scholarly journals DNA Stability Contrasts with Chromosome Variability in Allium Fistulosum Calli

2014 ◽  
Vol 56 (1) ◽  
pp. 66-72 ◽  
Author(s):  
Patryk Mizia ◽  
Dagmara Kwolek ◽  
Tomasz Ilnicki
Keyword(s):  
Genetic Similarity ◽  
Chromosomal Instability ◽  
Selection Pressure ◽  
Dna Polymorphism ◽  
Rapd Analysis ◽  
Polymorphic Band ◽  
Genetic Changes ◽  
Band Frequency ◽  
Callus Lines

Abstract RAPD analysis was applied to assess the degree of DNA polymorphism in A. fistulosum calli of high chromosomal instability. Nineteen of 24 randomly selected RAPD primers revealed scorable polymorphism between calli and seeds (reference material). Polymorphic band frequency was 55/237 in seeds and 36/233 in calli; variability on the DNA level was thus lower in calli than in seeds (15.4% vs. 23.2% of band positions). UPGMA analysis of Jaccard's coefficients confirmed the genetic similarity of the analyzed cultures. The most distinctive DNA changes in calli involved coincident loss of original bands or the appearance of novel bands. Seven such changes (4 losses, 3 gains) were observed. Our results suggest that changes on the chromosomal level and on the DNA level occurred independently of each other and that different callus lines underwent similar genetic changes during culture, presumably due to strong selection pressure effected by standard in vitro conditions.

scholarly journals Keragaman genetik Anggrek Grammatophyllum scriptum asal biji dari hasil Kultur In Vitro berdasarkan penanda RAPD

Cassowary ◽  
2021 ◽  
Vol 4 (1) ◽  
pp. 10-18
Author(s):  
Zarima Wibawati ◽  
Amelia Sarungallo ◽  
Barahima Abbas
Keyword(s):  
Tissue Culture ◽  
Molecular Markers ◽  
Genetic Distance ◽  
In Vitro Culture ◽  
Rapd Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Polymorphic Band ◽  
Genetic Character ◽  
Orchid Seed

Propagation through tissue culture by using orchid seed as explants will produce a lot of orchid plants. This study aims to measure the genetic character of orchid plantlets that were regenerated from seeds which have been resulted from in vitro culture. The genetic character of the original orchid plants produced from in vitro culture was determined using Random Amplified Polymorphic DNA (RAPD) molecular markers. The results showed that the primers used in the RAPD analysis showed a polymorphic band pattern of 14 DNA bands, with sizes between 500 bp - 8000 bp. The genetic distance of Grammatophyllum scriptum orchids that was regenerated from seeds is between 0.229 and 0.649.  The progenis produced from in vitro culture were clustered into seven groups at a dissimilarity coefficient of 45%.

scholarly journals INDUCING GENETIC VARIABILITY OF BLACK PEPPER (Piper nigrum L.) by IRRADIATION

2020 ◽  
Vol 10 (4) ◽  
pp. 166
Author(s):  
NURLIANI BERMAWIE
Keyword(s):  
Genetic Variation ◽  
Genetic Variability ◽  
Rapd Analysis ◽  
Black Pepper ◽  
Piper Nigrum ◽  
Genetic Changes ◽  
Band Size ◽  
Almost All ◽  
Dna Size

