Genetic transformation of 9 in vitro clones of Alnus and Betula by Agrobacterium tumefaciens

1988 ◽  
Vol 7 (4) ◽  
pp. 229-232 ◽  
Author(s):  
John Mackay ◽  
Armand Séguin ◽  
Maurice Lalonde
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Genetic Transformation

scholarly journals Transformasi Genetik Tanaman Kentang (Solanum tuberosum L.) dengan Gen acvB Menggunakan Vektor Agrobacterium tumefaciens

2021 ◽  
Vol 11 (1) ◽  
pp. 63
Author(s):  
DARWIN SILALAHI ◽  
I GEDE PUTU WIRAWAN ◽  
MADE SRITAMIN
Keyword(s):  
Solanum Tuberosum ◽  
Agrobacterium Tumefaciens ◽  
Treatment Group ◽  
Genetic Transformation ◽  
Solanum Tuberosum L ◽  
Control Group ◽  
Shoot Height ◽  
Specific Pcr ◽  
Convenient Technique

Agrobacterium tumefaciens Mediated Genetic Transformation of acvB Gene in Potato (Solanum tuberosum L.). Genetic transformations are now routinely applied to plant mediated by Agrobacterium tumefaciens as the most convenient technique. This study aimed to prove the success of A. tumefaciens mediated genetic transformation in potato. A. tumefaciens LBA (pBI 121) and explant of potato shoot were used in this study. Explants were grown in vitro on Murashige and Skoog media. Transformation was implemented using smear technique by smearing A. tumefaciens to injured explant. Experimental groups consisted of two groups: control group which did not receive transformation treatment and treatment group receiving transformation treatment. Explant growth was observed through the presence of shoots, branches and the shoot height. Explants in the treatment group resulted in a higher number of shoots, branches, and shoot heights compared to control. Phenol compounds appear in explant epidermal tissue, indicating the wounds produced by A. tumefaciens infection, thus the gene predicted to be transformed. Identification by PCR is needed to prove the existence of the acvB gene in potato plants genome, using acvB specific PCR primer as the marker, such as (5?-CCCT CTAG AGAC CCGC GCCA AGGCG-3?) and (5?CGCG TCGA CCTT GTCG GAAAG -3?) with 540-bp in base pair size produced.

scholarly journals Genetic transformation of the Brazilian BR 451 maize variety by the Agrobacterium tumefaciens method

2017 ◽  
Vol 47 (11) ◽  
Author(s):  
Marilia Rodrigues de Silva ◽  
Dielli Aparecida Didoné ◽  
Cássia Canzi Ceccon ◽  
Vinícius de Oliveira Almeida ◽  
Magali Ferrari Grando
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Genetic Transformation ◽  
Growth Regulators ◽  
Hybrid Control ◽  
Stress Effect ◽  
Embryogenic Calli ◽  
Maize Variety ◽  
In Vitro Response ◽  
Regeneration In Vitro

ABSTRACT: The asexually gene introduction by genetic engineering has brought enormous possibilities to innovate plant breeding. However, principally because of the low in vitro response, genetic transformation has been restricted to only certain genotypes of agronomically significant species. With the objective of establishing a protocol for genetically transforming the Brazilian BR 451 maize variety through Agrobacterium tumefaciens, it was studied the capacity of plant regeneration in vitro from embryogenic calli cultivated in three regeneration media, each having different growth regulators. It was also evaluated the temperature stress effect on the transformation of the immature embryos with A. tumefaciens EHA 101 containing the plasmid pTF102 with uidA and bar genes. The BR 451 variety embryos and those of the Hi-II hybrid (control) were exposed to three treatments applied as they were being infected with the agrobacteria (a) infection at 25°C; (b) infection at 40°C; (c) pretreatment at 40°C for 5 seconds followed by infection at 25°C. Transformation was determined by uidA gene expression and through the callus resistant to the herbicide Bialaphos® formation. Embryos infected at 40°C showed a higher degree of genetic transformation in the Hi-II, although the same was not noted in BR 451. When growth regulators were added to the culture medium the number of regenerated BR 451 plants showed no increase.

In vitro regeneration and Agrobacterium tumefaciens-mediated genetic transformation of D. lotus ( Diospyros lotus L.)

