scholarly journals The frequency of T-DNA integration during the genetic transformation of tobacco (Nicotiana tabacum L.)

2018 ◽  
Vol 22 ◽  
pp. 262-266
Author(s):  
A. G. Komisarenko ◽  
S. I. Mykhalska ◽  
O. O. Khrystan
Keyword(s):  
Nicotiana Tabacum ◽  
Transgenic Plants ◽  
Transformation Frequency ◽  
Tobacco Plants ◽  
Double Stranded Rna ◽  
Dna Integration ◽  
Agrobacterium Mediated Transformation ◽  
Nicotiana Tabacum L ◽  
Is Integration

Aim. Investigation of the is integration frequency of the genes and their components (elements of genetic constructions) in transgenic tobacco-regenerating plants. Methods. Agrobacterium-mediated transformation of tobacco (Nicotiana tabacum L.) in vitro. Results. There were obtained tobacco plants with integrated target and selective genes. It was found that a complete copy of T-DNA or its certain elements were embedded. Conclusions. The possibility of the integration of complete and incomplete DNA copies during in vitro Agrobacterium-mediated transformation of the Nicotiana tabacum were established. This event were observed both during manipulations with pBi2E plasmid with the double-stranded RNA PHD gene suppressor and pBi-OAT plasmid with OAT gene. The transformation frequency with the complete integration of the genetic construction was 91 %. The integration frequencies for various plasmids were: LBA4404 pBi2E – 78 %; AGLO pBi2E – 100 %; AGLO pBi-OAT – 94 %. Keywords: Nicotiana tabacum L., Agrobacterium-mediated transformation in vitro, transgenic plants, T-DNA.

scholarly journals Topical Application of Double-Stranded RNA Targeting 2b and CP Genes of Cucumber mosaic virus Protects Plants against Local and Systemic Viral Infection

Plants ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 963
Author(s):  
Maria C. Holeva ◽  
Athanasios Sklavounos ◽  
Rajendran Rajeswaran ◽  
Mikhail M. Pooggin ◽  
Andreas E. Voloudakis
Keyword(s):  
Cucumber Mosaic Virus ◽  
Mosaic Virus ◽  
Topical Application ◽  
Plant Virus ◽  
Plant Protection ◽  
Post Inoculation ◽  
Tobacco Plants ◽  
Double Stranded Rna

Cucumber mosaic virus (CMV) is a destructive plant virus with worldwide distribution and the broadest host range of any known plant virus, as well as a model plant virus for understanding plant–virus interactions. Since the discovery of RNA interference (RNAi) as a major antiviral defense, RNAi-based technologies have been developed for plant protection against viral diseases. In plants and animals, a key trigger of RNAi is double-stranded RNA (dsRNA) processed by Dicer and Dicer-like (DCL) family proteins in small interfering RNAs (siRNAs). In the present study, dsRNAs for coat protein (CP) and 2b genes of CMV were produced in vitro and in vivo and applied onto tobacco plants representing a systemic solanaceous host as well as on a local host plant Chenopodium quinoa. Both dsRNA treatments protected plants from local and systemic infection with CMV, but not against infection with unrelated viruses, confirming sequence specificity of antiviral RNAi. Antiviral RNAi was effective when dsRNAs were applied simultaneously with or four days prior to CMV inoculation, but not four days post inoculation. In vivo-produced dsRNAs were more effective than the in vitro-produced; in treatments with in vivo dsRNAs, dsRNA-CP was more effective than dsRNA-2b, while the effects were opposite with in vitro dsRNAs. Illumina sequencing of small RNAs from in vivo dsRNA-CP treated and non-treated tobacco plants revealed that interference with CMV infection in systemic leaves coincides with strongly reduced accumulation of virus-derived 21- and 22-nucleotide (nt) siRNAs, likely generated by tobacco DCL4 and DCL2, respectively. While the 21-nt class of viral siRNAs was predominant in non-treated plants, 21-nt and 22-nt classes accumulated at almost equal (but low) levels in dsRNA treated plants, suggesting that dsRNA treatment may boost DCL2 activity. Taken together, our findings confirm the efficacy of topical application of dsRNA for plant protection against viruses and shed more light on the mechanism of antiviral RNAi.

scholarly journals Antifungal Prenylated Diphenyl Ethers from Arthrinium arundinis, an Endophytic Fungus Isolated from the Leaves of Tobacco (Nicotiana tabacum L.)

