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 Tissues of immature and mature embryos as morphogenic competent explants for genetic transformation of wheat

2020 ◽  
Vol 26 ◽  
pp. 233-238
Author(s):  
S. I. Mykhalska ◽  
A. G. Komisarenko ◽  
V. M. Kurchii
Keyword(s):  
Triticum Aestivum ◽  
Winter Wheat ◽  
Molecular Genetic ◽  
Triticum Aestivum L ◽  
Callus Cultures ◽  
Molecular Genetic Analysis ◽  
Pcr Analysis ◽  
Agrobacterium Mediated Transformation ◽  
Mature Embryos

Aim. To analyze the efficiency of using tissues of immature and mature embryos for Agrobacterium-mediated transformation in vitro of new breeding-valuable genotypes of winter wheat (Triticum aestivum L.) for the purpose of their genetic improvement. Methods. Culture of in vitro, extraction and electrophoresis of DNA, PCR analysis. Results. The efficiency of induction of callusogenesis and regeneration of winter wheat shoots was analyzed. The morphogenetic response of callus cultures obtained from different explants under Agrobacterium-mediated transformation in vitro was investigated. Molecular genetic analysis of wheat regenerants for transgenes was performe. Conclusions. The tissues of immature and mature embryos of novel breeding-valuable wheat genotypes are competent explants for Agrobacterium-mediated transformation in vitro. In this case, the tissues of two daily sprouts of mature wheat germ are characterized by higher morphogenetic parameters, which helps to obtain a greater percentage of genetically modified variants. Keywords: Triticum aestivum L., in vitro, Agrobacterium-mediated transformation, immature embryos, mature embryos.

scholarly journals Genetic transformation of new perspective winter wheat genotypes in vitro

2020 ◽  
Vol 26 ◽  
pp. 270-275
Author(s):  
L. V. Slivka ◽  
O. V. Dubrovna
Keyword(s):  
Transgenic Plants ◽  
Genetic Transformation ◽  
Winter Wheat ◽  
Target Gene ◽  
Callus Cultures ◽  
Genetic Construct ◽  
Wheat Genotypes ◽  
Agrobacterium Mediated Transformation ◽  
New Perspective

Aim. Optimization of conditions for genetic transformation of new perspective winter wheat genotypes and production of transgenic plants. Methods. Agrobacterium-mediated transformation in vitro culture using callus cultures. Results. The influence of the optical density of cells of agrobacterial suspension, the concentration of the antibiotic cefotaxime, the duration of coculture on the frequency of obtaining kanamycin-resistant regenerants of new winter wheat genotypes by genetic transformation of callus cultures were investigated. By Agrobacterium-mediated transformation of morphogenic calluses of new perspective winter wheat genotypes were obtained plant-regenerants in the genome of which revealed the complete incorporation of a genetic construct containing oat and nptII transgenes. Conclusions. Agrobacterium-mediated transformation of callus cultures of new perspective winter wheat genotypes was optimized, and transgenic plants with the target gene of ornithine-δ-aminotransferase were obtained. Keywords: Triticum aestivum, Agrobacterium-mediated transformation, callus cultures, ornithine-δ-aminotransferase gene.

scholarly journals Genetic transformation of promising genotypes of winter bread wheat by in planta method

2021 ◽  
Vol 28 ◽  
pp. 106-111
Author(s):  
L. V. Slivka ◽  
O. V. Dubrovna
Keyword(s):  
Triticum Aestivum ◽  
Transgenic Plants ◽  
Genetic Transformation ◽  
Bread Wheat ◽  
Optical Density ◽  
Triticum Aestivum L ◽  
In Planta ◽  
Agrobacterium Mediated Transformation ◽  
Winter Bread Wheat ◽  
Inoculation Medium

Aim. Optimization of conditions and genetic transformation of new promising genotypes of winter bread wheat (Triticum aestivum L.) by in planta method. Methods. Agrobacterium-mediated transformation by the in planta method using strain AGL0 and vector construct pBi-OAT. Results. The influence of air temperature, optical density of cells of agrobacterial suspension, inoculation day and composition of inoculation medium on the frequency of obtaining transgenic plants of new promising genotypes of winter wheat was studied. The dependence of the frequency of obtaining transgenic plants on environmental conditions, in particular temperature, has been established. It was found that the temperature regime of 20-22 °C provided the largest number (4.4%) of wheat transformants, and when the temperature is reduced to 16-18 °C there is a decrease in the efficiency of T-DNA transfer into the plant genome and the lowest frequency of transformation is observed. Conclusions. The largest number of transformants was obtained using a inoculation medium without sucrose, the optical density of cells of the agrobacterial suspension of 0.4 op.od. and inoculation on the third day after castration of ears. Keywords: Triticum aestivum L., Agrobacterium-mediated transformation, ornithine-δ-aminotransferase gene.

scholarly journals Agrobacterium-mediated in planta genetic transformation of winter wheat (Triticum aestivum L.)

