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PubMed1975

Q-, C-, and G-banding patterns in the germ-line and somatic chromosomes of Miastor sp. (Diptera: Cecidomyidae).

Q-, C-, and G-banding patterns are described for the germ-line and somatic chromosomes of Miastor sp. The chromosome number in the germ line is 36, which is 3 less than that previously reported for the same cultured line 18 years ago. There are 8 chromosome groups: 8 large acrocentrics, 11 large and medium submetacentrics, 4 medium metacentrics, 4 medium subtelocentrics, 6 medium telocentrics, one medium acrocentric, one small submetacentric, and one small telocentric. All 8 large acrocentrics have a Q-band and an interstitial C-band in the short arm, suggesting that these chromosomes are at least partially homologous (or homeologous) and that the germ line originally was polyploid. Only one other germ-line chromosome, the small submetacentric, has a Q-band and an interstitial C-band. In somatic cells there are 8 chromosomes, 28 chromosomes having been eliminated during early cleavage. Three of the four pairs of somatic chromosomes are heteromorphic, despite the indications of a polyploid origin of the germ-line chromosomes and despite the presence of somatic pairing. Furthermore, there is a great deal of restriction on which chromosomes are retained in somatic cells, and it may be that the same 8 chromosomes are retained by all embryos. The G-banding pattern is not as clear as the other two banding patterns. The findings are discussed in relation to observations in other cecidomyids, and a model for the evolution of the chromosome system of Miastor is presented.

Animals↗
PubMed1983

Implication of Triticum searsii as the B-genome donor to wheat using DNA hybridizations.

In vitro DNA:DNA hybridizations and hydroxyapatite thermal chromatography were employed to help identify the species ancestral to the B genome of the polyploid wheats. We hybridized 3H-Triticum aestivum DNA to the unlabeled DNAs of T. urartu, T. speltoides, T. sharonensis, T. bicorne, T. longissimum, and T. searsii, 3H-Labeled DNA of T. urartu was hybridized with the DNA of a synthetic tetraploid. AADD. The heteroduplex thermal stabilities indicated that T. searsii was most closely related to T. aestivum (ABD) and that the genome of T. urartu was more closely related to the A genome than the B genome. The degree of reassociation which may have occurred between the six diploid species and the D genome of T. aestivum was evaluated by hybridizing 3H-T. tauschii DNA with the DNAs of the diploids. The results indicated that T. urartu had the least sequence homology to T. tauschii, the D-genome donor lending additional support to the conclusion that T. urartu is related to the A genome. Thus, it is highly probable that T. searsii is the B-genome donor to the polyploid wheats or a major chromosome donor if the B genome is, in fact, polyphyletic in origin.

DNA↗
PubMed1984

Additional evidence implicating Triticum searsii as the B-genome donor to wheat.

In vitro DNA:DNA hybridizations and hydroxyapatite thermal-elution chromatography were employed to identify the diploid wheat species ancestral to the B genome of Triticum turgidum. 3H-T. turgidum DNA was hybridized to the unlabeled DNAs of T. urartu, T. speltoides, T. sharonensis, T. bicorne, T. longissimum, and T. searsii. 3H-Labeled DNAs of T. monococcum and a synthetic tetraploid AADD were hybridized with unlabeled DNAs of T. urartu and T. searsii to determine the relationship of the A genome of polyploid wheat and T. urartu. The heteroduplex thermal stabilities indicated that T. searsii was most closely related to the B genome of T. turgidum (AB) and that the genome of T. urartu and the A genome have a great deal of base-sequence homology. Thus, it appears that T. searsii is the B-genome donor to polyploid wheat or a major chromosome donor if the B genome is polyphyletic in origin.

Biological Evolution↗
PubMed1986

DNA-flow fluorescence--cytometry of ependymomas. Report on ten surgically removed tumours.