Molecules ◽  
2018 ◽  
Vol 23 (12) ◽  
pp. 3179 ◽  
Author(s):  
Peng Zhang ◽  
Xin Li ◽  
Xiao-Long Yuan ◽  
Yong-Mei Du ◽  
Bin-Gui Wang ◽  
...  
Keyword(s):  
Nicotiana Tabacum ◽  
Cell Line ◽  
Endophytic Fungus ◽  
In Vitro Cytotoxicity ◽  
Ionization Mass ◽  
Monocytic Cell ◽  
Monocytic Cell Line ◽  
Nicotiana Tabacum L ◽  
Diphenyl Ethers

An endophytic fungus Arthrinium arundinis TE-3 was isolated and purified from the fresh leaves of cultivated tobacco (Nicotiana tabacum L.). Chemical investigation on this fungal strain afforded three new prenylated diphenyl ethers (1−3) as well as three known analogues (4−6). Structure elucidation of the isolated compounds was carried out by analysis of 1D and 2D nuclear magnetic resonance (NMR) and high-resolution electrospray ionization mass spectroscopy (HRESIMS) spectra, as well as by comparison of those data with literature data. The absolute configuration of the stereogenic center at C-8 in 1 was assigned by comparison of the experimental and calculated ECD spectra. Compounds 1 and 2 showed selective antifungal activity against Mucor hiemalis with minimum inhibitory concentration (MIC) values of 8 and 4 μg/mL, respectively. Compounds 5 and 6 exhibited inhibitory activity against Alteraria alternata with an MIC value of 8 μg/mL. In the cytotoxic assay, 2, 5, and 6 displayed moderate in vitro cytotoxicity against the human monocytic cell line (THP-1 cell line), with IC50 values of 40.2, 28.3, and 25.9 μM, respectively. This study indicated that endophytic fungi possess great potential for exploring new bioactive secondary metabolites.

scholarly journals Effect of arginine, putrescine and spermidine on the polyamine, proline and chlorophyll content of tobacco (Nicotiana tabacum L.)

2021 ◽  
pp. 39-43
Author(s):  
Ákos Mendel ◽  
László Kovács ◽  
Erzsébet Kiss
Keyword(s):  
Nicotiana Tabacum ◽  
Salt Tolerance ◽  
Lower Class ◽  
Arginine Decarboxylase ◽  
Polyamine Metabolism ◽  
Spermine Synthase ◽  
Nicotiana Tabacum L ◽  
Exogenous Addition ◽  
Precursor Molecules

Polyamines, such as spermidine (Spd) spermine (Spm) and their direct precursor, the diamine putrescine (Put) are vital and essential aliphatic amines which are also present in plants. Although ethylene and polyamines are also involved in fruit ripening, the genes coding them must also take part in other biosynthetic pathways. In the ethylene and polyamines play an important role in development of salt stress tolerance, and in responses for biotic and abiotic stresses. Exogenous application of all three main polyamines (Put, Spd, Spm) increase salt tolerance of plants, but, accordingly to previous experiments, spermidine has the main effect on the enhancement of salt tolerance. Nicotiana tabacum L. plants were grown in vitro on MS medium, the treatments were as follows: arginine (150 mg l-1), putrescine (10 mg l-1), spermidine (10 mg l-1). Proline, chlorophyll a, b and polyamine contents were measured. The obtained results show that the arginine decarboxylase and the spermidine synthase genes involved in polyamine metabolism, cannot be enhanced by exogenous addition of their precursor molecules. On the contrary, the spermine synthase gene has a positive effect to the lower-class forms of polyamines.

scholarly journals Receiving of genetic-modified wheat plants with heterolous ornitin-δ-aminotransferase gene

2018 ◽  
Vol 22 ◽  
pp. 222-227
Author(s):  
O. M. Honcharuk ◽  
O. V. Dubrovna
Keyword(s):  
Triticum Aestivum ◽  
Transgenic Plants ◽  
Bread Wheat ◽  
Binary Vector ◽  
Triticum Aestivum L ◽  
Callus Cultures ◽  
Pcr Analysis ◽  
Genetic Construct ◽  
Agrobacterium Mediated Transformation

