scholarly journals Glyphosate selection of maize transgenic callus lines among genotypes of Ukrainian plant breeding

1970 ◽  
pp. 237-242
Author(s):  
I. O. Nitovska ◽  
I. K. Komarnytskyi ◽  
B. V. Morgun
Keyword(s):  
Plant Breeding ◽  
Transgenic Maize ◽  
Pcr Analysis ◽  
Agrobacterium Mediated Transformation ◽  
Maize Germplasm ◽  
Maize Transformation ◽  
Callus Lines ◽  
Selection Of ◽  
Maize Callus

Aim. Glyphosate selection has a number of advantages over other commonly used selectable markers for maize. There is some natural variability within maize germplasm for degree of sensitivity to glyphosate. We investigated the selective effect of glyphosate for production transgenic maize callus after Agrobacterium-mediated transformation among geno-types of Ukrainian plant breeding. Methods. Agrobacterium-mediated transformation, glyphosate selection in vitro, and PCR analysis were used to obtain transgenic maize callus and to confirm its status. Results. An efficient selectable marker system for production transgenic maize callus lines tolerant to herbicide glyphosate was proposed. Calluses of four maze genotypes of Ukrainian plant breeding and pCB135 vector containing CP4epsps gene were used in Agrobacterium-mediated transformation experiments. Three callus maize lines of DK267×PLS61 genotype containing CP4epsps gene were obtained. Conclusions. The use of glyphosate as a selective agent after Agrobacterium-mediated transformation proved to be effective for transgenic maize callus lines production containing the gene CP4epsps. The success of Agrobacterium-mediated transformation of maize callus strongly depended on the genotype of source ma-terial. Keywords: Agrobacterium-mediated maize transformation, CP4epsps gene, glyphosate selection, PCR analysis.

scholarly journals Glyphosate selection of maize transformants containing CP4epsps gene

2020 ◽  
Vol 26 ◽  
pp. 239-244
Author(s):  
I. O. Nitovska ◽  
B. V. Morgun ◽  
O. Ye. Abraimova ◽  
T. M. Satarova
Keyword(s):  
Transgenic Plants ◽  
Dna Analysis ◽  
Selective Agent ◽  
Regeneration Rate ◽  
Agrobacterium Mediated Transformation ◽  
Plant Dna ◽  
Morphogenic Callus ◽  
Selection Of ◽  
Maize Callus

Aim. To study the selection conditions of maize transformants containing the CP4epsps gene using glyphosate as a selective agent. Methods. Tissue culture in vitro, Agrobacterium-mediated transformation, selection of transgenic plants, isolation of total plant DNA, analysis of plant DNA by polymerase chain reaction (PCR). Results. The morphogenic maize callus of immature embryos of the hybrid (PLS61)R2×PLS61 was produced, which had a high regeneration rate (up to 95%), that persisted over long cultivation. Agrobacterium mediated transformation of the morphogenic callus and selection of the transgenic material using glyphosate yielded maize transformants containing the CP4epsps gene at a frequency of 1%. Conclusions. Maize genotype (PLS61)R2×PLS61 is promising for studies on the maize genetic transformation, in particular for the production of transgenic maize resistant to glyphosate herbicide. The use of morphogenic maize callus (PLS61)R2×PLS61 and glyphosate as a selective agent at a concentration of 0.1 mM and 0.25 mM in media for callusogenesis and 0.01 mM in the medium for regeneration was effective for the selection of transgenic plants with the gene CP4epsps. Keywords: Zea mays L., morphogenic callus, Agrobacterium-mediated transformation, PCR, genetic engineering.

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 INTRODUCTION OF EXPLANTS AND REPRODUCTION ON NUTRIENT MEDIUM OF DONOR MATERIAL IN VITRO VARIETIES OF CALLISTEPHUS CHINENSIS (L.) NESS. FOR ITS FURTHER USE IN LANDSCAPING

2020 ◽  
Vol 1 (383) ◽  
pp. 89-96
Author(s):  
V. Polishchuk ◽  
S. Turchina ◽  
A. Balabak ◽  
I. Kozachenko ◽  
V. Mamchur ◽  
...  
Keyword(s):  
Plant Breeding ◽  
Vegetative Propagation ◽  
Nutrient Medium ◽  
Cell Biology ◽  
Plant Biotechnology ◽  
Forest Steppe ◽  
Physical Parameters ◽  
Adaptation Period ◽  
Selection Of

