Identification and phenotypic description of new wheat – six-rowed winter barley disomic additions

To increase the allelic variation in wheat–barley introgressions, new wheat–barley disomic addition lines were developed containing the 2H, 3H, 4H, 6H, and 7H chromosomes of the six-rowed Ukrainian winter barley ‘Manas’. This cultivar is agronomically much better adapted to Central European environmental conditions than the two-rowed spring barley ‘Betzes’ previously used. A single ‘Asakaze’ × ‘Manas’ wheat × barley hybrid plant was multiplied in vitro and one backcross plant was obtained after pollinating 354 regenerant hybrids with wheat. The addition lines were selected from the self-fertilized seeds of the 16 BC2 plants using genomic in situ hybridization. The addition lines were identified by fluorescence in situ hybridization using repetitive DNA probes (HvT01, GAA, pTa71, and Afa family), followed by confirmation with barley SSR markers. The addition lines were grown in the phytotron and in the field, and morphological parameters (plant height, fertility, tillering, and spike characteristics) were measured. The production of the disomic additions will make it possible to incorporate the DNA of six-rowed winter barley into the wheat genome. Addition lines are useful for genetic studies on the traits of six-rowed winter barley and for producing new barley dissection lines.

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Development and molecular cytogenetic identification of new winter wheat – winter barley (‘Martonvásári 9 kr1’ – ‘Igri’) disomic addition lines

Genome ◽

10.1139/g06-134 ◽

2007 ◽

Vol 50 (1) ◽

pp. 43-50 ◽

Cited By ~ 20

Author(s):

É. Szakács ◽

M. Molnár-Láng

Keyword(s):

In Situ Hybridization ◽

Winter Wheat ◽

Fluorescent In Situ Hybridization ◽

Triticum Aestivum L ◽

Winter Barley ◽

Addition Lines ◽

Hordeum Vulgare L ◽

Disomic Addition Lines ◽

Simple Sequence

This paper describes a series of winter wheat – winter barley disomic addition lines developed from hybrids between winter wheat line Triticum aestivum L. ‘Martonvásári 9 kr1’ and the German 2-rowed winter barley cultivar Hordeum vulgare L. ‘Igri’. The barley chromosomes in a wheat background were identified from the fluorescent in situ hybridization (FISH) patterns obtained with various combinations of repetitive DNA probes: GAA–HvT01 and pTa71–HvT01. The disomic addition lines 2H, 3H, and 4H and the 1HS isochromosome were identified on the basis of a 2-colour FISH with the DNA probe pairs GAA–pAs1, GAA–HvT01, and pTa71–HvT01. Genomic in situ hybridization was used to confirm the presence of the barley chromosomes in the wheat genome. The identification of the barley chromosomes in the addition lines was further confirmed with simple-sequence repeat markers. The addition lines were also characterized morphologically.

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Identification of new winter wheat – winter barley addition lines (6HS and 7H) using fluorescence in situ hybridization and the stability of the whole ‘Martonvásári 9 kr1’ – ‘Igri’ addition set

Genome ◽

10.1139/g09-085 ◽

2010 ◽

Vol 53 (1) ◽

pp. 35-44 ◽

Cited By ~ 20

Author(s):

É. Szakács ◽

M. Molnár-Láng

Keyword(s):

In Situ Hybridization ◽

Winter Wheat ◽

Fluorescence In Situ Hybridization ◽

Morphological Characterization ◽

Winter Barley ◽

Addition Lines ◽

Disomic Addition Lines ◽

The Stability

A previous paper reported the development of disomic addition lines (2H, 3H, 4H, and 1HS isochromosomic) from hybrids between the winter wheat ‘Martonvásári 9 kr1’ and the two-rowed winter barley cultivar ‘Igri’. The present paper describes the isolation of two new additions, the 7H disomic and 6HS ditelosomic additions, using fluorescence in situ hybridization with the repetitive DNA probes Afa-family and HvT01. The identification of the barley chromosomes in the wheat genome was confirmed with simple sequence repeat markers. The morphological characterization of the new addition lines is also discussed. Studies of the genetic stability of the whole set (2H, 3H, 4H, 7H, 1HS iso, 6HS) of ‘Martonvásári 9 kr1’ – ‘Igri’ additions revealed that the most stable disomic additions are 2H and 3H and the most unstable line is the 1HS isochromosomic addition.

