Genomic in situ hybridization in plants with small genomes is feasible and elucidates the chromosomal parentage in interspecific Arabidopsis hybrids

Genome ◽  
2004 ◽  
Vol 47 (5) ◽  
pp. 954-960 ◽  
Author(s):  
Hoda B.M Ali ◽  
Martin A Lysak ◽  
Ingo Schubert
Keyword(s):  
In Situ Hybridization ◽  
Genomic In Situ Hybridization ◽  
Closely Related Species ◽  
Meiotic Chromosomes ◽  
Hybrid Plants ◽  
Synthetic Hybrids ◽  
High Concentrations ◽  
Arabidopsis Suecica ◽  
Allotetraploid Species

Genomic in situ hybridization (GISH) is a useful tool to analyse natural polyploids, hybrid plants, and their backcross progenies as to their origin, genomic composition, and intergenomic rearrangements. However, in angiosperms with very small genomes (<0.6 pg/1 C), often only heterochromatic regions were found to be labeled. We have modified the GISH technique to label entire mitotic and meiotic chromosomes of Arabidopsis thaliana (2n = 10) and closely related species with very small genomes by using high concentrations of DNA (7.5–15 µg per probe per slide) or 5 µg of probe and long hybridization times (>60 h). According to our GISH data, Cardaminopsis carpatica (2n = 16) is most likely the diploid ancestor of the autotetraploid Arabidopsis arenosa (2n = 32). Furthermore, within the allotetraploid species Arabidopsis suecica (2n = 26), it was possible to elucidate the origin of chromosomes contributed by the parental species A. thaliana and A. arenosa for a specimen with 2n = 26 or a deviating chromosome number.Key words: genomic in situ hybridization (GISH), Arabidopsis, Brassicaceae, allopolyploids, synthetic hybrids.

Genomic in situ hybridization analysis of a trigenomic hybrid involving Solanum and Lycopersicon species

Genome ◽  
2001 ◽  
Vol 44 (2) ◽  
pp. 299-304 ◽  
Author(s):  
S N Haider Ali ◽  
Dirk Jan Huigen ◽  
M S Ramanna ◽  
Evert Jacobsen ◽  
Richard GF Visser
Keyword(s):  
In Situ Hybridization ◽  
Chromosome Doubling ◽  
Genomic In Situ Hybridization ◽  
Potato Genome ◽  
Meiotic Chromosomes ◽  
Lycopersicon Species ◽  
Bridge Species ◽  
Genomic Probes ◽  
Homoeologous Chromosomes

A 4x potato (+) tomato fusion hybrid (2n = 4x = 48) was successfully backcrossed with a diploid Lycopersicon pennellii (2n = 2x = 24). Genomic in situ hybridization (GISH) on somatic and meiotic chromosomes confirmed that the progenies were triploids (2n = 3x = 36) and possessed three different genomes: potato, tomato, and L. pennellii. Therefore, they have been called trigenomic hybrids. Total genomic probes of both Lycopersicon species were found to hybridize mutually, whereas the potato genome was clearly differentiated. During metaphase I, bivalents were formed predominantly between tomato and L. pennellii chromosomes and the univalents of potato chromosomes were most common. Trivalents in all cases included homoeologous chromosomes of potato, tomato, and L. pennellii. However, the triploids were totally sterile as determined from extensive crossing. On chromosome doubling of triploids by shoot regeneration from callus, hexaploids (2n = 6x = 72) were obtained. Despite exhibiting clear allohexaploid behaviour by forming 36 bivalents at meiosis, these were also completely sterile like their triploid counterparts. In spite of this drawback, the prospects of chromosome pairing between potato L. pennellii and Solanum genomes does open the possibilities for bringing the two genera close.Key words: trigenomic triploids, GISH, bridge species, potato (+) tomato fusion hybrids.

Molecular cytogenetic analysis of wheat–barley hybrids using genomic in situ hybridization and barley microsatellite markers

Genome ◽  
2003 ◽  
Vol 46 (2) ◽  
pp. 314-322 ◽  
Author(s):  
L Malysheva ◽  
T Sjakste ◽  
F Matzk ◽  
M Röder ◽  
M Ganal
Keyword(s):  
Molecular Markers ◽  
In Situ Hybridization ◽  
Microsatellite Markers ◽  
Genetic Material ◽  
Introgressive Hybridization ◽  
Intergeneric Hybrids ◽  
Genomic In Situ Hybridization ◽  
Genome Constitution ◽  
Hybrid Plants

