scholarly journals Formamide-Free Genomic in situ Hybridization Allows Unambiguous Discrimination of Highly Similar Parental Genomes in Diploid Hybrids and Allopolyploids

2015 ◽  
Vol 146 (4) ◽  
pp. 325-331 ◽  
Author(s):  
Tae-Soo Jang ◽  
Hanna Weiss-Schneeweiss
Keyword(s):  
In Situ Hybridization ◽  
Repetitive Dna ◽  
Species Complex ◽  
Genomic In Situ Hybridization ◽  
Genomic Evolution ◽  
Parental Chromosome ◽  
Unambiguous Discrimination ◽  
Modified Method ◽  
Prospero Autumnale

Polyploidy and hybridization play an important role in plant diversification and speciation. The application of genomic in situ hybridization (GISH) allows the identification of parental genomes in hybrids, thus elucidating their origins and allowing for analysis of their genomic evolution. The performance of GISH depends on the similarity of the parental genomes and on the age of hybrids. Here, we present the formamide-free GISH (ff-GISH) protocol applied to diploid and polyploid hybrids of monocots (Prospero, Hyacinthaceae) and dicots (Melampodium, Asteraceae) differing in similarity of the parental genomes and in chromosome and genome sizes. The efficiency of the new protocol is compared to the standard GISH protocol. As a result, ff-GISH allowed efficient labeling and discrimination of the parental chromosome sets in diploid and allopolyploid hybrids in Prospero autumnale species complex. In contrast, the standard GISH protocol failed to differentiate the parental genomes due to high levels of similar repetitive DNA. Likewise, an unambiguous identification of parental genomes in allotetraploid Melampodium nayaritense (Asteraceae) was possible after ff-GISH, whereas the standard GISH hybridization performance was suboptimal. The modified method is simple and non-toxic and allows the discrimination of very similar parental genomes in hybrids. This method lends itself to modifications and improvements and can also be used for FISH.

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.

Fluorescence in situ hybridization on plant extended chromatin DNA fibers for single-copy and repetitive DNA sequences

2011 ◽  
Vol 30 (9) ◽  
pp. 1779-1786 ◽  
Author(s):  
Kun Yang ◽  
Hecui Zhang ◽  
Richard Converse ◽  
Yong Wang ◽  
Xiaoying Rong ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Repetitive Dna ◽  
Dna Sequences ◽  
Single Copy ◽  
Repetitive Dna Sequences

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

A rapid procedure for the isolation of C0t-1 DNA from plants

Genome ◽  
1997 ◽  
Vol 40 (1) ◽  
pp. 138-142 ◽  
Author(s):  
Michael S. Zwick ◽  
Robert E. Hanson ◽  
M. Nurul Islam-Faridi ◽  
David M. Stelly ◽  
Rod A. Wing ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Repetitive Dna ◽  
Copy Number ◽  
Genomic Dna ◽  
Mammalian Species ◽  
Repetitive Dnas ◽  
Rapid Procedure ◽  
Dna Elements ◽  
Repetitive Dna Elements

In situ hybridization (ISH) for the detection of single- or low-copy sequences, particularly large DNA fragments cloned into YAC or BAC vectors, generally requires the suppression or "blocking" of highly-repetitive DNAs. C0t-1 DNA is enriched for repetitive DNA elements, high or moderate in copy number, and can therefore be used more effectively than total genomic DNA to prehybridize and competitively hybridize repetitive elements that would otherwise cause nonspecific hybridization. C0t-1 DNAs from several mammalian species are commercially available, however, none is currently available for plants to the best of our knowledge. We have developed a simple 1-day procedure to generate C0t-1 DNA without the use of specialized equipment.Key words: C0t-1 DNA, in situ hybridization, BACs, plants.

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.

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