Specificity of PCR and in situ hybridization assays designed for detection of Marteilia sydneyi and M. refringens

Parasitology ◽  
2002 ◽  
Vol 125 (2) ◽  
pp. 131-141 ◽  
Author(s):  
S. N. KLEEMAN ◽  
F. LE ROUX ◽  
F. BERTHE ◽  
R. D. ADLARD
Keyword(s):  
Related Species ◽  
Pcr Primers ◽  
Dna Probe ◽  
Specific Detection ◽  
Closely Related Species ◽  
Tissue Sections ◽  
Saccostrea Glomerata ◽  
Polymerase Chain ◽  
Species Specific

Primers and DNA probes designed for use in the specific detection of the paramyxean parasites Marteilia sydneyi and Marteilia refringens were tested for their potential to cross-react with closely related species in Polymerase Chain Reaction (PCR) and in situ hybridization. PCR primers and a DNA probe designed within the ITS1 rRNA of M. sydneyi were specific for M. sydneyi when compared with related species of Marteilia and Marteilioides. PCR primers designed within the 18S rRNA of M. refringens were specific in the detection of this species in PCR while a DNA probe (named Smart 2) designed on the same gene cross-reacted with M. sydneyi in tissue sections of Saccostrea glomerata as well as Marteilioides sp. infecting Striostrea mytiloides. Though not species specific, the Smart 2 probe provided a stronger signal in detection of all stages of M. sydneyi than the ITS1 probe. The ITS probe is proposed for use as a confirmatory diagnostic tool for M. sydneyi.

scholarly journals A Comparative Study of 5S rDNA Non-Transcribed Spacers in Elaeagnaceae Species

Plants ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 4
Author(s):  
Oleg S. Alexandrov ◽  
Olga V. Razumova ◽  
Gennady I. Karlov
Keyword(s):  
Polymerase Chain Reaction ◽  
Molecular Markers ◽  
Dna Markers ◽  
5S Rdna ◽  
Chain Reaction ◽  
Closely Related Species ◽  
Pcr Products ◽  
Polymerase Chain ◽  
Species Specific ◽  
Shepherdia Canadensis

5S rDNA is organized as a cluster of tandemly repeated monomers that consist of the conservative 120 bp coding part and non-transcribed spacers (NTSs) with different lengths and sequences among different species. The polymorphism in the 5S rDNA NTSs of closely related species is interesting for phylogenetic and evolutional investigations, as well as for the development of molecular markers. In this study, the 5S rDNA NTSs were amplified with universal 5S1/5S2 primers in some species of the Elaeagnaceae Adans. family. The polymerase chain reaction (PCR) products of five Elaeagnus species had similar lengths near 310 bp and were different from Shepherdia canadensis (L.) Nutt. and Sh. argentea (Pusch.) Nutt. samples (260 bp and 215 bp, respectively). The PCR products were cloned and sequenced. An analysis of the sequences revealed that intraspecific levels of NTS identity are high (approximately 95–96%) and similar in the Elaeagnus L. species. In Sh. argentea, this level was slightly lower due to the differences in the poly-T region. Moreover, the intergeneric and intervarietal NTS identity levels were studied and compared. Significant differences between species (except E. multiflora Thunb. and E. umbellata Thunb.) and genera were found. Herein, a range of the NTS features is discussed. This study is another step in the investigation of the molecular evolution of Elaeagnaceae and may be useful for the development of species-specific DNA markers in this family.

scholarly journals Repeated evolution of asymmetric genitalia and right-sided mating behavior in theDrosophila nannopteraspecies group

2019 ◽  
Author(s):  
Andrea Acurio ◽  
Flor T. Rhebergen ◽  
Sarah Paulus ◽  
Virginie Courtier-Orgogozo ◽  
Michael Lang
Keyword(s):  
Related Species ◽  
Current Data ◽  
The Other ◽  
Genital Morphology ◽  
Closely Related Species ◽  
Outgroup Species ◽  
Repeated Evolution ◽  
Drosophila Pachea ◽  
Species Specific ◽  
Group Species

AbstractBackgroundMale genitals have repeatedly evolved left-right asymmetries, and the causes of such evolution remain unclear. TheDrosophila nannopteragroup contains four species, among which three exhibit left-right asymmetries of distinct genital organs. In the most studied species,Drosophila pachea, males display asymmetric genital lobes and they mate right-sided on top of the female. Copulation position of the other species is unknown.ResultsTo assess whether the evolution of genital asymmetry could be linked to the evolution of one-sided mating, we examined phallus morphology and copulation position inD. pacheaand closely related species. The phallus was found to be symmetric in all investigated species exceptD. pachea, which display an asymmetric phallus with a right-sided gonopore, andD. acanthoptera, which harbor an asymmetrically bent phallus. In all examined species, males were found to position themselves symmetrically on top of the female, except inD. pacheaandD. nannoptera, where males mated right-sided, in distinctive, species-specific positions. In addition, the copulation duration was found to be increased innannopteragroup species compared to closely related outgroup species.ConclusionOur study shows that gains, and possibly losses, of asymmetry in genital morphology and mating position have evolved repeatedly in thenannopteragroup. Current data does not allow us to conclude whether genital asymmetry has evolved in response to changes in mating position, or vice versa.

