Transformation of hybrid black poplar with selective and reporter genes affects leaf proteome, yet without indication of a considerable environmental hazard

Kutsokon, N. K.; Danchenko, Maksym; Škultéty, Ľudovít; Kleman, Juraj; Rashydov, Namik M.

doi:10.1007/s11738-020-03072-6

Cited by 2 publications

(2 citation statements)

References 73 publications

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“…The disruption or alteration of gene expression due to the integration site of the transgene has been shown in plants [ 58 ] and fungi [ 59 ]. In particular, changes in the leaf proteome were observed in poplar with the uidA gene in vitro [ 60 ]. Transgenic Populus deltoides plants containing empty vectors unexpectedly demonstrated modified bud set, bud flush, and growth parameters after transfer from the greenhouse to the field [ 3 ].…”

Section: Discussionmentioning

confidence: 99%

Impact of Intron and Retransformation on Transgene Expression in Leaf and Fruit Tissues of Field-Grown Pear Trees

Lebedev

2023

IJMS

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Stable and high expression of introduced genes is a prerequisite for using transgenic trees. Transgene stacking enables combining several valuable traits, but repeated transformation increases the risk of unintended effects. This work studied the stability and intron-mediated enhancement of uidA gene expression in leaves and different anatomical parts of pear fruits during field trials over 14 years. The stability of reporter and herbicide resistance transgenes in retransformed pear plants, as well as possible unintended effects using high-throughput phenotyping tools, were also investigated. The activity of β-glucuronidase (GUS) varied depending on the year, but silencing did not occur. The uidA gene was expressed to a maximum in seeds, slightly less in the peel and peduncles, and much less in the pulp of pear fruits. The intron in the uidA gene stably increased expression in leaves and fruits by approximately twofold. Retransformants with the bar gene showed long-term herbicide resistance and exhibited no consistent changes in leaf size and shape. The transgenic pear was used as rootstock and scion, but grafted plants showed no transport of the GUS protein through the graft in the greenhouse and field. This longest field trial of transgenic fruit trees demonstrates stable expression under varying environmental conditions, the expression-enhancing effect of intron and the absence of unintended effects in single- and double-transformed woody plants.

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Section: Discussionmentioning

confidence: 99%

Impact of Intron and Retransformation on Transgene Expression in Leaf and Fruit Tissues of Field-Grown Pear Trees

Lebedev

2023

IJMS

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“…Poplars are also trees of great scientific importance due to ease of vegetative propagation, short rotation cycle, small genome size, and the ability to regenerate in vitro. These important features, combined with the complete sequencing of the genome of Populus trichocarpa, made poplar a model plant offering molecular tools for basic research in genetics, physiology, and biochemistry of trees (Han et al 2000;Tuskan et al 2006;Kutsokon et al 2020). In recent decades, short-rotation plantations of fastgrowing trees for biomass production have been increasingly established worldwide (Wullschleger et al 2002;Aylott et al 2008;Kutsokon et al 2017;Moravčíková et al 2017).…”

Section: Introductionmentioning

confidence: 99%

In vitro culture initiation and regeneration of two highly productive clones of poplar

Khoma

Khudolieieva

Rashydov

et al. 2022

Nova Biotechnol. Chim.

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In vitro shoot regeneration is a fast and reliable method for propagation of valuable tree genotypes and important step in genetic manipulations. This study aimed to establish an optimal in vitro culture initiation and regeneration protocol for highly productive poplar clones ‘Novoberlinska-3’ (Populus pyramidalis × P. laurifolia) and ‘Volosystoplidna’ (P. trichocarpa). In vitro culture was initiated either on MS (IM-MS) or WPM (IM-WPM) medium both containing BAP (0.2 mg.L-1) and IBA (0.1 mg.L-1). Results demonstrated that the best initiation medium for the clone ‘Volosystoplidna’ was IM-WPM on which 93.3 % of explants survived, while explants from ‘Novoberlinska-3’ better survived on IM-MS. After establishing both genotypes into aseptic culture, regeneration experiments were started using two types of explants, leaf and petioles, planted on callus induction medium containing 2iP (1.02 mg.L-1) and NAA (1.86 mg.L-1). Furthermore, microshoots were placed on shoot induction medium supplemented with 0.04 mg.L-1 thidiazuron, and then transferred on shoot elongation medium with 0.2 mg.L-1 BAP. Regeneration protocol showed better performance of poplar ‘Novoberlinska-3’, but it was also efficient for ‘Volosystoplidna’. Plant regeneration from leaf explants in the clone ‘Novoberlinska-3’ showed the highest regeneration percentage (92.3 %) and the number of shoots per explant (3.1), which significantly exceeded other explants. Our results showed a significant difference between the survivability of two clones during culture initiation. The differences in regeneration rates between the clones as well as leaf and petiole explants were also determined. Obtained aseptic cultures of highly productive poplar hybrids will be used in our further studies, especially for genetic transformation.

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Transformation of hybrid black poplar with selective and reporter genes affects leaf proteome, yet without indication of a considerable environmental hazard

Cited by 2 publications

References 73 publications

Impact of Intron and Retransformation on Transgene Expression in Leaf and Fruit Tissues of Field-Grown Pear Trees

Impact of Intron and Retransformation on Transgene Expression in Leaf and Fruit Tissues of Field-Grown Pear Trees

In vitro culture initiation and regeneration of two highly productive clones of poplar

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