Enhancement of Agrobacterium-Mediated Transformation Frequency in Potato Slices

2008 ◽  
Vol 51 (4) ◽  
pp. 342-343 ◽  
Author(s):  
Dong-Il Shin
Keyword(s):  
Transformation Frequency ◽  
Agrobacterium Mediated Transformation ◽  
Potato Slices

scholarly journals Improvement of Soybean Agrobacterium-Mediated Transformation Efficiency by Adding Glutamine and Asparagine into the Culture Media

2018 ◽  
Vol 19 (10) ◽  
pp. 3039 ◽  
Author(s):  
Li Chen ◽  
Yupeng Cai ◽  
Xiujie Liu ◽  
Weiwei Yao ◽  
Chen Guo ◽  
...  
Keyword(s):  
Culture Media ◽  
Genetically Modified ◽  
Global Scale ◽  
Transformation Frequency ◽  
Genetically Modified Soybean ◽  
Soybean Transformation ◽  
Agrobacterium Mediated Transformation ◽  
Key Factor ◽  
Probable Role ◽  
Better Than

As a genetically modified crop, transgenic soybean occupies the largest global scale with its food, nutritional, industrial, and pharmaceutical uses.Efficient transformation is a key factor for the improvement of genetically modified soybean. At present, the Agrobacterium-mediated method is primarily used for soybean transformation, but the efficiency of this method is still relatively low (below 5%) compared with rice (above 90%). In this study, we examined the influence of l-glutamine and/or l-asparagine on Agrobacterium-mediated transformation in soybean and explored the probable role in the process of Agrobacterium-mediated transformation. The results showed that when the amino acids l-glutamine and l-asparagine were added separately or together to the culture medium, the shoot induction frequency, elongation rate, and transformation frequency were improved. The combined effects of l-glutamine and l-asparagine were better than those of l-glutamine and l-asparagine alone. The 50 mg/L l-glutamine and 50 mg/L l-asparagine together can enhance the transformation frequency of soybean by attenuating the expression level of GmPRs (GmPR1, GmPR4, GmPR5, and GmPR10) and suppression of the plant defense response. The transgene was successfully transmitted to the T1 generation. This study will be useful in genetic engineering of soybean.

scholarly journals Agrobacterium-mediated transformation of potato with human lactoferrin gene

2019 ◽  
Vol 25 ◽  
pp. 184-189
Author(s):  
A. Yu. Buziashvili ◽  
A. I. Yemets
Keyword(s):  
Fungal Pathogens ◽  
Transgenic Potato ◽  
Transformation Frequency ◽  
Potato Cultivars ◽  
Human Lactoferrin ◽  
Chain Reaction ◽  
Agrobacterium Mediated Transformation ◽  
Lactoferrin Gene ◽  
Polymerase Chain ◽  
Resistance To Phytopathogens

Aim. Obtaining of the new Ukrainian potato cultivars with complex resistance to phytopathogens. Methods. Agrobacterium-mediated transformation of potato cultivars Vernisage and Zarevo with human lactoferrin gene possessing antimicrobial activity was carried out. For transformation, supervirulent A. tumefaciens strain EHA 105 with hLf gene and gene for the resistance to kanamycine was used. Selection was carried out for 3 months on MSK-S1 and MSK-S2 media supplemented with 100 mg/l kanamycine. Integration of lactoferrin gene in transgenic plant lines was confirmed by polymerase chain reaction with specific for hLf gene primers. Results. After selection, potato lines with the resistance to kanamycine were obtained. The transgenic potato lines with integrated into their genomes hLf gene were obtained. Integration of lactoferrin gene was confirmed for 1 line of Zarevo cultivar and 3 lines of Vernisage cultivar. Transformation frequency was of 6.8% for cv. Vernisage and 6.25% for cv. Zarevo. Conclusions. Selected potato lines would be used in further studies on their resistance to bacterial and fungal pathogens. Keywords: human lactoferrin gene hLf, Solanum tuberosum, Agrobacterium-mediated transformation, transgenic plants.

scholarly journals Sonication, Vacuum Infiltration and Thiol Compounds Enhance the Agrobacterium-Mediated Transformation Frequency of Withania somnifera (L.) Dunal

PLoS ONE ◽  
2015 ◽  
Vol 10 (4) ◽  
pp. e0124693 ◽  
Author(s):  
Ganeshan Sivanandhan ◽  
Gnajothi Kapil Dev ◽  
Jeevaraj Theboral ◽  
Natesan Selvaraj ◽  
Andy Ganapathi ◽  
...  
Keyword(s):  
Withania Somnifera ◽  
Transformation Frequency ◽  
Vacuum Infiltration ◽  
Thiol Compounds ◽  
Agrobacterium Mediated Transformation

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.

scholarly journals Perturbation of H3K27me3-Associated Epigenetic Processes Increases Agrobacterium-Mediated Transformation