Karyotype Characterisation of Two Australian Dragon Lizards (Squamata: Agamidae: Amphibolurinae) Reveals Subtle Chromosomal Rearrangements Between Related Species with Similar Karyotypes

2020 ◽  
Vol 160 (10) ◽  
pp. 610-624
Author(s):  
Shayer M.I. Alam ◽  
Stephen D. Sarre ◽  
Arthur Georges ◽  
Tariq Ezaz
Keyword(s):  
Sex Chromosomes ◽  
Related Species ◽  
Chromosomal Rearrangements ◽  
In Situ Hybridisation ◽  
Fluorescence In Situ Hybridisation ◽  
Closely Related Species ◽  
Pogona Vitticeps ◽  
Molecular Cytogenetic Techniques ◽  
Heterogametic Sex

Agamid lizards (Squamata: Agamidae) are karyotypically heterogeneous. Among the 101 species currently described from Australia, all are from the subfamily Amphibolurinae. This group is, with some exceptions, karyotypically conserved, and all species involving heterogametic sex show female heterogamety. Here, we describe the chromosomes of 2 additional Australian agamid lizards, <i>Tympanocryptis lineata</i> and <i>Rankinia diemensis</i>. These species are phylogenetically and cytogenetically sisters to the well-characterised <i>Pogona vitticeps,</i> but their sex chromosomes and other chromosomal characteristics are unknown. In this study, we applied advanced molecular cytogenetic techniques, such as fluorescence in situ hybridisation (FISH) and cross-species gene mapping, to characterise chromosomes and to identify sex chromosomes in these species. Our data suggest that both species have a conserved karyotype with <i>P. vitticeps</i> but with subtle rearrangements in the chromosomal landscapes. We could identify that <i>T. lineata</i> possesses a female heterogametic system (ZZ/ZW) with a pair of sex microchromosomes, while <i>R. diemensis</i> may have heterogametic sex chromosomes, but this requires further investigations. Our study shows the pattern of chromosomal rearrangements between closely related species, explaining the speciation within Australian agamid lizards of similar karyotypes.

scholarly journals Intracellular amplification of proviral DNA in tissue sections using the polymerase chain reaction.

1992 ◽  
Vol 40 (3) ◽  
pp. 333-341 ◽  
Author(s):  
K P Chiu ◽  
S H Cohen ◽  
D W Morris ◽  
G W Jordan
Keyword(s):  
Polymerase Chain Reaction ◽  
Cultured Cells ◽  
Chain Reaction ◽  
Tissue Sections ◽  
Mouse Mammary Tumor ◽  
Tumor Virus ◽  
Polymerase Chain ◽  
Thermal Cycler ◽  
Cellular Dna

We developed a new method to amplify cell DNA in situ using the polymerase chain reaction (PCR). Proviral sequences of mouse mammary tumor virus (MMTV) contained in cultured cells and tissue sections were amplified intracellularly using a thermal cycler. Two techniques were employed to maintain the localization of the amplified DNA. First, complementary tails at the 5' ends of the oligonucleotide primers resulted in the synthesis of high molecular weight concatamers containing the target sequences. Second, the PCR was carried out in a thin film of agarose solidified over the tissue sections. The specifically amplified and localized DNA was then detected by in situ hybridization (ISH). Our results demonstrate that (a) DNA in tissue sections can serve as the target for the polymerase chain reaction in situ, (b) cell morphology is maintained, and (c) a target of 167 BP can be specifically detected in individual cells. This technique should be generally applicable to amplifying cellular DNA targets in tissue sections for detection in situ.

Altitudinal Variation in, and Morphological Divergence between, Three Related Species of Grasshopper, Praxibulus sp., Kosciuscola cognatus and K. usitatus (Orthoptera:Acrididae)

1980 ◽  
Vol 28 (1) ◽  
pp. 103 ◽  
Author(s):  
NA Campbell ◽  
JM Dearn
Keyword(s):  
Related Species ◽  
Character Displacement ◽  
Altitudinal Variation ◽  
Multivariate Statistical ◽  
Closely Related Species ◽  
Genetic Changes ◽  
Parallel Change ◽  
Variable Character ◽  
Multivariate Statistical Approach ◽  
Species Specific

Morphological variation between and within the closely related species Praxibuius sp.. Kosciuscola cognatus and K. usiratus has been examined along three independent altitudinal transects, by a multivariate statistical approach. The analyses, which were restricted to males. show that there is complete morphological separation between the three species. Moreover. there are species-specific patterns of character correlation which are consistent and relatively invariant within species, and do not exhibit altitudinal variation. The results suggest that there exist both distinct invariant species-specific character patterns and variable character patterns showing intraspecific variation. It is concluded that speciation in these grasshoppers could have involved genetic changes quite distinct from those involved in local intraspecific adaptation. Two further results are: first. evidence has been obtained for character displacement between Kosciuscola cognaius and Praxibulus sp. in an area of extensive sympatry: second. populations of K. cognatus along one transect, with a karyotype intermediate between typical K. cognatus and X usiiatus, show a parallel change in morphology towards that characteristic of K. usiiatus.

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