A PRELIMINARY STUDY ON GENOTYPE-INDEPENDENT AGROBACTERIUM-MEDIATED TRANSFORMATION METHOD TO OBTAIN GENETICALLY ENGINEERED APRICOT PLANTS

The primary problem in producing fuel ethanol through microorganism fermentation with lignocellulose is the strain. We constructed a URA3-directed low copy integration-expression plasmid pZMYBX1 and rDNA-directed high copy integration-expression plasmid pZMYX2. Using the lithium acetate transformation method, we co-transformed the linearized plasmid pZMYBX1 (StuI) and pZMYX2 (HpaI) into theS. cerevisiaecells. Ultimately, we obtain three recombinants: HDY-ZMYWBG1, HDY-ZMYWBG2 and HDY-ZMYWBG3. The ethanol yield for HDY-ZMYWBG1 and HDY-ZMYWBG3 are 0.368 g/g and 0.365 g/g, respectively, which are higher than the 0.330 g/g yield for W5. This findings show that the xylose metabolic pathway could be introduced into theS. cerevisiaeto produce an alternative strain for the production of biological ethanol from lignocellulose substrate.

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Evaluation of transgenic canola plants under field conditions

Genome ◽

10.1139/g92-010 ◽

1992 ◽

Vol 35 (1) ◽

pp. 58-63 ◽

Cited By ~ 14

Author(s):

M. Arnoldo ◽

C. L. Baszczynski ◽

G. Bellemare ◽

G. Brown ◽

J. Carlson ◽

...

Keyword(s):

Kanamycin Resistance ◽

Genetically Engineered ◽

Field Conditions ◽

Enzyme Assays ◽

Nptii Gene ◽

Neomycin Phosphotransferase ◽

Agrobacterium Mediated Transformation ◽

Transgenic Canola ◽

Progeny Analysis ◽

Transformation Vectors

Eleven independent transgenic canola (Brassica napus ssp. oleifera L. cv. Westar and Regent) lines were evaluated in the field. The plants carried a neomycin phosphotransferase (NPTII) gene for kanamycin resistance that was introduced via Agrobacterium-mediated transformation. NPTII enzyme assays, Southern blot hybridizations and progeny analysis, confirmed the stable, heritable integration and expression of the introduced NPTII gene. A number of agronomic characteristics evaluated under field conditions, including maturity, yield, and oil and protein content, were all statistically comparable between the transformed and nontransformed plants. These results indicate that canola can be genetically engineered successfully, and that the Agrobacterium-based transformation system employed does not induce any adverse effects on the intrinsic agronomic and qualitative traits critical to the agricultural industry.Key words: transgenic field trial, canola, Agrobacterium-mediated transformation, vectors.

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Tissue culture-based Agrobacterium-mediated and in planta transformation methods

Czech Journal of Genetics and Plant Breeding ◽

10.17221/177/2016-cjgpb ◽

2017 ◽

Vol 53 (No. 4) ◽

pp. 133-143 ◽

Cited By ~ 8

Author(s):

M. Niazian ◽

S.A. Sadat Noori ◽

P. Galuszka ◽

S.M.M. Mortazavian

Keyword(s):

Tissue Culture ◽

Transformation Method ◽

Gene Transformation ◽

In Planta Transformation ◽

In Planta ◽

Agrobacterium Mediated Transformation ◽

Advantages And Disadvantages ◽

Culture Independent ◽

Transformation Methods ◽

Mutant Lines

Gene transformation can be done in direct and indirect (Agrobacterium-mediated) ways. The most efficient method of gene transformation to date is Agrobacterium-mediated method. The main problem of Agrobacterium-method is that some plant species and mutant lines are recalcitrant to regeneration. Requirements for sterile conditions for plant regeneration are another problem of Agrobacterium-mediated transformation. Development of genotype-independent gene transformation method is of great interest in many plants. Some tissue culture-independent Agrobacterium-mediated gene transformation methods are reported in individual plants and crops. Generally, these methods are called in planta gene transformation. In planta transformation methods are free from somaclonal variation and easier, quicker, and simpler than tissue culture-based transformation methods. Vacuum infiltration, injection of Agrobacterium culture to plant tissues, pollen-tube pathway, floral dip and floral spray are the main methods of in planta transformation. Each of these methods has its own advantages and disadvantages. Simplicity and reliability are the primary reasons for the popularity of the in planta methods. These methods are much quicker than regular tissue culture-based Agrobacterium-mediated gene transformation and success can be achieved by non-experts. In the present review, we highlight all methods of in planta transformation comparing them with regular tissue culture-based Agrobacterium-mediated transformation methods and then recently successful transformations using these methods are presented.