2018 ◽  
Vol 22 ◽  
pp. 293-298
Author(s):  
S. I. Mykhalska ◽  
A. G. Komisarenko ◽  
V. M. Kurchii ◽  
O. M. Tishchenko
Keyword(s):  
Triticum Aestivum ◽  
Transgenic Plants ◽  
Genetic Transformation ◽  
Winter Wheat ◽  
Natural Frequency ◽  
Triticum Aestivum L ◽  
Transformation Method ◽  
In Planta ◽  
Agrobacterium Mediated Transformation ◽  
Pollination Methods

Aim. To optimize the agrobacterium-mediated method of winter wheat transformation (Triticum aestivum L.); to select the conditions and period of inoculation to effectively transfer the genes during pollination. Methods. Agrobacterium-mediated in planta genetic transformation of winter wheat (Triticum aestivum L.) during pollination. Results. The conditions for agrobacterium-mediated transformation method of winter wheat during natural (frequency pollination was 1 %) and non-natural (frequency pollination was 4 %) pollination were defined. Conclusions. The possibility of integrating transgenes into the genome of winter wheat plants by the method of Agrobacterium-mediated transformation in planta in the process of forced and natural pollination is demonstrated. It is found that the transformation efficiency to a large extent depends on the plant genotype and the method of carrying out the transformation procedure. The selection of transgenic plants under water deficit conditions allowed to identify the plants with functional transgene. The signs of functioning transgene have been remaining in the next generation of genetically modified winter wheat. Keywords: Triticum aestivum L., Agrobacterium-mediated transformation in planta, transgenic plants, seeds.

scholarly journals Annotation and Molecular Characterisation of the TaIRO3 and TaHRZ Iron Homeostasis Genes in Bread Wheat (Triticum aestivum L.)

Genes ◽  
2021 ◽  
Vol 12 (5) ◽  
pp. 653
Author(s):  
Oscar Carey-Fung ◽  
Jesse T. Beasley ◽  
Alexander A. T. Johnson
Keyword(s):  
Triticum Aestivum ◽  
Bread Wheat ◽  
Iron Homeostasis ◽  
Zinc Finger Protein ◽  
Expression Profiles ◽  
Diploid Species ◽  
Triticum Aestivum L ◽  
Fe Deficiency ◽  
Pcr Analysis ◽  
Fe Homeostasis

Effective maintenance of plant iron (Fe) homoeostasis relies on a network of transcription factors (TFs) that respond to environmental conditions and regulate Fe uptake, translocation, and storage. The iron-related transcription factor 3 (IRO3), as well as haemerythrin motif-containing really interesting new gene (RING) protein and zinc finger protein (HRZ), are major regulators of Fe homeostasis in diploid species like Arabidopsis (Arabidopsis thaliana) and rice (Oryza sativa L.), but remain uncharacterised in hexaploid bread wheat (Triticum aestivum L.). In this study, we have identified, annotated, and characterised three TaIRO3 homoeologs and six TaHRZ1 and TaHRZ2 homoeologs in the bread wheat genome. Protein analysis revealed that TaIRO3 and TaHRZ proteins contain functionally conserved domains for DNA-binding, dimerisation, Fe binding, or polyubiquitination, and phylogenetic analysis revealed clustering of TaIRO3 and TaHRZ proteins with other monocot IRO3 and HRZ proteins, respectively. Quantitative reverse-transcription PCR analysis revealed that all TaIRO3 and TaHRZ homoeologs have unique tissue expression profiles and are upregulated in shoot tissues in response to Fe deficiency. After 24 h of Fe deficiency, the expression of TaHRZ homoeologs was upregulated, while the expression of TaIRO3 homoeologs was unchanged, suggesting that TaHRZ functions upstream of TaIRO3 in the wheat Fe homeostasis TF network.

scholarly journals Effects of NaCl Stress on Callus Proliferation and Plant Regeneration from Mature Embryos of Bread Wheat (Triticum aestivum L.) Cultivars Mahon Demias and Hidhab

1970 ◽  
Vol 17 (1) ◽  
pp. 19-27 ◽  
Author(s):  
L Benderradji ◽  
H Bouzerzou ◽  
A Djekoun ◽  
A Benmahammed
Keyword(s):  
Triticum Aestivum ◽  
Bread Wheat ◽  
Salinity Stress ◽  
Callus Induction ◽  
Selection Process ◽  
Triticum Aestivum L ◽  
Nacl Stress ◽  
Mature Embryos ◽  
Callus Proliferation

