Transgenic Plants: Gene Constructs, Vector and Transformation Method

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.

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In Vitro Regeneration and Genetic Transformation of Cucumis metuliferus through Cotyledon Organogenesis

HortScience ◽

10.21273/hortsci.46.4.616 ◽

2011 ◽

Vol 46 (4) ◽

pp. 616-621 ◽

Cited By ~ 4

Author(s):

Yu-Tsung Lin ◽

Chia-Wei Lin ◽

Chien-Hung Chung ◽

Mei-Hsiu Su ◽

Hsiu-Yin Ho ◽

...

Keyword(s):

Transgenic Plants ◽

In Vitro Regeneration ◽

Polymerase Chain Reaction Analysis ◽

Transformation Method ◽

Culture Conditions ◽

Watermelon Mosaic Virus ◽

Adventitious Buds ◽

Cotyledon Explants ◽

Shoot Primordia

This study was undertaken to establish for the first time an efficient regeneration and transformation system for Cucumis metuliferus line PI292190, which is the source of a well-defined resistant gene, Wmv, that provides resistance against Papaya ringspot virus type P (PRSV-P) and PRSV-W (formerly known as Watermelon mosaic virus 1, WMV-1). Different combinations of growth regulators were evaluated for the regeneration of cotyledon explants. Adventitious buds or shoot primordia were obtained within 3 to 4 weeks on regeneration medium. After shoot development, adventitious buds or shoot primordia were transferred to elongation medium for 3 to 4 weeks and these shoots were subcultured onto rooting medium for another 1 to 2 weeks. Under optimal culture conditions, a total of 7 to 10 weeks was necessary to obtain C. metuliferus plantlets from cotyledons. Furthermore, transgenic plants were successfully obtained using an Agrobacterium tumefaciens-mediated transformation method as shown by polymerase chain reaction analysis and histochemical β-glucuronidase (GUS) assay. A total of nine transgenic plants were developed from 360 cotyledon explants, giving a transformation frequency of 2.5%.

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Optimization of the parameters of Agrobacterium-mediated transformation of bar-shaped millet

BULLETIN of the L N Gumilyov Eurasian National University BIOSCIENCE Series ◽

10.32523/2616-7034-2020-133-4-32-43 ◽

2020 ◽

pp. 32-43

Author(s):

A.T. Zhumabek ◽

◽

Y.M. Ramankulov ◽

S.A. Manabayeva ◽

◽

...

Keyword(s):

Transgenic Plants ◽

Optical Density ◽

Panicum Virgatum ◽

Virulence Gene ◽

Transformation Method ◽

Bacterial Suspension ◽

Embryogenic Calli ◽

Agrobacterium Mediated Transformation ◽

Agrobacterial Transformation ◽

Panicum Virgatum L

An efficient Agrobacterium-mediated transformation method has been developed and optimized for switchgrass (Panicum virgatum L.). The aim of this study is to optimize the conditions for agrobacterial transformation of upland switchgrass cultivars Alamo, Forestburg, Pathfinder, Shawnee, and Trailblazer. For genetic transformation, the authors have used embryogenic calli of switchgrass cultivars. As a result of the study the effect of the virulence gene inducer, surfactant, optical density of bacterial suspension and co-cultivation time, it has been revealed that the presence of acetosyringone with silwet-L77 in the medium, bacterial suspension at an optical density of 0.6 and 8 days of co-cultivation with bacteria is optimal for Agrobacterium– mediated transformation of embryogenic calli of switchgrass. The results obtained can serve as a basis for the creation of transgenic plants of switchgrass.

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Transgenic Miscanthus lutarioriparius that co-expresses the Cry 2Aa# and Bar genes

Canadian Journal of Plant Science ◽

10.1139/cjps-2018-0168 ◽

2019 ◽

Vol 99 (6) ◽

pp. 841-851

Author(s):

Yanan Xia ◽

Jun Xu ◽

Junyi Duan ◽

Qingbo Liu ◽

Hongmei Huang ◽

...

