Exploring the origin of the D genome of oat by fluorescence in situ hybridization

Genome ◽  
2014 ◽  
Vol 57 (9) ◽  
pp. 469-472 ◽  
Author(s):  
Xiaomei Luo ◽  
Haiqin Zhang ◽  
Houyang Kang ◽  
Xing Fan ◽  
Yi Wang ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Avena Sativa ◽  
Genetic Improvement ◽  
Similar Distribution ◽  
Diploid Progenitor ◽  
D Genome ◽  
Genome Donor ◽  
Sequential Experiments

Further understanding of the origin of cultivated oat would accelerate its genetic improvement. In particular, it would be useful to clarify which diploid progenitor contributed the D genome of this allohexaploid species. In this study, we demonstrate that the landmarks produced by fluorescence in situ hybridization (FISH) of species of Avena using probes derived from Avena sativa can be used to explore the origin of the D genome. Selected sets of probes were hybridized in several sequential experiments performed on exactly the same chromosome spreads, with multiple probes of cytological preparations. Probes pITS and A3-19 showed there might be a similar distribution of pITS between the Ac and D genomes. These results indicated that the Ac genome is closely related to the D genome, and that Avena canariensis (AcAc) could be the D-genome donor of cultivated oat.

Fluorescence in situ hybridization mapping of Avena sativa L. cv. SunII and its monosomic lines using cloned repetitive DNA sequences

Genome ◽  
2002 ◽  
Vol 45 (6) ◽  
pp. 1230-1237 ◽  
Author(s):  
M L Irigoyen ◽  
C Linares ◽  
E Ferrer ◽  
A Fominaya
Keyword(s):  
In Situ Hybridization ◽  
Dna Sequences ◽  
Avena Sativa ◽  
Meiotic Chromosome ◽  
D Genome ◽  
Fish Mapping ◽  
Intergenomic Translocations ◽  
A Genome ◽  
C Genome

Fluorescent in situ hybridization (FISH) employing multiple probes was used with mitotic or meiotic chromosome spreads of Avena sativa L. cv. SunII and its monosomic lines to produce physical chromosome maps. The probes used were Avena strigosa pAs120a (which hybridizes exclusively to A-genome chromosomes), Avena murphyi pAm1 (which hybridizes exclusively to C-genome chromosomes), A. strigosa pAs121 (which hybridizes exclusively to A- and D-genome chromosomes), and the wheat rDNA probes pTa71 and pTa794. Simultaneous and sequential FISH employing two-by-two combinations of these probes allowed the unequivocal identification and genome assignation of all chromosomes. Ten pairs were found carrying intergenomic translocations: (i) between the A and C genomes (chromosome pair 5A); (ii) between the C and D genomes (pairs 1C, 2C, 4C, 10C, and 16C); and (iii) between the D and C genomes (pairs 9D, 11D, 13D, and 14D). The existence of a reciprocal intergenomic translocation (10C–14D) is also proposed. Comparing these results with those of other hexaploids, three intergenomic translocations (10C, 9D, and 14D) were found to be unique to A. sativa cv. SunII, supporting the view that 'SunII' is genetically distinct from other hexaploid Avena species and from cultivars of the A. sativa species. FISH mapping using meiotic and mitotic metaphases facilitated the genomic and chromosomal identification of the aneuploid chromosome in each monosomic line. Of the 18 analyzed, only 11 distinct monosomic lines were actually found, corresponding to 5 lines of the A genome, 2 lines of the C genome, and 4 lines of the D genome. The presence or absence of the 10C–14D interchange was also monitored in these lines.Key words: Avena sativa, monosomics, FISH mapping, genomic identification, intergenomic translocations.

A complex arrangement of genes at a starch branching enzyme I locus in the D-genome donor of wheat

Genome ◽  
1997 ◽  
Vol 40 (4) ◽  
pp. 465-474 ◽  
Author(s):  
S. Rahman ◽  
M. Morell ◽  
R. Appels ◽  
S. Abrahams ◽  
D. Abbott ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Branching Enzyme ◽  
Starch Branching Enzyme ◽  
D Genome ◽  
Triticum Tauschii ◽  
Putative Protein ◽  
Genome Donor ◽  
Opposite Strand ◽  
Enzyme I

Genomic DNA fragments from Triticum tauschii (D-genome donor to wheat) carrying starch branching enzyme I (SBE I) type genes have been characterized. One fragment contains one complete gene and two partial genes in 16 kb of DNA. One of the partial genes is oriented in the opposite strand to the other two. The gene that is complete was sequenced. Its structure corresponds closely to that of rice in that exons 3–8 are retained at similar sizes and spacings. A cDNA closely corresponding to the complete gene was isolated and characterized; it codes for a putative protein that represents a novel type of SBE I, as it is shorter at the 3′ end than the forms reported so far in other plants. A second genomic fragment contains a different SBE I gene. There appear to be approximately 10 copies of SBE I type genes in wheat (approximately 5 in T. tauschii) and most of them have been assigned to group 7 chromosomes. In situ hybridization indicates that a major locus for the genes is located at the distal end of the short arm of chromosome 7D.Key words: starch, branching enzyme genes, wheat, Triticum tauschii, in situ hybridization.