The relevance of the research topic. On the recent methods of biotechnology are increasingly used in plant breeding and seed production. Herbaceous plants such as strawberries, potatoes, a vegetable, some medicinal and others are capable of vegetative propagation the traditional methods of culture, successfully introduced in both in vitro and can achieve a high rate of reproduction. Modern plant biotechnology – the sum of the technologies developed in molecular and cell biology of plants – a new stage in the development of the technology of plant breeding. With these improved characteristics may occur at the level of individual genes and individual genes that determine a specific trait, can be identified. They may be the final selection, they can be isolated, insert, delete, or modify the genotype or variety. Goal. Identify the features of the manifestation of economically valuable features and decorative properties of Callistephus chinensis and the inclusion of the best varieties in the biotech link, their adaptation to the conditions of the Forest-Steppe of Ukraine and their further use in landscaping. Methods. Laboratory – determination of seed germination; mathematical and statistical - for processing the reliability of the obtained research results. Results. The nutrient medium for growing plant tissues and cells, by analogy with the medium for culturing animal tissues, should contain all that the tissues in the plant organism receive from xylem and phloem currents of substances. However, in practice it has been found that vegetable juices cannot serve as a complete nutrient medium for growing isolated tissues and cells. This manifests the specificity of the receipt, transportation and especially the redistribution of nutrients in the plant. Based on the analysis, research was conducted to study the possibility of mass off-season vegetative propagation of plants of Callistephus chinensis in vitro. Practical recommendations on the selection of sterilizer, sterilization, nutrient medium and for the adaptation period of the best genotypes of this culture have been developed. As a result of the conducted researches the methods of selection of the initial plant material of Callistephus chinensis (Callistephus Chinensis (L.) NEES) and its surface sterilization, modification of existing aseptic culture methods have been studied and mastered. The morphogenetic potential of explants from different plant organs was investigated and selection of nutrient medium and study of the influence of plant growth regulators and physical parameters on the process of morphogenesis was carried out. The features of regeneration of isolated explants depending on the composition of the nutrient medium and selection of conditions for obtaining self-clones of Callistephus chinensis (Callistephus Chinensis (L.) NEES) were studied. Key words: in vitro, plant biotechnology, Callistephus Chinensis, nutrient medium, rhizogenesis.

scholarly journals In vitro selection of winter triticale for salt resistance

1970 ◽  
pp. 247-251
Author(s):  
S. V. Pykalo ◽  
O. V. Dubrovna ◽  
O. A. Demydov
Keyword(s):  
Salt Stress ◽  
Cell Lines ◽  
In Vitro Selection ◽  
High Survival ◽  
Winter Triticale ◽  
Step Type ◽  
Callus Lines ◽  
Selection Of

Aim. To obtain of cell lines and plant-regenerants of winter triticale resistant for salt stress the in vitro selection was carried out. Methods. In order to select resistant to salt stress forms of triticale the efficiency of using direct and step-type in vitro selection with application of selective system based on sodium chloride has been investigated. Results. The direct and step-type in vitro selection was conducted and the selection of callus lines of triticale being resistant to simu-lated salinity was carried out. As a result, from line 38/1296 and variety Obriy respectively, 5 and 4 resistant callus lines were identified that had a high survival rate on the selective medium with 1.2 % NaCl and maintained morphogenetic potential. From the resistant lines plant regenerants were induced and their rearing, rooting and transfer to in vivo condi-tions were optimized. Conclusions. A step-type in vitro selection was more effective, because resulted from the selec-tion more resistant callus forms were identified. First cell lines of winter triticale with resistance to salt stress were de-rived.Keywords: Triticale, in vitro selection, callus, salt stress, resistance.

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 In Vitro Organogenesis of a Slipper Orchid, Paphiopedilum ‘Alma Gavaert’

2018 ◽  
Vol 10 (4) ◽  
pp. 607-613 ◽  
Author(s):  
Jen-Tsung CHEN
Keyword(s):  
Continuous Selection ◽  
Callus Line ◽  
Root Explants ◽  
In Vitro Organogenesis ◽  
Vegetative Tissues ◽  
Slipper Orchid ◽  
Regeneration Efficiency ◽  
Callus Lines ◽  
Selection Of