The distribution of DNA is estimated from flow cytometric histograms in surgical specimens of ten ependymomas of different location and varying anaplasia. In three cerebral tumours grade I--II, including one ependymoma of the 4th ventricle, only limited elevation of the 4 C maxima was a prominent feature, corresponding to the microscopical frequency of typical mitoses. Four grade-III ependymomas showed aneuploid or polyploid histograms with stem lines. One frontal tumour was classified as "transitional" because of more numerous mitoses and abnormally elevated S and G2 + M phases, which increased in tissue culture. A correlation between the degree of anaplasia with the DNA pattern was difficult to pursue in two spinal ependymomas obviously lacking microscopical mitoses: Both--one a so-called tanycytic variant of grade I--II, and the other probably a metastasis from a cerebellar tumour--had a clear polyploid DNA histogram with a strikingly increased proliferation index, similar to the more malignant tumours of grade III. Also flow DNA measurements probably allow the decoding of heterogenous mixtures of tumour cells which are not always benign in ependymomas of lower grades of anaplasia microscopically.

Adolescent↗
PubMed1992

DNA image cytometry: a prognostic tool in rectal cancer?

In 68 patients the DNA content of tumor cells was measured by image cytometry after resection of the rectum because of cancer. In the DNA histogram a differentiation between diploid (n = 19), polyploid (n = 24), hypotriploid (n = 17), and hypertriploid (n = 8) tumors was possible. The best relapse-free survival time was found in patients with diploid tumors. The prognosis worsened from polyploid to hypotriploid and was worse in hypertriploid tumors. Testing for a prognostic advantage of diploid over aneuploid tumors without adjustment for additional factors simply by means of the log-rank statistic gave a (one-sided) P of 0.1013. In a multivariate analysis the degree of differentiation turned out most important. Again, an appropriate test for prognostic relevance of DNA content failed to be significant (P = 0.3264).

Adult↗
PubMed1977

In vitro development of plants from microspores of rice.

Rice (Oryza sativa L., 2n=24) anthers containing microspores in the early-uninucleate to first-mitosis stages were induced successfully to develop into plants in vitro through an intermediary step of callus formation. Callus initiation occurred with highest frequency in anthers containing mid-uninucleate imcrospores. The callus derived from different stages of microspore development differed in the potential to differentiate into plants. The plants regenerated from pollen callus were predominantly haploid or diploid; polyploid and aneuploid plants were relatively infrequent. The first division of the uninucleate microspores was asymmetrical, resulting in the formation of large vegetative and small generative nuclei. The vegetative nucleus divided repeatedly and assumed the major role in the formation of callus, whereas the generative nucleus degenerated rapidly. Simultaneous division of the two nuclei was observed in a few pollen grains. Nuclear fusion during the very initial stages of pollen development was postulated to account for the occurrence of the diploid and polyploid plants.

Cell Differentiation↗
PubMed1990

DNA ploidy and proliferative activity of human pulmonary epithelium.

DNA ploidy and distribution has been determined in normal and abnormal bronchial, bronchiolar and alveolar epithelium from 22 patients, aged between 0 and 85 years, 9 of whom had received chemotherapy for malignant disease. The DNA ploidy was diploid in all the specimens examined. The S + G2/M fraction was significantly greater in diseased than normal bronchial trees. In the bronchial epithelium, mean values +/- the standard deviation (SD) were 5.5 +/- 2.2% vs 1.1 +/- 0.6%, in bronchiolar epithelium 4.6 +/- 1.6% vs 1.0 +/- 0.9% and in alveolar epithelium 4.6 +/- 1.6% vs 0.8 +/- 0.5%. The highest S + G2/M value of 8.9% was obtained from inflamed bronchial epithelium. Polyploid cells up to the octaploid range occurred infrequently but their incidence was slightly increased to between 0.16% and 0.9% in diseased lungs and in patients who had received chemotherapeutic drugs. It was concluded that (1) non-cancerous drugs. It was concluded that (1) non-cancerous pulmonary epithelium is diploid, that (2) pulmonary epithelium shows steady-state renewal at all ages and polyploid cells are rare under normal conditions and that (3) the S + G2/M fraction increases up to approximately 10% in reactive proliferative states.

Adolescent↗
PubMedDate not supplied

Characterization of three cloned cell lines from a N-nitrosobis(2-hydroxypropyl)amine-induced transplantable hamster pancreatic ductal adenocarcinoma.