Repeated DNA sequences isolated by microdissection. I. Karyotyping of barley (Hordeum vulgare L.)

Genome ◽  
1995 ◽  
Vol 38 (6) ◽  
pp. 1082-1090 ◽  
Author(s):  
Winfried Busch ◽  
Regina Martin ◽  
Reinhold G. Herrmann ◽  
Uwe Hohmann
Keyword(s):  
In Situ Hybridization ◽  
Hordeum Vulgare ◽  
Fluorescence In Situ Hybridization ◽  
Dna Sequences ◽  
Fish Analysis ◽  
Hordeum Vulgare L ◽  
Agropyron Elongatum ◽  
D Genome ◽  
In Situ Experiments

We report on microdissection, cloning and sequence, and Southern and fluorescence in situ hybridization (FISH) analysis of one moderately and one highly amplified repetitive DNA element, pHvMWG2314 and pHvMWG2315, respectively, isolated from barley (Hordeum vulgare L.) chromosome arm 3HL. The pHvMWG2315 sequence hybridizes to all 14 telomeric or subtelomeric regions of the barley chromosomes as determined by FISH. The 50 different hybridization sites that include intercalary signals allow the discrimination of all 14 chromosome arms and the construction of a karyotype of barley. The tandemly repeated subtelomeric element of 331 bp exists in all Triticeae species tested (H. vulgare, Agropyron elongatum, Secale cereale, Triticum tauschii, T. turgidum, and T. aestivum). It is AT rich (66%), exhibits 84% sequence homology to subfragments of the D genome "specific" 1-kb element pAsl of T. tauschii and 75% homology to the interspersed genome-specific DNA sequence pHcKB6 from H. chilense. The repetitive sequence pHvMWG2314 is moderately amplified in barley and highly amplified in hexaploid wheat. The in situ experiments revealed no distinct signals on barley chromosomes, indicating a dispersed character for the sequence. The significance of the results for the identification of chromosomes and chromosome aberrations in FISH experiments are discussed.Key words: karyotype, fluorescence in situ hybridization, FISH, DNA sequencing.

RNAs radiate from gene to cytoplasm as revealed by fluorescence in situ hybridization

1995 ◽  
Vol 108 (7) ◽  
pp. 2565-2572 ◽  
Author(s):  
R.W. Dirks ◽  
K.C. Daniel ◽  
A.K. Raap
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Distribution Patterns ◽  
Transcription Unit ◽  
Similar Distribution ◽  
Small Nuclear Rna ◽  
Barr Virus ◽  
Nuclear Rna ◽  
Nuclear Signal

Genes for Epstein-Barr virus, human cytomegalovirus immediate early antigen and luciferase are abundantly transcribed in Namalwa, rat 9G and X1 cells, respectively. The EBV transcripts and HCMV-IE transcripts are extensively spliced, while in the luciferase transcript only a small intron sequence has to be spliced out. EBV transcripts are hardly localized in the cytoplasm while the luciferase and HCMV-IE transcripts are present in the cytoplasm and translated into proteins. We have correlated these characteristics with nuclear RNA distribution patterns as seen by fluorescence in situ hybridization. Transcripts of the HCMV-IE transcription unit were shown to be present in a main nuclear signal in the form of a track or elongated dot and as small nuclear RNA signals that radiate from this site towards the cytoplasm. A similar distribution pattern of small RNA signals was observed for transcripts of the luciferase gene, whereas the main nuclear signal was always observed as a dot and never as a track or elongated dot. In Namalwa cells, EBV transcripts were only present as track-like signals. The results suggest that when the extent for splicing is high, unspliced or partially spliced mRNAs begin to occupy elongated dot or track-like domains in the vicinity of the gene. When the extent of splicing is low, splicing is completed co-transcriptionally, leading to a bright dot-like signal. The presence of small nuclear spots in addition to the main signal correlates with cytoplasmic mRNA expression. The small spots most likely represent, therefore, mRNAs in transport to the cytoplasm.

