Evaluation of Factors Affecting In Planta Gene Editing Efficiency in Wheat (Triticum aestivum L.)

2021 ◽  
Author(s):  
Wayne Xu ◽  
Zhen Yao ◽  
Xiben Wang ◽  
Andriy Bilichak
Keyword(s):  
Triticum Aestivum ◽  
Gene Editing ◽  
Triticum Aestivum L ◽  
In Planta ◽  
Factors Affecting ◽  
Editing Efficiency

Development of simple and efficient in Planta transformation method for wheat (Triticum aestivum L.) using Agrobacterium tumefaciens

2006 ◽  
Vol 102 (3) ◽  
pp. 162-170 ◽  
Author(s):  
Putu Supartana ◽  
Tsutomu Shimizu ◽  
Masahiro Nogawa ◽  
Hidenari Shioiri ◽  
Tadashi Nakajima ◽  
...  
Keyword(s):  
Triticum Aestivum ◽  
Agrobacterium Tumefaciens ◽  
Triticum Aestivum L ◽  
Transformation Method ◽  
In Planta Transformation ◽  
In Planta

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 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 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 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 Factors Affecting the Radiosensitivity of Hexaploid Wheat to γ-Irradiation: Radiosensitivity of Hexaploid Wheat (Triticum aestivum L.)

PLoS ONE ◽  
2016 ◽  
Vol 11 (8) ◽  
pp. e0161700 ◽  
Author(s):  
Bing Han ◽  
Jiayu Gu ◽  
Linshu Zhao ◽  
Huijun Guo ◽  
Yongdun Xie ◽  
...  
Keyword(s):  
Triticum Aestivum ◽  
Hexaploid Wheat ◽  
Triticum Aestivum L ◽  
Γ Irradiation ◽  
Factors Affecting
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