<p>Genetic variability of black pepper germplasm in Indonesia is low. To broaden genetic variability, newly growth shoot tips from in vitro culture of black pepper var. LDL were y irradiated with doses 0, 0.3 0.6, 0.9, 1.2 and 1.5 krad. The treatments were designed in a complete block with five replications. The irradiaed plantlets were grown on MS medium. Response of the variety is described by recording an increase in leaves, shoots and node, numbers, plantlet height, and morphological abnormality in the irst vegetative mutation generation (MVI) and the second vegetative mutation generation (MV2). Ater 6 weeks, the plantlets were sub cultured and the leaves of MV2 were used for RAPD analysis. Six random primers were used for the study, i.e. OPC-01 (TTCGAGC- CAG), OPC-02 (GTGAGGCGTC), OPC-04 (CCGCATCTAC), OPC-05 (GATGACCGCC), OPC-06 (GAACGGACTC) and Abi 117.17 (GCTC- GTCAAC). The results showed that the lowest averages value on the increase of leaves, shoots, nodes and plantlets height al MVI are resulted at dose 1.5 krad, whereas dose 0.3 krad increases averages value on shoots and plantlet height. The highest percentage of abnormal leaves is resulted at dose 1.2 krad. Ater subculture, the MV2 plantlets showed higher averages value for almost all parameters observed than the untreated plantlets. The number of score able bands varied from 2-5 bands with molecular weight 0.4-12 kb. Thirty three bands were detected from the six primers, with OPC-01, OPC-04 and OPC-06 showed polymorphisms with 8 (24%) polymorphic bands. In OPC-01 one band with DNA size 1 -1.5 kb was absence rom the treated plants at dose 0.9-1.5 krad, while with OPC- 04, one band size 1 5 kb present only at 1.2 krad and with OPC-06 one band size 12 kb absence from 0.6 and 0.9 krad, and 3-5 bands size 1.5, 1.8 and bands with size 3-12 kb disappeared at dose 1.2 and 1.5 krad. The appearance and disappearance of bands may be related to the genetic changes due to y irradiation, and further exploration may be needed to ind how much genetic variation induced by irradiation in ield and the relationships with the changes in plant characters.</p><p>Key words: Piper nigrum L., mutation, irradiation, RAPD, genetic variation</p><p> </p><p><strong>ABSTRAK </strong></p><p><strong>Peningkatan keragaman genetik tanaman lada (Piper nigrum L.) dengan iradiasi sinar gamma</strong></p><p>Keragaman genetik plasma nutfah lada sempit, untuk meningkatkan keragaman genetik, mata tunas yang tumbuh dari biakan lada varietas LDL diradiasi dengan sinar y dengan dosis 0, 0.3 0.6, 0.9, 1.2, dan 1.5 krad. Perlakuan menggunakan rancangan acak lengkap dengan lima ulangan. Tunas hasil radiasi ditanam pada media MS. Respon tanaman terhadap perlakuan iradiasi dilakukan dengan mengamati peningkatan jumlah daun, tunas, buku, tinggi tanaman dan morfologi pada planlet hasil perbanyakan vegetatif generasi pertama (MVI) dan kedua setelah iradiasi (MV2). Tunas hasil perbanyakan sub-kultur setelah iradiasi (MV2) dianaltsa keragaman genetiknya dengan RAPD (Randomly Ampliied Polymorphic DNA) menggunakan enam primer acak, yaitu OPC-01 (TTCGAGCCAG), OPC-02 (GTGAGGCGTC), OPC-04 (CCGCATCTAC), OPC-05 (GATGACCGCC), OPC-06 (GAACGGACTC) dan Abi 117.17 (GCTCGTCAAC). Hasil penelitian menunjukkan bahwa iradiasi menyebabkan perubahan yang nyata pada planlet generasi pertama setelah perbanyakan vegetatif (MVI) terutama pada jumlah buku dan tinggi tanaman. tetapi tidak berbeda nyata untuk penambahan jumlah daun dan<br /><br />166 <br /><br />tunas. Nilai rata-rata penambahan jumlah daun, tunas, buku dan tinggi planlet terendah ditunjukkan oleh perlakuan iradiasi pada dosis 1.5 krad, sedangkan pada iradiasi 0.3 krad meningkatkan nilai rata-rata jumlah tunas dan tinggi planlet. Persentase daun abnormal diperoleh pada perlakuan 1.2 krad. Setelah sub-kultur, planlet generasi kedua setelah perbanyakan vegetatif (MV2) yang tumbuh menunjukkan nilai rata-rata yang lebih tinggi dari normal pada semua parameter. Persentase daun variegata pada MVI diperoleh dari perlakuan 1.2 krad tetapi pada MV2 diperoleh dari perlakuan 0.6 krad. Jumlah pita DNA yang terampliikasi berkisar antara 2-5 dengan berat molekul 0.4-12 kb. Tiga puluh tiga pita tcrdetcksi, 8 (24 %) pita diantaranya polimorfik, yang berasal dari primer OPC-01, OPC-04 dan OPC-06. Pada OPC-01 satu pita dengan ukuran 1-1.5 kb hilang dari perlakuan 0.9-1.5 krad, sementara pada OPC-04, satu pita dengan ukuran 1.5 kb muncul hanya pada perlakuan 1.2 krad dan pada OPC-06 satu pita 12 kb hilang dari perlakuan 0-6 dan 0.9 krad, 3-5 pita dengan ukuran 1.5 kb, 1.8 kb dan antara 3-12 kb hilang dari perlakuan 1.2 dan 1.5 krad. Hilang dan munculnya pita kemungkinan berhubungan dengan perubahan genetik akibat radiasi sinar y dan penelitian lanjut perlu dilakukan untuk menge¬ tahui tingkat keragaman yang ditimbulkan akibat iradiasi di lapang dan hubungannya dengan perubahan sifat terutama sifat yang mengunlungkan<br /><br />Kara kunci: Piper nigrum L„ Lada, mutasi, radiasi, RAPD, variasi genetik</p>