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Reconstruction and validation of three different binary vectors suitable for generation of genetically engineered Helicoverpa protected crops

Indian Journal of Genetics and Plant Breeding (The) ◽

10.31742/ijgpb.79.1.15 ◽

2019 ◽

Vol 79 (01) ◽

Author(s):

Nayana Hazarika ◽

Rashmi Rekha Boruah ◽

Pratap Jyoti Handique ◽

Sumita Acharjee ◽

Bidyut Kumar Sarmah

Keyword(s):

Transgenic Crops ◽

Transformation Method ◽

Small Subunit ◽

Genetically Engineered ◽

Agrobacterium Strain ◽

Binary Vectors ◽

Tobacco Leaves ◽

Cry1ac Gene ◽

Assembly Method ◽

Cry1ac Protein

Availability of a suitable plant transformation binary vector is necessary for the generation of transgenic crops with an adequate expression of transgenic proteins. Therefore, three binary vectors were constructed viz., pBK204, pBK205, and pBK206 harboring either a truncated or a full-length version of a Cry1Ac gene for the generation of Helicoverpa protected crops. Two different promoters viz., Arabidopsis Rubisco small subunit (AtSSU) gene promoter or CaMV35S promoters were used to regulate the various versions of Cry1Ac gene. The binary vectors were reconstructed either by the Gibson assembly method and others by ligating the restriction enzyme digested fragments. The reconstructed binary vectors were mobilized into Agrobacterium strain AGL1 and validated by Agrobacterium infiltration assays of Nicotiana benthamiana. The amount of Cry1Ac protein accumulated in the Agroinfiltrated tobacco leaves was quantified using the quantitative ELISA assay. The expression of the Cry1Ac protein in the tobacco leaves ranged from 0.25 to 0.26 µg /g fresh weight (FW) when transformed with these three constructs. Thus, the vectors constructed in this study appeared to be suitable for generation of Helicoverpa resistant transgenic crops by Agrobacterium-mediated genetic transformation method.

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

Faktori eksperimental'noi evolucii organizmiv ◽

10.7124/feeo.v22.964 ◽

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.

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An efficient Agrobacterium-mediated transformation method using hypocotyl as explants for Brassica napus

Molecular Breeding ◽

10.1007/s11032-020-01174-0 ◽

2020 ◽

Vol 40 (10) ◽

Author(s):

Cheng Dai ◽

Yuqing Li ◽

Long Li ◽

Zhuolin Du ◽

Shengli Lin ◽

...

Keyword(s):

Brassica Napus ◽

Transformation Method ◽

Agrobacterium Mediated Transformation

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Development of an Agrobacterium-mediated transformation method for Taxus suspension cultures

In Vitro Cellular & Developmental Biology - Plant ◽

10.1007/s11627-017-9876-8 ◽

2018 ◽

Vol 54 (1) ◽

pp. 36-44 ◽

Cited By ~ 3

Author(s):

Sarah A. Wilson ◽

Patricia Keen ◽

Michelle C. McKee ◽

Nicole Raia ◽

Joyce Van Eck ◽

...

Keyword(s):

Transformation Method ◽

Suspension Cultures ◽

Agrobacterium Mediated Transformation

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Effects of Medium Composition and Genetic Background on Agrobacterium-Mediated Transformation Efficiency of Lentinula edodes

Genes ◽

10.3390/genes10060467 ◽

2019 ◽

Vol 10 (6) ◽

pp. 467 ◽

Cited By ~ 2

Author(s):

Lianlian Yan ◽

Ruiping Xu ◽

Yan Zhou ◽

Yuhua Gong ◽

Shenghong Dai ◽

...

Keyword(s):

Filamentous Fungi ◽

Lentinula Edodes ◽

Transformation Efficiency ◽

Edible Mushroom ◽

Transformation Method ◽

Medium Composition ◽

Agrobacterium Mediated Transformation ◽

Genomics Research ◽

Wild Strains ◽

Positive Frequency

The establishment of genetic transformation method is crucial for the functional genomics research in filamentous fungi. Although the transformation method has been developed in several types of fungi, a highly efficient and convenient transformation system is desperately needed in Lentinula edodes. Present work established the Agrobacterium-mediated transformation (ATMT) of basidiomycete L. edodes in both monokaryon and dikaryon mycelia by using constructed binary plasmid pCAMBIA-1300-GFP. Then, the transformation efficiency of ATMT was evaluated by using different mediums for recipient incubation and different varieties of L. edodes. The results showed that in dikaryon strain W1, the positive hygromycin-resistant transformants was observed in all medium with the positive frequency of selected transformants that ranged from 0 to 30%. While in the monokaryon strain W1-26, only the millet medium group obtained positive transformants with a positive frequency of 75.48%. Moreover, three dikaryotic wild strains (YS55, YS3334, and YS3357) and two dikaryotic cultivated strains (W1 and S606) showed the highest transformation efficiency, with 32.96% of the germination frequency, and 85.12% of positive frequency for hygromycin-resistant transformants. This work demonstrated that Agrobacterium-mediated transformation was successfully performed in L. edodes, and the genotype of recipients as well as the medium for mycelial incubation were suggested to play key roles in determining the transformation efficiency. These findings may provide new avenues for the genetic modification of edible mushroom and may extend the cognition of DNA-mediated transformation in filamentous fungi.

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