• Title/Summary/Keyword: GISH

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Chromosome Analysis Using GISH and FISH of Interspecific Hybrids between Allium cepa L. and A. fistulosum L. (GISH와 FISH를 이용한 양파와 파간 종간교잡계통의 염색체 분석)

  • Kim, Cheol-Woo;Lee, Eul-Tai;Kim, Hwa-young;Choi, In-Hu;Bang, Jin-Ki;Koo, Dal-Hoe;Bang, Jae-Wook
    • Korean Journal of Breeding Science
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    • v.41 no.4
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    • pp.468-473
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    • 2009
  • Fluorescence in situ hybridization (FISH) and genomic in situ hybridization (GISH) were used for chromosome analysis of hybrids (2n=16) between onion (Allium cepa L., 2n=2X=16) and welsh onion (A. fistulosum L., 2n=2X=16). 5S rDNA, 45S rDNA, and tandemly repeated DNA (TSD) sequence were used as probes for FISH analysis. A. fistulosum specific DNA probe of telomeric repeats and A. fistulosum DNA were used for GISH analysis. In the analysis of meiotic chromosome GISH revealed that hybrids have 7 bivalants and 2 univalents chromosome and 2 univalents were derived from A. fistulosum chromosomes. In somatic chromosomes of hybrid each 8 chromosomes were derived from A. cepa and A. fistulosum, respectively. FISH signal of 45S rDNA probe in A. fistulosum was detected at secondary constriction of chromosomes, while FISH signal in A. cepa was observed in both secondary constriction and telomere of chromosomes. TDS signals in A. fistulosum chromosomes were detected at all subtelomeric of 8 chromosomes and also in 2 pericentromeric of the chromosomes, whereas TDS signals in A. cepa were observed only in subtelomeric in all chromosomes. The pattern of TDS signal in hybrid chromosomes was similar to those of A. fistulosum chromosomes.

Production and Molecular Cytogenetic Identification of Wheat-Barley Hybrids and Translocations

  • Lang, Marta-Molnar;Gabriella Linc;Jozsef Sutka
    • Journal of Plant Biotechnology
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    • v.1 no.1
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    • pp.8-12
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    • 1999
  • New winter wheat winter barley hybrids were produced (Mv9 kr1 Igri, Mv9 kr1 Osnova, Asakazekomugi Manas). The wheat-barley hybrids showed entire male sterility and were multiplied in tissue culture. Chromosome configurations were studied with GISH in meiosis in the Mv9 krl x Igri hybrid and in its progenies multiplied in vitro. Chromosome pairing between wheat and barley has been observed in some cells in the hybrids multiplied in vitro. Backcross plants with 43 and 44 chromosomes were selected with the aim of developing new wheat-barley addition lines. Wheat-barley translocations were demonstrated with GISH in backcross progenies originating from in vitro regenerated wheat (Triticum aestivum L. cv. Chinese Spring) x barley (Hordeum vulgare L. cv. Betzes) hybrids. Five different translocations were observed. Sequential N-banding and GISH analyses were performed to further identify the translocations. The N-banding pattern of the Robertsonian translocation suggests that this chromosome consists of the short arm of barley chromosome 4H translocated to the long arm of 2B of wheat. Plants with four different homozygous translocations were selected from the following BC2F3 generation.

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Production and Characterizations of Somatic Hybrids between Brassica campestris L. ssp pekinensis and Brassica of oleracea L. var capitata

  • Lian, Yu-Ji;Lim, Hak-Tae
    • Journal of Plant Biotechnology
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    • v.3 no.1
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    • pp.33-38
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    • 2001
  • Protoplasts isolated from inbred lines of Brassica oleracea L. var capitata (cabbage) and Brassica campestris L. ssp. pekinensis (Chinese cabbage) were fused by PEG-mediated method, and somatic hybrid cells were differentiated into plants. for the identification of somatic hybrid plants, ploidy level, plant morphology, and cytological analysis were performed. All of the regenerated plants derived from fused protoplasts were shown to be 2X-4X, or higher ploidy level, presumably due to somatic hybridization or chromosome doubling. The morphology of leaves, petioles, and flowers showed an intermediate phenotype between Chinese cabbage and cabbage. Chromosome numbers in these somatic hybrids ranged mostly from 33 to 38. According to Genomic in situ hybridization (GISH) pattern, signals from both fusion parents of B.campestris or B.oleracea were detected in different colors when chromosomes of putative somatic hybrids were observed.

