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Genomic affinity between Oryza sativa and Oryza brachyantha as revealed by in situ hybridization and chromosome pairing
FM Abbasi, AH Shah, F Perveen, M Afzal, M Sajid, R Masood, F Nawaz
African Journal of Biotechnology , 2010,
Abstract: Genomic affinity between Oryza sativa (2n = 24 AA), and Oryza brachyantha (2n = 24 FF) was assessed by using three strategies: genomic in situ hybridization (GISH), meiotic chromosome pairing, pollen and spikelet sterility. The chromosome pairing was examined in pollen mother cells of O. brachyantha, O.sativa and the hybrid between O. sativa and O. brachyantha. The hybrid was highly sterile with no pollen stain ability. Both parents showed regular meiosis with normal chromosome pairing. The F1 hybrid exhibited limited chromosome pairing. On an average, 0-2 bivalents and 20-24 univalents were recorded at metaphase-1 and 0 - 1 univalent at diakinesis. The most frequent configuration was two bivalents and twenty univalent. The meiosis was highly irregular showing unequal distribution of chromosomes at anaphase, formation of multipolar bodies and variation in the cell cycle of both genomes. GISH revealed unequivocal discrimination of O. brachyantha chromosomes as appeared red from O. sativa chromosomes that fluoresced yellow. No cross hybridization was examined between the labeled genomic DNA of O. brachyantha and the chromosomes of O. sativa. Mitotic chromosomes of O. brachyantha and O. sativa, in the hybrid, were discriminated by GISH. High sterility in this hybrid could be due to abnormal meiosis and lack of pairing.
An Improved Genomic In situ Hybridization (GISH) Method to Differentiate Chromosomes from Closely Related Genomes  [PDF]
N. Iqbal,S.M. Reader,P.D.S. Caligari,T.E. Miller
Pakistan Journal of Biological Sciences , 2002,
Abstract: Previously reported genomic in situ hybridization protocols failed to differentiate Aegilops uniaristata chromosomes in a polyploid wheat background. Different protocols and hybridization temperatures were tested to optimize the differentiation of N genome A. uniaristata chromosomes from those of the A, B and D genomes of wheat. A combination of pre-hybridization of cytological preparations, pre-annealing of the genomic probe with blocking DNA and hybridization at elevated temperatures proved successful. A genomic in situ hybridization protocol for the differentiation of chromosomes from very closely related genomes in a polyploid background is reported.
Localizing introgression on the chromosome of rice by genomic in situ hybridization (GISH)
FM Abbasi, SH Shah, A Gul, A Majid, M Afzal, R Mujaddad, AH Shah, R Masood, N Shafqat, I Bukhari, A Ali
African Journal of Biotechnology , 2010,
Abstract: Genomic in situ hybridization was used to detect introgressed segment from Oryza australinesis onto the chromosomes of introgression line derived from the hybrid O. sativa x O. australinesis. Genomic DNA from Oryza australinesis was labeled with biotin and hybridized to the homologous sequences on the O. sativa chromosomes. The probe hybridization fluoresced green and non labeled O. sativa chromosomes appeared red or blue due to counterstaining with propidium iodide (PI) or 4,6-diamidino-2-phenylindole (DAPI). This differential painting of chromosomes unequivocally detected the introgressed segment. Among the 200 cells analyzed, 6.5% of the cells showed hybridization signal. Signal appeared on one chromosome in 5%, on two homologous chromosomes in 1% and on sister chromatids in 0.5% of the cells. Hybridization was seen on the short arm of the chromosome 12 of the introgression line.
Development and characterization of interspecific hybrids between Oryza sativa and O. latifolia by in situ hybridization
ChuanDeng Yi,ShuZhu Tang,Yong Zhou,GuoHua Liang,ZhiYun Gong,MingHong Gu
Chinese Science Bulletin , 2008, DOI: 10.1007/s11434-008-0399-x
Abstract: Oryza sativa and O. latifolia belong to the AA and CCDD genomes of Oryza, respectively. In this study, interspecific hybrids of these species were obtained using the embryo rescue technique. Hybrid panicle traits, such as long awns, small grain, exoteric large purple stigma, grain shattering and dispersed panicles, resemble that of the paternal parent, O. latifolia, whereas there is obvious heterosis in such respects as plant height, tillering ability and vegetative vigor. Chromosome pairing and the genomic components of the hybrid were subsequently investigated using genomic in situ hybridization (GISH) and fluorescent in situ hybridization (FISH) analysis. Based on the mitotic metaphase chromosome numbers of the root tips investigated, the hybrid is a triploid with 36 chromosomes. The genomic constitution of the hybrid is ACD. In the meiotic metaphase I of the hybrid pollen mother cell, poor chromosome pairing was identified and most of the chromosomes were univalent, which resulted in complete male sterility in the hybrid.
The Selection of Secondary Tritipyrum Genotypes by Genomic In Situ Hybridization (GISH)  [cached]
S Falahati por,H SHahsavand hasani,A Baghizadeh,GH Karimzadeh
Journal of Science and Technology of Agriculture and Natural Resources , 2009,
Abstract: The genomic in situ hybridization (GISH) has been used to identify euploidy and aneuploidy in segregation generations of various plants. In this study, the GISH with minor modifications including, slide preparation of putative secondary Tritipyrum (F2) root meristemic cells, labeled genomic DNA of Thinopyrum bessarabicum by fluorescein 12-dUTP nucleotide as probe, genomic DNA of Thinopyrum bessarabicum for in situ hybridization on root meristemic cells of F2 (2n=6x=42, AABBDEb) and unlabeled Chinese Spring cultivar in pre hybridization, was carried out for the first time in Iran. The results not only indicated the various Eb chromosomes in putative 6x secondary Tritipyrum plants, but also showed different numbers of A, B and D chromosomes. The range of aneuoploidy in F2 genotypes was from %30 to %66.7, which could be due to various numbers of Eb and D chromosomes in each genotype. The selfing or back crossing of F2 plants with bread wheat varieties could lead to chromosomal stability and aneoploidy reduction in secondary Tritipyrum genotypes.
Comparative Analysis of Oryza latifolia Genome with C0t-1 DNA and Genomic DNA of Oryza sativa
利用栽培稻C_0t-1 DNA和基因组DNA比较分析宽叶野生稻基因组