Assessment of the genotoxic potential of tetrachlorvinphos insecticide by cytokinesis-block micronucleus and sister chromatid exchange assays

2021 ◽  
pp. 096032712110361
Author(s):  
Hayal Cobanoglu ◽  
Akin Cayir
Keyword(s):  
Sister Chromatid ◽  
Sister Chromatid Exchange ◽  
Chromosomal Instability ◽  
Human Peripheral Blood ◽  
Peripheral Blood Lymphocytes ◽  
Proliferation Index ◽  
Dna Damages ◽  
Genotoxic Potential ◽  
Index Level

Tetrachlorvinphos is an organophosphate that is classified as a carcinogen in humans by several authorities. Due to very limited data regarding the genotoxic potential, we aimed to comprehensively investigate in vitro genotoxic potential of tetrachlorvinphos. We performed our study by applying the cytokinesis-block micronucleus cytome and sister chromatid exchange (SCE) assays to human peripheral blood lymphocytes. We evaluated micronucleus (MN) and SCE frequencies and cytokinesis-block proliferation index in both exposed and non-exposed lymphocytes. We also calculated the chromosomal instability level in response to exposure by combining the results of MN and SCE. We found that MN frequency did not increase with exposure to tetrachlorvinphos (0–50 µg/ml). In contrast, we observed that SCE frequencies significantly increased with exposure to ≥5 µg/ml tetrachlorvinphos. Furthermore, exposure to tetrachlorvinphos at concentrations of 50 µg/ml induced a significant increase in chromosomal instability level ( p < 0.05). Cytokinesis-block proliferation index level did not significantly decrease in response to tetrachlorvinphos exposure. Our findings reveal that tetrachlorvinphos resulted in different DNA damages that were measured by two assays. Furthermore, our findings suggested that exposure to tetrachlorvinphos increased chromosomal instability that is a hallmark of many malignancies. We conclude that although tetrachlorvinphos does not significantly increase the MN level, the significant increase of both SCE and CIN frequencies indicates the genotoxic potential of tetrachlorvinphos in human peripheral lymphocytes. Additionally, tetrachlorvinphos is not cytotoxic in the range of tested concentrations.

