Utilization of sucrose during cocultivation positively affects Agrobacterium-mediated transformation efficiency in sugar beet (Beta vulgaris L.)

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 T 0 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 T 0 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.

Prediction of the concentration of plant growth regulators for somatic embryogenesis and regeneration of Hyoscyamus niger using Box–Behnken design of response surface methodology

“…Therefore, optimization of the somatic embryogenesis protocol is the rst and most important step in gene transformation studies, but it is a laborious, time-consuming, and complex process (Hesami and Jones 2020, . In in vitro cultures, somatic embryogenesis processes such as dedifferentiation, re-differentiation, or differentiation are caused by genetic and environmental factors (Gürel et al 2019, Joung et al 2020). Therefore, it will be di cult to predict various cases, including the rate of callus formation, and the rate of embryogenesis, as well as the optimization of the factors involved in them through conventional mathematical methods (Hesami and Jones 2020, Salehi et al 2020).…”

In Vitro Multiple Shoot Induction From Embryonic Axes of Annual Herbaceous Legume Faba Bean (Vicia faba L.)

“…This in conjunction with surrounding temperature and humidity helped the plants to grow flourish and acclimatize easily. Establishment of reliable acclimatization of tissue cultured plantlets (Goncalves et al, 1998;Gürel et al, 2019) permitted saving of time. This can also help by use of tissue culture for breeding, transformation and functional genomic studies.…”

Gamma Radiation Effect on Agrobacterium tumefaciens-Mediated Gene Transfer in Potato (Solanum tuberosum L.)

“…Agrobacterium-mediated transformation method is the most proficient and commonly used plant transformation method among different gene transfer techniques. However, there are a number of variables that affect the success of Agrobacteriummediated genetic transformation [1]. The success of genetic transfer efficiency with A. tumefaciens depends on, Agrobacterium strain used, bacteria concentration, antibiotic types and concentration used for in vitro selection, inoculation time, and temperature [2].…”