Direct and sensitive fluorescence in situ hybridization of 45S rDNA on tomato chromosomes

Genome ◽  
1994 ◽  
Vol 37 (6) ◽  
pp. 1062-1065 ◽  
Author(s):  
Jie Xu ◽  
E. D. Earle
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Chromosome 2 ◽  
45S Rdna ◽  
Plant Chromosomes ◽  
Signal Sensitivity ◽  
Direct Fluorescence ◽  
Hybridization Protocol ◽  
Tomato Genotypes

We describe a direct and sensitive fluorescence in situ hybridization protocol for plant chromosomes. We labelled 45S rDNA with fluorescein-12-dUTP and hybridized to somatic chromosomes of four tomato genotypes. This technique does not require posthybridization immunocytochemical amplifications. The improved signal sensitivity with this technique allowed identification of new rDNA loci on three pairs of chromosomes, in addition to the previously known locus on chromosome 2. We discuss favorable features of direct fluorescence in situ hybridization for chromosomes fixed on a slide and chromosomes or cells in suspension.Key words: direct fluorescence in situ hybridization, 45S rDNA, tomato chromosomes.

scholarly journals A populational survey of 45S rDNA polymorphism in the Jefferson salamander Ambystoma jeffersonianum revealed by fluorescence in situ hybridization (FISH)

2009 ◽  
Vol 55 (2) ◽  
pp. 145-149 ◽  
Author(s):  
Ke Bi ◽  
James P. Bogart ◽  
Jinzhong Fu
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Dispersal Ability ◽  
Habitat Isolation ◽  
45S Rdna ◽  
Isolated Populations ◽  
Ambystoma Jeffersonianum ◽  
Rdna Sites ◽  
Rdna Polymorphism

Abstract The chromosomal localization of 45S ribosomal RNA genes in Ambystoma jeffersonianum was determined by fluorescence in situ hybridization with 18S rDNA fragment as a probe (FISH-rDNA). Our results revealed the presence of rDNA polymorphism among A. jeffersonianum populations in terms of number, location and FISH signal intensity on the chromosomes. Nine rDNA cytotypes were found in ten geographically isolated populations and most of them contained derivative rDNA sites. Our preliminary study provides strong indication of karyotypic diversification of A. Jeffersonianum that is demonstrated by intraspecific variation of 45S rDNA cytotypes. rDNA cytotype polymorphism has been described in many other caudate amphibians. We predict that habitat isolation, low dispersal ability and decline of effective population size could facilitate the fixation and accumulation of variable rDNA cytotypes during their chromosome evolution.

scholarly journals Catalyzed Reported Deposition-Fluorescence In Situ Hybridization Protocol To Evaluate Phagotrophy in Mixotrophic Protists

2005 ◽  
Vol 71 (11) ◽  
pp. 7321-7326 ◽  
Author(s):  
Juan M. Medina-Sánchez ◽  
Marisol Felip ◽  
Emilio O. Casamayor
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Signal Amplification ◽  
Cell Attachment ◽  
Cell Loss ◽  
Alexa Fluor ◽  
Hybridization Protocol ◽  
Card Fish ◽  
First Time

ABSTRACT We describe a catalyzed reported deposition-fluorescence in situ hybridization (CARD-FISH) protocol particularly suited to assess the phagotrophy of mixotrophic protists on prokaryotes, since it maintains cell and plastid integrity, avoids cell loss and egestion of prey, and allows visualization of labeled prey against plastid autofluorescence. This protocol, which includes steps such as Lugol's-formaldehyde-thiosulfate fixation, agarose cell attachment, cell wall permeabilization with lysozyme plus achromopeptidase, and signal amplification with Alexa-Fluor 488, allowed us to detect almost 100% of planktonic prokaryotes (Bacteria and Archaea) and, for the first time, to show archaeal cells ingested by mixotrophic protists.