Emergence of ceftazidime/avibactam resistance in KPC-3-producing Klebsiella pneumoniae in vivo

2019 ◽  
Vol 74 (11) ◽  
pp. 3211-3216 ◽  
Author(s):  
Stephan Göttig ◽  
Denia Frank ◽  
Eleonora Mungo ◽  
Anika Nolte ◽  
Michael Hogardt ◽  
...  
Keyword(s):  
Combination Therapy ◽  
Klebsiella Pneumoniae ◽  
Selection Pressure ◽  
Galleria Mellonella ◽  
Phenotypic Characterization ◽  
Infection Model ◽  
Development Of Resistance ◽  
Time Kill

Abstract Objectives The β-lactam/β-lactamase inhibitor combination ceftazidime/avibactam is active against KPC-producing Enterobacterales. Herein, we present molecular and phenotypic characterization of ceftazidime/avibactam resistance in KPC-3-producing Klebsiella pneumoniae that emerged in vivo and in vitro. Methods Sequence analysis of blaKPC-3 was performed from clinical and in vitro-generated ceftazidime/avibactam-resistant K. pneumoniae isolates. Time–kill kinetics and the Galleria mellonella infection model were applied to evaluate the activity of ceftazidime/avibactam and imipenem alone and in combination. Results The ceftazidime/avibactam-resistant clinical K. pneumoniae isolate revealed the amino acid change D179Y in KPC-3. Sixteen novel mutational changes in KPC-3 among in vitro-selected ceftazidime/avibactam-resistant isolates were described. Time–kill kinetics showed the emergence of a resistant subpopulation under selection pressure with either imipenem or ceftazidime/avibactam. However, combined selection pressure with imipenem plus ceftazidime/avibactam prevented the development of resistance and resulted in bactericidal activity. Concordantly, the G. mellonella infection model revealed that monotherapy with ceftazidime/avibactam is prone to select for resistance in vivo and that combination therapy with imipenem results in significantly better survival. Conclusions Ceftazidime/avibactam is a valuable antibiotic against MDR and carbapenem-resistant Enterobacterales. Based on time–kill kinetics as well as an in vivo infection model we postulate a combination therapy of ceftazidime/avibactam and imipenem as a strategy to prevent the development of ceftazidime/avibactam resistance in KPC-producing Enterobacterales in vivo.

Effects of in-Office and at-Home Bleaching on Human Enamel and Dentin: An in vitro Application of Fourier Transform Infrared Study

2008 ◽  
Vol 62 (11) ◽  
pp. 1274-1279 ◽  
Author(s):  
Feride Severcan ◽  
Kurtulus Gokduman ◽  
Ayca Dogan ◽  
Sukran Bolay ◽  
Saadet Gokalp
Keyword(s):  
Fourier Transform ◽  
Structural Changes ◽  
Protein Denaturation ◽  
Random Coil ◽  
Protein Ratio ◽  
Band Frequency ◽  
Human Enamel ◽  
Ft Ir ◽  
At Home

In-office and at-home bleaching techniques are widely used methods for the whitening of teeth. However, the safety of these techniques has not been clarified yet. The aim of the current study is to investigate the in-office- and at-home-bleaching-induced structural and quantitative changes in human enamel and dentin at the molecular level, under in vitro conditions. The Fourier transform mid-infrared (mid-FT-IR) spectroscopic technique was used to monitor bleaching-induced structural changes. Band frequency and intensity values of major absorptions such as amide A, amide I, phosphate (PO4), and carbonate (CO3−2) bands, for treatment groups and control, were measured and compared. The results revealed that both procedures have negligible effects on dentin constituents. In office-bleached enamel, in addition to demineralization, a decrease in protein and polysaccharide concentrations, mineral-to-protein ratio, and the strength of hydrogen bonds around NH groups, as well as a change in protein secondary structure were observed. The protein structure changed from β-sheet to random coil, which is an indication of protein denaturation. However, no significant variations were observed for at-home bleached enamel. The control, at-home, and in-office bleached enamel samples were differentiated with a high accuracy using cluster analysis based on FT-IR data. This study revealed that office bleaching caused deleterious alterations in the composition and structure of enamel that significantly affected the crystallinity and mineralization of the tissue. Therefore, at-home bleaching seems to be much safer than in-office bleaching in terms of molecular variations.