In the present investigation, genomic in situ hybridization (GISH) and barley microsatellite markers were used to analyse the genome constitution of wheat–barley hybrids from two backcross generations (BC1 and BC2). Two BC1 plants carried 3 and 6 barley chromosomes, respectively, according to GISH data. Additional chromosomal fragments were detected using microsatellites. Five BC2 plants possessed complete barley chromosomes or chromosome segments and six BC2 plants did not preserve barley genetic material. Molecular markers revealed segments of the barley genome with the size of one marker only, which probably resulted from recombination between wheat and barley chromosomes. The screening of backcrossed populations from intergeneric hybrids could be effectively conducted using both genomic in situ hybridization and molecular microsatellite markers. GISH images presented a general overview of the genome constitution of the hybrid plants, while microsatellite analysis revealed the genetic identity of the alien chromosomes and chromosomal segments introgressed. These methods were complementary and provided comprehensive information about the genomic constitution of the plants produced.Key words: wheat–barley hybrids, introgressive hybridization, recombination, molecular markers, genomic in situ hybridization (GISH).

Pre-annealing of total genomic DNA probes for simultaneous genomic in situ hybridization

Genome ◽  
1995 ◽  
Vol 38 (4) ◽  
pp. 814-816 ◽  
Author(s):  
K. Anamthawat-Jónsson ◽  
S. M. Reader
Keyword(s):  
In Situ Hybridization ◽  
Dna Sequences ◽  
Genomic Dna ◽  
Dna Probes ◽  
Genomic In Situ Hybridization ◽  
Whole Process ◽  
Meiotic Chromosomes ◽  
Cereal Species ◽  
Working Day

We used pre-annealing of differently labelled total genomic DNA probes to perform simultaneous genomic in situ hybridization on mitotic and meiotic chromosomes of interspecific hybrids between plant species of the Tribe Triticeae. The species origin of chromosomes was demonstrated by a two-colour fluorescence after in situ hybridization with directly labelled probes incorporating fluorescein (visualized green) and rhodamine (visualized red). The pre-annealing blocked out common DNA sequences between the different genomes, hence increasing species specificity of the probes. The method is simple and rapid because the hybridization takes only about 2 h, including the pre-annealing step, and hence the whole process can be accomplished easily within a working day making it suitable for routine analysis of chromosomes and genomes.Key words: pre-annealing, genomic in situ hybridization, total genomic DNA probe, cereal species.

Genome organization of Brassica napus and Lesquerella fendleri and analysis of their somatic hybrids using genomic in situ hybridization

Genome ◽  
1998 ◽  
Vol 41 (5) ◽  
pp. 691-701 ◽  
Author(s):  
Marina Skarzhinskaya ◽  
Jan Fahleson ◽  
Kristina Glimelius ◽  
Armand Mouras
Keyword(s):  
In Situ Hybridization ◽  
Brassica Napus ◽  
Somatic Hybrids ◽  
Chromosome Translocation ◽  
Genomic In Situ Hybridization ◽  
Lesquerella Fendleri ◽  
Hybrid Plants ◽  
Transformed Root Cultures ◽  
Asymmetric Fusion

Construction of Brassica napus (2n = 38) and Lesquerella fendleri (2n = 12) karyotypes revealed that B. napus contains 10 pairs of metacentric, 5 pairs of submetacentric, and 4 pairs of subtelocentric chromosomes, while L. fendleri contains 6 pairs of metacentric chromosomes that are, on average, 1.5 times longer than those of B. napus. By analysing the karyotypes and performing genomic in situ hybridization (GISH), the chromosome complements of somatic hybrids produced between Brassica napus (+) Lesquerella fendleri were studied. Protoplasts fused with no pretreatment, symmetric fusions, resulted in hybrids containing L. fendleri chromosomes in numbers varying from 2 chromosomes to 2 chromosome complements. In the asymmetric fusion experiments, in which L. fendleri protoplasts were irradiated before fusion, plants with from 38 to more than 76 chromosomes were obtained. In the hybrids with 38 chromosomes, the presence of L. fendleri chromosomes was not detected by GISH analysis, even though L. fendleri DNA was revealed by Southern blotting. Intra- and inter-genomic recombinations were observed in hybrids from both symmetric and asymmetric fusions. The rearrangements were more prevalent in plants resulting from asymmetric fusions. Modifications of the phenotype and reduced fertility of the hybrid plants were correlated with increased dosages of the L. fendleri genome. At least one genome complement of L. fendleri was required to express its morphological features.Key words: Agrobacterium rhizogenes transformed root cultures, Brassica napus, Lesquerella fendleri, somatic hybrids, chromosome translocation.

