GENETIC TRANSFORMATION AND REGENERATION OF COMMON BEAN (Phaseolus vulgaris L.) USING Agrobacterium SYSTEM

Eissa, E. A.

doi:10.21608/ejgc.2013.10461

Cited by 4 publications

(5 citation statements)

References 58 publications

(47 reference statements)

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“…The results showed that MS medium supplemented with 1 mg/l BA and 0.1 mg/l NAA was the optimal for shoot induction from meristematic ring of cotyledonary node explants in both bean varieties. A similar finding was reported by Eissa et al (2002) and Eissa (2013b), who showed that MS medium containing 1 mg/l BA and 0.1 mg/l NAA was the optimal for shoot regeneration in the two varieties. This in vitro culture system carries a high potential for regeneration and transformation of P. vulgaris to improve this important recalcitrant species.…”

Section: Results and Discussion Establishment Of A Regeneration Systesupporting

confidence: 90%

“…Stable transformants were then selected by culturing the explants on medium containing 100 mg/l kanamycin. Results showed that kanamycin at a concentration of 100 mg/l, totally suppressed the growth of the cotyledonary node tissues as reported by Eissa (2013b). These results provide strong confirmation for a kanamycin resistant gene transferred to common bean plant cells.…”

Section: Establishment Of a Transformation System And Production Of Dsupporting

confidence: 83%

“…Cultures were maintained in a growth chamber at 24-25°C with a 16/8 h light/dark cycle at white light. The cotyledonary node meristematic tissues of the two common bean varieties "Fönix" and "Maxidor" were chosen as explants for the genetic transformation experiment because it produces high number of shoot regeneration (Eissa 2013b).…”

Section: Materials and Methods Plant Materials Seeds Sterilization Mmentioning

confidence: 99%

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Genetic Transformation Via Biolistic Gene Gun Method and Regeneration of Common Bean

Eissa¹

2013

EJPB

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The objective of the present research was to utilize biotechnology to improve agronomical traits such as drought stress tolerance in common bean (Phaseolus vulgaris L.) as important recalcitrant crop species via a particle gun "Genebooster TM " for the bombardment of plant tissues with DNA. The process involves the high velocity acceleration of microprojectiles carrying foreign DNA, penetration of the cell wall and membrane by microprojectiles, and delivery of the DNA into plant cells. The initial phase of the two common bean varieties "Fönix" and "Maxidor" focused on the development of a highly regenerable tissue culture procedure amenable to the particle gun physical method. The results showed that MS medium supplemented with 1 mg/l BA and 0.1 mg/l NAA were the optimal for shoot induction from meristematic ring of cotyledonary node explants in the both bean varieties. The author has developed a tissue culture protocol that enables the "meristematic ring" that develops below axillary shoots on nodal bean explants to produce shoot buds, and developed an electric-discharge particle acceleration procedure to produce transgenic plants in the two common bean varieties. The plasmid pFF19K, harboring the kanamycin resistance gene nptII as a selectable marker and pFF19-mtlD harboring the mtlD gene which confers mannitol (drought stress) tolerance were used for adapting transformation in the two common bean varieties. The results indicated that the successful delivery of plasmids DNA and efficient gene transfer into common bean meristematic tissues included 20 bar nitrogen pressure, 135 mm shooting distance, 1.1 µm diameter size of microprojectiles, and twice bombarding of the target tissues. Multiple shoots are then generated and screened to recover transgenic plants at a rate of 7.6% and 8.4% in "Fönix" and "Maxidor" bean varieties, respectively. Transgenic plants were recovered using both kanamycin (100 mg/l) and mannitol (1.2 M/l) screening to introduce the neomycin phosphotransferase II (nptII) and the mannitol-1-phosphate dehydrogenase (mtlD) genes into the two common bean varieties. The introduced nptII and mtlD genes have been shown to confer strong resistance and tolerance in transgenic beans to kanamycin and mannitol stress applications. The transgenic bean plants were healthy and the method has proven to be applicable to common bean crop. To my knowledge, this is the first report on the production of common bean transgenic plants with mtlD gene using biolistic gun-mediated gene transfer system.