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

  • Belandres, Hadassah Roa;Waminal, Nomar Espinosa;Hwang, Yoon-Jung;Park, Beom-Seok;Lee, Soo-Seong;Huh, Jin Hoe;Kim, Hyun Hee
    • Horticultural Science & Technology
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    • v.33 no.1
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    • pp.83-92
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    • 2015
  • 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 $5.01{\mu}m$. 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 a re discussed. The results p resented h erein will b e usef ul f or f urther g enomic s tudy o f xBrassicoraphanus lines and their improvement as promising new breeding varieties.

Multi-dimensional analyses of plant chromosomes and genomes.

  • Fukui, Kiichi;Ohmido, Nobuko;Wako, Toshiyuki
    • Proceedings of the Botanical Society of Korea Conference
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    • 1998.07a
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    • pp.61-70
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    • 1998
  • Genome and chromosome analyses in plants using fluorescence in situ hybridization (FISH) and immuno-staining (IMS) methods are reviewed by presenting the recent results obtained by the Chromosome Link, a group of chromosome and genome researchers. FISH is now effective to detect unique nucleotide sequences with 153 bp on the extended DNA fibers. Genomic in situ hybridization (GISH) also allows painting plant chromosomes of different genomes. GISH is quite effective to detect the genomic differentiation in the individual chromosomes within a nucleus. Three dimensional (3D) analyses are now available by confocal microscopy and a deconvolution system. These techniques are invaluable to visualize both the structural and functional dynamics within a nucleus. 3D-FISH revealed the spatial differentiation of different genomees within a nucleus. 3D-FISH also proved structural partition of centromeric and telomeric domains within a barely nucleus. The dynamic acetylation of histone H4 at the specific regions of a genome during a cell cycle is also analyzed using 3D-IMS. It is anticipated that these methods will provide us powerful tools to understand the structural and functional significance of plant chromosomes and genomes.

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Progeny Analysis of Hybrid Lilies Crossed Between Several Genotypes and Tetraploid Oriental-Asiatic Hybrids (체세포 염색체가 배가된 4배체 Oriental-Asiatic(OA) hybrid 유래 종간잡종 백합의 후대검정)

  • Chung, Mi-Young;Chung, Jae-Dong;Van Tuyl, Jaap M.;Lim, Ki-Byung
    • Korean Journal of Breeding Science
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    • v.42 no.1
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    • pp.100-108
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    • 2010
  • This study was carried out to confirm the chromosome constitution and homoeologous recombination of progenies derived from various cross combination using tetraploid OA interspecific hybrid originated from mitotic chromosome doubling. Based on the chromosome analysis of progenies crossed reciprocally, there were only triploid progenies when crossed with diploid Asiatics as male or female parent. While only tetraploid progenies were produced when crossed tetraploid Asiatics or tetraploid OA hybrid with tetraploid OA hybrid, respectively. However, two types of progenies, that is, diploid and triploid plants, were produced from cross combinations between diploid Oriental hybrid and tetraploid OA hybrid. From the GISH analysis of OA hybrid, it was confirmed that diploid $F_1$ OA hybrid was consisted of 24 chromosomes (12 Oriental and 12 Asiatics) showing authentic OA hybrid. On the other hand, it was notified that triploid plants (3x=36) were consisted of 24 Asiatics lily chromosomes and 12 Oriental lily chromosomes by analysis of backcross progenies derived from either $A{\times}OA$ or $OA{\times}A$ crosses. In cross between tetraploid OA and OA, all the progenies were tetraploid with equal number of chromosomes without any homoeologous recombination, i.e. each 24 chromosomes of Oriental and Asiatics. In 2x-4x ($O{\times}OA$) cross combination, some progenies had 2x=24 chromosomes originated from only Oriental hybrid, and other progenies had 3x=36 chromosomes derived from 24 chromosomes of Oriental hybrid and 12 chromosomes of Asiatic hybrid. Only tetraploid Asiatics chromosomes without any Oriental one were produced in all the progenies from 4x-4x ($AA{\times}OA$) cross combination.