Synthesis and cytological characterization of trigeneric hybrids of durum wheat with and without Ph1

Genome ◽  
2004 ◽  
Vol 47 (6) ◽  
pp. 1173-1181 ◽  
Author(s):  
Prem P Jauhar ◽  
M Doğramaci ◽  
T S Peterson
Keyword(s):  
In Situ Hybridization ◽  
Chromosome Pairing ◽  
Genetic Recombination ◽  
Genomic In Situ Hybridization ◽  
Homoeologous Pairing ◽  
Alien Gene Transfer ◽  
Chromosome 5B ◽  
Alien Gene ◽  
Trigeneric Hybrids

Wild grasses in the tribe Triticeae, some in the primary or secondary gene pool of wheat, are excellent reservoirs of genes for superior agronomic traits, including resistance to various diseases. Thus, the diploid wheatgrasses Thinopyrum bessarabicum (Savul. and Rayss) Á. Löve (2n = 2x = 14; JJ genome) and Lophopyrum elongatum (Host) Á. Löve (2n = 2x = 14; EE genome) are important sources of genes for disease resistance, e.g., Fusarium head blight resistance that may be transferred to wheat. By crossing fertile amphidiploids (2n = 4x = 28; JJEE) developed from F1 hybrids of the 2 diploid species with appropriate genetic stocks of durum wheat, we synthesized trigeneric hybrids (2n = 4x = 28; ABJE) incorporating both the J and E genomes of the grass species with the durum genomes A and B. Trigeneric hybrids with and without the homoeologous-pairing suppressor gene, Ph1, were produced. In the absence of Ph1, the chances of genetic recombination between chromosomes of the 2 useful grass genomes (JE) and those of the durum genomes (AB) would be enhanced. Meiotic chromosome pairing was studied using both conventional staining and fluorescent genomic in situ hybridization (fl-GISH). As expected, the Ph1-intergeneric hybrids showed low chromosome pairing (23.86% of the complement), whereas the trigenerics with ph1b (49.49%) and those with their chromosome 5B replaced by 5D (49.09%) showed much higher pairing. The absence of Ph1 allowed pairing and, hence, genetic recombination between homoeologous chromosomes. Fl-GISH analysis afforded an excellent tool for studying the specificity of chromosome pairing: wheat with grass, wheat with wheat, or grass with grass. In the trigeneric hybrids that lacked chromosome 5B, and hence lacked the Ph1 gene, the wheat–grass pairing was elevated, i.e., 2.6 chiasmata per cell, a welcome feature from the breeding standpoint. Using Langdon 5D(5B) disomic substitution for making trigeneric hybrids should promote homoeologous pairing between durum and grass chromosomes and hence accelerate alien gene transfer into the durum genomes.Key words: alien gene transfer, chiasma (xma) frequency, chromosome pairing, fluorescent genomic in situ hybridization (fl-GISH), homoeologous-pairing regulator, specificity of chromosome pairing, wheatgrass.

Genome constitutions ofRoegneriaalashanica,R.elytrigioides,R.magnicaespesandR.grandis(Poaceae: Triticeae) via genomic in-situ hybridization

2010 ◽  
Vol 28 (2) ◽  
pp. 206-211 ◽  
Author(s):  
Hai-Qing Yu ◽  
Chun Zhang ◽  
Chun-Bang Ding ◽  
Hai-Qin Zhang ◽  
Yong-Hong Zhou
Keyword(s):  
In Situ Hybridization ◽  
Genomic In Situ Hybridization

Genomic in situ hybridization in Brassica amphidiploids and interspecific hybrids

1997 ◽  
Vol 95 (8) ◽  
pp. 1320-1324 ◽  
Author(s):  
R. J. Snowdon ◽  
W. Köhler ◽  
W. Friedt ◽  
A. Köhler
Keyword(s):  
In Situ Hybridization ◽  
Interspecific Hybrids ◽  
Genomic In Situ Hybridization

Analysis of Oriental × Trumpet (OT) Lilium hybrids by genomic in situ hybridization based on ploidy level

2017 ◽  
pp. 253-258
Author(s):  
F. Ramzan ◽  
A. Younis ◽  
K.B. Lim ◽  
S.H. Bae ◽  
M.J. Kwon ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Ploidy Level ◽  
Genomic In Situ Hybridization

A novel genome of C and the first autotetraploid species in the Setaria genus identified by genomic in situ hybridization

2009 ◽  
Vol 56 (6) ◽  
pp. 843-850 ◽  
Author(s):  
Yongqiang Wang ◽  
Hui Zhi ◽  
Wei Li ◽  
Haiquan Li ◽  
Yongfang Wang ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Genomic In Situ Hybridization
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