An in vitro experiment with two cultivars, Mahon Demias and Hidhab of bread wheat (Triticum aestivum L.) exhibited appreciable callus induction but differed significantly in the capacity of calli proliferation and regeneration under salinity stress; even though Mahon Demias appeared relatively more tolerant than Hidhab. The results indicated the need to optimize a robust protocol for callus induction, maintenance and regeneration before selection process for tolerance to salinity is embarked upon.Key words: Bread wheat, Salinity stress, Callus proliferation, Mature embryoDOI = 10.3329/ptcb.v17i1.1117Plant Tissue Cult. & Biotech. 17(1): 19-27, 2007 (June)

scholarly journals Effect of Ceftriaxone and Timentin antibiotics on morphogensis in the in vitro culture of bread wheat Triticum aestivum L.

2016 ◽  
Vol 32 (5) ◽  
pp. 367-376 ◽  
Author(s):  
M. A. Bannikova ◽  
I. R. Gorbatyuk ◽  
I. S. Hnatiuk ◽  
L. V. Malysheva-Otto ◽  
V. P. Duplij
Keyword(s):  
Triticum Aestivum ◽  
In Vitro Culture ◽  
Bread Wheat ◽  
Triticum Aestivum L

scholarly journals Optimization of Agrobacterium-mediated transformation of perspective winter wheat genotypes in vitro

2020 ◽  
Vol 26 ◽  
pp. 190-195
Author(s):  
O. V. Dubrovna ◽  
L. V. Slivka
Keyword(s):  
Triticum Aestivum ◽  
Genetic Transformation ◽  
Winter Wheat ◽  
Optical Density ◽  
Callus Cultures ◽  
Optimal Parameters ◽  
Wheat Genotypes ◽  
Agrobacterium Mediated Transformation ◽  
New Perspective

Aim. Optimization of conditions for genetic transformation of new perspective winter wheat genotypes. Methods. Agrobacterium-mediated transformation in culture in vitro. Results. The influence of the optical density of cells of the agrobacterial suspension, the concentration of the antibiotic cefotaxime, the duration of coculture on the frequency of obtaining kanamycin-resistant regenerants of new winter wheat genotypes by genetic transformation of callus cultures were investigated using LBA4404 and AGL0 strains. It is shown that depending on the strain the most optimal is the concentration of agrobacteria 0.2-03 OD, duration of coculture for 2-3 days and the use of cefotaxime at a concentration of 250-500 mg/L. Conclusions. The optimal parameters for conducting Agrobacterium-mediated transformation of callus cultures of new perspective winter wheat genotypes were selected. Keywords: Triticum aestivum, Agrobacterium-mediated transformation, callus cultures.

scholarly journals Study of transgene expression in Triticum aestivum L. after Agrobacterium-mediated in planta transformation

2019 ◽  
Vol 16 (2) ◽  
pp. 159-165
Author(s):  
N. A. Zhalii ◽  
M. O. Bannikova ◽  
M. O. Plugatar ◽  
L. H. Velikozhon ◽  
A. M. Taranenko ◽  
...  
Keyword(s):  
Triticum Aestivum ◽  
Winter Wheat ◽  
Genetically Modified Organisms ◽  
Triticum Aestivum L ◽  
Pcr Analysis ◽  
Test Results ◽  
In Planta ◽  
Genetic Construct ◽  
Pcr Method

Aim. Detection of sequences of target transgenes nptII and bar in the genome of probable transformants of bread winter wheat Triticum aestivum L. cultivars Zymoiarka and Podolianka obtained as a result of Agrobacterium-mediated transformation in planta and determination of their expression level. Methods. Polymerase chain reaction (PCR) method was used independently and in combination with reverse transcription (RT-PCR), electrophoresis of DNA in agarose gel. Tolerance to the herbicide was evaluated in the physiological test. Results Through PCR analysis, the sequence of nptII transgene was detected in 30 samples of 145 analyzed, the frequency of transformation was 20.7 %. The sequence of the gene bar was observed in 85 experimental plants, and the frequency of transformation was 15.6 %. mRNAs of both transgenes were detected, indicating their transcriptional activity and stable expression. Conclusions PCR analysis allowed to detect nptII transgenic signal in 20.7 % of plants, while the presence of the bar gene was detected in 15.6 % of cases, indicating a higher efficiency of this genetic construct. The transcription is shown in all the specimens studied for both transgene. According to the results of the physiological test, 25 % of plants containing the gene bar showed resistance to the Basta® herbicide.Keywords: genetically modified organisms, transgenic plants, biotechnological cultures, bread winter wheat, genetic engineering.

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