2018 ◽  
Vol 236 ◽  
pp. 229-237 ◽  
Author(s):  
Xuehan Li ◽  
Zhenying Jiang ◽  
Yanying Shen ◽  
Feihong Li ◽  
Xinyi Yu ◽  
...  
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Genetic Transformation ◽  
In Vitro Regeneration ◽  
Diospyros Lotus

scholarly journals Genetic transformation of apical meristematic shoots in the banana cultivar ‘Williams’

Bionatura ◽  
2021 ◽  
Vol 6 (1) ◽  
pp. 1462-1465
Author(s):  
Liliana Villao ◽  
José Flores ◽  
Efrén Santos-Ordóñez
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Genetic Transformation ◽  
Export Market ◽  
Luciferase Reporter ◽  
Luciferase Reporter Gene ◽  
Meristematic Tissue ◽  
Embryogenic Cell ◽  
In Vitro Plants

Bananas and plantains (Musa spp.) are among the most critical socioeconomic crops globally, being a staple food for millions of people in the tropics and an essential component for the export market, including the subtropics. Besides conventional breeding, genetic improvement of bananas and plantains could be performed through genetic engineering and new breeding techniques. Furthermore, plant tissue culture is essential for these technologies, including developing embryogenic cell suspensions and in vitro plants. The transient and stable genetic transformation could be performed from in vitro plants, shortening Musa transgenic lines development compared to genetic transformation while using embryogenic cell suspension. In this study, a genetic transformation protocol was established from banana apical meristems for the ‘Williams’ cultivar (genotype AAA). The protocol was based on the co-cultivation of the explants (whole in vitro plants or bisected meristematic tissues derived from in vitro plants) with Agrobacterium tumefaciens strain LBA4404 harboring two binary vectors denominated pLVCIBE1 (cassette: MabHIPP promoter::luc2::Tnos, P35S::hpt::Tnos) and pLVCIBE2 (cassette: P35S::luc2::Tnos, P35S::hpt::Tnos), independently. The stable genetic transformation was obtained by subculturing in vitro banana plants in selection medium (12.5µg/mL of hygromycin) for 8 weeks from bisected meristematic tissue transformation. Genetic transformation was confirmed in vivo with the use of the luciferase reporter gene system. Furthermore, PCR was performed on DNA extracted from leaves of regenerated transgenic in vitro plants after 8 weeks of selection, confirming stable genetic transformation. Therefore, genetic transformation was achieved in the apical meristematic tissue of in vitro banana plants with co-cultivation of Agrobacterium tumefaciens.

scholarly journals Agrobacterium tumefaciens mediated transformation of the aquatic carnivorous plant Utricularia gibba

2020 ◽  
Author(s):  
Araceli Oropeza-Aburto ◽  
Sergio Alan Cervantes-Perez ◽  
Victor A Albert ◽  
Luis Rafael Herrera-Estrella
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Genetic Transformation ◽  
In Vitro Selection ◽  
Carnivorous Plant ◽  
Carnivorous Plants ◽  
35S Promoter ◽  
Transformation Protocol ◽  
Utricularia Gibba

Abstract Background The genus Utricularia belongs to Lentibulariaceae, the largest family of carnivorous plants, which includes terrestrial, epiphytic and aquatic species. The development of specialized structures that evolved for carnivory is a feature of this genus that has been of great interest to biologists since Darwin‘s early studies. Utricularia gibba is itself an aquatic plant with sophisticated bladder traps having one of the most complex suction mechanisms for trapping prey. However, the molecular characterization of the mechanisms that regulate trap development and the biophysical processes involved in prey trapping are still largely unknown due to the lack of a simple and reproducible gene transfer system. Results Here, we report the establishment of a simple, fast and reproducible protocol for genetic transformation of U. gibba based on the T-DNA of Agrobacterium tumefaciens . An in vitro selection system using Phosphinotricin as a selective agent was established for U. gibba . Plant transformation was confirmed by histochemical GUS assays and PCR and qRT-PCR analyses. We report on the expression pattern of the 35S promoter and of the promoter of a trap-specific ribonuclease gene in transgenic U. gibba plants. Conclusions The genetic transformation protocol reported here is an effective method for studying developmental biology and functional genomics of this genus of carnivorous plants and advances the utility of U. gibba as a model system to study developmental processes involved in trap formation.

scholarly journals An Efficient Method for the Genetic Transformation of Acmella oleracea L. (Spilanthes acmella Linn.) with Agrobacterium tumefaciens

Plants ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 198
Author(s):  
Valentina Maggini ◽  
Priscilla Bettini ◽  
Fabio Firenzuoli ◽  
Patrizia Bogani
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Secondary Metabolites ◽  
Genetic Transformation ◽  
Bioactive Compound ◽  
Raw Material ◽  
Nptii Gene ◽  
Spilanthes Acmella ◽  
In Vitro Plant ◽  
High Level