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Molecular cytogenetic characterization ofAegilops biuncialisand its use for the identification of 5 derived wheat – Aegilops biuncialisdisomic addition lines

Genome ◽

10.1139/g05-062 ◽

2005 ◽

Vol 48 (6) ◽

pp. 1070-1082 ◽

Cited By ~ 48

Author(s):

Annamária Schneider ◽

Gabriella Linc ◽

István Molnár ◽

Márta Molnár-Láng

Keyword(s):

Triticum Aestivum ◽

In Situ Hybridization ◽

Fluorescence In Situ Hybridization ◽

Genomic In Situ Hybridization ◽

Addition Lines ◽

Aegilops Umbellulata ◽

Aegilops Comosa ◽

Disomic Addition Lines ◽

M Genome

The aim of the experiments was to produce and identify different Triticum aestivum – Aegilops biuncialis disomic addition lines. To facilitate the exact identification of the Ae. biuncialis chromosomes in these Triticum aestivum – Ae. biuncialis disomic additions, it was necessary to analyze the fluorescence in situ hybridization (FISH) pattern of Ae. biuncialis (2n = 4x = 28, UbUbMbMb), comparing it with the diploid progenitors (Aegilops umbellulata, 2n = 2x = 14, UU and Aegilops comosa, 2n = 2x = 14, MM). To identify the Ae. biuncialis chromosomes, FISH was carried out using 2 DNA clones (pSc119.2 and pAs1) on Ae. biuncialis and its 2 diploid progenitor species. Differences in the hybridization patterns of all chromosomes were observed among the 4 Ae. umbellulata accessions, the 4 Ae. comosa accessions, and the 3 Ae. biuncialis accessions analyzed. The hybridization pattern of the M genome was more variable than that of the U genome. Five different wheat – Ae. biuncialis addition lines were produced from the wheat – Ae. biuncialis amphiploids produced earlier in Martonvásár. The 2M, 3M, 7M, 3U, and 5U chromosome pairs were identified with FISH using 3 repetitive DNA clones (pSc119.2, pAs1, and pTa71) in the disomic chromosome additions produced. Genomic in situ hybridization (GISH) was used to differentiate the Ae. biuncialis chromosomes from wheat, but no chromosome rearrangements between wheat and Ae. biuncialis were detected in the addition lines.Key words: Triticum aestivum, Aegilops biuncialis, fluorescence in situ hybridization, genomic in situ hybridization, wheat – Aegilops biuncialis addition lines.

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Characterization of an Intermediate Filament Protein from the Platyhelminth, Dugesia japonica

Protein and Peptide Letters ◽

10.2174/0929866526666191025102902 ◽

2020 ◽

Vol 27 (5) ◽

pp. 432-446

Author(s):

Akiko Yamamoto ◽

Ken-ichiro Matsunaga ◽

Toyoaki Anai ◽

Hitoshi Kawano ◽

Toshihisa Ueda ◽

...

Keyword(s):

In Situ Hybridization ◽

Immunohistochemical Analysis ◽

Protein Characterization ◽

Intermediate Filament Protein ◽

Tail Domain ◽

Animal Cells ◽

Dugesia Japonica ◽

Function Relationship

Background: Intermediate Filaments (IFs) are major constituents of the cytoskeletal systems in animal cells. Objective: To gain insights into the structure-function relationship of invertebrate cytoplasmic IF proteins, we characterized an IF protein from the platyhelminth, Dugesia japonica, termed Dif-1. Method: cDNA cloning, in situ hybridization, immunohistochemical analysis, and IF assembly experiments in vitro using recombinant Dif-1, were performed for protein characterization. Results: The structure deduced from the cDNA sequence showed that Djf-1 comprises 568 amino acids and has a tripartite domain structure (N-terminal head, central rod, and C-terminal tail) that is characteristic of IF proteins. Similar to nuclear IF lamins, Djf-1 contains an extra 42 residues in the coil 1b subdomain of the rod domain that is absent from vertebrate cytoplasmic IF proteins and a nuclear lamin-homology segment of approximately 105 residues in the tail domain; however, it contains no nuclear localization signal. In situ hybridization analysis showed that Djf-1 mRNA is specifically expressed in cells located within the marginal region encircling the worm body. Immunohistochemical analysis showed that Djf-1 protein forms cytoplasmic IFs located close to the microvilli of the cells. In vitro IF assembly experiments using recombinant proteins showed that Djf-1 alone polymerizes into IFs. Deletion of the extra 42 residues in the coil 1b subdomain resulted in the failure of IF formation. Conclusions: Together with data from other histological studies, our results suggest that Djf- 1 is expressed specifically in anchor cells within the glandular adhesive organs of the worm and that Djf-1 IFs may play a role in protecting the cells from mechanical stress.

