Production and identification of wheat – Agropyron cristatum (1·4P) alien translocation lines

The P genome of Agropyron Gaertn., a wild relative of wheat, contains an abundance of desirable genes that can be utilized as genetic resources to improve wheat. In this study, wheat – Aegilops cylindrica Host gametocidal chromosome 2C addition lines were crossed with wheat – Agropyron cristatum (L.) Gaertn. disomic addition line accession II-21 with alien recombinant chromosome (1·4)P. We successfully induced wheat – A. cristatum alien chromosomal translocations for the first time. The frequency of translocation in the progeny was 3.75%, which was detected by molecular markers and genomic in situ hybridization (GISH). The translocation chromosomes were identified by dual-color GISH /fluorescence in situ hybridization (FISH). The P genomic DNA was used as probe to detect the (1·4)P chromosome fragment, and pHvG39, pAs1, or pSc119.2 repeated sequences were used as probes to identify wheat translocated chromosomes. The results showed that six types of translocations were identified in the three wheat – A. cristatum alien translocation lines, including the whole arm or terminal portion of a (1·4)P chromosome. The (1·4)P chromosome fragments were translocated to wheat chromosomes 1B, 2B, 5B, and 3D. The breakpoints were located at the centromeres of 1B and 2B, the pericentric locations of 5BS, and the terminals of 5BL and 3DS. In addition, we obtained 12 addition–deletion lines that contained alien A. cristatum chromosome (1·4)P in wheat background. All of these wheat – A. cristatum alien translocation lines and addition–deletion lines would be valuable for identifying A. cristatum chromosome (1·4)P-related genes and providing genetic resources and new germplasm accessions for the genetic improvement of wheat. The specific molecular markers of A. cristatum (1·4)P chromosome have been developed and used to track the (1·4)P chromatin.

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Development and Characterization of Wheat-Agropyron cristatum Introgression Lines Induced by Gametocidal Genes and Wheat ph1b Mutant

Agronomy ◽

10.3390/agronomy11020277 ◽

2021 ◽

Vol 11 (2) ◽

pp. 277

Author(s):

Alejandro Copete-Parada ◽

Carmen Palomino ◽

Adoración Cabrera

Keyword(s):

Genetic Resources ◽

Agropyron Cristatum ◽

Homologous Gene ◽

Aegilops Cylindrica ◽

Induction Frequency ◽

Structural Aberrations ◽

Translocation Lines ◽

New Germplasm

The P genome of Agropyron cristatum Gaertn. contains many desirable genes that can be utilized as genetic resources to improve wheat. In this research, we used both the gametocidal chromosome 2Cc and the pairing homologous gene (Ph1b) mutant to induce structural aberrations and translocations between wheat and the 4P, 5P, and 6P genome chromosomes. By using the two approaches, a total of 19 wheat-A. cristatum translocations have been identified, in which 13 were induced by the Triticum aestivum cv. Chinese Spring (CS) ph1b mutant (CS ph1b) and six were induced by gametocidal chromosome 2Cc from Aegilops cylindrica Host. The wheat-4P, -5P and -6P A. cristatum translocations were characterized by in situ hybridization and by a set of conserved orthologous set (COS) molecular markers. The aberrations included centromeric translocations, terminal translocations, dicentric translocations, and deletions. The average induction frequency of chromosome structural aberrations was 10.9% using gametocidal 2Cc chromosome and 8.8% using ph1b mutant. The highest frequency obtained was for chromosome 4P using both approaches. All the wheat-A. cristatum translocation lines obtained were valuable for identifying A. cristatum chromosome 4P, 5P, and 6P related genes. In addition, these lines provided genetic resources and new germplasm accessions for the genetic improvement of wheat.

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Recovery of a telomere-truncated chromosome via a compensating translocation in maize

Genome ◽

10.1139/g10-108 ◽

2011 ◽

Vol 54 (3) ◽

pp. 184-195 ◽

Cited By ~ 9

Author(s):

Robert T. Gaeta ◽

Tatiana V. Danilova ◽

Changzeng Zhao ◽

Rick E. Masonbrink ◽

Morgan E. McCaw ◽

...

Keyword(s):

In Situ Hybridization ◽

Fluorescent In Situ Hybridization ◽

Transgene Expression ◽

De Novo ◽

Extra Chromosome ◽

Single Gene ◽

B Chromosomes ◽

Chromosome 1 ◽

Chromosome 6

Maize-engineered minichromosomes are easily recovered from telomere-truncated B chromosomes but are rarely recovered from A chromosomes. B chromosomes lack known genes, and their truncation products are tolerated and transmitted during meiosis. In contrast, deficiency gametes resulting from truncated A chromosomes prevent their transmission. We report here a de novo compensating translocation that permitted recovery of a large truncation of chromosome 1 in maize. The truncation (trunc-1) and translocation with chromosome 6 (super-6) occurred during telomere-mediated truncation experiments and were characterized using single-gene fluorescent in situ hybridization (FISH) probes. The truncation contained a transgene signal near the end of the broken chromosome and transmitted together with the compensating translocation as a heterozygote to approximately 41%–55% of progeny. Transmission as an addition chromosome occurred in ~15% of progeny. Neither chromosome transmitted through pollen. Transgene expression (Bar) cosegregated with trunc-1 transcriptionally and phenotypically. Meiosis in T1 plants revealed eight bivalents and one tetravalent chain composed of chromosome 1, trunc-1, chromosome 6, and super-6 in diplotene and diakinesis. Our data suggest that de novo compensating translocations allow recovery of truncated A chromosomes by compensating deficiency in female gametes and by affecting chromosome pairing and segregation. The truncated chromosome can be maintained as an extra chromosome or together with the super-6 as a heterozygote.

