Efficient particle bombardment-mediated transformation of Cuban soybean (INCASoy-36) using glyphosate as a selective agent

Soto, Natacha; Delgado, Celia; Hernández, Y.; Rosabal, Yamilka; Ferreira, Aleines; Pujol, Merardo; Aragão, F. J. L.; Enríquez, G. A.

doi:10.1007/s11240-016-1099-x

Cited by 10 publications

(11 citation statements)

References 43 publications

(71 reference statements)

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“…The soybean [Glycine max (L.)] was of the variety DT-84 from Vietnam. Embryonic axes of mature seeds were used as explants for their bombardment-mediated transformation, according to Soto et al (2017).…”

Section: Plant Materialsmentioning

confidence: 99%

“…The pCP4EPSPS-DEF vector carrying the cp4epsps gene and the NmDef02 defensin gene isolated from Nicotiana megalosiphon was the vector system used for the transformation. The cassette with p35S/NmDef02/tnos obtained by Portieles et al (2010) was cloned into the pCP4EPSPS binary vector (Soto et al, 2017) to generate the pCP4EPSPS-DEF vector (Figure 1A). This was done at the soybean biotechnology laboratory at CIGB, Havana.…”

Section: Vector Constructionmentioning

confidence: 99%

“…The controls used were explants derived from cultivar DT-84, which were cultured under the same conditions and without selective agent. The regenerated shoots were excised and transferred to the same medium without selecting for rooting, as described by Soto et al (2017). Plantlets were transferred to pots containing a mixture of organic material and zeolite (50/50) in an acclimatized greenhouse at 26-27 • C to produce seeds.…”

Section: Transformation Selection and Plant Regenerationmentioning

confidence: 99%

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Field Resistance to Phakopsora pachyrhizi and Colletotrichum truncatum of Transgenic Soybean Expressing the NmDef02 Plant Defensin Gene

et al. 2020

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Fungal diseases lead to significant losses in soybean yields and a decline in seed quality; such is the case of the Asian soybean rust and anthracnose caused by Phakopsora pachyrhizi and Colletotrichum truncatum, respectively. Currently, the development of transgenic plants carrying antifungal defensins offers an alternative for plant protection against pathogens. This paper shows the production of transgenic soybean plants expressing the NmDef02 defensin gene using the biolistic delivery system, in an attempt to improve resistance against diseases and reduce the need for chemicals. Transgenic lines were assessed in field conditions under the natural infections of P. pachyrhizi and C. truncatum. The constitutive expression of the NmDef02 gene in transgenic soybean plants was shown to enhance resistance against these important plant pathogens. The quantification of the P. pachyrhizi biomass in infected soybean leaves revealed significant differences between transgenic lines and the non-transgenic control. In certain transgenic lines there was a strong reduction of fungal biomass, revealing a less severe disease. Integration and expression of the transgenes were confirmed by PCR, Southern blot, and qRT-PCR, where the Def1 line showed a higher relative expression of defensin. It was also found that the expression of the NmDef02 defensin gene in plants of the Def1 line did not have a negative effect on the nodulation induced by Bradyrhizobium japonicum. These results indicate that transgenic soybean plants expressing the NmDef02 defensin gene have a substantially enhanced resistance to economically important diseases, providing a sound environmental approach for decreasing yield losses and lowering the burden of chemicals in agriculture.

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Section: Plant Materialsmentioning

confidence: 99%

Section: Vector Constructionmentioning

confidence: 99%

Section: Transformation Selection and Plant Regenerationmentioning

confidence: 99%

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Field Resistance to Phakopsora pachyrhizi and Colletotrichum truncatum of Transgenic Soybean Expressing the NmDef02 Plant Defensin Gene

et al. 2020

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“…Previous research has shown the use of glyphosate as a selection agent in the genetic transformation of plants from several species like rice, maize, cotton and soybean, using concentrations in a range from 0.5 mM to 10 mM of the growth medium (Latif et al, 2015;Ren et al, 2015;Soto et al, 2017). Most reports of genetic transformation of N. tabacum to confer tolerance to glyphosate use antibiotics to select transformed plants (Wang et al, 2003;Yan et al, 2011;Peng et al, 2012), but there is evidence of the use of glyphosate to select transformants in this species (Akbarzadeh et al, 2010).…”

Section: In Silico Design Of Expression Cassettesmentioning

confidence: 99%

“…Despite losing some of the initial explants due to A. tumefaciens contamination, the transformation efficiency for each of the expression cassettes was 16.6% (E-IGP), 16.2% (E-IGP2) and 28% (E2). Glyphosate has been used successfully as a selective agent for transformed plants in cotton (Latif et al, 2015), soybean (Soto et al, 2017), maize (Ren et al, 2015), and other species. We obtained transformed N. benthamiana plants using this selection approach, but it was with a low transformation efficiency, possibly due to the effect of the herbicide on the regeneration capacity of the explants.…”

Section: Molecular Testsmentioning

confidence: 99%

Freedom to operate analysis, design and evaluation of expression cassettes that confer tolerance to glyphosate

2020

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Tolerance to the herbicide glyphosate is the most extended feature in commercial transgenic events released worldwide and it is an example of the successful use of genetic modification to improve weed control in crops. Glyphosate-tolerant genotypes have been developed by multinational corporations using patent- protected technologies. For some of these events, all associated patents have expired and, therefore, have become a good target for the development of herbicide-tolerant agbiogenerics by national research institutions, using local crop varieties. As a first step in this process, we present the design (in silico) of three expression cassettes with the purpose of using them in the transformation of Colombian soybean (Glycine max) varieties to confer them tolerance to glyphosate. We transformed Nicotiana benthamiana as a model to validate the functionality of the expression cassettes and detected the expression of the transgene by RT-PCR. Additionally, a Freedom to Operate analysis of the sequences used in the expression cassettes suggests that their commercial use in Colombia does not infringe third party rights. This analysis must be updated and validated by intellectual property experts prior to commercialization.

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Improved direct transformation via particle bombardment of split-immature embryo explants in soybean (Glycine max)

Chennareddy¹,

Cicak²,

Mall³

et al. 2018

Plant Cell Tiss Organ Cult

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Efficient particle bombardment-mediated transformation of Cuban soybean (INCASoy-36) using glyphosate as a selective agent

Cited by 10 publications

References 43 publications

Field Resistance to Phakopsora pachyrhizi and Colletotrichum truncatum of Transgenic Soybean Expressing the NmDef02 Plant Defensin Gene

Field Resistance to Phakopsora pachyrhizi and Colletotrichum truncatum of Transgenic Soybean Expressing the NmDef02 Plant Defensin Gene

Freedom to operate analysis, design and evaluation of expression cassettes that confer tolerance to glyphosate

Improved direct transformation via particle bombardment of split-immature embryo explants in soybean (Glycine max)

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