Aim. Receiving of genetically modified plants of bread wheat with heterologous ornithine‑δ‑aminotransferase gene. Methods. Agrobacterium-mediated transformation of callus cultures in vitro, PCR-analysis. Results. By Agrobacterium-mediated transformation of the morphogenic calluses of bread wheat (Triticum aestivum L.) using the AGLO strain containing the binary vector pBi-OAT with the target ornithine-δ-aminotransferase (oat) and selective neomycinphosphotransferase II (nptII), transgenic plants-regenerators have been obtained. Conclusions. As a result of the genetic transformation of Zimoyarka variety, 12 wheat regenerants were obtained in the genome which revealed a complete integration of the genetic construct containing the oat and nptII transgenes. Keywords: Triticum aestivum L., Agrobacterium-mediated transformation, ornithine‑δ‑aminotransferase gene, PCR-analysis.

scholarly journals Lead Phytoremediation Potential of Wild Type and Transgenic Tobacco Plants

2021 ◽  
Vol 5 (1) ◽  
pp. 168-182
Author(s):  
Hatice DAGHAN ◽  
Veli UYGUR ◽  
Abdullah EREN
Keyword(s):  
Nicotiana Tabacum ◽  
Transgenic Tobacco ◽  
Transgenic Tobacco Plants ◽  
Wild Type ◽  
Tobacco Plants ◽  
Nicotiana Tabacum L ◽  
Lead Phytoremediation

Genetiği değiştirilmiş bitkiler, kurşunun (Pb) kökten yer üstü kısımlarına translokasyonunu geliştirmek için büyük bir potansiyele sahip olabilir. Transgenik olmayan ( Nicotiana tabacum L. cv. Petit Havana SR1) ve transgenik (p-cV-ChMTII GFP) tütün bitkileri tarafından Pb alımının sağlanması araştırmak için Çin hamsteri metalotiyonin II gezen bir kap deneyi yapıldı . Transgenik ve transgenik olmayan tütün bitkileri, 0, 1000, 2500, 5000 mg Pb kg- 1 ile Pb (NO 3 ) 2 olarak işlenmiş topraklarda yetiştirildi. Kelimede bir büyüme bölümünde 6 hafta boyunca çiçeklenme aşamasına kadar.Bitkilerin büyümesi, klorofil içeriği, mineral besin elementleri ve düşük glutatyon (GSH) bezleri, bitkilerin Pb alım potansiyeli ile birlikte incelenmiştir. Hem transgenik hem de transgenik olmayan bitkiler için Pb uygulamasındaki artışa bağlı olarak yer üstü biyokütle çevrildi aşamalı bir düşüş gözlendi. Yaprak besinlerinin bulaştığı, aşırı Pb işlemlerinden olumsuz etkilenmiştir, bunlardan en büyük düşüşü. Sürgün Pb yüksek derecesi 76.0 mg kg kadar ulaşan -1 transgenik ve 70.9 mg kg -1 transgenik olmayan bitkilerde. Pb alımı, p-cV-ChMTII GFP'nin tütün bitkisine aktarılmasıyla iyileştirildi; ancak, Pb fitoremediasyonunda yeterli değildi. 

scholarly journals Growth and Division of Single Cells of Higher Plants in Vitro

1960 ◽  
Vol 43 (4) ◽  
pp. 841-851 ◽  
Author(s):  
Ludwig Bergmann
Keyword(s):  
Nicotiana Tabacum ◽  
Higher Plants ◽  
Single Cells ◽  
Tissue Cultures ◽  
Phaseolus Vulgaris L ◽  
Isolated Cells ◽  
Nicotiana Tabacum L ◽  
Agar Layer ◽  
Petri Dishes

The cultivation of single cells of Nicotiana tabacum L. var. "Samsun" and Phaseolus vulgaris L. var. "Early Golden Cluster" on a thin agar layer in Petri dishes is described. Under these conditions about 20 per cent of the cells divided repeatedly and established tissue clones which could be isolated and maintained as growing tissue cultures. It was possible also to follow the successive divisions of isolated cells and to observe their behavior during cytogenesis under the microscope.

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