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The genome of the olive fruit fly Bactrocera oleae: localization of molecular markers by in situ hybridization to the salivary gland polytene chromosomes

Genome ◽

10.1139/gen-42-4-744 ◽

1999 ◽

Vol 42 (4) ◽

pp. 744-751 ◽

Cited By ~ 13

Author(s):

Anna Zambetaki ◽

Antigone Zacharopoulou ◽

Zacharias G. Scouras ◽

Penelope Mavragani-Tsipidou

Keyword(s):

Salivary Gland ◽

Molecular Markers ◽

In Situ Hybridization ◽

Polytene Chromosomes ◽

Fruit Fly ◽

Bactrocera Oleae ◽

Olive Fruit Fly ◽

Olive Fruit

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The genome of the Mediterranean fruitflyceratitis capitata: Localization of molecular markers by in situ hybridization to salivary gland polytene chromosomes

Chromosoma ◽

10.1007/bf00582839 ◽

1992 ◽

Vol 101 (7) ◽

pp. 448-455 ◽

Cited By ~ 49

Author(s):

A. Zacharopoulou ◽

M. Frisardi ◽

C. Savakis ◽

A. S. Robinson ◽

P. Tolias ◽

...

Keyword(s):

Salivary Gland ◽

Molecular Markers ◽

In Situ Hybridization ◽

Polytene Chromosomes ◽

The Mediterranean

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High-resolution cytogenetic mapping of the short arm of chromosome 1 with newly isolated 411 cosmid markers by fluorescence in situ hybridization: the precise order of 18 markers on 1p36.1 on prophase chromosomes and “stretched” DNAs

Genomics ◽

10.1016/0888-7543(95)80116-4 ◽

1995 ◽

Vol 25 (1) ◽

pp. 114-123 ◽

Cited By ~ 19

Author(s):

Takeshi Ariyama ◽

Johji Inazawa ◽

Toshihiko Ezaki ◽

Yusuke Nakamura ◽

Akira Horii ◽

...

Keyword(s):

In Situ Hybridization ◽

High Resolution ◽

Fluorescence In Situ Hybridization ◽

Chromosome 1 ◽

Cytogenetic Mapping

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Fluorescence in situ hybridization in diluted and flow cytometrically sorted boar spermatozoa using specific DNA direct probes labelled by nick translation

Reproduction ◽

10.1530/rep.0.1260317 ◽

2003 ◽

pp. 317-325 ◽

Cited By ~ 16

Author(s):

I Parrilla ◽

JM Vazquez ◽

M Oliver-Bonet ◽

J Navarro ◽

J Yelamos ◽

...

Keyword(s):

In Situ Hybridization ◽

Fluorescence In Situ Hybridization ◽

Y Chromosome ◽

Internal Control ◽

Dna Probes ◽

Chromosome 1 ◽

Nick Translation ◽

Fluorescent Signal ◽

Sperm Separation

Successful evaluation of X- and Y-chromosome-bearing sperm separation technology using flow cytometry-cell sorter is of great importance. Fluorescence in situ hybridization (FISH), which allows for the detection of specific nucleic acid sequences on morphologically preserved spermatozoa, is an ideal method for quantitatively and qualitatively assessing the purity of sorted sperm samples. In this study specific pig DNA direct probes for small regions of chromosomes 1 and Y were used. Chromosome 1 was labelled in green and used as internal control to detect a lack of hybridization, whereas chromosome Y was labelled in red. Nick translation was used as the labelling method for the preparation of these probes. Spermatozoa, unsorted and sorted for high and low Y-chromosome purity from ejaculates of five boars, were fixed on slides and two-colour direct FISH was performed for chromosomes 1 and Y. About 500 non-sorted and 200 sorted spermatozoa per sample were scored. The proportion of Y-chromosome-bearing spermatozoa was determined by the presence of a red fluorescent signal on the sperm head and the proportion of X-chromosome-bearing spermatozoa was determined by subtraction. The efficiency of the hybridization procedure was established as near 98% on sorted and unsorted samples. The results of this study confirm that direct FISH using specific pig DNA probes labelled by nick translation provides a useful tool for laboratory validation of sperm separation by flow sorting technology. Moreover, the ease of nick translation and the quality of the fluorescent signal obtained using this method makes this procedure the most appropriate method for labelling pig DNA probes to be used for direct FISH on pig spermatozoa.