To investigate characteristics of pancreatic carcinoma growth behavior, the cloned cell lines, HPD1NR, HPD2NR, and HPD3NR, were established from a transplantable hamster ductal adenocarcinoma induced by N-nitrosobis(2-hydroxypropyl)amine (BHP). All three clones showed similar epithelial cell morphology and grew as sheets in culture with no differences in doubling times, ranging from 23-28 h. Mutation in the c-Ki-ras exon 1 was detected in common. The modal chromosome numbers were also found to be similar at 60, 62, and 60-62 in the less than tetraploid cells in the three clones. In contrast, a clear difference in frequencies of tetraploid or polyploid cells at 24.7, 22.5, and 75.5% in HPD1NR, HPD2NR, and HPD3NR, respectively, was evident. Tumorigenic potency evaluated by transplanting individual clones revealed HPD3NR to display pronouncedly less growth in syngeneic hamsters. The results suggest that increase in frequency of tetraploid or polyploid cells might be associated with a decreased in vivo growth potential of hamster pancreatic ductal adenocarcinomas, and suggest that these clones might become a valuable tool for understanding in vivo growth mechanisms of cancer cells.

Animals↗
PubMed2002

Molecular control of megakaryopoiesis and thrombopoiesis.

Megakaryopoiesis and subsequent thrombopoiesis occur through complex biologic steps: megakaryocyte precursors that developed from hematopoietic stem cells initially proliferate, then differentiate into mature polyploid megakaryocytes, and finally release platelets. Although a number of growth factors can augment megakaryopoiesis in vitro, thrombopoietin is a physiologic and the most potent regulator of megakaryopoiesis in vitro and in vivo. Thrombopoietin induces the growth of megakaryocyte precursors through activation of multiple signaling cascades, including Ras/mitogen-activated protein kinase (MAPK), signal transducers and activators of transcription 5 (STAT5), phosphatidylinositol 3-kinase (PI3-K)/Akt, and protein kinase C, whereas it induces megakaryocytic maturation primarily through the Ras/MAPK pathway. During the maturation step, megakaryocytes undergo polyploidization characterized by repeated rounds of DNA replication without concomitant cell division. During these rounds of replication, cytokinesis is neglected because of the down-regulated expression of AIM-1, and DNA replication occurs through the increased expression of D-type cyclins. As for transcriptional regulation during megakaryopoiesis, GATA-1 plays a central role in the lineage commitment of hematopoietic stem cells toward erythroid/megakaryocytic lineage and subsequent maturation. p45 NF-E2 is essential for platelet release from terminally differentiated megakaryocytes. At present, mutations of GATA-1, AML1, and HOXA11 genes have been found in hereditary diseases accompanying thrombocytopenia among humans.

Animals↗
PubMed1998

[Intratumoral heterogeneity of DNA content in lung adenocarcinoma].

Quantitative microspectrophotometry was performed in 10 resected lung adenocarcinomas for the purpose of studying intratumoral heterogeneity of DNA content. Histologically, there were 3 stage I, 2 stage IIIA, 2 stage IIIB, and 3 stage IV cases, consisting of 3 well, 4 moderately and 3 poorly differentiated cases. The tumors were cut at the greatest dimension and fine needle aspirations were performed from 5 separate areas (central, cranial, caudal, inward, and outward). One of the imprint smears was submitted for cytology and the other for microspectrophotometry using Feulgen staining. Although G0G1 and G2M phase cell populations varied in each specimen, all 5 specimens showed the same DNA histogram in 9 cases. These were 4 aneuploid, 4 polyploid (euploid but not diploid) and only one diploid tumor. Two stem lines were observed in the remaining single case. One of 5 specimens showed aneuploid, the other 4 specimens showed polyploid. Namely, intratumoral heterogeneity was observed in one case. Ther was no relationship between intratumoral heterogeneity tumor size or differentiation. Most lung adenocarcinomas showed DNA stability. Fine needle aspiration specimens obtained from one area of lung adenocarcinoma represent the DNA contents of the entire tumor.