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Combination of Direct Viable Count and Fluorescent In Situ Hybridization (DVC-FISH) as a Potential Method for Identifying Viable Vibrio parahaemolyticus in Oysters and Mussels

Foods ◽

10.3390/foods10071502 ◽

2021 ◽

Vol 10 (7) ◽

pp. 1502

Author(s):

Jorge García-Hernández ◽

Manuel Hernández ◽

Yolanda Moreno

Keyword(s):

In Situ Hybridization ◽

Vibrio Parahaemolyticus ◽

Fluorescent In Situ Hybridization ◽

Potential Method ◽

Viable Count ◽

Hybridization Technique ◽

Fish Technique ◽

Viable Cells

Vibrio parahaemolyticus is a human food-borne pathogen with the ability to enter the food chain. It is able to acquire a viable, non-cultivable state (VBNC), which is not detected by traditional methods. The combination of the direct viable count method and a fluorescent in situ hybridization technique (DVC-FISH) makes it possible to detect microorganisms that can present VBNC forms in complex samples The optimization of the in vitro DVC-FISH technique for V. parahaemolyticus was carried out. The selected antibiotic was ciprofloxacin at a concentration of 0.75 μg/mL with an incubation time in DVC broth of 5 h. The DVC-FISH technique and the traditional plate culture were applied to detect and quantify the viable cells of the affected pathogen in artificially contaminated food matrices at different temperatures. The results obtained showed that low temperatures produced an important logarithmic decrease of V. parahaemolyticus, while at 22 °C, it proliferated rapidly. The DVC-FISH technique proved to be a useful tool for the detection and quantification of V. parahaemolyticus in the two seafood matrices of oysters and mussels. This is the first study in which this technique has been developed to detect viable cells for this microorganism.

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Human neutrophilic and eosinophilic granulocytes display different levels of c-fos proto-oncogene expression: an in situ hybridization study.

Journal of Histochemistry & Cytochemistry ◽

10.1177/35.8.3298425 ◽

1987 ◽

Vol 35 (8) ◽

pp. 837-842 ◽

Cited By ~ 9

Author(s):

H Kreipe ◽

H J Radzun ◽

K Heidorn ◽

C Mäder ◽

M R Parwaresch

Keyword(s):

In Situ Hybridization ◽

Neutrophilic Granulocytes ◽

Blood Monocytes ◽

In Vitro Differentiation ◽

Monocytic Differentiation ◽

Eosinophilic Granulocytes ◽

Fos Expression ◽

High Level

The cellular homologue of the retroviral oncogene v-fos has been shown to be involved in cell differentiation of hematopoietic cells. By use of the human promyelocyte cell line HL-60, several in vitro differentiation studies suggested a selective activation of c-fos during monocytic differentiation of myeloid precursor cells. In contrast to these observations, we found high levels of c-fos mRNA in purified normal human granulocytes, whereas c-fos was only faintly expressed in blood monocytes. In situ hybridization revealed that the high level of c-fos expression is restricted to neutrophilic granulocytes, whereas c-fos transcription is not detectable in eosinophilic granulocytes. These results indicate that in vitro differentiation systems can be misleading and may not reflect the in vivo situation. The high level of c-fos expression in neutrophilic granulocytes may be caused by superinduction due to the reduced capacity for protein synthesis in these cells.

