Recent Developments in Generation of Marker-Free Transgenic Plants

Singh, Rupesh Kumar; Sharma, Latika; Bohra, Nitin; Anandhan, Sivalingam; Ruiz‐May, Eliel; Quiroz-Figueroa, Francisco Roberto

doi:10.1007/978-981-13-9624-3_6

Cited by 3 publications

(5 citation statements)

References 95 publications

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“…Particle bombardment can deliver the preferred DNA into both nuclear and organellar genomes. For instance, chloroplasts in higher plants and chloroplasts as well as mitochondria in algae were successfully transformed by the biolistic genetic transformation approach [ 151 , 152 , 153 , 154 , 155 ]. Another difference between both methods for transformation is that Agrobacterium-mediated transformation can integrate from one to three copies of donor DNA, while the biolistic method can integrate multiple copies [ 156 ].…”

Section: Molecular Toolsmentioning

confidence: 99%

Biotechnological Advances to Improve Abiotic Stress Tolerance in Crops

Villalobos-López

Arroyo-Becerra

Quintero-Jiménez

et al. 2022

IJMS

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The major challenges that agriculture is facing in the twenty-first century are increasing droughts, water scarcity, flooding, poorer soils, and extreme temperatures due to climate change. However, most crops are not tolerant to extreme climatic environments. The aim in the near future, in a world with hunger and an increasing population, is to breed and/or engineer crops to tolerate abiotic stress with a higher yield. Some crop varieties display a certain degree of tolerance, which has been exploited by plant breeders to develop varieties that thrive under stress conditions. Moreover, a long list of genes involved in abiotic stress tolerance have been identified and characterized by molecular techniques and overexpressed individually in plant transformation experiments. Nevertheless, stress tolerance phenotypes are polygenetic traits, which current genomic tools are dissecting to exploit their use by accelerating genetic introgression using molecular markers or site-directed mutagenesis such as CRISPR-Cas9. In this review, we describe plant mechanisms to sense and tolerate adverse climate conditions and examine and discuss classic and new molecular tools to select and improve abiotic stress tolerance in major crops.

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Section: Molecular Toolsmentioning

confidence: 99%

Biotechnological Advances to Improve Abiotic Stress Tolerance in Crops

Villalobos-López

Arroyo-Becerra

Quintero-Jiménez

et al. 2022

IJMS

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“…Generation of selection marker-free transgenic plants is a goal of plant breeding, since marker genes pose safety concerns regarding the expression of resistance marker genes that could cause antibiotic resistance in plant pathogens, potentially posing a threat to humans and animals (Xu et al, 2020). Therefore, it is crucial to develop transgenic plants with no selectable markers, and the independent transgene segregation after cotransformation is a suitable strategy for the compliment of this objective (Pérez-Bernal et al, 2017;Singh et al, 2019). One aspect that makes co-transformation advantageous is that it does not require additional genetic manipulation for removal of the selectable marker gene, as required for example in the Cre/loxPbased approach (Chong-Pérez et al, 2013).…”

Section: Transgenic Analysis Through Three Successive Progeniesmentioning

confidence: 99%

“…The integration of a transgene into the plant genome is a random and complex event depending on the transgene itself and the host genome, and its expression may be stable or variable regardless of the method of gene transfer (Ahuja and Fladung, 2014). Determining the expression and inheritance of the transgenes is of paramount importance for the introduction of genetically modified plants in agriculture, since the main role of plant biotechnology is to improve the crops and, simultaneously, reduce the possible risk threats (Singh et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

“…Both Mendelian and non-Mendelian transgene inheritance has been described through several generations of crop plants (Ahuja and Fladung, 2014). Once transgenes are integrated into the host plant cells, events can occur that lead to opposite results: the transference of transgenes to further progenies through sexual generations while retaining the expression stability, or their loss, inactivation or silencing (Singh et al, 2019). Transgene integration sites exhibit different levels of structural complexity in the host DNA, and the primary transformants revealing consistent transgene expression may not give rise to progeny with the same characteristics (Betts et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

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Inheritance of NmDef02 synthetic defensin gene and hpt selectable marker in four successive generations of transgenic rice (Oryza sativa, cv. J-104)

Pérez-Bernal¹,

Delgado²,

Abreu³

et al. 2021

Int. J. Curr. Res. Biosci. Plant Biol.

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The evaluation of the inheritance and stability of the transgenes is essential for the application of transgenic plants in agriculture. In this work, we studied the inheritance of two transgenes in T1, T2, T3 and T4 rice progenies. Transgenic rice plants (cv. J-104) was obtained by biolistic method using a synthetic defensin gene (NmDef02) and hpt as selectable marker gene, co-transformed in a 4:1 proportion, respectively. Regenerated plants were acclimated under natural conditions. The study started from the primary transformants that fulfilled the agronomic characters reported by the experts for the J-104 rice cultivar in the maturation stage. The integration and relative expression of NmDef02 in T1 plants was verified by Southern blot and qRT-PCR, respectively. The inheritance of transgenes over four generations was analyzed by PCR. The following transgene combinations were identified: NmDef02(+)hpt(+), NmDef02(+)hpt(-) and NmDef02(-)hpt(-). The most advantageous combination was NmDef02(+)hpt(-), which corresponded to the marker-free plants harboring the gene of interest. The inheritance of NmDef02 was confirmed in T1 and T2 progenies, but in some T3 and T4 lines the loss of this gene was verified. This inheritance analysis provided information concerning the transgenic population, but the mechanisms that destabilize the inheritance of the gene of interest will be the goal of future research.

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“…For enabling upgradability of plant biosystems design, it would be desirable to consider marker-free plant transformation systems, in which the selectable marker gene can be excised from the plant genome after transformation (Figure 4 (e)). Selectable marker genes can be self-excised in plants using various approaches mediated by site-specific recombinase [ 70 – 73 ], zinc finger nuclease [ 74 ], and CRISPR [ 75 , 76 ].…”

Section: Theoretical Approaches and Principles Of Plant Biosystems De...mentioning

confidence: 99%

Plant Biosystems Design Research Roadmap 1.0

et al. 2020

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Human life intimately depends on plants for food, biomaterials, health, energy, and a sustainable environment. Various plants have been genetically improved mostly through breeding, along with limited modification via genetic engineering, yet they are still not able to meet the ever-increasing needs, in terms of both quantity and quality, resulting from the rapid increase in world population and expected standards of living. A step change that may address these challenges would be to expand the potential of plants using biosystems design approaches. This represents a shift in plant science research from relatively simple trial-and-error approaches to innovative strategies based on predictive models of biological systems. Plant biosystems design seeks to accelerate plant genetic improvement using genome editing and genetic circuit engineering or create novel plant systems through de novo synthesis of plant genomes. From this perspective, we present a comprehensive roadmap of plant biosystems design covering theories, principles, and technical methods, along with potential applications in basic and applied plant biology research. We highlight current challenges, future opportunities, and research priorities, along with a framework for international collaboration, towards rapid advancement of this emerging interdisciplinary area of research. Finally, we discuss the importance of social responsibility in utilizing plant biosystems design and suggest strategies for improving public perception, trust, and acceptance.

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Recent Developments in Generation of Marker-Free Transgenic Plants

Cited by 3 publications

References 95 publications

Biotechnological Advances to Improve Abiotic Stress Tolerance in Crops

Biotechnological Advances to Improve Abiotic Stress Tolerance in Crops

Inheritance of NmDef02 synthetic defensin gene and hpt selectable marker in four successive generations of transgenic rice (Oryza sativa, cv. J-104)

Plant Biosystems Design Research Roadmap 1.0

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