Explant position effect on gene transformation to flax (Linum usitatissimum L.) via Agrobacterium tumefaciens

Beyaz, Ramazan

doi:10.18054/pb.v119i3.5020

Cited by 4 publications

(1 citation statement)

References 9 publications

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“…As the single major criterion for effective transgenesis is effective totipotency and regeneration, recalcitrance of flax to tissue culture-based regeneration has been the major bottleneck. Hence, there has always been a dearth of successful flax transformation strategies despite several attempts toward standardization and optimization of regeneration as well as non-tissue culture-based approaches ( Lamblin et al, 2007 ; Beranova et al, 2008 ; Bleho et al, 2012 ; Bastaki and Cullis, 2014 , 2019 ; Beyaz et al, 2016a , b , 2017 ; Mandal et al, 2018 ; Aycan et al, 2019 ; Majumder et al, 2020 ).…”

Section: Introductionmentioning

confidence: 99%

An Apical Meristem-Targeted in planta Transformation Method for the Development of Transgenics in Flax (Linum usitatissimum): Optimization and Validation

et al. 2021

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Efficient regeneration of explants devoid of intrinsic somaclonal variations is a cardinal step in plant tissue culture, thus, a vital component of transgenic technology. However, recalcitrance of economically important crops to tissue culture-based organogenesis ensues a setback in the use of transgenesis in the genetic engineering of crop plants. The present study developed an optimized, genotype-independent, nonconventional tissue culture-independent in planta strategy for the genetic transformation of flax/linseed. This apical meristem-targeted in planta transformation protocol will accelerate value addition in the dual purpose industrially important but recalcitrant fiber crop flax/linseed. The study delineated optimization of Agrobacterium tumefaciens-mediated transformation and stable T-DNA (pCambia2301:GUS:nptII) integration in flax. It established successful use of a stringent soilrite-based screening in the presence of 30 mg/L kanamycin for the identification of putative transformants. The amenability, authenticity, and reproducibility of soilrite-based kanamycin screening were further verified at the molecular level by GUS histochemical analysis of T0 seedlings, GUS and nptII gene-specific PCR, genomic Southern hybridization for stable integration of T-DNA, and expression analysis of transgenes by sqRT-PCR. This method resulted in a screening efficiency of 6.05% in the presence of kanamycin, indicating amenability of in planta flax transformation. The strategy can be a promising tool for the successful development of transgenics in flax.

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