Genetic Transformation of Banana and Plantain (Musa spp.) via Particle Bombardment

Sági, László; Panis, Bart; Remy, Serge; Schoofs, H.; Smet, Karen De; Swennen, Rony; Cammue, Bruno P. A.

doi:10.1038/nbt0595-481

Cited by 150 publications

(61 citation statements)

References 28 publications

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“…Agrobacterium-mediated transformation has been the most accessible tool with which to introduce heterologous DNA and manipulate gene expression in plants (Ribas et al 2011). Therefore, a reliable transformation and regeneration system with a low degree of chimerism and high regeneration frequency is a prerequisite (May et al 1995;Sági et al 1995). There are several factors that affect the transformation efficiency, such as Agrobacterium density, infection time, co-cultivation duration and conditions (Yong et al 2006;Vrbová et al 2013), and temperature (Li et al 2003).…”

Section: Introductionmentioning

confidence: 99%

Stable integration of mgfp5 transgenes following Agrobacterium-mediated transformation in Boesenbergia rotunda cell suspension culture

Wong

Zoolkefli

Karim

et al. 2015

Frontiers in Life Science

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Boesenbergia rotunda, a herb in the ginger family, contains numerous beneficial compounds, such as flavonoids, flavones and cyclohexenyl chalcone derivatives, that have great potential for pharmaceutical applications. However, the low concentration of the bioactive compounds limits their commercial application. In this study, a simple and reliable Agrobacterium-mediated transformation protocol for B. rotunda cell suspension culture was successfully developed. The minimal inhibitory concentration and natural tolerance of the selective agent, hygromycin, against the cells were 20 mg l −l and 30 mg l −l in liquid media and solid media, respectively. The highest number of transformed regenerants (18 ± 0.00 per ml settled cell volume) was recorded when cells were infected with Agrobacterium tumefaciens harbouring pCAMBIA1304 for 10 min and co-cultivated for 2 days. Prolonged infection time ( > 10 min) and co-cultivation period ( > 2 days), however, did not increase the transformation efficiencies. The results clearly show that infection and co-cultivation periods strongly influenced the transformation efficiency in ginger. The transformed cells were recovered and showed green fluorescent signals under ultraviolet excitation. An intense blue colour was observed in the transformed cells after β-glucuronidase (GUS) histochemical staining, further confirming the functionality of the GUS enzymes in the regenerants. Polymerase chain reaction analysis of 3-, 6-, 9-and 12-month-old transformed cells confirmed that the protocol enabled stable integration of the mgfp5 gene. Moreover, the comparatively high number of transformed regenerants in this study made it possible to generate a large number of transgenic cells in a short period, which would be useful for high-throughput functional screening of enzymes involved in the biosynthetic pathways of bioactive compounds.

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Section: Introductionmentioning

confidence: 99%

Stable integration of mgfp5 transgenes following Agrobacterium-mediated transformation in Boesenbergia rotunda cell suspension culture

Wong

Zoolkefli

Karim

et al. 2015

Frontiers in Life Science

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“…Southern hybridisation of some of the PCR products with bar gene probe confirmed the identity of the amplified products (gel results not shown for T 0 plants). Similar confirmatory assays have been done for cowpea, banana and plantain (Kononowicz et al, 1997;Sagi et al, 1995). The amplification of DNA samples from GUS negative plants and the subsequent confirmation by Southern analysis proves that the plants were transformed with the exogenous DNA integrated into the genome.…”

Section: Molecular Evaluation Of T 0 Plantsmentioning

confidence: 54%

Stable gene transformation in cowpea (Vigna unguiculata L. walp.) using particle gun method

Ikea¹,

Ingelbrecht²,

Uwaifo³

et al. 2003

Afr. J. Biotechnol.

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We investigated the possibility of transforming and obtaining transgenic cowpea (Vigna unguiculata L Walp) plants using the particle bombardment process. Meristematic explants that could give rise to whole fertile plants were used in transformation experiments with reporter and selectable marker genes driven by a 35S CaMV promoter. Conditions for optimal delivery of DNA to explants were established based on transient gus expression assays two days after bombardment. The size of microcarriers, microflight distance and helium pressure significantly affected transient expression of reporter genes. A total of 1692 explants were bombarded with DNA-coated particles and placed on 3 mg/l bialaphos selective medium. Only 12 regenerated shoots produced seeds eventually, and all were Gus negative even though

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“…Protoplasts were isolated from ECS according to Panis et al (1993) and transformed by electroporation as described by Sági et al (1994). Particle bombardment of ECS was performed according to Sági et al (1995a) while Agrobacterium-mediated transformation was done as described by May et al (1995).…”

Section: Methodsmentioning

confidence: 99%

“…At present, three systems appear to be applicable to transformation of banana and plantain: (i) particle bombardment using a biolistic gun device which has been optimized for transformation of ECS (Sági et al, 1995a), (ii) introduction of DNA into regenerable, ECS-derived protoplasts by electroporation (Sági et al, 1994), and (iii) Agrobacteriummediated transformation of in vitro meristems (May et al, 1995). This paper summarizes the results obtained with various transformation techniques developed or adapted in our laboratory.…”

Section: Introductionmentioning

confidence: 99%

Production of Transgenic Banana and Plantain

Sági¹,

Remy²,

Cammue³

et al. 2000

Acta Hortic.

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Genetic transformation of banana and plantain has been performed in our laboratory by (i) particle bombardment of embryogenic cell suspensions (ECS), (ii) electroporation of protoplasts isolated from ECS, and (iii) cocultivation of meristematic tissues with Agrobacterium tumefaciens. So far, the following cultivars have been transformed successfully: Williams (AAA), Three Hand Planty (AAB), Bluggoe (ABB), Cardaba (ABB), Monthan (ABB) and Tani (BB). Transient expression of the gusA reporter gene has been observed in all these cultivars following at least one of the three transformation techniques. Transgenic plants have been regenerated after particle bombardment of Bluggoe, Three Hand Planty and Williams, representing the three economically important genomic groups in Musa. Molecular and biochemical analysis has confirmed integration and expression of the transgenes in the banana and plantain genome. At present, more than 200 independent transgenic lines, generated from more than 1000 shoots, are growing in the greenhouse. Practical applications of molecular breeding of banana and plantain to create resistance to pathogens and pests are in progress.

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Genetic Transformation of Banana and Plantain (Musa spp.) via Particle Bombardment

Cited by 150 publications

References 28 publications

Stable integration of mgfp5 transgenes following Agrobacterium-mediated transformation in Boesenbergia rotunda cell suspension culture

Stable integration of mgfp5 transgenes following Agrobacterium-mediated transformation in Boesenbergia rotunda cell suspension culture

Stable gene transformation in cowpea (Vigna unguiculata L. walp.) using particle gun method

Production of Transgenic Banana and Plantain

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