Simultaneous painting of three genomes in hexaploid wheat by BAC-FISH

Genome ◽  
2004 ◽  
Vol 47 (5) ◽  
pp. 979-987 ◽  
Author(s):  
Peng Zhang ◽  
Wanlong Li ◽  
Bernd Friebe ◽  
Bikram S Gill
Keyword(s):  
Triticum Aestivum ◽  
In Situ Hybridization ◽  
Transposable Elements ◽  
Fluorescence In Situ Hybridization ◽  
Hexaploid Wheat ◽  
D Genome ◽  
Bac Clones ◽  
Dispersed Repeat ◽  
Bac Fish

Fluorescence in situ hybridization (FISH) is widely used in the physical mapping of genes and chromosome landmarks in plants and animals. Bacterial artificial chromosomes (BACs) contain large inserts, making them amenable for FISH mapping. In our BAC-FISH experiments, we selected 56 restriction fragment length polymorphism (RFLP)-locus-specific BAC clones from the libraries of Triticum monococcum and Aegilops tauschii, which are the A- and D-genome donors of wheat (Triticum aestivum, 2n = 6x = 42), respectively. The BAC clone 676D4 from the T. monococcum library contains a dispersed repeat that preferentially hybridizes to A-genome chromosomes, and two BAC clones, 9I10 and 9M13, from the Ae. tauschii library contain a dispersed repeat that preferentially hybridizes to the D-genome chromosomes. These repeats are useful in simultaneously discriminating the three different genomes in hexaploid wheat, and in identifying intergenomic translocations in wheat or between wheat and alien chromosomes. Sequencing results show that both of these repeats are transposable elements, indicating the importance of transposable elements, especially retrotransposons, in the genome evolution of wheat.Key words: bacterial artificial chromosome (BAC), fluorescence in situ hybridization (FISH), transposable elements (TEs), wheat, Triticum aestivum.

Simultaneous discrimination of the three genomes in hexaploid wheat by multicolor fluorescence in situ hybridization using total genomic and highly repeated DNA probes

Genome ◽  
1993 ◽  
Vol 36 (3) ◽  
pp. 489-494 ◽  
Author(s):  
Yasuhiko Mukai ◽  
Yumiko Nakahara ◽  
Maki Yamamoto
Keyword(s):  
Triticum Aestivum ◽  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Common Wheat ◽  
Chinese Spring ◽  
Genomic Dna ◽  
D Genome ◽  
B Genome ◽  
Multicolor Fluorescence

Common wheat, Triticum aestivum, is an allohexaploid species consisting of three different genomes (A, B, and D). The three genomes were simultaneously discriminated with different colors. Biotinylated total genomic DNA of the diploid A genome progenitor Triticum urartu, digoxigenin-labeled total genomic DNA of the diploid D genome progenitor Aegilops squarrosa, and nonlabeled total genomic DNA of one of the possible B genome progenitors Ae. speltoides were hybridized in situ to metaphase chromosome spreads of Triticum aestivum cv. Chinese Spring. For detection, only two fluorochromes, fluorescein and rhodamine, were used. The A, B, and D genomes were simultaneously detected by their yellow, brown, and orange fluorescence, respectively. The genomic fluorescence in situ hybridization pattern of chromosome 4A of cv. Chinese Spring wheat showed that the distal 32% of the long arm was derived from a B genome chromosome. Furthermore, by using two highly repeated sequence probes, pSc 119.2 and pAsl, and two fluorochromes simultaneously, we were able to identify all B and D genome chromosomes and chromosomes 1A, 4A, and 5A of wheat.Key words: common wheat, in situ hybridization, multicolor fluorescence.

Genetic diversity of Asian and European common wheat lines assessed by fluorescence in situ hybridization

Genome ◽  
2021 ◽  
Author(s):  
Xiu Yang ◽  
Binwen Tan ◽  
Yulu Yang ◽  
Xiaohui Zhang ◽  
Wei Zhu ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Common Wheat ◽  
Genetic Relationships ◽  
Structural Variations ◽  
D Genome ◽  
Western Asia ◽  
Wheat Lines

Understanding the genetic diversity of wheat is important for wheat breeding and improvement. However, there have been limited attempts to evaluate wheat diversity using fluorescence in situ hybridization (FISH). In this study, the chromosomal structures of 149 wheat accessions from 13 countries located between the latitudes of 30° and 45°N, the principal growing region for wheat, were characterized using FISH with pTa535 and pSc119.2 probes. The ranges of the numbers of FISH types in the A-, B-, and D-genomes were 2–8, 3–7, and 2–4, respectively, and the average numbers in the A- and B-genomes were greater than in the D-genome. Chromosomal translocations were detected by these probes, and previously undescribed translocations were also observed. Using the FISH, the genetic relationships among the 149 common wheat lines were divided into three groups (G1, G2, and G3). G1 mainly consisted of Southern European lines, G2 consisted of most lines from Japan and some lines from Western Asia, China, and Korea, and G3 consisted of the other lines from Southern Europe and most of the lines from Western Asia, China, and Korea. FISH karyotypes of wheat chromosomes distinguished chromosomal structural variations, revealed the genetic diversity among wheat varieties. Furthermore, these results provide valuable information for the further genetic improvement of wheat in China.

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