Activity characterization of the plantain promoter from the heavy metal-associated isoprenylated plant gene (MabHIPP) using the luciferase reporter gene

Bananas, one of the most valued fruits worldwide, are produced in more than 135 countries in the tropics and subtropics for local consumption and export due to their tremendous nutritional value and ease of access. The genetic improvement of commercial crops is a crucial strategy for managing pests or other diseases and abiotic stress factors. Although conventional breeding has developed new hybrids with highly productive or agronomic performance characteristics, in some banana cultivars, due to the high level of sterility, the traditional breeding strategy is hampered. Therefore, modern biotechniques have been developed in a banana for genetic improvement. In vitro, culture techniques have been a basis for crop micropropagation for elite banana varieties and the generation of methods for genetic modification. This review includes topics of great interest for improving bananas and their products worldwide, from their origins to the different improvement alternatives. Keywords. Banana, genetic improvement, pest management, diseases, abiotic stress factors.

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“…3). Green fluorescent protein (GFP) and luciferase (LUC) is also highly used as plant reporter genes [43][44][45][46] .…”

Section: Methods Of Genetic Transformationmentioning

confidence: 99%

Genetic improvement in Musa through modern biotechnological methods

Villao

Chávez

Pacheco

et al. 2023

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“…122 This can be further utilized for the detection of recombinant antigens fused with reporter genes such as fluorescent proteins (mCherry, yellow fluorescent protein (YFP), cyan fluorescent protein (CFP), and GFP), beta-glucuronidase (GUS), and luciferase (LUC). [123][124][125] Critically, the transgene design also allows the addition of molecular adjuvants, such as LTB, CTB, or flagellin, which is advantageous to increase immunogenicity and uptake of recombinant antigens. 84 Lastly, the selectable markers, such as genes coding hygromycin phosphotransferase, neomycin phosphotransferase, or a genes for resistance to herbicide bialaphos, [126][127][128] represent an additional part of the transformation construct, critical for the consequent selection of transformed plants, and targeted organelles.…”

Section: Preparation Of Transgenic Plantsmentioning

confidence: 99%

“…Furthermore, depending on the production strategy, the antigen‐coding gene can be fused with signal peptides, determining the localization and secretion of the desired protein 122 . This can be further utilized for the detection of recombinant antigens fused with reporter genes such as fluorescent proteins (mCherry, yellow fluorescent protein (YFP), cyan fluorescent protein (CFP), and GFP), beta‐glucuronidase (GUS), and luciferase (LUC) 123–125 . Critically, the transgene design also allows the addition of molecular adjuvants, such as LTB, CTB, or flagellin, which is advantageous to increase immunogenicity and uptake of recombinant antigens 84 .…”

Section: Molecular Farming Of Fish Vaccinesmentioning

confidence: 99%

Molecular farming: Expanding the field of edible vaccines for sustainable fish aquaculture

Mičúchová

Piačková

Frébort

et al. 2022

Reviews in Aquaculture

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Vaccines represent one of the most promising strategies for mitigating economic losses imposed by infectious diseases to global aquaculture. While the majority of commercial vaccines employ injection as the main route of delivery, a substantial body of research explores other avenues for the stimulation of protective immunity. Oral vaccines, representing the most favourable and convenient solution for the aquaculture industry, have been at the forefront of scientific interest for the last couple of decades. The orchestrated effort resulted in identifying the main roadblocks on the path to an ideal edible vaccine and provided several ingenious solutions improving the activation of immunity, ensuring the stability of administered antigens and their uptake upon the passage through the stomach. In the presented review, we describe advances in the available knowledge of the processes prerequisite for developing protective mucosal vaccines and focus readers' attention on the untapped potential of plant‐based production systems in this effort. We propose that these approaches not only meet production demands but also fulfil all requirements of an oral vaccine, addressing all the obstacles, previously solved via separate strategies, in one platform. Thus, combined with the available knowledge of molecular adjuvants and produced for a fraction of the cost, plant‐based production, so‐called molecular farming, is an ideal candidate for vaccine development, paving the road to sustainable aquaculture.

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“…In this study, the Agrobacterium tumefaciens strain LBA404 was used, and the binary vectors pLVCIBE1 and pLV-CIBE2 were developed containing the gene cassettes PMabHI-PP::luc2::Tnos and P35S::luc2::Tnos, respectively 9 . The selectable marker gene used was the hpt for hygromycin B resistance, fused to the CaMV 35S promoter.…”

Section: Agrobacterium Strains and Plasmidsmentioning

confidence: 99%

Genetic transformation of apical meristematic shoots in the banana cultivar ‘Williams’

Villao

Flores

Santos-Ordóñez

2021

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Bananas and plantains (Musa spp.) are among the most critical socioeconomic crops globally, being a staple food for millions of people in the tropics and an essential component for the export market, including the subtropics. Besides conventional breeding, genetic improvement of bananas and plantains could be performed through genetic engineering and new breeding techniques. Furthermore, plant tissue culture is essential for these technologies, including developing embryogenic cell suspensions and in vitro plants. The transient and stable genetic transformation could be performed from in vitro plants, shortening Musa transgenic lines development compared to genetic transformation while using embryogenic cell suspension. In this study, a genetic transformation protocol was established from banana apical meristems for the ‘Williams’ cultivar (genotype AAA). The protocol was based on the co-cultivation of the explants (whole in vitro plants or bisected meristematic tissues derived from in vitro plants) with Agrobacterium tumefaciens strain LBA4404 harboring two binary vectors denominated pLVCIBE1 (cassette: MabHIPP promoter::luc2::Tnos, P35S::hpt::Tnos) and pLVCIBE2 (cassette: P35S::luc2::Tnos, P35S::hpt::Tnos), independently. The stable genetic transformation was obtained by subculturing in vitro banana plants in selection medium (12.5µg/mL of hygromycin) for 8 weeks from bisected meristematic tissue transformation. Genetic transformation was confirmed in vivo with the use of the luciferase reporter gene system. Furthermore, PCR was performed on DNA extracted from leaves of regenerated transgenic in vitro plants after 8 weeks of selection, confirming stable genetic transformation. Therefore, genetic transformation was achieved in the apical meristematic tissue of in vitro banana plants with co-cultivation of Agrobacterium tumefaciens.

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Activity characterization of the plantain promoter from the heavy metal-associated isoprenylated plant gene (MabHIPP) using the luciferase reporter gene

Cited by 12 publications

References 19 publications

Genetic improvement in Musa through modern biotechnological methods

Genetic improvement in Musa through modern biotechnological methods

Molecular farming: Expanding the field of edible vaccines for sustainable fish aquaculture

Genetic transformation of apical meristematic shoots in the banana cultivar ‘Williams’

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