A Cassava vein mosaic virus promoter cassette induces high and stable gene expression in clonally propagated transgenic cassava (Manihot esculenta Crantz)

Oyelakin, Oladele; Opabode, Jelili T.; Raji, Adebola; Ingelbrecht, Ivan

doi:10.1016/j.sajb.2014.11.011

Cited by 11 publications

(24 citation statements)

References 28 publications

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“…The observed strong and constitutive expressions in vascular tissues of petioles, stems and tubers and in leaf mesophyll tissues and vascular stele of roots of GUS positive cassava plants were driven by the CsVMV promoter. Similarly, CsVMV has been reported to drive constitutive expressions of the GUS gene in rice, tobacco and cassava (Verdaguer et al, 1996;Oyelakin et al, 2015). Furthermore, we observed GUS expression in the primary roots and tubers of transgenic cassava, which confirms an earlier report on TME 12 transgenic cassava (Oyelakin et al, 2015).…”

Section: Discussionsupporting

confidence: 91%

“…However, a chimeric expression of the GUS was observed on regenerated plants obtained from somatic embryos derived from immature leaf lobes, while a uniform expression was observed on regenerated plants derived from axillary buds. A chimeric expression of the GUS in cassava has been previously reported (Ingelbrecht et al, 2010;Oyelakin et al, 2015). In our study, the presence of GUS genetic material was confirmed by PCR amplification in regenerated plants derived from somatic embryos obtained from explants of both, axillary buds and immature leaf lobes; however, a uniform GUS activity was detected in plants regenerated from axillary buds only.…”

Section: Discussionsupporting

confidence: 78%

“…The transgenic cassava plants used for the present study carried a T-DNA vector with CsVMV-GUS and CaMV 35S-NPTII cassettes transcribing in the opposite direction introduced into the cassava genome through Agrobacterium tumefaciens strains LB4404 (Oyelakin et al, 2015). From transgenic cassava plants at the sixth cycle of vegetative propagation, primary, secondary and cyclic somatic embryogeneses were successfully established using explants of axillary buds and immature leaf lobes.…”

Section: Discussionmentioning

confidence: 99%

“…The transgenic cassava plants carried a T-DNA vector with CsVMV-GUS and CaMV 35S-Faithful transmission and expression of transgenes via somatic embryos of transgenic cassava plants at the sixth cycle 67 NPTII cassettes transcribing in the opposite direction (Oyelakin et al, 2015 …”

Section: Plant Materials and Growth Conditionsmentioning

confidence: 99%

“…Although somatic embryogenesis has been developed in a large number of cassava cultivars and the system has been widely used for producing transgenic plants for molecular biology, functional genomics and in biotechnology for plant trait improvement (Zhang et al, 2005;Ihemere et al, 2006;Oyelakin et al, 2015), the production of transgenic somatic embryos and regeneration of plants stably transformed from transgenic cassavas has not been investigated. Additional understanding of the faithful transmission of transgenes in cassava is essential to improve the efficacy of transgene expression in this economically important crop.…”

Section: Introductionmentioning

confidence: 99%

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Faithful transmission and expression of transgenes via somatic embryos of transgenic cassava plants at the sixth cycle of vegetative propagation