Adenocarcinoma↗
PubMed2002

Characterisation of polymorphic microsatellite markers from Aegilops tauschii and transferability to the D-genome of bread wheat.

Microsatellites were isolated from a Aegilops tauschii (the D-genome donor of bread wheat) library enriched for various motifs. Primers generated from the flanking region of the microsatellites were used successfully to amplify the corresponding loci in the D genome of bread wheat. Additional amplification sometimes also occurred from the A and B genomes. The majority of the microsatellites contained (GA)(n) and (GT)(n) motifs. GA and GT repeats appeared to be both more abundant in this library and more polymorphic than other types of repeats. The allele number for both types of dinucleotide repeats fitted a Poisson distribution. Deviance analysis showed that GA and GT were more polymorphic than other motifs in bread wheat. Within each motif type (di-, tri- and tetra-nucleotide repeats), repeat number has no influence on polymorphism. The microsatellites were mapped using the Triticum aestivum Courtot x Chinese Spring mapping population. A total of 100 markers was developed on this intraspecific map, mainly on the D genome. For polyploid species, isolation of microsatellites from an ancestral diploid donor seems to be an efficient way of developing markers for the corresponding genome in the polyploid plant.

Journal Article↗
PubMed2002

Studies of the transferability of microsatellites derived from Triticum tauschii to hexaploid wheat and to diploid related species using amplification, hybridization and sequence comparisons.

Hexaploid wheat ( Triticum aestivum L em Thell) is derived from a complex hybridization procedure involving three diploid species carrying the A, B and D genomes, respectively. We recently isolated microsatellites from a T. tauschii library enriched for various motifs and evaluated the transferability of these markers to several diploid species carrying the A, B or D genomes. All of the primer pairs amplifying more than one locus on bread wheat and half of those giving D-genome-specific loci gave an amplification product on A-and/or B-diploid species. All of the markers giving a single amplification product for T. tauschii and no amplification on the other diploid species were D-genome-specific at the hexaploid level. The non-specific microsatellite markers (which gave an amplification product on diploid species carrying the A, B or D genome) gave either a complex amplification pattern on bread wheat (with several bands) or generated a single band which mapped to the D genome. Southern blot hybridizations with probes corresponding to the microsatellite flanking regions gave a signal on all diploid and hexaploid species, whatever the specificity of the microsatellite. The patterns observed on bread wheat were generally in accordance with those observed for diploid species, with slight rearrangements. This suggests that the specificity of microsatellite markers is probably due to mutations in microsatellite flanking regions rather than sequence elimination during polyploidization events and that genome stringency is higher at the polyploid than at the diploid level.

Journal Article↗
PubMed2002

Identification of chromosome arms influencing expression of the HMW glutenins in wheat.