The aim of the present study was to improve the regeneration efficiency of callus lines in a slipper orchid, Paphiopedilum ‘Alma Gavaert’. Three kinds of vegetative tissues, root, stem and leaf segments, were used as explants to induce callogenesis; out of these, only root explants formed callus and was subcultured in the presence of 5 mg/L dicamba and 5 mg/L 2,4-D combined with 1 or 2 mg/L TDZ. The resulting four callus lines, assigned as 5Di1T, 5Di2T, 5D1T and 5D2T, respectively, were used to test the effect of NAA to BA ratios on re-differentiation, wherein the highest number of shoots (approximately 2 shoots/0.1 g callus clump) were obtained in callus line 5D2T at ratios of 0.001 and 0.002. A largely improvement of shoot regeneration efficiency was obtained by continuous selection of callus lines which derived from different explant positions. Eventually, six callus lines, including 5D2T-T6-G5 to 5D2T-T6-G10, were able to produce approximately 10 times of shoots per callus clump when compared with the parental callus line 5D2T.

scholarly journals Agrobacterium-mediated Genetic Transformation of Lentil (Lens culinaris Medik.) with Chitinase Gene followed by In vitro Flower and Pod Formation

2019 ◽  
Vol 29 (1) ◽  
pp. 99-109
Author(s):  
Subroto K Das ◽  
Kishwar Jahan Shethi ◽  
MI Hoque ◽  
RH Sarker
Keyword(s):  
Genetic Transformation ◽  
Lens Culinaris ◽  
Target Gene ◽  
Gene Selection ◽  
Chitinase Gene ◽  
Pcr Analysis ◽  
Agrobacterium Strain ◽  
Half Strength ◽  
Selection Of

To investigate the integration of chitinase gene in lentil (Lens culinaris Medik.) namely, BARI masur-4 (BM-4), BARI masur-5 (BM-5) and BARI masur-6 (BM-6) through Agrobacterium-mediated genetic transformation was performed using Agrobacterium strain EHA 105 harboring bar (resistant to phosphinotrycin) and chitinase (gene of interest) gene. Selection of transformed shoots was carried out by gradually increasing the concentration of phosphinotrycin (PPT) up to 2.0 mg/l. Transgenic lentil shoots were produced with an overall frequency of 0.36 in case of BM-4 and BM-6 and 0.34 in case of BM-5, respectively. Most of the selected shoots developed in vitro flowers and pods following their sub-culture on half strength of MS supplemented with 20 mg/l IBA, 0.5 mg/l NAA with 50 mg/l ticarcillin. Seedlings germinated from the seeds were successfully transferred to soil for the development of further progeny. Stable integration of target gene was confirmed through PCR analysis. Plant Tissue Cult. & Biotech. 29(1): 99-109, 2019 (June)

scholarly journals AGROBACTERIUM-MEDIATED TRANSFORMATION OF TWO TOMATO CULTIVARS (LYCOPERSICON ESCULENTUM MILL.) CV. SANDRA AND ROCKY

2021 ◽  
Vol 52 (3) ◽  
pp. 745-755
Author(s):  
G. H. Danial ◽  
D. A. Ibrahim ◽  
G. Q. Song
Keyword(s):  
Marker Gene ◽  
Kanamycin Resistance ◽  
Transformation Frequency ◽  
Pcr Analysis ◽  
Nptii Gene ◽  
Agrobacterium Mediated Transformation ◽  
Nos Terminator ◽  
Transgenic Tomato Plants ◽  
Tomato Cultivars

An efficient protocol for Agrobacterium-mediated transformation of tomato cultivars Sandra and Rocky was conducted to examine the possibility of producing transgenic tomato plants cultivars harbouring the nptII gene, conferring kanamycin resistance. To achieve this aim, tomato cotyledon explants were transformed using EHA105 Agrobacterium tumefaciens strain harboring the binary vectors pBI121 which contains Gus gene, and neomycin phosphotransferase II (nptII) as selectable marker gene under the control of a CaMV35S promoter and nopaline synthase (nos) Terminator. Transformant detection was carried out in three distinct ways. First antibiotic selection, Kanamycin at a concentration of100 mgl-1 found to be efficient for this purpose. Second histochemical GUS assay revealed the presence of blue colored zones in a number of shoots and leaves for both in vitro and the greenhouse-grown transgenic plants. Third PCR analysis indicated positive result by showing the fragment for nptII gene in tested transformants, while was absent in non-transgenic control (wild type). On the other hand, the results showed that Sandra cultivar was more efficient for regeneration and subsequently transformation frequency than Rocky cultivar, which record 26.66% of transformation frequency compared with 11.57% in Rocky cultivar.

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