A robust genetic transformation protocol to obtain transgenic shoots of Solanum tuberosum L. cultivar ‘Kufri Chipsona 1’

Kaur, Amanpreet; Guleria, Shivani; Reddy, M. Sudhakara; Kumar, Anil

doi:10.1007/s12298-019-00747-4

Cited by 9 publications

(4 citation statements)

References 63 publications

(80 reference statements)

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“…We used Kanamycin (50 mg/L) in the selection medium for shoot formation, while Deng et al [ 25 ] used Hygromycin B (40 mg/L) in Oryza sativa. The transformation efficiency after screening with NPTII primers was found to be 44% in Kufri Jyoti putative mutants, which was considerably higher in comparison to 10–16% in Kufri Chipsona 1 cultivar [ 40 ].…”

Section: Discussionmentioning

confidence: 99%

Generation of Asynaptic Mutants in Potato by Disrupting StDMC1 Gene Using RNA Interference Approach

Kumar

Siddappa

Bhardwaj

et al. 2023

Life

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Fixing the genomic composition and multiplication through true potato seed (TPS) is an important challenge in autotetraploid potato. Disrupted meiotic cDNA (DMC1) is a meiotic gene that plays a central role in DNA recombination through crossing over in meiosis. Using the Arabidopsis DMC1 (AtDMC1) gene sequence, we retrieved Solanum tuberosum DMC1(StDMC1) from the diploid potato genome, and subsequently, sense and antisense regions of the StDMC1 gene were amplified in potato cv. Kufri Jyoti. The sense and antisense fragments were confirmed by Sanger-sequencing and cloned in the pRI101 vector. Agrobacterium-mediated transformation of the RNAi construct resulted in 44% transformation efficiency, and a total of 137 mutant lines were obtained. These mutant lines were further validated through pollen viability testing, and selected lines were used for gene expression analysis. The acetocarmine-based pollen staining showed reduced pollen viability ranging from 14 to 21% in four DMC1 mutant lines (DMC4-37, DMC4-41, DMC6-20, and DMC6-21), as compared to the Kufri Jyoti control plants, which on average exhibited 78% pollen viability. The phenotypic data was supported by the reduced expression of the StDMC1 gene in these four mutant lines compared to the control Kufri Jyoti. The results confirmed the generation of StDMC1 knockdown lines. This is the first report of StDMC1 mutant line generation in tetraploid potatoes and will be a step forward in generating non-recombinant mutants through sexual reproduction in potatoes.

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Section: Discussionmentioning

confidence: 99%

Generation of Asynaptic Mutants in Potato by Disrupting StDMC1 Gene Using RNA Interference Approach

Kumar

Siddappa

Bhardwaj

et al. 2023

Life

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“…However, despite being successful for the diploid materials, it has low e ciency. In this methodology, we did not apply the preculture procedures that are normally applied in some protocols (Kaur et al 2020). These pre-culture procedures require about one week to implement.…”

Section: Discussionmentioning

confidence: 99%

A convenient Agrobacterium tumefaciens mediated transgenic system for diploid and tetraploid potatoes

Xu,

Qi,

Chang

et al. 2023

Preprint

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As the fourth staple food crop in the world, potatoes can provide enough energy, protein, and nutrients necessary for humans. However, the negative effects of climate change and population growth call for improved potato yields and resilience. Genetic engineering is a convenient way to improve potato varieties and create new germplasm resources. Some important traits may be obtained through transgenic technology, such as drought, heat, cold tolerance, and increasing the concentration of micronutrients in tubers. Therefore, Agrobacterium tumefaciens mediated transformation is a stable and widely used method for genetic improvement. In this study, a convenient transgenic system on two diploid potatoes and one popular tetraploid potato variety (Desiree) were transformed using this process. A simple and easy complete transgenic system was built by obtaining positive transformed seedlings in 40 days, and the positive transgenic efficiencies of diploid and tetraploid recipient materials were 3.75–8.45% and 70%, respectively. This fast transgenic system provides a powerful tool for supporting basic research on functional genes and creating new materials for genetic engineering breeding. The new genetic transformation system has the advantages of greater time efficiencies and fewer operation steps which can accelerate the process of candidate gene verification.