2017 ◽  
Vol 30 (1) ◽  
pp. 35-44 ◽  
Author(s):  
Hidekazu Iwakawa ◽  
Benjamin C. Carter ◽  
Brett C. Bishop ◽  
Joe Ogas ◽  
Stanton B. Gelvin
Keyword(s):  
Gene Expression ◽  
Critical Role ◽  
Histone H3 ◽  
Transformation Frequency ◽  
Agrobacterium Mediated Transformation ◽  
Core Technology ◽  
Basic Plant ◽  
Lysine Residues ◽  
Novel Strategy

Agrobacterium-mediated transformation is a core technology for basic plant science and agricultural biotechnology. Improving transformation frequency is a major goal for plant transgenesis. We previously showed that T-DNA insertions in some histone genes decreased transformation susceptibility, whereas overexpression of several Arabidopsis H2A and H4 isoforms increased transformation. Overexpression of several histone H2B and H3 isoforms had little effect on transformation frequency. However, overexpression of histone H3-11 (HTR11) enhanced transformation. HTR11 is a unique H3 variant that lacks lysine at positions 9 and 27. The modification status of these lysine residues in canonical H3 proteins plays a critical role in epigenetic determination of gene expression. We mutated histone H3-4 (HTR4), a canonical H3.3 protein that does not increase transformation when overexpressed, by replacing either or both K9 and K27 with the amino acids in HTR11 (either K9I, K27Q, or both). Overexpression of HTR4 with the K27Q but not the K9I substitution enhanced transformation. HTR4K27Q was incorporated into chromatin, and HTR4K27Q overexpression lines exhibited deregulated expression of H3K27me3-enriched genes. These results demonstrate that mutation of K27 in H3.3 is sufficient to perturb H3K27me3-dependent expression in plants as in animals and suggest a distinct epigenetic role for histone HTR11. Further, these observations implicate manipulation of H3K27me3-dependent gene expression as a novel strategy to increase transformation susceptibility.

scholarly journals The frequency of T-DNA integration during the genetic transformation of tobacco (Nicotiana tabacum L.)

2018 ◽  
Vol 22 ◽  
pp. 262-266
Author(s):  
A. G. Komisarenko ◽  
S. I. Mykhalska ◽  
O. O. Khrystan
Keyword(s):  
Nicotiana Tabacum ◽  
Transgenic Plants ◽  
Transformation Frequency ◽  
Tobacco Plants ◽  
Double Stranded Rna ◽  
Dna Integration ◽  
Agrobacterium Mediated Transformation ◽  
Nicotiana Tabacum L ◽  
Is Integration

Aim. Investigation of the is integration frequency of the genes and their components (elements of genetic constructions) in transgenic tobacco-regenerating plants. Methods. Agrobacterium-mediated transformation of tobacco (Nicotiana tabacum L.) in vitro. Results. There were obtained tobacco plants with integrated target and selective genes. It was found that a complete copy of T-DNA or its certain elements were embedded. Conclusions. The possibility of the integration of complete and incomplete DNA copies during in vitro Agrobacterium-mediated transformation of the Nicotiana tabacum were established. This event were observed both during manipulations with pBi2E plasmid with the double-stranded RNA PHD gene suppressor and pBi-OAT plasmid with OAT gene. The transformation frequency with the complete integration of the genetic construction was 91 %. The integration frequencies for various plasmids were: LBA4404 pBi2E – 78 %; AGLO pBi2E – 100 %; AGLO pBi-OAT – 94 %. Keywords: Nicotiana tabacum L., Agrobacterium-mediated transformation in vitro, transgenic plants, T-DNA.

Agrobacterium-Mediated Transformation Of Plant Cells

2018 ◽  
Author(s):  
Christine Kronfoth ◽  
◽  
Peter Grayson ◽  
Keyword(s):  
Plant Cells ◽  
Agrobacterium Mediated Transformation

scholarly journals Optimizing Transformation Frequency of Cryptococcus neoformans and Cryptococcus gattii Using Agrobacterium tumefaciens

2021 ◽  
Vol 7 (7) ◽  
pp. 520
Author(s):  
Jianmin Fu ◽  
Nohelli E. Brockman ◽  
Brian L. Wickes
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Yeast Cell ◽  
Transformation Efficiency ◽  
Transformation Frequency ◽  
Yeast Cells ◽  
Agar Concentration ◽  
Genetic Tool ◽  
Number Of Factors ◽  
Cryptococcus Spp ◽  
Transformation Systems

The transformation of Cryptococcus spp. by Agrobacterium tumefaciens has proven to be a useful genetic tool. A number of factors affect transformation frequency. These factors include acetosyringone concentration, bacterial cell to yeast cell ratio, cell wall damage, and agar concentration. Agar concentration was found to have a significant effect on the transformant number as transformants increased with agar concentration across all four serotypes. When infection time points were tested, higher agar concentrations were found to result in an earlier transfer of the Ti-plasmid to the yeast cell, with the earliest transformant appearing two h after A. tumefaciens contact with yeast cells. These results demonstrate that A. tumefaciens transformation efficiency can be affected by a variety of factors and continued investigation of these factors can lead to improvements in specific A. tumefaciens/fungus transformation systems.

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