Acmella oleracea L. is an important medicinal plant, commonly known as the toothache plant. It is a rich source of secondary metabolites used for the treatment of different human disorders. The demand for Acmella oleracea L. has increased due to its putative health benefits (in terms of both biomass quantity and bioactive compound purification). In vitro plant cultures have allowed the rapid increase of raw material availability through the use of suitable regeneration and multiplication systems. On the other hand, there is a general lack of methods for Acmella genetic transformation as a promising new technological approach for the improvement of secondary metabolites. In this work, an efficient transformation protocol has been established using the Agrobacterium tumefaciens LBA4404 strain bearing the binary vector pBI121 containing the NPTII gene for the resistance to kanamycin. Plant genetic transformation has been verified by direct polymerase chain reaction and GUS assay on regenerants. Transformation efficiency has been affected by the high level of the selection agent kanamycin. To our knowledge, this is the first report on the genetic transformation of A. oleracea, paving the way to further studies to improve in vitro plant growth and secondary metabolite production.

scholarly journals Genetic transformation of Paulownia elongata S. Y. Hu., mediated by Agrobacterium tumefaciens and biolistic system

2021 ◽  
Vol 14 (2) ◽  
Author(s):  
Carlos R. Castillo-Martínez ◽  
Ma. Alejandra Gutiérrez-Espinosa ◽  
Jorge Cadena-Iñiguez ◽  
Marco T. Buenrostro-Nava ◽  
VALERIA MARTÍNEZ SIAS
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Genetic Transformation ◽  
Pcr Analysis ◽  
Optimum Dose ◽  
Indirect Organogenesis ◽  
Gus Reporter ◽  
Selection Gene ◽  
Transformation Systems ◽  
Paulownia Elongata

Objective: The most appropriate conditions for genetic transformation through direct (bioballistic) and indirect (Agrobacterium tumefaciens) transformation systems in Paulownia elongata were established. Design/methodology/approach: Starting from in vitro propagation through both direct and indirect organogenesis, internodal stem segments with 0.5 to 1 cm length were determined as the best explant. The optimum dose for selection media was determined to be 15 mg L-1 of kanamycin. It was possible to obtain transgenic plants under both transformation systems. In the case of Agrobacterium tumefaciens, two hours of incubation, 48 h of co-cultivation, and optical density of 0.9 were used; while for bioballistics, the best conditions were 120 PSI of shot pressure, shot height at level 6 (16 cm), and vacuum pressure of 22 Hg mm, with particle inflow gun system (PIG). Results: Both systems produced complete transformants, chimeras, as well as those confirmed by histochemical X-GLUC and PCR analysis, producing a total of 14 positive plants by A. tumefaciens transformation from 26 trials and ten positive plants by the bioballistic system from 30 trials; a construction with chitinase and glucanase, NPT II selection gene and the GUS reporter gene were used. Findings/conclusions: So far, this has been the first report including integration of chitinase and glucanase genes.

scholarly journals Agrobacterium tumefaciens mediated transformation of the aquatic carnivorous plant Utricularia gibba

2020 ◽  
Author(s):  
Araceli Oropeza-Aburto ◽  
Sergio Alan Cervantes-Perez ◽  
Victor A Albert ◽  
Luis Rafael Herrera-Estrella
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Genetic Transformation ◽  
In Vitro Selection ◽  
Carnivorous Plant ◽  
Carnivorous Plants ◽  
35S Promoter ◽  
Transformation Protocol ◽  
Utricularia Gibba

Abstract Background The genus Utricularia belongs to Lentibulariaceae, the largest family of carnivorous plants, which includes terrestrial, epiphytic and aquatic species. The development of specialized structures that evolved for carnivory is a feature of this genus that has been of great interest to biologists since Darwin‘s early studies. Utricularia gibba is itself an aquatic plant with sophisticated bladder traps having one of the most complex suction mechanisms for trapping prey. However, the molecular characterization of the mechanisms that regulate trap development and the biophysical processes involved in prey trapping are still largely unknown due to the lack of a simple and reproducible gene transfer system. Results Here, we report the establishment of a simple, fast and reproducible protocol for genetic transformation of U. gibba based on the T-DNA of Agrobacterium tumefaciens . An in vitro selection system using Phosphinotricin as a selective agent was established for U. gibba . Plant transformation was confirmed by histochemical GUS assays and PCR and qRT-PCR analyses. We report on the expression pattern of the 35S promoter and of the promoter of a trap-specific ribonuclease gene in transgenic U. gibba plants. Conclusions The genetic transformation protocol reported here is an effective method for studying developmental biology and functional genomics of this genus of carnivorous plants and advances the utility of U. gibba as a model system to study developmental processes involved in trap formation.

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