Opabode¹,

Akinyemiju²

2016

bta

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The transmission of transgenes via somatic embryos from one vegetative cycle to another in cassava (Manihot esculenta Crantz) has not been well studied to date. This study examined somatic embryogenesis and regeneration from transgenic cassava plants expressing the β -glucuronidase gene (GUS) under the control of a cassava vein mosaic promoter (CsVMV) at the sixth cycle of vegetative propagation. Primary, secondary and cyclic somatic embryos were induced from an axillary bud and immature leaf lobe explants. Plantlet regeneration via shoot organogenesis was examined in the cotyledons of somatic embryos. Histochemical, polymerase chain reaction (PCR) and reverse transcriptase PCR analyzes were used to confirm the presence of the GUS gene in regenerated plants. The frequencies of primary, secondary and cyclic somatic embryos from immature leaf lobes were significantly greater (P < 0.05) than those of axillary buds. The transient expression of the GUS gene was detected in all types of somatic embryos from both axillary bud and immature leaf lobe explants. Shoot induction from cotyledons of somatic embryos produced from axillary buds was 27.0% greater than that of leaf lobes, with selection performed with the use of kanamycin. In regenerated transgenic plants (grown from somatic embryos obtained from immature leaf lobes), a non-uniform expression of GUS was observed in three chimeric lines assessed by histochemical and molecular analyzes. In regenerated transgenic plants obtained from somatic embryos from axillary buds, uniform and high expression of GUS was observed in one line in all tested tissues and in most cell types of leaves, stems, petioles, roots and tubers, thus showing the faithful transmission of the GUS transgene under the control of CsVMV promoter via a somatic embryo.

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Section: Discussionsupporting

confidence: 91%

Section: Discussionsupporting

confidence: 78%

Section: Discussionmentioning

confidence: 99%

Section: Plant Materials and Growth Conditionsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Faithful transmission and expression of transgenes via somatic embryos of transgenic cassava plants at the sixth cycle of vegetative propagation

Opabode¹,

Akinyemiju²

2016

bta

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Synthetic Salicylic acid inducible recombinant promoter for translational research

Deb

Dey

2019

Journal of Biotechnology

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White LED intensities during co-cultivation affect the Agrobacterium-mediated soybean (Glycine max) transformation using mature half seeds as explants

Shi,

Li,

Rojas-Pierce

et al. 2024

PLoS ONE

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The transition of light fixture from fluorescent light to light-emitting diodes (LEDs) in growth chambers prompts a reevaluation of current practices in plant biotechnology. Agrobacterium-mediated transformation is crucial for genetic engineering and genome editing in soybean (Glycine max). The critical co-cultivation step of soybean transformation occurs under light condition. Current protocols for co-cultivation in soybean transformation lack a standard for light intensity. In the present study, the objective is to investigate the effect of light intensity during co-cultivation on soybean transformation efficiency. Five light intensities were implemented during five days of co-cultivation: 50, 100, 150, 190 μmol∙m−2∙s−1 of white LEDs in addition to 100 μmol∙m−2∙s−1 of fluorescent light. After co-cultivation, all the explants underwent shoot induction and elongation with selection pressure, rooting and acclimation under uniform condition. The experiment was conducted with two selectable markers, hppdPf-4Pa and bar, separately, investigating whether the potential light effects vary due to the marker-associated pathways. The positive PCR analysis of rooted in vitro plants suggested successful transformation events achieved under both selectable markers across all light treatments ranging from 2.4% to 6.9%. Increasing LED light intensity during co-cultivation resulted in different transformation efficiencies between the two selectable markers. Results indicated that increasing the light intensity during co-cultivation led to a linear increase in transformation efficiency when shoot regeneration was under 4-Hydroxyphenylpyruvate dioxygenase (HPPD) inhibitor selection. No difference in transformation efficiency was detected among the treatments under glufosinate selection. Furthermore, when selection occurred with HPPD inhibitor, variation of transformation efficiency was also observed between fluorescent light and white LED at 100 μmol∙m−2∙s−1. The results highlight the significance and potential applications of investigating the impact of light on transformation efficiency.

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A Cassava vein mosaic virus promoter cassette induces high and stable gene expression in clonally propagated transgenic cassava (Manihot esculenta Crantz)

Cited by 11 publications

References 28 publications

Faithful transmission and expression of transgenes via somatic embryos of transgenic cassava plants at the sixth cycle of vegetative propagation

Faithful transmission and expression of transgenes via somatic embryos of transgenic cassava plants at the sixth cycle of vegetative propagation

Synthetic Salicylic acid inducible recombinant promoter for translational research

White LED intensities during co-cultivation affect the Agrobacterium-mediated soybean (Glycine max) transformation using mature half seeds as explants

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