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“…Despite the great number of potato varieties available, few cultivars are commercialized, chosen for their viability to be marketed and stored ( Yang et al, 2015 ). Cultivars ‘Desiree’ and ‘Bintje’ were early favorites for transformation assays but nowadays, most potato cultivars are amenable to tissue culture and nearly all commercially important varieties can be successfully transformed with modifications of the standard protocols ( Vinterhalter, 2008 ; Han et al, 2015 ; Halterman et al, 2016 ; Nadakuduti et al, 2018 ; Bruce and Shoup Rupp, 2019 ; Kaur et al, 2020 ). In 1988, it was reported the development of a genotype-independent method to transform four potato cultivars (‘Bintje’, ‘Berolina’, ‘Desiree’, and ‘Russet Burbank’) using leaf discs ( De Block, 1988 ).…”

Section: Main Parameters Influencing Transformation Efficiencymentioning

confidence: 99%

State of the Art of Genetic Engineering in Potato: From the First Report to Its Future Potential

et al. 2022

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Potato (Solanum tuberosum L.) is a crop of world importance that produces tubers of high nutritional quality. It is considered one of the promising crops to overcome the challenges of poverty and hunger worldwide. However, it is exposed to different biotic and abiotic stresses that can cause significant losses in production. Thus, potato is a candidate of special relevance for improvements through conventional breeding and biotechnology. Since conventional breeding is time-consuming and challenging, genetic engineering provides the opportunity to introduce/switch-off genes of interest without altering the allelic combination that characterize successful commercial cultivars or to induce targeted sequence modifications by New Breeding Techniques. There is a variety of methods for potato improvement via genetic transformation. Most of them incorporate genes of interest into the nuclear genome; nevertheless, the development of plastid transformation protocols broadened the available approaches for potato breeding. Although all methods have their advantages and disadvantages, Agrobacterium-mediated transformation is the most used approach. Alternative methods such as particle bombardment, protoplast transfection with polyethylene glycol and microinjection are also effective. Independently of the DNA delivery approach, critical steps for a successful transformation are a rapid and efficient regeneration protocol and a selection system. Several critical factors affect the transformation efficiency: vector type, insert size, Agrobacterium strain, explant type, composition of the subculture media, selective agent, among others. Moreover, transient or stable transformation, constitutive or inducible promoters, antibiotic/herbicide resistance or marker-free strategies can be considered. Although great efforts have been made to optimize all the parameters, potato transformation protocols are highly genotype-dependent. Genome editing technologies provide promising tools in genetic engineering allowing precise modification of targeted sequences. Interestingly, transient expression of genome editing components in potato protoplasts was reported to generate edited plants without the integration of any foreign DNA, which is a valuable aspect from both a scientific and a regulatory perspective. In this review, current challenges and opportunities concerning potato genetic engineering strategies developed to date are discussed. We describe their critical parameters and constrains, and the potential application of the available tools for functional analyses or biotechnological purposes. Public concerns and safety issues are also addressed.

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A robust genetic transformation protocol to obtain transgenic shoots of Solanum tuberosum L. cultivar ‘Kufri Chipsona 1’

Cited by 9 publications

References 63 publications

Generation of Asynaptic Mutants in Potato by Disrupting StDMC1 Gene Using RNA Interference Approach

Generation of Asynaptic Mutants in Potato by Disrupting StDMC1 Gene Using RNA Interference Approach

A convenient Agrobacterium tumefaciens mediated transgenic system for diploid and tetraploid potatoes

State of the Art of Genetic Engineering in Potato: From the First Report to Its Future Potential

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