complex is active for DNA replication-coupled chromatin assembly. Furthermore, we have established that the putative 75-kDa polypeptide of dCAF-1 is a C-terminally truncated form of p105 that does not coexist in dCAF-1 complexes containing the p105 subunit. The analysis of native and recombinant dCAF-1 revealed an interaction between dCAF-1 and theDrosophila anti-silencing function 1 (dASF1) component of replication-coupling assembly factor (RCAF). The binding of dASF1 to dCAF-1 is mediated through the p105 subunit of dCAF-1. Consistent with the interaction between dCAF-1 p105 and dASF1 in vitro, we observed that dASF1 and dCAF-1 p105 colocalized in vivo inDrosophila polytene chromosomes. This interaction between dCAF-1 and dASF1 may be a key component of the functional synergy observed between RCAF and dCAF-1 during the assembly of newly synthesized DNA into chromatin.

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