Agrobacterium-mediated transformation of Phaseolus vulgaris L. using indirect organogenesis

Collado, Raúl; Bermúdez-Caraballoso, Idalmis; García, Lourdes R.; Veitía, Novisel; Torres, Damaris; Romero, Carlos; Martirena-Ramírez, Amanda; Angenon, Geert

doi:10.1016/j.scienta.2015.06.046

Cited by 13 publications

(14 citation statements)

References 29 publications

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“…Optimum bacteria concentration is a relevant parameter for a transformation protocol (Collado et al, 2015). An OD600nm lower than 0.5 will not allow for genetic transformation and higher concentrations kill the explants.…”

Section: Discussionmentioning

confidence: 99%

“…There have been several efforts to conduct genetic transformation via A. tumefaciens (Franklin, Trieu, Cassidy, Dixon, & Nelson, 1993;Zhang, Coyne, & Mitra, 1997), with the efficiency of 2.8-to 28.6 % (Espinosa-Huerta, Quintero-Jiménez, Cabrera-Becerra, & Mora-Avilés, 2013;Collado et al, 2015). Regeneration of plants from transformed explants is difficult in genetic transformation protocols, and for recalcitrant species like common bean is one of the unsolved aspects (Liu, Park, Kanno, & Kameya, 2005;Amugune et al, 2011;Collado et al, 2015). Recently the green nodular compact calli were used as explants to obtain transformed plants, based on the indirect organogenesis regeneration pathway (Collado et al, 2015(Collado et al, , 2016).…”

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confidence: 99%

“…Regeneration of plants from transformed explants is difficult in genetic transformation protocols, and for recalcitrant species like common bean is one of the unsolved aspects (Liu, Park, Kanno, & Kameya, 2005;Amugune et al, 2011;Collado et al, 2015). Recently the green nodular compact calli were used as explants to obtain transformed plants, based on the indirect organogenesis regeneration pathway (Collado et al, 2015(Collado et al, , 2016).…”

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confidence: 99%

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Agrobacterium tumefaciens-mediated transformation of common bean (Phaseolus vulgaris) var. Brunca

Solís-Ramos

Ortiz-Pavón

Andrade‐Torres³

et al. 2019

RBT

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Common bean is a crop recalcitrant to in vitro regeneration and therefore it lacks an efficient transformation protocol that can be reproduced using A. tumefaciens. The main goal of this study was to establish a protocol for A. tumefaciens mediated transformation of Phaseolus vulgaris var. Brunca by marker genes (gusA and nptII) together with the gene for trehalose-6-phosphate synthase from Saccharomyces cerevisiae (TPS1) used in other species to increase tolerance to abiotic stress. The β-glucuronidase activity was detected in 45 % of the LBA4404 ElectroMAX® pCAMBIA1301 infected explants. Transformed explants regenerated new shoots after four to five months period in a kanamycin rich media. Surviving plants were evaluated by PCR and presented an 0.5 % efficiency of transformation. The established protocol for genetic transformation of common bean has two additional advantages with respect to previous reports: (1) it allows for obtaining transformed regenerants and (2) the genetic transformation was stable for the selective gene.

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Section: Discussionmentioning

confidence: 99%

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confidence: 99%

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confidence: 99%

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Agrobacterium tumefaciens-mediated transformation of common bean (Phaseolus vulgaris) var. Brunca

Solís-Ramos

Ortiz-Pavón

Andrade‐Torres³

et al. 2019

RBT

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“…In fact, it has been reported that the EHA105-mediated transformation of primary and proliferative calli of common bean cv. CIAP7247F resulted in transgenic plants [20].…”

Section: Meristem Tissues Are a Reliable Source Of Regenerable Callimentioning

confidence: 99%

“…Genetic engineering is a powerful tool to introduce genes from sources that are inaccessible through sexual hybridization [12][13][14]. Thus, substantial efforts have been made to develop reliable transformation methods for engineering common beans with various traits [13,[15][16][17][18][19][20][21]. To date, no genetically engineered (GE) common bean has been commercialized, despite of the regulatory approval for the transgenic "Embrapa 5.1" common bean for golden mosaic virus (BGMV) resistance in Brazil in 2011 [22][23][24].…”

Section: Introductionmentioning

confidence: 99%

Induction of competent cells for Agrobacterium tumefaciens-mediated stable transformation of common bean (Phaseolus vulgaris L.)

et al. 2020

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Stable transformation of common bean (Phaseolus vulgaris L.) has been successful, to date, only using biolistic-mediated transformation and shoot regeneration from meristemcontaining embryo axes. In this study, using precultured embryo axes, and optimal co-cultivation conditions resulted in a successful transformation of the common bean cultivar Olathe using Agrobacterium tumefaciens strain EHA105. Plant regeneration through somatic embryogenesis was attained through the preculture of embryo axes for 12 weeks using induced competent cells for A. tumefaciens-mediated gene delivery. Using A. tumefaciens at a low optical density (OD) of 0.1 at a wavelength of 600 nm for infection and 4-day co-cultivation, compared to OD 600 of 0.5, increased the survival rate of the inoculated explants from 23% to 45%. Selection using 0.5 mg L-1 glufosinate (GS) was effective to identify transformed cells when the bialaphos resistance (bar) gene under the constitutive 35S promoter was used as a selectable marker. After an 18-week selection period, 1.5%-2.5% inoculated explants, in three experiments with a total of 600 explants, produced GS-resistant plants through somatic embryogenesis. The expression of bar was confirmed in first-and secondgeneration seedlings of the two lines through reverse polymerase chain reaction. Presence of the bar gene was verified through genome sequencing of two selected transgenic lines. The induction of regenerable, competent cells is key for the successful transformation, and the protocols described may be useful for future transformation of additional Phaseolus germplasm.

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Genetic transformation of common beans (Phaseolus vulgaris L.) through Agrobacterium tumefaciens carrying Cry1Ab gene

2022

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Agrobacterium-mediated transformation of Phaseolus vulgaris L. using indirect organogenesis

Cited by 13 publications

References 29 publications

Agrobacterium tumefaciens-mediated transformation of common bean (Phaseolus vulgaris) var. Brunca

Agrobacterium tumefaciens-mediated transformation of common bean (Phaseolus vulgaris) var. Brunca

Induction of competent cells for Agrobacterium tumefaciens-mediated stable transformation of common bean (Phaseolus vulgaris L.)

Genetic transformation of common beans (Phaseolus vulgaris L.) through Agrobacterium tumefaciens carrying Cry1Ab gene

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