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Section: Results and Discussion Establishment Of A Regeneration Systesupporting

confidence: 90%

Section: Establishment Of a Transformation System And Production Of Dsupporting

confidence: 83%

Section: Materials and Methods Plant Materials Seeds Sterilization Mmentioning

confidence: 99%

See 1 more Smart Citation

Genetic Transformation Via Biolistic Gene Gun Method and Regeneration of Common Bean

Eissa¹

2013

EJPB

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“…Stable transformants were then selected by culturing the explants on medium containing 100 mg/l kanamycin. Results showed that kanamycin at a concentration of 100 mg/l, totally suppressed the growth of the cotyledonary node tissues as reported by Eissa (2013b). Following several sub cultures on MS medium containing kanamycin and mannitol, most of the shoots obtained from biolistic gun bombarded turned yellow, browned and died.…”

Section: Optimization Of Transformation Conditions For C Roseussupporting

confidence: 73%

Genetic Improvement of Catharanthus roseus L. for Abiotic Stress Tolerance using HVA-1 Gene Via Bombardment Approach

نجم¹

2021

Annals of Agricultural Science, Moshtohor

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The main aim of this study was to introduce HVA1 gene into Catharanthus roseus callus by using PDS (Particle delivery system). In vitro seed germination of C. roseus was achieved using vigor seeds. The shootlets induction medium was MS containing 2.0 mg/l benzyl aminopurine l −1 . Somatic embryogenesis was obtained by culturing various parts of explants (stem, root and leaves disc) on MS containing 2, 4-D at 3.0 mg/l and subcultured on MS medium containing coconut juice 3%. Genetically transformed C. roseus somatic embryos were obtained after bombardment of embryonic calli, by pAB1 plasmid harboring HVA-1 gene, salt resistance, and Bar gene, herbicide resistance, coated gold particles 1.0micron, 1100 and 1350 Psi with 130 mm with HAV1 gene. The data showed that the transgenic line exhibited different levels of salt tolerance as expressed by the performance of plants dry weight, proline content and drought tolerance test. HVA-1 gene enhanced vinca salt tolerance hence enhancing the production of the important secondary metabolites from transgenic plants. The results of the present study can be seen as a step towards development of salinity tolerant transgenic vinca genotypes.

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“…La efectividad de los dos sistemas se selección usando los genes nptII y bar en nudos cotiledonales de P. vulgaris transformados vía A. tumefaciens ha sido descrita (EISSA, 2013). Esta autora empleó kanamicina (50 mg l -1 ) y glufosinato de amonio (1,0 mg L -1 ) como agentes selectivos.…”

Section: Efecto De La Selección Con Geneticina Y Glufosinato De Amoniounclassified

Regeneración de brotes a partir de tejido transformado en (Phaseolus vulgaris) Fabaceae cultivares 'ICA Pijao' y 'BAT 93'

et al. 2022

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El cultivo de Phaseolus vulgaris L. se encuentra muy difundido en el mundo. La transformación genética vía Agrobacterium tumefaciens constituye una alternativa en esta especie para la obtención de cultivares con tolerancia o resistencia a condiciones ambientales desfavorables. Sin embargo, la condición cultivar dependiente y la baja regeneración del tejido transformado en los protocolos establecidos para P. vulgaris, han limitado el empleo de esta tecnología a un grupo reducido de cultivares. El objetivo de este trabajo fue regenerar brotes a partir de tejido transformado en P. vulgaris cv. 'ICA Pijao’ y 'BAT 93’. Se determinó el efecto de la concentración de la suspensión bacteriana, la infiltración por vacío, el periodo de co-cultivo sobre la regeneración de brotes. Además, se determinó la efectividad de la geneticina y el glufosinato de amonio durante la etapa de selección en la regeneración de brotes a partir de explantes inoculados con A. tumefaciens. Los explantes ½ NC-1 inoculados con una suspensión bacteriana ajustada a una densidad óptica de 0,1, sin infiltración con vacío y mantenidos por tres días en co-cultivo presentaron la mejor respuesta de regeneración. La geneticina y el glufosinato de amonio fueron efectivos para la selección de brotes. La regeneración de brotes a partir de tejido transformado vía A. tumefaciens en P. vulgaris cv. 'ICA Pijao' y 'BAT 93', no descrita en la literatura científica, resulta importante para su posterior aplicación en programas de mejoramiento genéticos de esta especie.

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GENETIC TRANSFORMATION AND REGENERATION OF COMMON BEAN (Phaseolus vulgaris L.) USING Agrobacterium SYSTEM

Cited by 4 publications

References 58 publications

Genetic Transformation Via Biolistic Gene Gun Method and Regeneration of Common Bean

Genetic Transformation Via Biolistic Gene Gun Method and Regeneration of Common Bean

Genetic Improvement of Catharanthus roseus L. for Abiotic Stress Tolerance using HVA-1 Gene Via Bombardment Approach

Regeneración de brotes a partir de tejido transformado en (Phaseolus vulgaris) Fabaceae cultivares 'ICA Pijao' y 'BAT 93'

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