Phylogenetic relationships of five morphological groups of hexaploid wheat (Triticum aestivum L. em Thell.) based on RAPD analysis

Genome ◽  
2000 ◽  
Vol 43 (4) ◽  
pp. 724-727 ◽  
Author(s):  
Wenguang Cao ◽  
G Scoles ◽  
P Hucl ◽  
R N Chibbar
Keyword(s):  
Hexaploid Wheat ◽  
Phylogenetic Relationships ◽  
Genetic Similarity ◽  
Rapd Analysis ◽  
Genetic Relationships ◽  
Random Amplified Polymorphic Dna ◽  
Triticum Aestivum L ◽  
Similarity Coefficients ◽  
Morphological Classification ◽  
Wild Wheat

The genetic relationships among the five groups of hexaploid wheat: common, spelta, macha, vavilovii, and semi-wild wheat (SWW) are not clear. Random amplified polymorphic DNA (RAPD) analysis was used to assess phylogenetic relationships among these five morphological groups of hexaploid wheat. RAPD data were analyzed using the NTSYS-PC computer program to generate Jaccard genetic similarity coefficients. A dendrogram based on RAPD analysis grouped 15 accessions into five distinct clusters. These results are in agreement with those based on morphological classification, suggesting that common wheat is most closely related to SWW, followed by spelta, vavilovii, and macha.Key words: RAPD, macha, spelta, vavilovii, semi-wild wheat, phylogenetic relationships.

scholarly journals The SARS-CoV-2 Omicron (B.1.1.529) variant exhibits altered pathogenicity, transmissibility, and fitness in the golden Syrian hamster model

2022 ◽  
Author(s):  
Shuofeng Yuan ◽  
Zi-Wei Ye ◽  
Ronghui Liang ◽  
Kaiming Tang ◽  
Anna Jinxia Zhang ◽  
...  
Keyword(s):  
Syrian Hamster ◽  
Neutralizing Antibodies ◽  
Selection Pressure ◽  
Immune Selection ◽  
Hamster Model ◽  
Golden Syrian Hamster ◽  
New Variant ◽  
Contact Transmission

The newly emerging SARS-CoV-2 Omicron (B.1.1.529) variant first identified in South Africa in November 2021 is characterized by an unusual number of amino acid mutations in its spike that renders existing vaccines and therapeutic monoclonal antibodies dramatically less effective. The in vivo pathogenicity, transmissibility, and fitness of this new Variant of Concerns are unknown. We investigated these virological attributes of the Omicron variant in comparison with those of the currently dominant Delta (B.1.617.2) variant in the golden Syrian hamster COVID-19 model. Omicron-infected hamsters developed significantly less body weight losses, clinical scores, respiratory tract viral burdens, cytokine/chemokine dysregulation, and tissue damages than Delta-infected hamsters. The Omicron and Delta variant were both highly transmissible (100% vs 100%) via contact transmission. Importantly, the Omicron variant consistently demonstrated about 10-20% higher transmissibility than the already-highly transmissible Delta variant in repeated non-contact transmission studies (overall: 30/36 vs 24/36, 83.3% vs 66.7%). The Delta variant displayed higher fitness advantage than the Omicron variant without selection pressure in both in vitro and in vivo competition models. However, this scenario drastically changed once immune selection pressure with neutralizing antibodies active against the Delta variant but poorly active against the Omicron variant were introduced, with the Omicron variant significantly outcompeting the Delta variant. Taken together, our findings demonstrated that while the Omicron variant is less pathogenic than the Delta variant, it is highly transmissible and can outcompete the Delta variant under immune selection pressure. Next-generation vaccines and antivirals effective against this new VOC are urgently needed.

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