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Use of cross-species in-situ hybridization (ZOO-FISH) to assess chromosome abnormalities in day-6 in-vivo- or in-vitro-produced sheep embryos

Chromosome Research ◽

10.1007/s10577-007-1125-2 ◽

2007 ◽

Vol 15 (3) ◽

pp. 399-408 ◽

Cited By ~ 11

Author(s):

Gianfranco Coppola ◽

Basil Alexander ◽

Dino Di Berardino ◽

Elizabeth St John ◽

Parvathi K. Basrur ◽

...

Keyword(s):

In Situ Hybridization ◽

Chromosome Abnormalities

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Human Immunodeficiency Virus Infection of Early Passage Cervical Epithelial Cultures

International Journal of STD & AIDS ◽

10.1177/095646249300400608 ◽

1993 ◽

Vol 4 (6) ◽

pp. 342-345 ◽

Cited By ~ 5

Author(s):

S L Patrick ◽

T C Wright ◽

H E Fox ◽

H S Ginsberg

Keyword(s):

In Situ Hybridization ◽

Epithelial Cells ◽

Female Genital Tract ◽

Virus Production ◽

Heterosexual Transmission ◽

Early Passage ◽

Epithelial Cultures ◽

Cervical Epithelial Cells

Women are infected with HIV in increasing numbers; the predominant mode of spread is through heterosexual transmission. Little is known regarding the mechanism of HIV transit through the female genital tract. We investigated whether early passaage cervical epithelial cells could be directly infected with HIV-1LAI*. Virus production was measured using the reverse transcriptase (RT) assay and direct assay for syncytia-forming units. In-situ hybridization was performed on infected cervical cell cultures. Immunostaining was carried out using a monoclonal antibody to leukocyte common antigen (LCA). Virus was recovered in the supernatants of all infected cervical cultures. Localization of HIV infection using in-situ hybridization identified rare cells in the population which gave a strong signal. These infected cells had a lymphoid morphology and were also detected using immunostaining for LAC. Cervical epithelial cells were uninfected in this in vitro model; cells in this population which supported viral replication were most likely of the macrophage/monocyte lineage.

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Interleukin-6 and its receptor are expressed by human megakaryocytes: in vitro effects on proliferation and endoreplication

Blood ◽

10.1182/blood.v77.3.461.461 ◽

1991 ◽

Vol 77 (3) ◽

pp. 461-471 ◽

Cited By ~ 72

Author(s):

S Navarro ◽

N Debili ◽

JP Le Couedic ◽

B Klein ◽

J Breton-Gorius ◽

...

Keyword(s):

In Situ Hybridization ◽

Interleukin 6 ◽

Dot Blot ◽

Autocrine Loop ◽

Megakaryocytic Differentiation ◽

Normal Human ◽

In Vitro Effects ◽

A Minor

Abstract Interleukin-6 (IL-6) is a pleiotropic cytokine that plays an important role in the megakaryocytic differentiation. Recently, we have observed that IL-6 is synthesized by several human cell lines with megakaryocytic features. In this study, we have investigated whether a similar phenomenon occurs during normal megakaryocytic differentiation. Human megakaryocytes (MK) were obtained by culturing normal marrow in liquid culture with aplastic plasma (AP). First, an IL-6 secretion in bone marrow culture enriched in MK as well as in purified MK populations was demonstrated by a biologic assay. Second, IL-6 mRNA was detected in a purified population of MK by the polymerase chain reaction and dot blot analysis. IL-6 mRNA and protein were undetectable in platelets. Third, in situ hybridization procedure demonstrated the presence of IL-6 mRNA in individual immature MK. Fourth, IL-6 protein was detected in MK at the unicellular level by an immunoalkaline phosphatase technique using a monoclonal antibody against IL-6. Furthermore, the presence of IL-6 receptor (IL-6-R) on MK was demonstrated by in situ hybridization using an IL-6-R probe and in situ autoradiography after binding with [125I]-labeled recombinant IL-6. The IL-6 endogenously produced in liquid cultures containing normal human plasma or AP was subsequently neutralized. This resulted in a 50% decrease of the MK growth with a minor shift in the ploidy distribution toward lower values. In semisolid cultures the addition of anti-IL-6 antibodies led to a 42% decrease in colony number in cultures stimulated by IL-3 but not in other conditions of culture. These results suggest that normal human megakaryocytopoiesis might be regulated in part by an IL-6 autocrine loop.

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