Detection of transgenes in three genetically modified rice lines by fluorescence in situ hybridization

Park, Hye Mi; Jeon, Eun Jin; Waminal, Nomar Espinosa; Shin, Kong Sik; Kweon, Soon Jong; Park, Beom-Seok; Suh, Seok Cheol; Kim, Hyun Hee

doi:10.1007/s13258-010-0064-z

Cited by 7 publications

(3 citation statements)

References 29 publications

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“…Chromosome spreads were prepared with a drop of 45% acetic acid on the macerated root tips. Probe labeling and FISH procedures followed the protocol of Park et al (2010). FISH signals were visualized using a Nikon Eclipse 80i fluorescent microscope and photographed with a Canon digital camera.…”

Section: Fluorescent In Situ Hybridization (Fish)mentioning

confidence: 99%

Sequence Variation and Comparison of the 5S rRNA Sequences in Allium Species and their Chromosomal Distribution in Four Allium Species

et al. 2011

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The gene structure and sequence diversity of 5S rRNA genes were analyzed in 13 Allium species. While the lengths and sequences of the coding gene segments were conserved, the spacers were highly variable and could be characterized as either short (213-404 bp) or long (384-486 bp) spacers. The short spacers were further classified into five subtypes (SS-I to SS-V) and the long spacers into four subtypes (LS-I to LS-IV). The short spacers were more conserved than were the long spacers. There was a sequence duplication of 85 bp in SS-III that distinguished it from SS-II. The coding sequences of the 5S rRNA genes started with CGG and ended with either CCC or TCC. Both long and short spacers started with TTTT at their 5′-ends. However, the long spacers ended with a 3′-TGA sequence, whereas the short spacers terminated with various sequences, such as TTA, ATA, or TGA. GC content ranged from 27 to 41% in whole repeats, and the GC content in the long spacers was lower than in the short spacers. The nucleotide diversity in the coding regions was lower than in the spacers, and the nucleotide diversity in the coding regions did not correlate with that of the spacers. FISH analysis confirmed that each Allium species has either short spacers or long spacers. Although chromosomal locations of the 5S rRNA genes in Allium wakegi confirmed the allodiploid nature of A. cepa and A. fistulosum, spacer sequences revealed the absence of SS-II in A. cepa and in A. wakegi. The current study demonstrated that the 5S rRNA genes diverged in early stages in Allium species differentiation except of the allodiploid A. wakegi.

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Section: Fluorescent In Situ Hybridization (Fish)mentioning

confidence: 99%

Sequence Variation and Comparison of the 5S rRNA Sequences in Allium Species and their Chromosomal Distribution in Four Allium Species

et al. 2011

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“…Breeding and development of the species has been expected, and the molecular cytogenetic research based on chromosomal constitution should precede genomic research to get information of the chromosomal organization of a genome. FISH technique has been useful molecular cytogenetic tool which can localize specific DNA sequences on chromosomes making it possible to visualize chromosomal landscape of a genome and construct cytogenetic physical maps (Park et al 2010). Tandem repeat sequences or multigene families can be used as FISH probes for chromosome characterization (Waminal et al 2011).…”

Section: Discussionmentioning

confidence: 99%

Chromosome Karyotyping of Senna covesii and S. floribunda based on Triple?color FISH Mapping of rDNAs and Telomeric Repeats

Youn

Kim

2018

Plant Breed. Biotech.

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The genus Senna in the family Fabaceae is distributed and cultivated around the world and has been used as sources of medicine and commercial goods. Molecular cytogenetic study based on FISH technique is useful for breeding and genomic research of the species. Triple-color FISH karyotype analyses were carried out to understand the genome structure of two Senna species, S. covesii (A. Gray) H. S. Irwin & Barneby and S. floribunda (Cav.) H. S. Irwin & Barneby using 5S rDNA, 45S rDNA, and telomeric repeat probes. Three and one pairs of 45S and 5S rDNA signals were detected, respectively, in S. covesii, whereas one pair each of 45S and 5S rDNA signals, were detected in S. floribunda. Telomeric signals appeared at all chromosome termini in both species. This result provides basic cytogenetic information which will be useful for future breeding and genomic research of Senna species.

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“…Chromosome characterization and genome identification have advanced significantly with the development of molecular cytogenetics through fluorescence in situ hybridization (FISH) and genomic in situ hybridization (GISH) techniques (Capdeville et al, 2008;Fukui, 2005;Hwang et al, 2010Hwang et al, , 2012Levsky and Singer, 2003;Park et al, 2010). Indeed, these methods have been used to investigate genome structure and inter-genomic relationship of hybrid plants (Jellen et al, 1994;Kenton et al, 1993), allopolyploid species (Cao, 2003;Devi et al, 2005;Yang et al, 1999), and recombinant breeding lines (Hwang et al, 2012;Lou et al, 2010;Vasconcelos et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

FISH Karyotype and GISH Meiotic Pairing Analyses of a Stable Intergeneric Hybrid xBrassicoraphanus Line BB#5

Belandres¹,

Waminal²,

Hwang³

et al. 2015

HST

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xBrassicoraphanus line BB#5, a new synthetic intergeneric hybrid between Brassica rapa L. ssp. pekinensis and Raphanus sativus L. var. rafiphera induced by N-methyl-N-nitroso-urethane mutagenesis in microspore culture, shows high seed fertility and morphological uniformity. Dual-color fluorescence in situ hybridization (FISH) using 5S and 45S rDNA probes and genomic in situ hybridization (GISH) using B. rapa genomic DNA probe were carried out to analyze the chromosome composition and the meiosis pairing pattern compared to its parental lines. The somatic chromosome complement is 2n = 38, which consists of 17 metacentric and two submetacentric chromosomes with lengths of 2.18 to 5.01 µm. FISH karyotype analysis showed five and eight pairs of 5S and 45S rDNA loci. GISH meiosis pairing analysis showed that 19 complete bivalents were most frequent and accounted for 42% of the 100 pollen mother cells examined. Based on chromosome number, size, morphology, rDNA distribution, and meiosis pairing pattern, both parental genomes of B. rapa and R. sativus appear to exist in xBrassicoraphanus line BB#5, demonstrating its genome integrity. Such stable chromosome constitutions and meiotic pairing patterns in somatic and meiotic cells are very rare in natural and synthetic intergeneric hybrids. Chromosomal studies and genetic and phenotypic changes in allopolyploids are discussed. The results presented herein will be useful for further genomic study of xBrassicoraphanus lines and their improvement as promising new breeding varieties.

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Detection of transgenes in three genetically modified rice lines by fluorescence in situ hybridization

Cited by 7 publications

References 29 publications

Sequence Variation and Comparison of the 5S rRNA Sequences in Allium Species and their Chromosomal Distribution in Four Allium Species

Sequence Variation and Comparison of the 5S rRNA Sequences in Allium Species and their Chromosomal Distribution in Four Allium Species

Chromosome Karyotyping of Senna covesii and S. floribunda based on Triple?color FISH Mapping of rDNAs and Telomeric Repeats

FISH Karyotype and GISH Meiotic Pairing Analyses of a Stable Intergeneric Hybrid xBrassicoraphanus Line BB#5

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