WU Qi,QIN Rui,LI Gang,LIU Hong,
吴绮
,覃瑞,李刚,刘虹

植物科学学报 , 2010,
Abstract: C0t-1 DNA and genomic DNA(gDNA) of cultivated rice Oryza sativa were used as probes to analyze comparatively the Oryza latifolia genome by fluorescence in situ hybridization(FISH).Results showed that the hybridization signals of gDNA were more distinct on the chromosomes of O.latifolia than that of C0t-1 DNA.These hybridization signals were mostly distributed on centromeres and telomeres regions as well as those near centromeres under different stringencies.With increasing washing stringency,hybridization s...
Chromosomal imbalances detected in primary bone tumors by comparative genomic hybridization and interphase fluorescence in situ hybridization
Baruffi, Marcelo Razera;Engel, Edgard Edward;Squire, Jeremy Andrew;Tone, Luis Gonzaga;Rogatto, Silvia Regina;
Genetics and Molecular Biology , 2003, DOI: 10.1590/S1415-47572003000200001
Abstract: we applied a combination of comparative genomic hybridization (cgh) and fluorescence in situ hybridization (fish), to characterize the genetic aberrations in three osteosarcomas (os) and one ewing's sarcoma. cgh identified recurrent chromosomal losses at 10p14-pter and gains at 8q22.3-24.1 in os. interphase fish allowed to confirm 8q gain in two cases. a high amplification level of 11q12-qter was detected in one os. the ewing's sarcoma showed gain at 1p32-36.1 as the sole chromosome alteration. these studies demonstrate the value of molecular cytogenetic methods in the characterization of recurrent genomic alterations in bone tumor tissue.
Chromosomal imbalances detected in primary bone tumors by comparative genomic hybridization and interphase fluorescence in situ hybridization
Baruffi Marcelo Razera,Engel Edgard Edward,Squire Jeremy Andrew,Tone Luis Gonzaga
Genetics and Molecular Biology , 2003,
Abstract: We applied a combination of comparative genomic hybridization (CGH) and fluorescence in situ hybridization (FISH), to characterize the genetic aberrations in three osteosarcomas (OS) and one Ewing's sarcoma. CGH identified recurrent chromosomal losses at 10p14-pter and gains at 8q22.3-24.1 in OS. Interphase FISH allowed to confirm 8q gain in two cases. A high amplification level of 11q12-qter was detected in one OS. The Ewing's sarcoma showed gain at 1p32-36.1 as the sole chromosome alteration. These studies demonstrate the value of molecular cytogenetic methods in the characterization of recurrent genomic alterations in bone tumor tissue.
Assessment of genomic relationship between Oryza sativa and Oryza australinesis
FM Abbasi, H Ahmad, F Perveen, M Inamullah, M Sajid, DS Brar
African Journal of Biotechnology , 2010,
Abstract: The genomic relationship between Oryza sativa (2n = 24 AA) and Oryza australinesis (2n = 24 EE) has not been established. Genomic relationship between these two species was assessed by using three strategies: genomic in situ hybridization (GISH), meiotic chromosome pairing, pollen and spikelet sterility. The hybrid was produced between these two species at the International Rice Research Institute using embryo rescue technique. The chromosome pairing was examined in pollen mother cells of O. australinesis, O. sativa and the hybrid between O. sativa and O. australinesis. The hybrid was highly sterile with pollen stain ability being 0.05%. Both parents showed regular meiosis with normal chromosome pairing. The F1 hybrid exhibited limited chromosome pairing. On an average, 0 - 4 bivalents and 16 - 24 univalents were recorded at metaphase-1. The most frequent configuration was two bivalent and twenty univalent. The chromosomes of O. australiensis appeared larger and darkly stained. For genomic in situ hybridization, genomic DNA from O. australiensis was used as probe for the mitotic and meiotic chromosomes of the hybrid between O. sativa and O. australiensis. GISH revealed unequivocal discrimination of O. australiensis chromosomes that appeared yellow due to hybridization signal from O. sativa chromosomes that fluoresced red due to counterstaining with propidium iodide (PI). No cross hybridization was examined between the labeled genomic DNA of O. australiensis and the chromosomes of O. sativa. The paired chromosomes were discriminated as autosyndetic and allosyndetic pairing. Meiotic and mitotic chromosomes of the O. australiensis and O. sativa, in the hybrid were discriminated by GISH for the first time. Results showed that both genomes were highly divergent.
Comparative genome research between maize and rice using genomic in situ hybridization

NING Shunbin,JIN Weiwei,WANG Ling,DING Yi,SONG Yunchun,

科学通报(英文版) , 2001,
Abstract: Using the genomic DNAs of maize and rice as probes respectively, the homology of maize and rice genomes was assessed by genomic in situ hybridization. When rice genomic DNAs were hybridized to maize, all chromosomes displayed many multiple discrete regions, while each rice chromosome delineated a single consecutive chromosomal region after they were hybridized with maize genomic DNAs. The results indicate that the genomes of maize and rice share high homology, and confirm the proposal that maize and rice are diverged from a common ancestor.
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