Keyword(s):

Agrobacterium Tumefaciens ◽

Transgenic Plants ◽

Herbicide Tolerance ◽

Transformation Method ◽

Energy Crop ◽

Embryogenic Calli ◽

Transformation Protocol ◽

Agrobacterium Mediated Transformation ◽

Domestic Species ◽

Miscanthus Lutarioriparius

Miscanthus lutarioriparius, a domestic species of China, belongs to the genus of perennial rhizomatous Miscanthus and is a promising energy crop for biomass production. To establish an Agrobacterium tumefaciens transformation protocol and obtain transgenic plants co-expressing the Cry 2Aa# and Bar genes, an Agrobacterium-mediated transformation method for M. lutarioriparius was developed in this study. To inhibit tissue browning, optimized callus induction and regeneration media were developed. Embryogenic calli were efficiently induced from immature inflorescences on Murashige & Skoog (MS) medium containing 0.2 g L−1 citric acid. Agrobacterium tumefaciens EHA105 carrying pC3300 with the Cry 2Aa# and Bar genes were introduced into embryogenic calli. Six transgenic lines were selected by Basta screening and confirmed by genomic PCR. The herbicide tolerance of M. lutarioriparius transgenic plants was verified by spraying with glufosinate and expression of the Cry 2Aa# protein by ELISA. Therefore, an Agrobacterium-mediated transformation protocol for M. lutarioriparius was established and the co-expression of herbicide tolerance and Cry 2Aa# was successfully introduced with the system.

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Development of an efficient transformation method by Agrobacterium tumefaciens and high throughput spray assay to identify transgenic plants for woodland strawberry (Fragaria vesca) using NPTII selection

Plant Cell Reports ◽

10.1007/s00299-012-1366-1 ◽

2012 ◽

Vol 32 (3) ◽

pp. 329-337 ◽

Cited By ~ 6

Author(s):

Christopher J. Pantazis ◽

Sarah Fisk ◽

Kerri Mills ◽

Barry S. Flinn ◽

Vladimir Shulaev ◽

...

Keyword(s):

Agrobacterium Tumefaciens ◽

Transgenic Plants ◽

High Throughput ◽

Transformation Method ◽

Fragaria Vesca ◽

Efficient Transformation ◽

Woodland Strawberry

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Phenotypic Characterization of TransgenicMiscanthus sinensisPlants OverexpressingArabidopsisPhytochrome B

International Journal of Photoenergy ◽

10.1155/2014/501016 ◽

2014 ◽

Vol 2014 ◽

pp. 1-9 ◽

Cited By ~ 6

Author(s):

Ok-Jin Hwang ◽

Soo-Hyun Lim ◽

Yun-Jeong Han ◽

Ah-Young Shin ◽

Do-Soon Kim ◽

...

Keyword(s):

Transgenic Plants ◽

Herbicide Resistance ◽

Blot Analysis ◽

Red Light ◽

Transformation Method ◽

Phenotypic Characterization ◽

Selection Marker ◽

Miscanthus Sinensis ◽

Phytochrome B ◽

Agricultural Performance

Phytochromes are dimeric pigment proteins with reversible photochromism between red and far-red light-absorbing forms. They are photoreceptors that regulate various aspects of plant growth and development and have been used for biotechnological applications to improve agricultural performance of crops.Miscanthusspecies have been suggested as one of the most promising energy crops. In this paper,Arabidopsisphytochrome B(PHYB)gene was introduced intoMiscanthus sinensisusingAgrobacterium-mediated transformation method that we developed recently, with the herbicide resistance gene(BAR)as a selection marker. After putative transgenic plants were selected using the herbicide resistance assay, genomic integration of the transgene was confirmed by genomic PCR and Southern blot analysis, and transgene expression was validated by Northern blot analysis. Compared to nontransformed control plants, transgenic plants overexpressingPHYBshowed phenotypes with increased phytochrome B function, which includes increased chlorophyll content, decreased plant height, and delayed flowering. Therefore, these results suggest thatArabidopsisphytochrome B is functional inM. sinensisand provide a method to developMiscanthusvarieties with enhanced agricultural performance using phytochromes.

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