Higher axial-resolution and sensitivity pachytene fluorescence in situ hybridization protocol in tetraploid cotton

2009 ◽  
Vol 17 (8) ◽  
pp. 1041-1050 ◽  
Author(s):  
Kai Wang ◽  
Zaijie Yang ◽  
Changshen Shu ◽  
Jing Hu ◽  
Qiuyun Lin ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Tetraploid Cotton ◽  
Axial Resolution ◽  
Hybridization Protocol

Assignment1 of maltase glucoamylase (MGAM) to pig chromosome 2 (2q21) by fluorescence in situ hybridization and confirmation by genetic mapping

2000 ◽  
Vol 90 (3-4) ◽  
pp. 236-237 ◽  
Author(s):  
J.H. Calvo ◽  
N.L. Lopez-Corrales ◽  
R. Osta ◽  
T.M. Skinner ◽  
S.I. Anderson ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Genetic Mapping ◽  
Fluorescence In Situ Hybridization ◽  
Chromosome 2

Distribution of 45S rDNA in Modern Rose Cultivars (Rosa hybrida), Rosa rugosa, and Their Interspecific Hybrids Revealed by Fluorescence in situ Hybridization

2016 ◽  
Vol 149 (3) ◽  
pp. 226-235 ◽  
Author(s):  
Xiao-Liu Ding ◽  
Ting-Liang Xu ◽  
Jing Wang ◽  
Le Luo ◽  
Chao Yu ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Interspecific Hybrids ◽  
Evolutionary Dynamics ◽  
The Other ◽  
45S Rdna ◽  
Expected Number ◽  
Rdna Sites ◽  
Rdna Loci

To elucidate the evolutionary dynamics of the location and number of rDNA loci in the process of polyploidization in the genus Rosa, we examined 45S rDNA sites in the chromosomes of 6 modern rose cultivars (R. hybrida), 5 R. rugosa cultivars, and 20 hybrid progenies by fluorescence in situ hybridization. Variation in the number of rDNA sites in parents and their interspecific hybrids was detected. As expected, 4 rDNA sites were observed in the genomes of 4 modern rose cultivars, while 3 hybridization sites were observed in the 2 others. Two expected rDNA sites were found in 2 R. rugosa cultivars, while in the other 3 R. rugosa cultivars 4 sites were present. Among the 20 R. hybrida × R. rugosa offspring, 13 carried the expected number of rDNA sites, and 1 had 6 hybridization sites, which exceeded the expected number by far. The other 6 offspring had either 2 or 3 hybridization sites, which was less than expected. Differences in the number of rDNA loci were observed in interspecific offspring, indicating that rDNA loci exhibit instability after distant hybridization events. Abnormal chromosome pairing may be the main factor explaining the variation in rDNA sites during polyploidization.

scholarly journals Detection and Differentiation of Chlamydiae by Fluorescence In Situ Hybridization

2002 ◽  
Vol 68 (8) ◽  
pp. 4081-4089 ◽  
Author(s):  
Sven Poppert ◽  
Andreas Essig ◽  
Reinhard Marre ◽  
Michael Wagner ◽  
Matthias Horn
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Single Step ◽  
Detection Methods ◽  
The Novel ◽  
Antibody Staining ◽  
Uncultured Bacteria ◽  
Direct Fluorescence ◽  
Probe Set

ABSTRACT Chlamydiae are important pathogens of humans and animals but diagnosis of chlamydial infections is still hampered by inadequate detection methods. Fluorescence in situ hybridization (FISH) using rRNA-targeted oligonucleotide probes is widely used for the investigation of uncultured bacteria in complex microbial communities and has recently also been shown to be a valuable tool for the rapid detection of various bacterial pathogens in clinical specimens. Here we report on the development and evaluation of a hierarchic probe set for the specific detection and differentiation of chlamydiae, particularly C. pneumoniae, C. trachomatis, C. psittaci, and the recently described chlamydia-like bacteria comprising the novel genera Neochlamydia and Parachlamydia. The specificity of the nine newly developed probes was successfully demonstrated by in situ hybridization of experimentally infected amoebae and HeLa 229 cells, including HeLa 229 cells coinfected with C. pneumoniae and C. trachomatis. FISH reliably stained chlamydial inclusions as early as 12 h postinfection. The sensitivity of FISH was further confirmed by combination with direct fluorescence antibody staining. In contrast to previously established detection methods for chlamydiae, FISH was not susceptible to false-positive results and allows the detection of all recognized chlamydiae in one single step.

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