Abstract
Sugar beet (Beta vulgaris L.) is one of the most important industrial crops throughout world. With the availability of suitable genetic transformation technologies, the yield, quality, and stress tolerance of sugar beet could be improved significantly. However, low transformation efficiencies seriously limit the application of molecular technologies to the genetic improvement of sugar beet. With the aim of improving gene transfer techniques for sugar beet, the effect of different sucrose concentrations during cocultivation on the initial Agrobacterium-mediated transformation efficiencies in sugar beet was tested. To develop an efficient experimental system through which the effect of sucrose could be tested, first, a prolific regeneration system was optimized by testing the effect of different plant growth regulators on in vitro regeneration and rooting efficiencies from sugar beet cotyledonary node explants. The highest mean number of regenerated shoots per explant was obtained when the cotyledonary node explants excised from young seedlings were grown on MS medium supplemented with 1.0 mg/L 6-benzylaminopurine. Using this regeneration system, the effect of different concentrations of sucrose included in the cocultivation medium on the initial genetic transformation efficiencies observed in T0 plants was tested using an Agrobacterium tumefaciens strain carrying the pBin19/35S:GUS-INT construct. The inclusion of 4.5% sucrose in the cocultivation medium resulted in significantly higher transformation (34.09%) and expression efficiencies (22.72%), confirmed by polymerase chain reaction and β-glucuronidase assays, respectively, in regenerated T0 seedlings. If translated into stably inherited transformation efficiencies, these findings could contribute to the success of genetic transformation studies in sugar beet and other crops recalcitrant to Agrobacterium-mediated transformation.
Highlights
IntroductionSugar beet is one of the most important industrial plants widely cultivated in temperate climates of the world (Mishutkina et al, 2010; Lytvyn et al, 2014), and it is produced in 55 countries, with major producing countries being Russia, France, the USA, Germany, and Turkey
Industrial crops are cultivated annual or perennial plants that are used as raw material by various branches of industry and grow in various geographic regions including temperate, subtropical, and tropical latitudes (Vijayan et al, 2008; Ercisli et al, 2011; Ahmad et al, 2015; Cesur et al, 2018).Sugar beet is one of the most important industrial plants widely cultivated in temperate climates of the world (Mishutkina et al, 2010; Lytvyn et al, 2014), and it is produced in 55 countries, with major producing countries being Russia, France, the USA, Germany, and Turkey
Optimization of an in vitro plant regeneration system for sugar beet to test transformation efficiencies To develop an efficient regeneration system that could be used for the optimization of sugar beet transformation, cotyledonary node explants taken from sugar beet seedlings grown in sterile conditions were treated with different concentrations of BAP, GA3, or TDZ (Figure 1)
Summary
Sugar beet is one of the most important industrial plants widely cultivated in temperate climates of the world (Mishutkina et al, 2010; Lytvyn et al, 2014), and it is produced in 55 countries, with major producing countries being Russia, France, the USA, Germany, and Turkey. A number of biotic and abiotic stresses cause significant crop and quality losses in sugar beet (Lytvyn et al, 2014). Genetic transformation technologies offer great promise for improving stress tolerance in sugar beet and other crop plants through the introduction of genes conferring biotic and abiotic tolerance. Low transformation efficiencies remain a limiting factor in Agrobacterium-mediated genetic transformation of many crop plants. Despite the fact that the first successful sugar beet transformation was achieved at the end of the 20th century, low transformation and regeneration efficiencies still hinder the routine production of transgenic sugar beet plants (Mishutkina et al, 2010; Rivera et al, 2012; Pathi et al, 2013)
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