Overcoming sterility by Caffeine and Temperature in Oriental-Asiatic Interspecific Lily Hybrid (백합 Oriental-Asiatic 종간잡종의 임성 회복을 위한 카페인과 변온처리)

  • Park, Song Kyoung;Park, In Sook;Kim, Chang Kil;Jee, Sun Ok;Lim, Ki Byung
    • FLOWER RESEARCH JOURNAL
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    • v.17 no.4
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    • pp.279-284
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    • 2009
  • In order to recover fertility from sterile interspecific OA-1 $F_1$ hybrid (Oriental hybrid 'Mero Star' ${\times}$ Asiatic hybrid 'Connecticut King'), various concentrations (0.1, 0.3 and 0.5%) of caffeine were injected directly into flower buds and then confirmed the viability of OA-1 $F_1$ hybrid at the flowering time. After the caffeine treatment, fertilized $F_1$ hybrids were crossed as female with Asiatic hybrid 'Lanzarote' as male. Five plantlets were obtained from seven embryos of 16 pollinated flowers at 0.3% treatment of caffeine while 0.5% treatment of caffeine obtained one plant let and 0.1 % treatment of caffeine plantlet did not produce at all. Thus 0.3% of caffeine treatment was considered as optimum concentration to produce subsequent progenies and the OA-l $F_1$ hybrid treated with caffeine produced 51% of putative 2n gametes. Pollen germination of OA-2 ('Romero Star' ${\times}$ 'Lady Rosa') and OA-3 ('Expression' ${\times}$ 'Lady Rosa') was not differ between temperature treatment alone and in combination with caffeine and temperature treatment. In the reciprocal crosses of OA-1 and Asiatic hybrid 'Lanzarote' or Oriental hybrid 'Sorbonne', A ('Lanzarote') ${\times}$ OA-1 or OA-1 ${\times}$ A crosses showed better results than O ('Sorbonne') ${\times}$ OA-1 or OA-1 ${\times}$ a crosses in plant obtaining. All progenies obtained from A ${\times}$ OA-1 or OA-1 ${\times}$ A crosses were confirmed as triploids by GISH analysis.

Prospect of plant molecular cytogenetics in the 21st century

  • Mukai, Yasuhiko
    • Proceedings of the Korean Society of Life Science Conference
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    • 2003.10a
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    • pp.14-27
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    • 2003
  • The genomes of Arabidopsis and rice have been fully sequenced. Genomic sequencing provides global information about genome structure and organization. A comprehensive research account of our recent studies conducted on genome painting, comparative genomics and genome fusion is provided in order to project the prospects of plant cytogenetic research in post-genomics era. Genome analysis by GISH using genome painting is demonstrated as an excellent means suitable for visualization of a whole genome, since total genomic DNA representing the overall molecular composition of the genome is used as a probe. FISH on extended DNA fibers has been developed for high-resolution FISH and has contributed to determining the copy number and order of genes. We have also mapped a number of genes involving starch synthesis on wheat chromosomes by FISH and compared the position of these genes on linkage map of rice. Macro synteny between wheat and rice can be observed by comparing the location of these genes in spite of the fact that the size of DNA per chromosome differs by 20 fold in two. Moreover, to approach our goal towards making bread and udon noodles from rice flour in future by incorporating bread making and the noodle qualifies in rice, we have been successful in introducing large genomic DNA fragments containing agronomically important genes of wheat into a rice by successive introduction of large insert BAC clones, there by expanding genetic variability in rice. We call this method genome fusion.

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