The effect of the parameters of biolistic transformation of spring barley (Hordeum vulgare L.) on the level of transient expression of GFP reporter gene

Chernobrovkina, Mariya; Sidorov, E.; Baranov, I.A.; Харченко, П. Н.; Dolgov, Sergey

doi:10.1134/s1062359007060040

Cited by 5 publications

(4 citation statements)

References 28 publications

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“…In addition, several conditions affect stable transformation in biolistics, namely: mechanical damage, toxicity, and the adjustment of pressure, distance of flight, and type of particles. A balance between minor damage caused by particle bombardment and a high number of cells transiently expressing the transgene is desirable (Chernobrovkina et al 2007 ). Our results are not a product of optimization, but rather arise from the search for best particle bombardment conditions for nopal that pursuit high foci formation levels to ensure in part, a much higher number of stably transformed cells.…”

Section: Discussionmentioning

confidence: 99%

Factors affecting genetic transformation by particle bombardment of the prickly pear cactus (O. ficus-indica)

2019

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In the present study, a novel transformation protocol for Opuntia ficus-indica was generated by means of particle bombardment. The best conditions obtained were: 900 psi rupture disk pressure, 8 cm microprojectile travel distance, and 4 h of exposition to 0.2 M mannitol. For all experiments, gold particles coated with 1.0 µg/µL of pBI426 plasmid DNA were used. With all these conditions, a 23% of transformation efficiency in terms of regeneration in selection media (100 mg/L kanamycin) was obtained. Interestingly, the presence of both transgenes: nptII and uidA, by means of PCR and RT-PCR assays was detected. The regeneration percentage achieved in terms of stable integration for both genes was 10%. In addition, we also detected adequate amounts of β-glucuronidase activity by means of the GUS fluorometric assay. The procedure described in the present investigation reveals the feasibility of using nopal for the introduction, expression, and possible production of heterologous proteins.

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Section: Discussionmentioning

confidence: 99%

Factors affecting genetic transformation by particle bombardment of the prickly pear cactus (O. ficus-indica)

2019

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“…This distance is necessary for allowing the even spread of the DNA microcarrier onto the target tissue without causing damage to the tissues. 28,27,29 A highest GFP expression average of 142 spots (Fig. 1D) was observed when a 6-cm distance (Fig.…”

Section: Effect Of Distancementioning

confidence: 95%

Particle bombardment – mediated gene transfer and GFP transient expression in Setaria viridis

Mookkan

2018

Plant Signaling & Behavior

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Setaria viridis is one of the most important model grasses in studying monocot plant biology. Transient gene expression study is a very important tool in plant biotechnology, functional genomics, and CRISPR-Cas9 genome editing technology via particle bombardment. In this study, a particle bombardment-mediated protocol was developed to introduce DNA into Setaria viridis in vitro leaf explants. In addition, physical and biological parameters, such as helium pressure, distance from stopping screen to the target tissues, DNA concentration, and number of bombardments, were tested and optimized. Optimum concentration of transient GFP expression was achieved using 1.5 ug plasmid DNA with 0.6 mm gold particles and 6 cm bombardment distance, using 1,100 psi. Doubling the bombardment instances provides the maximum number of foci of transient GFP expression. This simple protocol will be helpful for genomics studies in the S. viridis monocot model.

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“…In principle, Agrobacterium-mediated transformation and particle bombardment are two transformation methods for most plants. Because of limitations and disadvantages of each method (4,(25)(26)(27), that vary even for different genotypes of one species, the existing of alternative methods can be helpful for improving molecular breeding programs. Agrobacteriummediated transformation has been previously described for African violet (1, 2); however, to the best of our knowledge, biolistic transformation method has not been reported yet.…”

Section: Discussionmentioning

confidence: 99%

Stable Transformation of the Saintpaulia ionantha by Particle Bombardment

Ghorbanzade

Ahmadabadi

2015

Iran J Biotech

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Background: A highly efficient genetic transformation system is essential for a successful genetic manipulation of the African violet (Saintpaulia ionantha Wendl.). Objectives: Developing a particle bombardment-based genetic transformation system for the African violet. Materials and Methods: A local cultivar of the African violet from Guilan province was used for transformation experiments. The pFF19G and pBin61-Ech42 vectors were used for transient and stable transformation experiments, respectively. The PCR and RT-PCR techniques were used to verify transgene presence and transcript levels in candidate transgenic lines, respectively. Results: Using leaf explants as target tissues, we transferred an endochitinase gene cDNA into African violet. Transgenic plants were regenerated on selection medium at a reasonable frequency (in average, one stable transgenic line per shot). Molecular analysis of transgenic plants by PCR and RT-PCR techniques confirmed successful integration and expression of transgene in several independent transgenic lines. Conclusions: Our results provide an efficient stable transformation system for genetic transformation of African violet.

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The effect of the parameters of biolistic transformation of spring barley (Hordeum vulgare L.) on the level of transient expression of GFP reporter gene

Cited by 5 publications

References 28 publications

Factors affecting genetic transformation by particle bombardment of the prickly pear cactus (O. ficus-indica)

Factors affecting genetic transformation by particle bombardment of the prickly pear cactus (O. ficus-indica)

Particle bombardment – mediated gene transfer and GFP transient expression in Setaria viridis

Stable Transformation of the Saintpaulia ionantha by Particle Bombardment

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