The high-molecular-weight (HMW) glutenin genes, located on the group 1L chromosome arms, are a major determinant for baking quality in wheat ( Triticum aestivum L.). In addition, the HMW glutenin genes provide a valuable model system for studying the evolution and regulation of orthologous and paralogous genes in polyploid species. The goal of this study was to identify loci that modify the expression of the HMW glutenins, and to map them to specific chromosome arms. Comparisons were made between endosperms with zero versus three (or three versus six) doses for each of the 42 chromosome arms of wheat. SDS-PAGE and scanning densitometry were used to quantify the protein expression levels of the four HMW glutenin genes in cv. Chinese Spring, for each of the dosage comparisons. Fifteen chromosome arms were found to have significant effects on Glu-B1-1, excluding the structural gene dosage effect: eight positive effects on 1AL, 2AS, 2BL, 2DS, 5DS, 6AL, 6DL, and 7AL and seven negative effects on 1BS, 1DS, 1DL, 4DL, 6BS, 6DS, and 7AS. Nineteen chromosome arms had significant effects on Glu-B1-2, excluding the structural gene dosage effect: eight positive effects on 1AL, 2AS, 2BS, 3AL, 4BL, 6DS, 7BL and 7DS and 11 negative effects on 1AS, 1BS, 1DS, 1DL, 2AL, 2BL, 3DS, 4BS, 4DL, 5BL, and 6BS. Twenty chromosome arms had significant effects on Glu-D1-1, excluding the structural gene dosage effect: 11 positive effects on 1AL, 1BL, 2BS, 2DS, 5BS, 5DS, 6AL, 6DS, 6DL, 7AL, and 7BL and nine negative effects on 1AS, 1BS, 1DS, 2BL, 4DL, 5BL, 5DL, 6BL, and 7DS. Twenty-five chromosome arms had significant effects on Glu-D1-2, excluding the structural gene dosage effect: 17 positive effects on 1BL, 2AS, 2BS, 2DS, 2DL, 3AS, 3AL, 3BS, 5AS, 5BS, 5DL, 6AL, 6DL, 7AL, 7BS, 7BL, and 7DL and eight negative effects on 1DS, 4DL, 5AL, 5BL, 6BS, 6BL, 6DS and 7DS. Of the 164 gene-chromosome arm tests performed, about 52% (85/164) showed no significant effects, and 48% (79/164) showed significant effects, excluding the structural gene dosage effects. Of the significant effects, 56% (44/79) were positive effects, and 44% (35/79) were negative effects. Comparisons of dosage effects on orthologous loci (both x-type or both y-type HMW glutenins) showed that orthologous HMW glutenin genes are largely influenced by the same regulatory systems. Less correlation was found for comparisons between paralogous genes, although considerable conservation was observed at this level as well. These observations suggest that after polyploidization, many of the duplicated orthologous regulatory loci were inactivated by mutation, thus consolidating control over the HMW glutenin genes. Possible candidates for orthologous regulatory genes were identified in maize and barley. This study represents the first comprehensive search of the wheat genome for regulators of the HMW glutenins.

Chromosome Mapping↗
PubMed2003

High-resolution organellar genome analysis of Triticum and Aegilops sheds new light on cytoplasm evolution in wheat.

We have utilised polymorphic chloroplast microsatellites to analyse cytoplasmic relationships between accessions in the genera Triticum and Aegilops. Sequencing of PCR products revealed point mutations and insertions/deletions in addition to the standard repeat length expansion/contraction which most likely represent ancient synapomorphies. Phylogenetic analyses revealed three distinct groups of accessions. One of these contained all the non- Aegilops speltoides S-type cytoplasm species, another comprised almost exclusively A, C, D, M, N, T and U cytoplasm-type accessions and the third contained the polyploid Triticum species and all the Ae. speltoides accessions, further confirming that Ae. speltoides or a closely related but now extinct species was the original B-genome donor of cultivated polyploid wheat. Successive decreases in levels of genetic diversity due to domestication were also observed. Finally, we highlight the importance of elucidating longer-term evolutionary processes operating at microsatellite repeat loci.

Base Sequence↗
PubMed2004

Substitutes for genome differentiation in tuber-bearing Solanum: interspecific pollen-pistil incompatibility, nuclear-cytoplasmic male sterility, and endosperm.

The cultivated potato, Solanum tuberosum L. (2n=4x=48), has a very large number of related wild and cultivated tuber-bearing species widely distributed in the Americas. These species, grouped in 16 taxonomic series, range from the diploid to the hexaploid level. Polyploid species are either disomic or polysomic, and sexual polyploidization via genetically controlled 2 n gametes has played a major role in their evolution. Species are separated in nature by geographical and ecological barriers. However, there are several examples of sympatric species that share the same niches but do not readily cross (i.e., the diploids S. commersonii and S. chacoense in certain areas of Argentina). External barriers alone are, therefore, not sufficient to explain species integrity. In addition, there is no strong evidence indicating that genome differentiation is important in the group. In this review we present evidence supporting the assertion that interspecific pollen-pistil incompatibility, nuclear-cytoplasmic male sterility, and the endosperm are major forces that strengthen the external hybridization barriers allowing, at the same time and under specific circumstances, a certain amount of gene exchange without jeopardizing the integrity of the species.

Diploidy↗
PubMed2025

Genomics-enabled dissection of sea wheatgrass genome for advancing wheat genetic resources.

