Setaria viridis floral-dip: A simple and rapid Agrobacterium-mediated transformation method

Abstract

Setaria viridis was recently described as a new monocotyledonous model species for C4 photosynthesis research and genetic transformation. It has biological attributes (rapid life cycle, small genome, diploid, short stature and simple growth requirements) that make it suitable for use as a model plant. We report an alternative method of S. viridis transformation using floral dip to circumvent the necessity of tissue culture phase for transgenic plant regeneration. S. viridis spikes at boot stage were selected to be immersed in Agrobacterium suspension. T1 seeds could be identified in 1.5–2 months after floral dipping. We demonstrated through molecular analysis and RFP expression that seeds and resulting plants from dipped inflorescences were transformed. Our results suggest the feasibility of S. viridis floral dip transformation as a time-saving and cost-effective compared with traditional methods. To our knowledge, this is the first report using floral dip in S. viridis as an Agrobacterium-mediated transformation method.

A Tightly Regulated Genetic Selection System with Signaling-Active Alleles of Phytochrome B

“…Whereas Brassica plants that flower and set seed in late spring may experience a short period of high temperature, the resulting expression of YHB might even enhance photosynthesis and delay senescence potentially leading to an overall benefit to crop yield. Successful in planta transformations were also reported from other eudicot and monocot plants, including Medicago truncatula (Trieu et al, 2000), soybean (Glycine max; Gao et al, 2007), tomato (Lycopersicon esculentum; Yasmeen et al, 2009), wheat (Zale et al, 2009), maize (Zea mays; Mu et al, 2012;Moiseeva et al, 2014), and Setaria viridis (Martins et al, 2015).…”

Agrobacterium tumefaciens-MEDIATED IN-PLANTA TRANSFORMATION OF INDONESIAN MAIZE USING pIG121Hm-Cs PLASMID CONTAINING nptII AND hpt GENES

“…The development of a simple and rapid transformation method for Setaria viridis as a C4 model for monocotyledonous plants was time-saving and cost-effective compared with traditional methods in plant genetic engineering (Martins et al 2015). Inplanta transformation techniques have been succeeded in buckwheat plant (Fagopyrum esculentum M.), mulberry (Morus alba L.) and kenaf (Hibiscus cannabinus L.) (Ping et al 2003;Kojima et al 2004).…”

“…The main step in A. tumefaciens method is based on co-incubation of plant vegetative organ or tissue (seeds, leaves, roots, stems or meristems) in bacterial-cell suspension (Chumakov 2007). These methods have been successfully used for transformation of important genes to many plants whose tissue culture system has already been established (Hiei et al 1994;Ishida et al 1996;Gelvin 2003;Carneiro and Carneiro 2013;Martins et al 2015;Jan et al 2016). Some advantages of transformation via A. tumefaciens have been reported such as higher transformation efficiency with a single copy gene and can be carried out with simple laboratory equipment and low-cost technique (Supartana et al 2005).…”

“…Transformants with a single copy gene are preferable as it is a easest analysis process and the segregation usually follows Mendelian inheritance pattern (Rahmawati 2006). But, the tissue culture-based transformation methods are time-consuming and lead some mutations from clonal variation that affect both qualitative and quantitative characters of plants (Martins et al 2015;Jan et al 2016).…”

“…Floral dip method has been used in genetic engineering strategy as it directly produces genetically modified transformant without the laborious tissue culturing procedures (Martins et al 2015). The production of a large number of uniform plants in a short time, less labor efforts and minimal reagent requirements are some of the main advantages of in-planta transformation system (Bent 2000).…”