scholarly journals Genetic and physiological analysis T1 biotechnological plants of winter wheat (Triticum aestivum L.)

2020 ◽  
Vol 26 ◽  
pp. 222-227
Author(s):  
A. G. Komisarenko ◽  
S. I. Mykhalska ◽  
V. M. Kurchii ◽  
O. O. Khrystan
Keyword(s):  
Triticum Aestivum ◽  
Winter Wheat ◽  
First Generation ◽  
Genetically Modified ◽  
Molecular Genetic ◽  
Triticum Aestivum L ◽  
Molecular Genetic Analysis ◽  
Pcr Analysis ◽  
Proline Dehydrogenase ◽  
In Planta

Aim. To investigate inheritance of transgenes in the first generation (T1) of winter wheat biotechnological plants (Triticum aestivum L.). Analyze the performance of T1 genetically modified plants with a double stranded RNA suppressor of the proline dehydrogenase (pdh) gene under normal growing conditions. Methods. PCR analysis, DNA electrophoresis; determination of indicators of the structure of the crop. Results. Molecular genetic analysis was performed and the performance indicators of control and T1 biotechnological plants were investigated. Conclusions. The first generation of genetically modified winter wheat plants resulting from Agrobacterium-mediated transformation in planta confirmed the inheritance of integrated genes. Among the transgenic variants identified plants that lack some fragments of the target gene required for partial suppression of the gene of proline dehydrogenase wheat. It is shown that at the optimal terms of growing biotechnological plants of wheat winter-annual of UK 322/17 and UK 209 h was characterized by the best indexes of structure of harvest as compared to an initial form, while the genetically changed plants of genotypes of UK 95/17 and UK 065 after the elements of the productivity did not differ from control plants. Keywords: Triticum aestivum L., biotechnological plants, T-DNA, proline dehydrogenase gene, structural analysis indicators.

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 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.

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 Study of transgen functionality in T2 biotechnological plants of winter wheat on the sign of osmoresistance

2021 ◽  
Vol 28 ◽  
pp. 88-93
Author(s):  
A. G. Komisarenko ◽  
S. I. Mykhalska ◽  
V. M. Kurchii
Keyword(s):  
Triticum Aestivum ◽  
Osmotic Stress ◽  
Winter Wheat ◽  
Protein Content ◽  
Water Deficit ◽  
Genetically Modified ◽  
Triticum Aestivum L ◽  
Quantitative Composition ◽  
Original Form ◽  
Growing Conditions

Aim. To investigate the functionality of the transgene in the seed generation (T2) of genetically modified winter wheat plants (Triticum aestivum L.) in terms of productivity in conditions of normal and insufficient water supply. Methods. Determination of yield structure and protein content. Results. The indicators of productivity of control and T2 biotechnological plants under normal growing conditions and water deficit are analyzed. The quantitative composition of protein in the conditions before and after osmotic stress and in the period of rehydration was determined. Conclusions. It is shown that under the action of water deficit there is a decrease in productivity for all studied plants. In this case, genetically modified plants had an advantage in the main elements of the crop structure over the original form under both analyzed growing conditions. It was found that under normal conditions of hydration, the amount of protein in the studied plants did not differ significantly. An increase in its content in control plants under the action of osmotic stress was recorded, which may indicate the synthesis of stress response proteins. Decreased protein content during rehydration may indicate the passage of normal metabolic processes. Keywords: Triticum aestivum L., biotechnological plants, proline dehydrogenase gene, structural analysis of yield, osmostability.

scholarly journals Naturally Occurring Ecdysteroids in Triticum aestivum L. and Evaluation of Fenarimol as a Potential Inhibitor of Their Biosynthesis in Plants

2021 ◽  
Vol 22 (6) ◽  
pp. 2855
Author(s):  
Anna Janeczko ◽  
Jana Oklestkova ◽  
Danuše Tarkowská ◽  
Barbara Drygaś
Keyword(s):  
Triticum Aestivum ◽  
Winter Wheat ◽  
Triticum Aestivum L ◽  
Growth Conditions ◽  
Animal Kingdom ◽  
Experimental Conditions ◽  
P450 Monooxygenase ◽  
Naturally Occurring ◽  
Regulatory Effects ◽  
Potential Use

Ecdysteroids (ECs) are steroid hormones originally found in the animal kingdom where they function as insect molting hormones. Interestingly, a relatively high number of these substances can also be formed in plant cells. Moreover, ECs have certain regulatory effects on plant physiology, but their role in plants still requires further study. One of the main aims of the present study was to verify a hypothesis that fenarimol, an inhibitor of the biosynthesis of ECs in the animal kingdom, also affects the content of endogenous ECs in plants using winter wheat Triticum aestivum L. as a model plant. The levels of endogenous ECs in winter wheat, including the estimation of their changes during a course of different temperature treatments, have been determined using a sensitive analytical method based on UHPLC-MS/MS. Under our experimental conditions, four substances of EC character were detected in the tissue of interest in amounts ranging from less than 1 to over 200 pg·g−1 FW: 20-hydroxyecdysone, polypodine B, turkesterone, and isovitexirone. Among them, turkesterone was observed to be the most abundant EC and accumulated mainly in the crowns and leaves of wheat. Importantly, the level of ECs was observed to be dependent on the age of the plants, as well as on growth conditions (especially temperature). Fenarimol, an inhibitor of a cytochrome P450 monooxygenase, was shown to significantly decrease the level of naturally occurring ECs in experimental plants, which may indicate its potential use in studies related to the biosynthesis and physiological function of these substances in plants.

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