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Rearrangement of chromosome 1 is a frequent finding in endometrial carcinoma An in situ hybridization study in nine endometrial carcinomas

Cancer Genetics and Cytogenetics ◽

10.1016/0165-4608(94)00226-2 ◽

1995 ◽

Vol 81 (2) ◽

pp. 109-114 ◽

Cited By ~ 4

Author(s):

Ralf Ketter ◽

Carl-Ludwig von Ballestrem ◽

Stefan Lampel ◽

Gerhard Seitz ◽

Klaus D. Zang ◽

...

Keyword(s):

In Situ Hybridization ◽

Endometrial Carcinoma ◽

Chromosome 1 ◽

Hybridization Study ◽

Frequent Finding ◽

Endometrial Carcinomas

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An evaluation of a new series of fluorescent dUTPs for fluorescence in situ hybridization.

Journal of Histochemistry & Cytochemistry ◽

10.1177/44.5.8627009 ◽

1996 ◽

Vol 44 (5) ◽

pp. 525-529 ◽

Cited By ~ 12

Author(s):

J Wiegant ◽

N Verwoerd ◽

S Mascheretti ◽

M Bolk ◽

H J Tanke ◽

...

Keyword(s):

In Situ Hybridization ◽

Fluorescence In Situ Hybridization ◽

Human Lymphocytes ◽

Single Copy ◽

Cyanine Dye ◽

Chromosome 1 ◽

Nick Translation ◽

Qualitative Comparison ◽

Copy Dna

Synthesis of fluorochrome-modified deoxyribonucleotides has been carried out mostly by linking the fluorochrome molecule to the C-5 position of dUTP via an allylamine spacer, similar to the modification of allylamine-dUTP with the haptens biotin and digoxigenin. Recently, a new series of fluorescent nucleotides has been prepared by using an alkynyl bridge between the uracil moiety and the fluorochrome. Here we report the qualitative and quantitative analysis of fluorescence in situ hybridization results obtained on interphase cells and chromosomes with a variety of highly repetitive and single-copy DNA probes that were modified by nick translation with such alkynyl dUTPs. A qualitative comparison was made of the alkynyl dUTPs conjugated to the fluorochromes fluorescein, the cyanine dye Cy3, tetramethylrhodamine, Lissamine and Texas Red. With the exception of tetramethylrhodamine, all fluorochromes performed satisfactorily. The cyanine dye Cy3 provided the highest sensitivity, i.e., cosmid and YAC probes could easily be visualized by conventional fluorescence microscopy. In a quantitative assay, different nick translation conditions were tested using a human chromosome 1 satellite III probe (pUC1.77) and alkynyl dUTPs labeled with fluorescein and Cy3. Using these two nucleotides, FISH signal intensities on interphase nuclei from human lymphocytes were quantitated by digital imaging microscopy. The strongest signals were obtained when during nick translation the ratio between dTTP and fluorescein-dUTP or Cy3-dUTP was 1:5.

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The genome composition of hexaploid Psammopyrum athericum and octoploid Psammopyrum pungens (Poaceae: Triticeae)

Genome ◽

10.1139/g02-115 ◽

2003 ◽

Vol 46 (1) ◽

pp. 164-169 ◽

Cited By ~ 8

Author(s):

Pernilla Ellneskog-Staam ◽

Björn Salomon ◽

Roland von Bothmer ◽

Kesara Anamthawat-Jónsson

Keyword(s):

In Situ Hybridization ◽

Genome Analysis ◽

Genomic In Situ Hybridization ◽

Agropyron Cristatum ◽

Genome Composition ◽

Genomic Constitution ◽

Hybridization Pattern ◽

Genomic Probe

The genomic constitution of two species in the genus Psammopyrum, i.e., Ps. athericum (2n = 6x = 42) and Ps. pungens (2n = 8x = 56), was studied by genomic in situ hybridization (GISH). In Ps. athericum, one diploid chromosome set hybridized to a genomic probe from Pseudoroegneria ferganensis (St genome), one diploid set to a probe from Agropyron cristatum (P genome), and one diploid set to a probe from Thinopyrum junceiforme (EbEe genomes) or Th. bessarabicum (Eb genome). Substituting the St-genome probe with an L-genome probe from Festucopsis serpentinii resulted in exactly the same hybridization pattern, suggesting a genomic constitution of EStP or ELP for Ps. athericum. The same probes used on Ps. pungens showed two diploid sets of chromosomes hybridizing to the St-genome probe, one diploid set hybridizing to the P-genome probe, and one diploid set hybridizing to the EbEe-genome probe. The L-genome probe hybridized to approximately 14 of the chromosomes that were labeled by the St-genome probe. Hence the genomic constitution for Ps. pungens is proposed to be EStStP or EStLP.Key Words: Psammopyrum athericum, Psammopyrum pungens, in situ hybridization, Elytrigia pycnantha, Elytrigia pungens, genome analysis.

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