Sand-Wounding of Shoot and Petiole Explants Enhances Transformation Efficiency in Sugar Beet ( Beta vulgaris L.) by Agro
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RESEARCH ARTICLE
Sand-Wounding of Shoot and Petiole Explants Enhances Transformation Efficiency in Sugar Beet (Beta vulgaris L.) by Agrobacterium-mediated Transformation Songul Gurel1
Received: 14 July 2020 / Accepted: 9 September 2020 Ó Society for Sugar Research & Promotion 2020
Abstract Genetic transformation of recalcitrant species, like sugar beet, has been always a challenging task. In this study, a selectable marker gene (phosphomannose isomerase) and a reporter gene (synthetic green fluorescent protein) were transferred to sugar beet via Agrobacteriummediated transformation by using sand-wounded shoot and petiole explants of two different diploid sugar beet genotypes. Considering the overall means of genotypes and explant types, a clear genotypic variation was evident as ELK345 produced a mean of 5.7% transient expression and 1.97% stable transformation efficiency while it was 4.3% and 1.28% for the genotype M1195. When explant types were compared independently from other parameters, it was shown that petiole explants were more productive than shoot explants, with an overall mean of 5.3% versus 4.7% and 1.77% versus 1.48% transient expressions and stable transformation efficiencies, respectively. On the other hand, when a comparison was made without taking into account the genotypes, the effect of wounding treatment on transient expression resulted in about 25% (4.7% vs. 5.9%) and 20% (4.3% vs. 5.1%) increases in petiole and shoot explants, respectively. Such a promoting effect of wounding was even more prominent when the stable transformation efficiencies were compared; petiole explants achieving almost a 60% increase (1.38% vs. 2.16%) while shoot explants doubled (0.98% vs. 1.97%) their efficiencies. The optimized protocol reported here can be employed for further increasing the transformation efficiency in recalcitrant plant species including sugar beet. & Songul Gurel [email protected] 1
Department of Biology, Faculty of Arts and Science, Bolu Abant Izzet Baysal University, 14030 Bolu, Turkey
The advantages of using GFP as a reporter and mannose as a positive selection are also discussed. Keywords Sugar beet Beta vulgaris L. Agrobacterium Transformation Wounding Sand
Introduction Sugar beet is one of the most important industrial crops in the world as sugar is economically produced only from two plant species; sugar cane and sugar beet. 78% of the world’s total sugar production is obtained from sugar cane while it is 22% from sugar beet (Licht 2019). From the time when the first successful gene transfer in sugar beet, which was restricted to the production of transgenic tissue or cells, substantial improvements have been made in the approaches using Agrobacterium tumefaciens (Lindsey and Gallois 1990; D’Halluin et al. 1992). However, the transformation success was low, and the method was specific to only a few genotypes. Early attempts in transforming sugar beet via Agrobacterium tumefaciens were not successful because of failure in obtaining plants from transferred callus (Krens et
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