Wheat production is challenged by biotic and abiotic stresses. Alien gene transfer is an effective approach to tackle such challenges. We previously showed that sea wheatgrass (SWG; Thinopyrum junceiforme (2n = 2x = 28; J1J2) is an untapped resource possessing resistance to an array of pests and abiotic stress. However, the transfer of these important traits has been hindered by the lack of genomic resources and a clear picture of its genome constitution. Using multi-color genomic in situ hybridization, we distinguished the SWG sub-genomes and corroborated that the J1 sub-genome is closely related to the E genome of Th. elongatum and the J genome of Th. bessarabicum and the J2 sub-genome to the V genome of Dasypyrum villosum. Meanwhile, we developed a draft SWG genome assembly and 127 SWG-specific DNA markers covering the 14 SWG chromosomes. Screening a population of 466 BC2F1 and BC2F2 individuals, derived from backcrosses of wheat-SWG amphiploid to wheat, by the SWG-specific markers led to selection of 72 plants putatively carrying one or two SWG chromosomes. The genome painting analysis of the 72 plants eventually identified a set of 37 wheat-SWG chromosome addition lines covering all the 14 pairs of SWG chromosomes and two compensating Robertsonian translocations (RobTs). While the wheat-SWG chromosome addition lines and RobTs are invaluable genetic resources for wheat improvement via chromosome engineering, our results showed the power of genome-specific markers in combination with genome painting in dissection of a polyploid genome and implicated the origin of a group of important polyploid grasses.

Triticum↗
PubMed2002

In vitro response of myelodysplastic megakaryocytopoiesis to megakaryocyte growth and development factor (MGDF).

To evaluate a potential therapeutic role for megakaryocyte growth and development factor (MGDF) in myelodysplastic syndromes (MDS) we compared the in vitro response of normal and myelodysplastic megakaryopoiesis to MGDF on bone marrow mononuclear cells from nine MDS patients and four healthy donors in a short term liquid culture system. The cells were incubated with MGDF alone (1 ng/ml, 10 ng/ml and 100 ng/ml) and in combination with 20 ng/ml stem cell factor, 2 U/ml erythropoietin (EPO) and 100 ng/ml interleukin-3 (IL-3). Cytospins were prepared after 4 days and 10 days for CD61 APAAP staining. In addition, the ploidy of propidium iodide stained CD61 positive cells were detected by flow cytometry. The CD61+ cell number significantly increased (13-fold after 4 days, 56-fold after 10 days of culture) when MGDF (100 ng/ml) was added to the normal bone marrow (NBM) cultures (P=0.0003; P=0.0001). In the MDS cultures the absolute number of endomitotic cells as well as the percentage of polyploid cells remained significantly lower than in NBM after 10 days (P=0.0001). The effect of MGDF on polyploidization of MDS cells was significantly dose dependent (P=0.0051). We found no correlation between peripheral platelet count, cellularity of the bone marrow, number of bone marrow megakaryocytes or FAB-subtype and response to MGDF. Additional administration of IL-3 resulted in a left-shift of megakaryopoiesis in both groups. Even though the response of myelodysplastic megakaryocytic progenitors to MGDF shows inter-individual variations, it is significantly impaired overall. Our data suggest that higher doses of MGDF may be able to ameliorate thrombocytopenia in a subgroup of MDS patients.

Aged↗
PubMed1999

Molecular basis to understand polyploidy.

Polyploidy which is defined as an increase in chromosome number by an exact multiple of the haploid chromosome number is a typical characteristic of plant species but can also be found in almost all organisms from protists to humans. In the plant kingdom, since the reproduction can occur vegetatively, polyploid cells have time to settle down from the evolution point of view as a stable genetic system. So, 30 to 70% of angiosperms, including many important crop plants, are estimated to have polyploidy in their lineages. The success of polyploid species in plants has been attributed to their ability to colonise a wider range of habitats and to survive better in unstable climates compared with their diploid progenitors, presumably due to increased heterozygosity and flexibility provided by the presence of additional alleles.

Animals↗