A simple and highly efficient Agrobacterium-mediated transformation protocol for Setaria viridis

Abstract

The production and use of sugarcane in Brazil is very important for bioenergy production and is recognized as one of the most efficient in the world. In our laboratory, Setaria viridis is being tested as a model plant for sugarcane. S. viridis has biological attributes (rapid life cycle, small genome, diploid, short stature and simple growth requirements) that make it suitable for use as a model system. We report a highly efficient protocol for Agrobacterium-mediated genetic transformation of S. viridis. The optimization of several steps in tissue culture allowed the rapid regeneration of plants and increased the rate of transformation up to 29%. This protocol could become a powerful tool for functional genomics in sugarcane.

The Status of Setaria viridis Transformation: Agrobacterium-Mediated to Floral Dip

“…Additional modifications of the Agrobacterium -mediated transformation methodology of S. viridis A10.1 were also reported by a second group (Martins et al, 2015b). Martins et al (2015b) followed similar methodology as reported by Van Eck et al (2017).…”

“…In addition to the hpt selectable marker gene, vectors used for the development of the S. viridis transformation methodology also contained the beta-glucuronidase (GUS) and green fluorescent protein (GFP) reporter genes (Martins et al, 2015b; Van Eck and Swartwood, 2015). …”

“…Martins et al (2015b) followed similar methodology as reported by Van Eck et al (2017). A comparison of the primary components that make up the various reported transformation methods (Martins et al, 2015b; Van Eck and Swartwood, 2015; Van Eck et al, 2017) is outlined in Table 1 .…”

“…Martins et al (2015b) followed similar methodology as reported by Van Eck et al (2017). A comparison of the primary components that make up the various reported transformation methods (Martins et al, 2015b; Van Eck and Swartwood, 2015; Van Eck et al, 2017) is outlined in Table 1 . The vectors used for transformation, more specifically the type of selectable marker gene and its promoter contained in the vector, were the key parameters that influenced transformation efficiency in the methods described by Martins et al (2015b).…”

Selection of reliable reference genes for RT-qPCR analysis during developmental stages and abiotic stress in Setaria viridis

“…It is a short, fast-growing, C4 metabolism plant, with its draft genome sequence recently available, making it suitable model plant for genetic and genomics studies12. Moreover, S. viridis is highly responsive to Agrobacterium tumefaciens -mediated genetic transformation, with a well-established transformation protocols13 and more recently, spike-dipping methods have also been proposed34. S. viridis could be used as a model plant for food and bioenergy grasses presenting C4 metabolism such as maize, sorghum, sugarcane and switchgrass.…”